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PubMed1991

The hypothalamic lateral tuberal nucleus in Alzheimer's disease.

The hypothalamic lateral tuberal nucleus was investigated in 5 young patients, aged 45 to 64 years, with Alzheimer's disease (AD) and in 5 age-matched control subjects. Combining conventional histopathological and immunocytochemical staining with neuronal counts, a peculiar form of neuronal pathology was characterized. Although neurons and neurites in the lateral tuberal nucleus of AD specimens were heavily stained by Alz-50, silver and thioflavine-S stains disclosed few neurofibrillary tangles or neurites. The numbers of neurons in the lateral tuberal nucleus of patients with AD (67,450; SEM = 5,050) were no different from those of control subjects (58,900; SEM = 2,450). In the AD patients, few plaques were present and were almost exclusively of the amorphous variety. We conclude that neurons in the lateral tuberal nucleus show an early stage of AD-related cytoskeletal pathology (Alz-50 positivity), but without plaques or neuronal death.

Alzheimer Disease↗
PubMed1994

Effects of denervation upon receptor cell survival and basal cell proliferation in tuberous electroreceptor organs of a weakly electric fish.

Weakly electric fish generate electric fields for the purposes of electrolocation and communication. These fields are detected by specialized receptor organs: the tuberous organs. In the present study we investigated the effects of denervation upon receptor cell survival and progenitor (basal) cell proliferation rate. The left, infraorbital, anterior lateral line nerve of brown ghosts (Apteronotus leptorhynchus) was sectioned, and the proximal stump was dipped in ricin to prevent regrowth. In groups of four, the animals were given two daily injections of the cell proliferation marker bromodeoxyuridine (BrdU) for 2 days at 1, 2, 3, or 4 weeks following denervation. At the completion of the BrdU injection schedule, a piece of cheek skin, rostroventral to the eye, was removed from the left (denervated) and the right (intact) sides and processed for light microscopy or immunocytochemistry. Our results show: (1) there is progressive receptor cell death and tuberous organ degeneration following denervation; (2) basal cell proliferation increases steadily with time after denervation and tuberous organ degeneration; and (3) despite denervation, some proliferating basal cells differentiate into receptor cells, but these new receptor cells eventually die. These results suggest that innervation is essential for tuberous electroreceptor cell survival and that the rate at which basal cells proliferate is regulated by receptor cell health, locally released factors, or both.

Animals↗
PubMed1994

Nuclear protein factors binding to a class I patatin promoter region are tuber-specific and sucrose-inducible.

Genes encoding patatin, the major storage protein of the potato tuber, are generally divided into two classes, class I and class II. The expression of the class I patatin genes is normally tuber-specific, but can be induced in leaves by high concentrations of sucrose. By employing electrophoretic mobility shift assays (EMSA), we have identified nuclear protein factors that interact specifically with the proximal portion of the class I patatin promoter that is required for tuber-specific and sucrose-inducible expression. The factors were detected in nuclear extracts prepared from potato tubers and sucrose-induced leaves, but not in extracts from leaves of normal potato plants. Four putative transcription factor-binding sites were localized using DNase I footprinting. Competitive EMSA was employed to show that the same protein factor binds to at least two of the sites (boxes D and M). Interestingly, these two binding sites are highly homologous to light-responsive elements present in genes for the ribulose-1,5-bisphosphate carboxylase small subunit.

Base Sequence↗
PubMed1989

Sucrose-regulated expression of a chimeric potato tuber gene in leaves of transgenic tobacco plants.

Patatin is a family of lipid acyl hydrolases that accounts for 30 to 40% of the total soluble protein in potato tubers. Class-I patatin genes encode 98 to 99% of the patatin mRNA in tubers, but are not normally expressed in other tissues. They are not totally 'tuber-specific'; however, since they can be induced to express at high levels in other tissues under conditions of sink limitation or in explants cultured on medium containing elevated levels of sucrose. To examine the evolution of the mechanisms that regulate patatin gene expression, we introduced a chimeric patatin-beta-glucuronidase (GUS) gene containing 2.5 kb of 5' flanking sequence from the Class-I potato patatin gene PS20 into tobacco plants. The construct was not expressed at significant levels in leaves of juvenile plants or plantlets cultured in vitro, but was expressed at high levels in explants cultured on medium containing 0.3 to 0.4 M sucrose. While there were differences in the expression of the chimeric gene between transgenic tobacco and potato plants, the pattern of sucrose induction was very similar. These results suggest that the mechanism that controls patatin gene expression in potato tubers evolved from a widely distributed mechanism in which gene expression is regulated by the level of available photosynthate.

Carbohydrates↗
PubMed1995

Isolation and characterisation of cDNA clones representing the genes encoding the major tuber storage protein (dioscorin) of yam (Dioscorea cayenensis Lam.).

cDNA clones encoding dioscorins, the major tuber storage proteins (M(r) 32,000) of yam (Dioscorea cayenesis) have been isolated. Two classes of clone (A and B, based on hybrid release translation product sizes and nucleotide sequence differences) which are 84.1% similar in their protein coding regions, were identified. The protein encoded by the open reading frame of the class A cDNA insert is of M(r) 30,015. The difference in observed and calculated molecular mass might be attributed to glycosylation. Nucleotide sequencing and in vitro transcription/translation suggest that the class A dioscorin proteins are synthesised with signal peptides of 18 amino acid residues which are cleaved from the mature peptide. The class A and class B proteins are 69.6% similar with respect to each other, but show no sequence identity with other plant proteins or with the major tuber storage proteins of potato (patatin) or sweet potato (sporamin). Storage protein gene expression was restricted to developing tubers and was not induced by growth conditions known to induce expression of tuber storage protein genes in other plant species. The codon usage of the dioscorin genes suggests that the Dioscoreaceae are more closely related to dicotyledonous than to monocotyledonous plants.

Amino Acid Sequence↗
PubMed1993

Constitutive or light-regulated expression of the rolC gene in transgenic potato plants has different effects on yield attributes and tuber carbohydrate composition.

Tetraploid potato clones, transgenic for the rolC gene of Agrobacterium rhizogenes under control of the light-inducible ribulose bisphosphate carboxylase small subunit promoter (rbcS-rolC), were compared, with respect to yield attributes and tuber carbohydrates, with transformed and untransformed controls and with 35S-rolC transgenic potato plants. In rbcS-rolC plants, the expression of the rolC gene was located mainly in leaves, while in 35S-rolC plant transcripts were detected as well in shoots and roots. Phenotypically, rbcS-rolC transgenic plants were found to be slightly reduced in plant size with a few more tillers than control plants. Photosynthetic rate and chlorophyll content were significantly lower in all rolC transgenic plants irrespective of the type of construct used. Tuber yield was not significantly different between controls and rbcS-rolC transgenic plants, but was reduced in the 35S-rolC transformants. Sucrose level was unchanged in all rolC clones investigated, whereas fructose content was significantly enhanced in 35S-rolC transformants, but not in the plants expressing the rolC gene in aerial plant parts only. In both types of rolC transgenic plants, glucose content was lower than in controls, resulting in a significant reduction of reducing sugar in tubers. The results suggest a hormonal influence on the carbohydrate composition of potato tubers.

Bacterial Proteins↗
PubMed1993

Jasmonic acid-inducible gene expression of a Kunitz-type proteinase inhibitor in potato tuber disks.

Messenger RNAs of a potato (Solanum tuberosum L.) Kunitz-type proteinase inhibitor(s) (PKPI) were present in potato disks excised from tubers stored for 14 months (old tubers) or 2 months (young tubers) after harvest, and disappeared during the aseptic culture. The PKPI mRNA accumulation was found to be induced in potato disks from the old tubers by the addition of jasmonic acid (JA) [3-oxo-2-(2'-cis-pentenyl)-cyclopentane-1-acetic acid].

Cyclopentanes↗
PubMed1978

[Tuberous cortical irregularity at the distal femoral metaphysis--an often malignancy pretending radiological finding (author's transl)].

Four juvenile patients are described who were referred to us because of a tuberous, irregularly demarcated cortical lesion at the dorsal surface of the distal femur metaphysis suspected to be a "malignant tumor". The histologic evaluation in three of these patients revealed a fibroplastic reaction of the periosteum with periosteal new bone formation and focal, partly also lacunar bone resorption by osteoclasts. During six months of observation there were no signs of further growing. No therapy was necessary. This tuberous cortical defect at the dorsal surface of the distal femur metaphysis has been found almost exclusively in children and juvenile persons. As causative agent an increased muscle power of the gastrocnemius muscles has been discussed at the site of its bone insertion. If the bone formative component prevails a tuberous proliferation is evident radiographically, however, if the fibroplastic reaction is predominant there is more evidence for a cortical defect. The resemblance of the tuberous cortical defects in the four patients discussed here with those cases described in the literature under various names is pointed out.

Adolescent↗
PubMed2002

Enhancement of metabolizing herbicides in young tubers of transgenic potato plants with the rat CYP1A1 gene.

A rat P450 monooxygenase gene ( CYP1A1) was introduced into potato plants to enhance the metabolism of the environmental contaminants in subterranean organs. The CYP1A1 gene was kept under the control of the potato patatin promoter to enhance tuber-specific expression. A total of 106 transgenic plants (PAT1A1 plants) were obtained following selection by a resistance test to kanamycin and PCR analysis. PAT1A1 plants treated with 10% exogenous sucrose showed a higher activity of monooxgenase in the leaves than the non-transgenic plants. This indicated that the activity enhanced by 10% sucrose was due to the patatin promoter containing the sucrose-inducted elements. One representative transgenic plant, Ag2197, was selected on the basis of monooxgenase activity in the leaves and Western blot analysis. Ag2197 was found to accumulate a large amount of CYP1A1 mRNA and protein in the developing tuber but not in the mature tuber. The residual herbicides, atrazine and chlortoluron, were analyzed in the micro-tubers of Ag2197 and non-transgenic plants. The amount of residual herbicides in Ag2197 was much lower than that in the non-transgenic plant, indicating that the transgenic plant metabolized the herbicides to a detoxified form. The transgenic plants produced in this study might be useful for the phytoremediation of chemical pollution in the soil.

Journal Article↗
PubMed2003

Evaluation of glycoalkaloids in tubers of genetically modified virus Y-resistant potato plants (var. Désirée) by non-aqueous capillary electrophoresis coupled with electrospray ionization mass spectrometry (NACE-ESI-MS).

The glycoalkaloid content of transgenic potatoes was evaluated by an optimised method based on non-aqueous capillary electrophoresis coupled on-line with electrospray ionization-mass spectrometry (NACE-ESI-MS). The potato material consisted of tubers from a conventional cv. Désirée and from three lines of modified plants resistant, intermediate and susceptible to infection by potato virus Y (PVY). The main glycoalkaloids were confirmed to be alpha-solanine and alpha-chaconine with parent ion masses m/z 852 and 868, respectively. In addition, an unknown minor peak at m/z 850.6 was found both in conventional (control) and susceptible line potato tubers. Such a compound exhibited an MS(2) spectrum with fragments ions at 704 and 396 m/z derived by loss of two ions, i.e. m/z 146 and 307, most likely corresponding to a rhamnose unit and a [glucose-(rhamnose)(2)] moiety, respectively. Up to 30-80-fold higher concentrations of total glycoalkaloids were found in the peel compared to flesh samples of all tubers examined. TGA content was nearly doubled in peel samples of resistant compared to control lines, and these levels were lower than the limit recommended for food safety, i.e. 20-60 mg of TGA per 100 g fresh weight. Moreover, it was established that tubers produced by virus-resistant clones are substantially equivalent in glycoalkaloid contents to those produced by conventional potato varieties.

Electrophoresis, Capillary↗
PubMed1998

Cytoskeletal alterations in the human tuberal hypothalamus related to argyrophilic grain disease.

The tuberal region of the human hypothalamus was examined for cytoskeletal changes related to argyrophilic grain disease (AGD). Hypothalamic sections of eight individuals afflicted with AGD and eight controls were cut serially in the frontal plane at 100 microm. The presence of argyrophilic AGD-related pathology was demonstrated utilizing the modified Gallyas silver iodide technique. Tau-positive cytoskeletal changes were stained by the phosphorylation-dependent antibody AT8. A characteristic pattern of tau-positive cytoskeletal alterations was revealed in the tuberal hypothalamus of AGD cases, while controls were devoid of such changes. The lateral tuberal nucleus was found to be particularly susceptible to AGD, demonstrating numerous tau-positive grains and neuronal cell bodies. Similar alterations were present to a moderate degree in the ventromedial nucleus. A previously unreported, conspicuous accumulation of tau-positive oligodendrocytes (coiled bodies) and interfascicular thread-like fibers was detected in the column of the fornix. Only sparse argyrophilic changes were noted in consecutive silver-stained sections, comprised mainly of accumulations of spindle-shaped grains within the lateral tuberal nucleus. Remarkably, a pronounced expression of AGD-related alterations was seen in the absence of hypothalamic changes related to other tau-positive cytoskeletal disorders, such as Alzheimer's disease. The present findings support the concept that AGD is a distinct neurodegenerative entity afflicting not only cortical but also subcortical predilection sites of the human brain.

Aged↗
PubMed2001

The contribution of plastidial phosphoglucomutase to the control of starch synthesis within the potato tuber.

The aim of this work was to evaluate the extent to which plastidial phosphoglucomutase (PGM) activity controls starch synthesis within potato (Solanum tuberosum L. cv. Desirée) tubers. The reduction in the activity of plastidial PGM led to both a correlative reduction in starch accumulation and an increased sucrose accumulation. The control coefficient of plastidial PGM on the accumulation of starch was estimated to approximate 0.24. The fluxes of carbohydrate metabolism were measured by investigating the metabolism of [U-14C]glucose in tuber discs from wild-type and transgenic plants. In tuber discs the control coefficient of plastidial PGM over starch synthesis was estimated as 0.36, indicating that this enzyme exerts considerable control over starch synthesis within the potato tuber.

Amino Acids↗
PubMed2001

The effect of exogenous sugars on the control of flux by adenosine 5'-diphosphoglucose pyrophosphorylase in potato tuber discs.

The aim of this work was to investigate the effect of exogenous sugars on the extent to which starch synthesis in potato ( Solanum tuberosum L.) is controlled by adenosine 5'-diphosphoglucose pyrophosphorylase (EC 2.7.7.27; AGPase). Tuber discs were incubated in the presence of a range of concentrations of glucose and sucrose, and metabolic fluxes measured following the supply of [U-14C]glucose and measurement of the specific radioactivity of the hexose phosphate pool. In the presence of glucose there was a marked increase in the flux through glucose-phosphorylating hexokinase, and at high concentrations of external glucose this led to a stimulation of the rate of starch and sucrose synthesis relative to those measured in the presence of sucrose. In the presence of glucose the ratio of the rate of starch synthesis to the rate of glycolysis was higher than in the presence of sucrose. Similar effects of glucose were observed at two stages of tuber development. We conclude that the presence of glucose perturbs the carbohydrate metabolism of tuber discs so that starch synthesis is favoured. In order to determine the extent to which AGPase controls flux, we measured fluxes in wild-type plants and transgenic plants with reduced AGPase activity as a result of the expression of a cDNA encoding the B subunit in the antisense orientation. In the presence of sucrose a reduction in AGPase activity had a greater impact on the rate of starch synthesis than in the presence of glucose. The flux control coefficient of AGPase over starch synthesis was higher in the presence of sucrose (0.7-0.9) than in the presence of glucose (0.4-0.6). Conversely, the impact of reduced AGPase activity on the rate of sucrose synthesis was lower in the presence of sucrose than glucose. In the presence of 200 mM sucrose the flux control coefficient of AGPase over the rate of sucrose synthesis was not significantly different from zero. This demonstrates that the nature of the sugar supplied to potato tuber discs can have a major influence on the distribution of control within metabolism. These data were also used to investigate the relationship between demand for ATP and the rate of hexose phosphate entry into glycolysis. A very strong correlation between ATP demand and glycolytic flux was demonstrated.

Carbohydrate Metabolism↗
PubMed2004

Changes in sugar content and activity of vacuolar acid invertase during low-temperature storage of potato tubers from six Japanese cultivars.

The storage of potato tubers at low temperatures leads to the accumulation of sugars in a process called "low-temperature sweetening." To understand this phenomenon, we measured the sugar contents and the activity of acid invertase over several months in tubers of six Japanese cultivars stored at 4 degrees C or 20 degrees C. At 20 degrees C, few changes in sugar contents took place in any of the tubers. On the other hand, when stored at 4 degrees C, three types of changes were observed among the cultivars: (1) increased levels of reducing sugars during storage; (2) a pattern similar to that of type 1, but with 4- to 6-fold lower levels of reducing sugars throughout storage; and (3) increased sucrose, but not reducing sugars. The activity of vacuolar acid invertase increased in the type-1 cultivars, whereas, in the type-2 and type-3 cultivars, the activities were very low during storage at 4 degrees C. Reverse transcription-polymerase chain reaction analysis of acid invertase showed that the transcripts of the enzyme accumulated in the tubers stored at 4 degrees C in the type-1 cultivars but not in type-3. These results suggest that the activity of vacuolar acid invertase is related to the types of changes that occurred in sugar content during low-temperature storage among the potato cultivars.

Cold Temperature↗
PubMed2006

In vitro availability of some essential minerals in commonly eaten processed and unprocessed Caribbean tuber crops.

The levels of three essential minerals Ca, Fe and Mg and the extent of their availability were assessed in four commonly eaten Caribbean tuber crops [dasheen (Xanthosoma spp.), Irish potato (Solanum tuberosum), sweet potato (Ipomoea batatas) and yellow yam (Dioscorea cayenensis)] in their processed and unprocessed states. Calcium was highest in cooked dasheen (5150+/-50 mg/kg) while Magnesium was highest in uncooked Irish potato (3600+/-200 mg/kg). There was no significant loss of calcium from the food samples upon cooking. All the uncooked food samples displayed higher levels minerals assessed compared to the cooked samples except for cooked Irish potato that recorded the level of iron (182.25+/-8.75 mg/kg). Availability of these minerals in the cooked and uncooked tubers crops upon digestion also showed a similar pattern. In conclusion, the consumption of these tuber crops in the Caribbean may not be responsible for the reported cases of iron deficiency in the region. However, the availability of minerals from these tuber crops when consumed with other foods (the usual practice in the Caribbean) needs further investigation.

Calcium↗
PubMed2005

Bitterness and toxicity in wild yam (Dioscorea spp.) tubers of Nepal.

Wild yams make a significant contribution to diets of tribal people in Nepal. However, these wild tubers are unpalatable, taste bitter, produce inflammation and show occasional toxicity. Four wild yam species, which are eaten after primary treatment by Nepali aborigines, were analyzed for bitter and toxic principles. Bitter components were identified as furanoid norditerpenes (diosbulbins A and B). Diosbulbins A and B were found in the range of 0.023-0.046 and 0.151-0.442 g kg(-1), respectively. Results demonstrated that diosbulbin B, with an average value of 0.314 g kg(-1), was the principal bitter compound as compared to diosbulbin A (0.037 g kg(-1)). The toxic alkaloid, dioscorine and histamine (an allergen) were not detected in these tubers, whereas cyanogens (as HCN equivalent) content were found ranging from 3.2 to 6.0 ppm. Our results revealed that Nepali wild yam tubers are not toxic varieties, as they do not contain either toxic dioscorine or histamine and cyanogens contents were satisfactorily below the safety limits. The inflammation and occasional toxicity observed could possibly be due to the presence of high level of oxalate in these tubers. Domestic cooking methods were found to be very efficient in removing bitterness, thus making the bitter yams palatable.

Consumer Product Safety↗
PubMed2005

Aconiti tuber increases plasma nitrite and nitrate levels in humans.

Some herbal medicines, including Aconiti tuber (Aconitum carmichaeli Debeaux, Ranunculaceae), have been recognized as being effective for the treatment of a "peripheral uncomfortable feeling of cold (hie)". We hypothesized that these compounds affect peripheral vascular function via the nitric oxide (NO) system, which leads to recovery from "hie". To answer this question, we investigated Aconiti tuber-induced changes in plasma levels of nitrite (p-NO2-) and nitrate (p-NO3-), final nitric oxide-oxidation products measurable in vivo. After written informed consent was obtained, patients suffering from "hie" were treated with several kinds of kampo (Japanese traditional herbal medicine), selected on the basis of traditional theory. Twenty-four patients took kampo formulas, some included Aconiti tuber (n=11; A-group) and others did not (n=13; C-group), for 4 weeks. p-NO2- and p-NO3- levels were measured before the start and after 4 weeks of treatment. In the A-group, the p-NO2- plus p-NO3- (p-NOx) level was significantly increased at 4 weeks (p=0.04), while that of the C-group was not. There was a statistically significant increase in the p-NOx level of the A-group as compared to the C-group (d.f.=1,22, F=9.38, p=0.006). The results suggest that Aconiti tuber may increase NO production in humans.

Aconitum↗
PubMed2004

Modulation of the cellulose content of tuber cell walls by antisense expression of different potato (Solanum tuberosum L.) CesA clones.

Four potato cellulose synthase (CesA) homologs (StCesA1, 2, 3 and 4) were isolated by screening a cDNA library made from developing tubers. Based on sequence comparisons and the fact that all four potato cDNAs were isolated from this single cDNA-library, all four StCesA clones are likely to play a role in primary cell wall biosynthesis. Several constructs were generated to modulate cellulose levels in potato plants in which the granule-bound starch synthase promoter was used to target the modification to the tubers. The StCesA3 was used for up- and down-regulation of the cellulose levels by sense (SE-StCesA3) and antisense (AS-StCesA3) expression of the complete cDNA. Additionally, the class-specific regions (CSR) of all four potato cellulose synthase genes were used for specific down-regulation (antisense) of the corresponding CesA genes (csr1, 2, 3 and 4). None of the transformants showed an overt developmental phenotype. Sections of tubers were screened for altered cell wall structure by Fourier Transform Infrared microspectroscopy (FTIR) and exploratory Principal Component Analysis (PCA), and those plants discriminating from WT plants were analysed for cellulose content and monosaccharide composition. Several transgenic lines were obtained with mainly decreased levels of cellulose. These results show that the cellulose content in potato tubers can be reduced down to 40% of the WT level without affecting normal plant development, and that constructs based on the CSR alone are specific and sufficient to down-regulate cellulose biosynthesis.

Cell Wall↗