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PubMed1994

Biosynthesis of defense-related proteins in transformed root cultures of Trichosanthes kirilowii Maxim. var japonicum (Kitam.).

We have established transformed ("hairy") root cultures from Trichosanthes kirilowii Maxim. var japonicum Kitam. (Cucurbitaceae) and four related species to study the biosynthesis of the ribosome-inactivating protein trichosanthin (TCN) and other root-specific defense-related plant proteins. Stable, fast-growing root clones were obtained for each species by infecting in vitro grown plantlets with Agrobacterium rhizogenes American Type Culture Collection strain 15834. Each species accumulated reproducibly a discrete protein pattern in the culture medium. Analysis of the extracellular proteins from T. kirilowii var japonicum root cultures showed differential protein accumulation in the medium during the time course of growth in batch cultures. Maximum protein accumulation, approaching 20 micrograms/mL, was observed at mid-exponential phase, followed by a degradation of a specific protein subset that coincided with the onset of stationary phase. Two major extracellular proteins and one intracellular protein, purified by ion-exchange and reverse-phase high-performance liquid chromatography, were identified as class III chitinases (EC 3.2.1.14) based on N-terminal amino acid sequence and amino acid composition homologies with other class III chitinases. The Trichosanthes chitinases also showed reactivity with a cucumber class III chitinase antiserum and chitinolytic activity in a glycol chitin gel assay. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot analysis of intracellular proteins showed that normal and transformed T. kirilowii var japonicum roots accumulated only low levels of TCN (approximately 0.5% total soluble protein). Storage roots from the plant displayed protein and antigen patterns different from root cultures and produced TCN as the dominant protein. Roots undergoing secondary growth and differentiation exhibited patterns similar to those of storage roots, including increased TCN levels, indicating that high production of TCN is associated with induction of secondary growth in roots.

Amino Acid Sequence↗
PubMed2000

Localization of a Nod factor-binding protein in legume roots and factors influencing its distribution and expression.

The roots of the legume Dolichos biflorus contain a lectin/nucleotide phosphohydrolase (Db-LNP) that binds to the Nod factor signals produced by rhizobia that nodulate this plant. In this study we show that Db-LNP is differentially distributed along the surface of the root axis in a pattern that correlates with the zone of nodulation of the root. Db-LNP is present on the surface of young and emerging root hairs and redistributes to the tips of the root hairs in response to treatment of the roots with a rhizobial symbiont or with a carbohydrate ligand. This redistribution does not occur in response to a non-symbiotic rhizobial strain or a root pathogen. Db-LNP is also present in the root pericycle where its level decreases upon initiation of nodule formation. Maximum levels of Db-LNP are found in 2-d-old roots, and the expression of this root protein is increased when the plants are grown in the absence of NO(3)(-) and NH(4)(+). These results support the possibility that Db-LNP is involved in the initiation of the Rhizobium legume symbiosis.

Bradyrhizobium↗
PubMed2000

Pericycle cell proliferation and lateral root initiation in Arabidopsis.

In contrast with other cells generated by the root apical meristem in Arabidopsis, pericycle cells adjacent to the protoxylem poles of the vascular cylinder continue to cycle without interruption during passage through the elongation and differentiation zones. However, only some of the dividing pericycle cells are committed to the asymmetric, formative divisions that give rise to lateral root primordia (LRPs). This was demonstrated by direct observation and mapping of mitotic figures, cell-length measurements, and the histochemical analysis of a cyclin-GUS fusion protein in pericycle cells. The estimated duration of a pericycle cell cycle in the root apical meristem was similar to the interval between cell displacement from the meristem and the initiation of LRP formation. Developmentally controlled LRP initiation occurs early, 3 to 8 mm from the root tip. Thus the first growth control point in lateral root formation is defined by the initiation of primordia in stochastic patterns by cells passing through the elongation and young differentiation zones, up to where lateral roots begin to emerge from the primary root. Therefore, the first growth control point is not restricted to a narrow developmental window. We propose that late LRP initiation is developmentally unrelated to the root apical meristem and is operated by a second growth control point that can be activated by environmental cues. The observation that pericycle cells divide and lateral root primordia form without intervening mitotic quiescence suggests that lateral organ formation in roots and shoots might not be as fundamentally different as previously thought.

Arabidopsis↗
PubMed2002

Regulation of root hair initiation and expansin gene expression in Arabidopsis.

The expression of two Arabidopsis expansin genes (AtEXP7 and AtEXP18) is tightly linked to root hair initiation; thus, the regulation of these genes was studied to elucidate how developmental, hormonal, and environmental factors orchestrate root hair formation. Exogenous ethylene and auxin, as well as separation of the root from the medium, stimulated root hair formation and the expression of these expansin genes. The effects of exogenous auxin and root separation on root hair formation required the ethylene signaling pathway. By contrast, blocking the endogenous ethylene pathway, either by genetic mutations or by a chemical inhibitor, did not affect normal root hair formation and expansin gene expression. These results indicate that the normal developmental pathway for root hair formation (i.e., not induced by external stimuli) is independent of the ethylene pathway. Promoter analyses of the expansin genes show that the same promoter elements that determine cell specificity also determine inducibility by ethylene, auxin, and root separation. Our study suggests that two distinctive signaling pathways, one developmental and the other environmental/hormonal, converge to modulate the initiation of the root hair and the expression of its specific expansin gene set.

Arabidopsis↗
PubMed2005

PGP4, an ATP binding cassette P-glycoprotein, catalyzes auxin transport in Arabidopsis thaliana roots.

Members of the ABC (for ATP binding cassette) superfamily of integral membrane transporters function in cellular detoxification, cell-to-cell signaling, and channel regulation. More recently, members of the multidrug resistance P-glycoprotein (MDR/PGP) subfamily of ABC transporters have been shown to function in the transport of the phytohormone auxin in both monocots and dicots. Here, we report that the Arabidopsis thaliana MDR/PGP PGP4 functions in the basipetal redirection of auxin from the root tip. Reporter gene studies showed that PGP4 was strongly expressed in root cap and epidermal cells. PGP4 exhibits apolar plasma membrane localization in the root cap and polar localization in tissues above. Root gravitropic bending and elongation as well as lateral root formation were reduced in pgp4 mutants compared with the wild type. pgp4 exhibited reduced basipetal auxin transport in roots and a small decrease in shoot-to-root transport consistent with a partial loss of the redirective auxin sink in the root. Seedlings overexpressing PGP4 exhibited increased shoot-to-root auxin transport. Heterologous expression of PGP4 in mammalian cells resulted in 1-N-naphthylthalamic acid-reversible net uptake of [3H]indole-3-acetic acid. These results indicate that PGP4 functions primarily in the uptake of redirected or newly synthesized auxin in epidermal root cells.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed1994

The effect of storage time on the degree of dye leakage of root-end filling materials.

The quality of the apical seal obtained by root-end filling materials generally has been assessed by the extent of dye penetration. Most investigators have allowed the root-end filling materials to set before they are tested. The purpose of this study was to evaluate if storage time had any influence on the amount of dye leakage. Seventy extracted single rooted human anterior teeth were used. After removal of their anatomical crowns, the roots were instrumented, filled with gutta-percha and Roth's sealer, and subjected to test conditions. The roots were coated with two coats of nail polish. Root ends were resected and class I cavities prepared. Sixty class I root-end cavities were filled with with either amalgam, super EBA, or IRM (20 each). Ten roots in each group were placed immediately into India ink for 48 h, the other 10 were stored for 24 h before placement into the ink. Ten roots (five per group) were used as positive (gutta-percha, no sealer) and negative (bone wax) controls. The roots were then demineralized, linear dye leakage was measured, and the data were evaluated statistically using ANOVA. Super EBA, and IRM showed significantly less dye leakage than amalgam (P < 0.0001). No significant difference existed between Super EBA and IRM. Storage time had no significant influence on the amount of dye leakage observed in this study.

Analysis of Variance↗
PubMed1996

Sterilization of infected root-canal dentine by topical application of a mixture of ciprofloxacin, metronidazole and minocycline in situ.

The aim of this study was to observe the potential of a mixture of ciprofloxacin, metronidazole and minocycline to kill bacteria in the deep layers of root canal dentine in situ. After the crowns of extracted teeth had been removed, the drug combination (0.5 mg of each drug), or sterile saline, as the control, was placed in the root canals which had been previously irrigated ultrasonically with G4M EDTA. The penetration and bactericidal efficacy were estimated by various procedures as follows. (1) A cell suspension of E. coli was placed into small cavities prepared parallel to the root canals on the cut planes of nine single-rooted teeth. The teeth were then entirely covered with blue inlay wax. At time 0, and at 5h, 24h and 48h after the drug combination had been applied, cells of E. coli were recovered from the cavities by washing the cavities several times with sterile saline solution, and were cultured on the surfaces of heart-infusion (HI) agar plates. Total colony-forming units were tuen counted. Bacterial recoveries decreased with time, and no bacteria were recovered 48 h after application of the drug combination, while bacteria survived in all cases with the controls, (2) After the drug combination or sterile saline had been placed into and sealed in the root canal with blue inlay wax, the teeth were placed into HI agar plates where cells of E. coli had been inoculated. After culturing, a clear zone caused by the inhibition of bacterial growth was observed around the teeth, but not in the control experiment. (3) After sampling infected root dentine of 12 freshly extracted teeth as positive controls, the drug combination (0.5 mg each) was placed in the root canals. No bacteria were recovered from the infected dentine of the root canal wall 24 h after application of the drug combination, except in one case in which a few bacteria were recovered. On the basis of these results, penetration through dentine and antibacterial efficacy of the drug combination can be expected against bacteria infecting the dentine of the root canal wall in situ when the drugs were placed in root canals which had been irrigated ultrasonically.

Anti-Bacterial Agents↗
PubMed2005

Calculus-like deposit on the apical external root surface of teeth with post-treatment apical periodontitis: report of two cases.

AIM: To report two cases in which calculus-like material was found on external root surfaces of (i) an extracted root and (ii) an apicected part of a root, both of which were removed due to post-treatment refractory apical periodontitis. SUMMARY: In each case, there was a fistulous tract, which did not heal after conventional root canal treatment. The first case did not heal even after apical surgery, and subsequent tooth extraction revealed calculus-like material on a root surface of complex anatomy. The second case showed radiographic signs of healing after apicectomy. Histology of the apical biopsy revealed a calculus-like material on the external surface of the root apex. It is suggested that the presence of calculus on the root surfaces of teeth with periapical lesions may contribute towards the aetiology of failure. KEY LEARNING POINTS: Biofilm on the external root surface has been implicated in the failure of apical periodontitis to heal, despite adequate root canal treatment. Calculus-like material was found, in two cases, on the root surface of teeth with post-treatment apical periodontitis, where the only communication externally was a sinus tract.

Adult↗
PubMed2005

The isthmuses of the mesial root of mandibular molars: a micro-computed tomographic study.

AIM: To investigate the prevalence of root canal isthmuses in the apical 5 mm of the mesial root of mandibular molars by means of micro-computed tomography (MCT) and to describe the morphology of the isthmuses. METHODOLOGY: Twenty extracted mandibular first molars that had fully formed roots were selected. The mesial roots were sectioned from the distal roots and subjected to MCT. The number of sections showing isthmuses in each of the apical 5 mm of the root canals was recorded. In each one of the apical 5 mm of the 20 roots examined, 40 sections were observed, giving a total of 800 sections observed in each group. Data were analysed as a contingency table using the chi-square statistic to test the null hypothesis that location of the sections in each of the apical 5 mm and presence of the isthmus were independent. RESULTS: Isthmuses were found to be present at all levels with prevalence figures between 17.25 and 50.25%. The chi-square test indicated a significant difference in the distribution of isthmuses with section (P = 0.001). It was found that sections in the first millimetre from the apex had fewer isthmuses than expected and that sections in the third millimetre from the apex had more isthmuses than expected under the null hypothesis. Calcifications were found to be present in most isthmuses, occasionally lateral canals originated from the central part of the isthmuses. CONCLUSIONS: Isthmuses were present in the vast majority of roots observed. The third millimetre from the apex showed more isthmuses than expected. The results of clinical and surgical endodontic procedures performed in the mesial root of mandibular molars may be affected by this aspect of the root canal anatomy.

Dental Pulp Cavity↗
PubMed2005

Survival of Enterococcus faecalis in root canals ex vivo.

AIM: The hypotheses tested in this study were that: (i) Enterococcus faecalis can survive long-term entombment in root filled teeth without additional nutrients, (ii) initial cell density influences the survival of E. faecalis in instrumented root canals and (iii) gelatinase-production capacity influences the survival of E. faecalis in root canals. METHODOLOGY: The root canals of 150 extracted single canal teeth were instrumented to apical size 60 and divided into six groups of 25. Within each group 10 canals were inoculated with either gelatinase-producing E. faecalis OG1-S and the other 10 with its gelatinase-defective mutant E. faecalis OG1-X. Five canals per group were kept as uninoculated controls. The root canals in groups 1 and 2 were inoculated with 10(6) bacteria, incubated for 48 h at 37 degrees C then filled with gutta-percha and zinc-oxide eugenol sealer. Root canals were inoculated with 10(6), 10(5), 10(4) and 10(3) bacteria in groups 3-6, respectively, and left unfilled. All teeth were sealed coronally with glass-ionomer cement. After 6- (groups 1, 3-6) and 12-month (group 2) incubation at 37 degrees C in 100% humidity, root fragments were analysed for presence of E. faecalis, using culture, polymerase chain reaction and histological methods. RESULTS: Viable E. faecalis was recovered from all root filled teeth and from 95-100% of unfilled inoculated teeth. Initial cell density and gelatinase production did not influence the recovery of viable E. faecalis (P > 0.05; chi-square test). Enterococcus faecalis 16S rRNA gene products were present in all inoculated teeth and absent in all noninoculated controls. Dentinal tubule infection was evident under light microscopy in sections from inoculated teeth after 48-h, 6- and 12-month incubation. CONCLUSIONS: Enterococcus faecalis inoculated into root canals maintained viability for 12-months ex vivo. The clinical implications are that viable E. faecalis entombed at the time of root filling could provide a long-term nidus for subsequent infection.

Adaptation, Physiological↗
PubMed2006

Cytotoxicity of Epiphany and Resilon with a root model.

AIM: To record the cytotoxicity of Resilon and Epiphany (Pentron clinical technologies, Wallingford, CT, USA) using a root model. METHODOLOGY: Thirty teeth with single roots were sectioned at the enamel-cementum junction, the root canals prepared and each root then sterilized before filling with the lateral condensation technique using one of three filling materials (n = 10 per group): Resilon and Epiphany, Sealite (Septodont, Pierre Rolland, Merignac, France) and gutta-percha, Roekoseal Automix (Coltène/Whaledent, Langenau, Germany) and gutta-percha. The roots were stored at 37 degrees C in an incubator to allow for setting of the root filling materials. The apices of the roots were dipped in 1 mL of MEM culture medium for 1, 2, 7 and 30 days renewing the medium every day. After 24 h contact between the medium and the filled roots, the medium was used to measure the cytotoxicity on mouse fibroblasts L 929 with the MTT assay that recorded the mitochondrial activity of the target cells. An additional test according to ISO 10993-5 standards was undertaken to compare Resilon and Epiphany. RESULTS: The root model showed no statistically significant differences between the sealers at 7 and 30 days (NS). Epiphany and Resilon were the most cytotoxic materials at 1 and 2 days (P < 0.001). Unlike Epiphany, Resilon was not cytotoxic when tested according to ISO 10993-5 standards. CONCLUSIONS: The cytotoxicity of Resilon + Epiphany, due mainly to Epiphany, decreased after 2 days to reach a level comparable with commonly used root canal sealers.

Analysis of Variance↗
PubMed2007

Calcium hydroxide barrier over the apical root-end of a type III dens invaginatus after endodontic and surgical treatment.

AIM: To report the simultaneous endodontic and surgical treatment of a tooth associated with Oehlers type III dens invaginatus and a persistent periapical lesion, which comprised root-end resection, root-end filling and application of a calcium hydroxide barrier placed on the resected dentine surface. SUMMARY: Three root canals were identified in a tooth with a type III dens invaginatus, which presented with a necrotic pulp, wide foraminal opening and extensive periapical lesion, and with a previous history of acute abscess, intracanal exudate and fistula. After root canal preparation followed by intracanal application of calcium hydroxide pastes, the clinical-pathological status persisted. After periapical curettage and root-end resection, the root canals were filled, followed by root-end filling with Sealer 26 mixed with zinc oxide powder to a clay-like consistency. Calcium hydroxide paste was then applied over the exposed dentinal surface forming a covering over the root apex. At the 20-month follow-up examination the patient had no symptoms and no fistula; advanced periapical bone repair was obvious on the radiograph. KEY LEARNING POINTS: Because of the variable morphology and extent of invagination, type III dens invaginatus represents a challenge for conventional treatment, often leading to the need for a surgical approach. Sealer 26 thickened with zinc oxide powder provided satisfactory clinical properties for use as a root-end filling material. Application of a calcium hydroxide barrier over the resected root-end is a potential treatment option to encourage tissue repair.

Adolescent↗
PubMed2004

Light induces phenylpropanoid metabolism in Arabidopsis roots.

Experiments have shown that many phenylpropanoid genes are highly expressed in light-grown Arabidopsis roots. Studies employing reporter gene constructs have indicated that the expression of these genes is localized not only to the lignifying root vasculature, but also to non-lignifying tissues, such as the root cortex, suggesting that the proteins encoded by these genes may be involved in aspects of phenylpropanoid metabolism other than lignification. Consistent with this hypothesis, roots of etiolated and soil-grown plants contain almost no soluble phenylpropanoids, but exposure to light leads to the accumulation of flavonoids, as well as high levels of coniferin and syringin (coniferyl and sinapyl-4-O-glycosides), compounds not previously reported to be accumulated in Arabidopsis. To elucidate the mechanism by which light induces root secondary metabolism, extracts of mutants defective in light perception and light responses were analyzed for phenylpropanoid content. The results of these assays showed that phytochrome (PHY)B and cryptochrome (CRY)2 are the primary photoreceptors involved in light-dependent phenylpropanoid accumulation, and that the hypocotyl elongated (HY5) transcription factor is also required for this response. The presence of phenylpropanoids in etiolated roots of cop (constitutively photomorphogenic)1, cop9, and det (de-etiolated)1 mutants indicate that the corresponding wild-type genes are required to repress root phenylpropanoid biosynthesis in the absence of light. Biochemical analysis of root cell walls and analysis of phenylpropanoid gene expression suggest that coniferin and syringin accumulation may be the result of both increased biosynthesis and decreased conversion of these compounds into other phenylpropanoid end products. Finally, our data suggest that the accumulation of coniferin, syringin, and flavonoids in Arabidopsis roots is a high-irradiance response (HIR), and suggest that comparative analysis of light- and dark-grown Arabidopsis roots may provide new insights into both phenylpropanoid biosynthesis and light signaling in plants.

Arabidopsis↗
PubMed1988

The influence of calcium and pH on growth in primary roots of Zea mays.

We investigated the interaction of Ca2+ and pH on root elongation in Zea mays L. cv. B73 x Missouri 17 and cv. Merit. Seedlings were raised to contain high levels of Ca2+ (HC, imbibed and raised in 10 mM CaCl2) or low levels of Ca2+ (LC, imbibed and raised in distilled water). In HC roots, lowering the pH (5 mM MES/Tris) from 6.5 to 4.5 resulted in strong, long-lasting growth promotion. Surprisingly, increasing the pH from 6.5 to 8.5 also resulted in strong growth promotion. In LC roots acidification of the medium (pH 6.5 to 4.5) resulted in transient growth stimulation followed by a gradual decline in the growth rate toward zero. Exposure of LC roots to high pH (pH shift from 6.5 to 8.5) also promoted growth. Addition of EGTA resulted in strong growth promotion in both LC and HC roots. The ability of EGTA to stimulate growth appeared not to be related to H+ release from EGTA upon Ca2+ chelation since, 1) LC roots showed a strong and prolonged response to EGTA, but only a transient response to acid pH, and 2) promotion of growth by EGTA was observed in strongly buffered solutions. We also examined the pH dependence of the release of 45Ca2+ from roots of 3-day-old seedlings grown from grains imbibed in 45Ca2+. Release of 45Ca2+ from the root into agar blocks placed on the root surface was greater the more acidic the pH of the blocks. The results indicate that Ca2+ may be necessary for the acid growth response in roots.

Buffers↗
PubMed2005

Forced retraction of spinal root injury enhances activation of p38 MAPK cascade in infiltrating macrophages.

Root-rupture injury is a type of preganglionic brachial plexus injury resulting from traction force, where a small section of the spinal root is usually left behind. We have established experimental models of both root-rupture injury with traction force and rhizotomy without traction force in rats and we examined the activation of microglia/ macrophages in both conditions. LGP107 and LGP96, which are rat homologs of lysosome-associated membrane proteins, were most useful as immunohistochemical markers of mononuclear phagocytes. The metabolic activation of macrophages was analyzed by immunohistochemistry with a series of antibodies against tumor necrosis factor-alpha (TNF-alpha), cathepsin B, p38 mitogen-activated protein kinase (MAPK), and mitogen-activated kinase kinase 3 (MKK3). Both root-rupture injury and rhizotomy rapidly induced the aggregation of numerous macrophages from the injured dorsal root to the dorsal funiculus and TNF-alpha was highly expressed by the macrophages in the injured dorsal root at 48 h. Activation of p38 MAPK was preferentially observed in the macrophages at the ruptured dorsal root; however, only slight activation of p38 MAPK was observed at the rhizotomized dorsal root. These findings suggest that traction injury of the spinal root might induce activation of the p38 MAPK cascade and production of TNF-alpha in the infiltrating macrophages, both of which might participate in aggravation of the root injury.

Animals↗
PubMed2005

Morphometric analysis of root shape.

Alterations in the root shape in plant mutants indicate defects in hormonal signalling, transport and cytoskeleton function. To quantify the root shape, we introduced novel parameters designated vertical growth index (VGI) and horizontal growth index (HGI). VGI was defined as a ratio between the root tip ordinate and the root length. HGI was the ratio between the root tip abscissa and the root length. To assess the applicability of VGI and HGI for quantification of root shape, we analysed root development in agravitropic Arabidopsis mutants. Statistical analysis indicated that VGI is a sensitive morphometric parameter enabling detection of weak gravitropic defects. VGI dynamics were qualitatively similar in auxin-transport mutants aux1, pin2 and trh1, but different in the auxin-signalling mutant axr2. Analysis of VGI and HGI of roots grown on tilted plates showed that the trh1 mutation affected downstream cellular responses rather than perception of the gravitropic stimulus. All these tests indicate that the VGI and HGI analysis is a versatile and sensitive method for the study of root morphology.

Arabidopsis↗
PubMed2006

Suites of root traits differ between annual and perennial species growing in the field.

Here, we tested whether root traits associated with resource acquisition and conservation differed between life histories (annuals, perennials) and families (Fabaceae, Asteraceae and Poaceae). Root topology, morphology, chemistry and mycorrhizal colonization were measured on whole root systems of 18 field-grown herbaceous species grown and harvested in central Argentina. Annuals differed from perennials in several root traits important in resource uptake and conservation. They exhibited higher specific root length (SRL), root nitrogen concentration (RNC) and mycorrhizal colonization but had lower root tissue density (RTD) than perennials. They did not differ in topology or construction cost. These differences were consistent among families. Families differed only in a few root traits known to be strongly associated with certain lineages such as topology and nitrogen concentration. There was a strong parallel between root traits and analogous leaf traits described in the literature for annuals and perennials. Our results suggest the existence at the root level of an acquisitive vs conservative syndrome consistent among families similar to that previously reported for above-ground traits.

Argentina↗
PubMed2006

Heavy metals in white lupin: uptake, root-to-shoot transfer and redistribution within the plant.

The translocation of manganese (Mn), nickel (Ni), cobalt (Co), zinc (Zn) and cadmium (Cd) in white lupin (Lupinus albus cv. Amiga) was compared considering root-to-shoot transport, and redistribution in the root system and in the shoot, as well as the content at different stages of cluster roots and in other roots. To investigate the redistribution of these heavy metals, lupin plants were labelled via the root for 24 h with radionuclides and subsequently grown hydroponically for several weeks. 54Mn, 63Ni and 65Zn were transported via the xylem to the shoot. 63Ni and 65Zn were redistributed afterwards via the phloem from older to younger leaves, while 54Mn remained in the oldest leaves. A strong retention in the root was observed for 57Co and 109Cd. Cluster roots contained higher concentrations of all heavy metals than noncluster roots. Concentrations were generally higher at the beginning of cluster root development (juvenile and immature stages). Mature cluster roots also contained high levels of 54Mn and 57Co, but only reduced concentrations of 63Ni, 65Zn and 109Cd.

Autoradiography↗