Maximum Label Research Articles

RESIDUAL EFFECTIVENESS OF PYRETHROID-TREATED FOLIAGE AGAINST ADULT AEDES ALBOPICTUS AND CULEX QUINQUEFASCIATUS IN SCREENED FIELD CAGES

The residual effectiveness of pyrethroid-treated foliage as a barrier against female Aedes albopictus and Culex quinquefasciatus was evaluated in large screened cage field tests. Individual potted southern wax myrtle, Myrica certifica, plants were treated with either Aqua Reslin 20 + 20 emulsifiable concentrate (EC) (20% active ingredient [AI] permethrin + 20% [AI] piperonyl butoxide), Permanone EC (10% [AI] permethrin), or Suspend suspension concentrate (SC) (4.75% [AI] deltamethrin) at maximum label rates. Generally, Aqua Reslin provided -83% overall reduction of mosquitoes during the week of treatment. After that time, mosquito reduction decreased to <50% and continued at this level for the remainder of the 12-wk study. Mosquito knockdown/mortality from excised Aqua Reslin-treated leaves revealed that this formulation quickly lost effectiveness after the initial week of treatment. Plants treated with Permanone provided =70% and 64% overall reduction during the week of, and first week after treatment, respectively. Knockdown/mortality from excised leaf bioassays of leaves treated with this formulation fell below 50% 2 wk after treatment where it remained for the rest of the study. Suspend generally provided 70 to approximately 80% overall mosquito reduction during the first 4 weeks after treatment and decreased to <50% thereafter. Residues on treated leaves provided >95% overall knockdown/mortality throughout the study and was not correlated with weekly reduction in the field cages. We believe this disparity was in part attributed to new plant growth. Newly formed leaves probably provided nontoxic surfaces for mosquito harborage.

Determination of long-chain fatty acids in bryophyte plants extracts by HPLC with fluorescence detection and identification with MS

A sensitive method for the determination of long-chain fatty acids (LCFAs) (>C20) using 1-[2-( p-toluenesulfonate)-ethyl]-2-phenylimidazole-[4,5-f]-9,10-phenanthrene (TSPP) as tagging reagent with fluorescence detection and identification with post-column APCI/MS has been developed. The LCFAs in bryophyte plant samples were obtained based on distillation extraction with 1:1 (v/v) chloroform/methanol as extracting solvent. TSPP could easily and quickly label LCFAs at 90 °C in the presence of K 2CO 3 catalyst in DMF. Eleven free LCFAs from the extracts of bryophyte plants were sensitively determined. Maximal labeling yields close to 100% were observed with a five-fold excess of molar reagent. Separation of the derivatized fatty acids exhibited a good baseline resolution in combination with a gradient elution on a reversed-phase Eclipse XDB-C 8 column. Calculated detection limits from 1.0 pmol injection, at a signal-to-noise ratio of 3, were 26.19–76.67 fmol. Excellent linear responses were observed with coefficients of >0.9996. Good compositional data were obtained from the analysis of the extracted LCFAs containing as little as 0.2 g of bryophyte plant samples. Therefore, the facile TSPP derivatization coupled with HPLC/APCI/MS analysis allowed the development of a highly sensitive method for the quantitation of trace levels of LCFAs from biological and natural environmental samples.

A sequential double labeling technique for studying changes in motoneuronal projections to muscle following nerve injury and reinnervation

The purpose of this study was to develop an anatomical technique that could directly demonstrate the motoneuron projections to the muscle both before injury and again following reinnervation. Investigation focused on the identification of a long-term retrograde fluorescent tracer that would label original motoneurons and persist long enough for reinnervating motoneurons to become labeled by a second fluorescent tracer. True Blue (TB) was evaluated as a potential long-term tracer, Fluoro-ruby (FR) and Fluoro-emerald (FE) were tested as potential short-term tracers in 45 adult Sprague-Dawley rats. In the initial phase of the study, TB was injected into the tibialis anterior (TA) muscle in 16 rats and sacrificed 1 week to 6 months later, to study its persistence. During the second stage, a short-term tracer was injected into the TA muscles bilaterally in 15 rats with survival time ranging from 4 to 28 days. Sequential double labeling was subsequently performed using the combination of TB and FR in 14 rats. The number and brightness of TB cells did not change over 6 months time, a period sufficient for complete reinnervation. FR and FE showed maximum labeling of motoneurons at 1 week after tracer application. In the double labeling study, we could easily distinguish double-labeled cells from those labeled only by TB or FR. These results suggest that sequential double labeling of TB and FR is a valuable method for long-term muscle reinnervation studies.

Heavy Water Labeling of Keratin as a Non-Invasive Biomarker of Skin Turnover In Vivo in Rodents and Humans

Measurement of skin turnover has been problematic in humans. Heavy water (2H2O) labeling has recently been developed as a safe, simple method to study in vivo kinetics of many biosynthetic processes, including DNA and protein synthesis. Here, we apply this approach to the measurement of 2H incorporation into skin keratin and show close agreement between keratin and keratinocyte turnover data in the epidermis of rodents. Elevated turnover rates of both keratin and keratinocytes were observed in the epidermis of the flaky skin mouse, although topical treatments effective in human psoriasis had no effect on either turnover rate in these mice. In humans, keratin turnover was monitored non-invasively by serial tape stripping during and after 2H2O labeling. Kinetic data were consistent with previous estimates of epidermal turnover, with a lag time of 18 days before label appeared at the skin surface and a transit time of 4-5 weeks. Variability in skin keratin turnover rates was present among healthy individuals. In summary, 2H2O labeling of skin keratin represents a non-invasive approach for assessing skin turnover dynamics in pre-clinical models and in human subjects.

New technology of technetium-99 labeled antimony trisulfide colloid intended for sentinel lymph node imaging

99mTc-Labeled antimony trisulfide (ATS) colloid is a radiopharmaceutical that can be useful in evaluation of the patients with breast cancer. The existing 99mTc-ATS preparation method involves a lengthy procedure of boiling on a water bath; the recommended quality control procedure is also rather laborious and time consuming. Using a microwave oven, the heating time necessary to provide a maximum labeling efficiency can be reduced to 15 sec. A new minipaper chromatography (MPC) system involving two developing solvents has been developed to analyze the radiochemical purity (RCP) of the labeled preparation. The conventional thin layer chromatography (TLC) system involving the use of an Al2O3 coated plate requires an average time of 38.4 ± 0.21 min (n = 30) for complete drying and development, whereas the new MPC system has an average development time of 4.1 ± 0.32 min (n = 20). In the RCP evaluation, the MPC and TLC methods give closely correlated results (r = 0.94). The combined use of the proposed microwave heating technique and the rapid MPC quality control system facilitate the emergency use of 99mTc-ATS colloid diagnostics.

Compact Labeling Scheme for Ancestor Queries

We consider the following problem. Given a rooted tree $T$, label the nodes of $T$ in the most compact way such that, given the labels of two nodes $u$ and $v$, one can determine in constant time, by looking only at the labels, whether $u$ is ancestor of $v$. The best known labeling scheme is rather straightforward and uses labels of length at most $2\log_2 n$ bits each, where $n$ is the number of nodes in the tree. Our main result in this paper is a labeling scheme with maximum label length $\log_2 n + \Oh(\sqrt{\log n})$. Our motivation for studying this problem is enhancing the performance of web search engines. In the context of this application each indexed document is a tree, and the labels of all trees are maintained in main memory. Therefore even small improvements in the maximum label length are important.

Preparation of 99mTc Labeled Vitamin C (Ascorbic Acid) and Biodistribution in Rats

The aim of this study was to label ascorbic acid with (99m)Tc and to investigate its radiopharmaceutical potential in rats. Ascorbic acid was labeled with (99m)Tc using the stannous chloride method. The radiochemical purity of [(99m)Tc]ascorbic acid ((99m)Tc-AA) was determined by RTLC, paper electrophoresis, and RHPLC methods. The labeling yield was found to be 93+/-5.0%. The maximum labeling yield of (99m)Tc-AA was determined at pH 5 and 25 degrees C. The biodistribution studies related to (99m)Tc-AA were done in male albino Wistar rats. (99m)Tc-AA, which has a specific activity of 13.02 GBq/mmol, was administered into the tail vein of the rats. The rats were sacrificed at 15, 30, 60, and 120 min after the injection by heart puncture under ether anaesthesia. The organs were weighed after removal. Their activities were counted using a Cd(Te) detector equipped with a RAD 501 count system. The %ID/g (% of injected dose per gram of tissue weight) in each organ and in blood was calculated. Maximum uptake of (99m)Tc-AA was observed in prostate and kidneys at the 60th min. (99m)Tc-AA may be a promising radiopharmaceutical for the imaging of prostate and kidneys.

Formulation of Technetium-99m Labeled Antimony Trisulfide Colloid, Intended for Sentinel Lymph Node Imaging, Using New Techniques

99mTc-Antimony trisulfide colloid is a radiopharmaceutical that can be useful in evaluation of patients with breast cancer. The current preparation method requires prolonged boiling on a water bath, and the recommended quality control procedure is cumbersome and time-consuming. With a microwave oven, the heating time necessary to provide the maximal labeling efficiency is reduced to 15 s. A new mini-paper chromatography (MPC) system has been developed to analyze the radiochemical purity (RCP) of the labeled preparation involving two different developing solvents. The recommended thin-layer chromatography (TLC) system involving the use of an Al2O3-coated plate requires an average time for drying and development of 38.4 × 0.21 min (n = 30) to complete, whereas the new MPC system has an average developing time of 4.1 ± 0.32 min (n = 20). For RCP values, the MPC and TLC data are well consistent (r = 0.94). The combined use of the microwave oven heating and the quick quality control system will facilitate the rapid emergency use of 99mTc-antimony trisulfide colloid.

Phytotoxicity of TriStar Formulations on Fully Colored Poinsettia Cultivars

Late season control of whitefly is problematic in many production ranges as systemic insecticides may not provide full season control. Most commercially available contact insecticides are not labeled for use on fully colored poinsettia, Euphorbia pulcherrima, plants due to potential phytotoxicity or residue on colored bracts. Recent formulation changes in TriStar make late season applications possible. This study assessed phytotoxicity and residue impacts of two formulations of TriStar on potted poinsettias. On 4 Aug. 2004, rooted cuttings of 47 poinsettia cultivars obtained from four commercial suppliers were transplanted into pots containing a soilless medium. A total of 235 cuttings were used to arrive at five pots per cultivar. Plants were grown using standard production techniques. On 11 Nov. 2004, all plants were fully colored and treated with TriStar 70 WSP or TriStar 30SG at maximum label rates. Phytotoxicity and residue levels were assessed 7 days later using a 1 to 9 visual scale. Overall the formulations had few negative impacts on poinsettias. Phytotociticy ratings were minimal for most cultivars; however, some cultivars, such as `Silverstar Red' expressed an elevated level of phytotoxicity. Dark colored cultivars showed more residue than light colored cultivars. The TirStar 30SG formulation had the least amount of residue. TriStar 30SG may be an acceptable insecticide for late season control of whitefly on poinsettia crops. Producers are cautioned to test cultivars for phytotoxicity before applying to an entire crop as some cultivars are sensitive to TriStar 30SG.

Optimization and Applications of CDAP Labeling for the Assignment of Cysteines

The aim of the study is to provide a methodology for assigning unpaired cysteine residues in proteins formulated in a variety of different conditions to identify structural heterogeneity as a potential cause for protein degradation. 1-Cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) was employed for cyanylating free cysteines in proteins and peptides. Subsequent basic cleavage of the peptide bond at the N-terminal side of the cyanylated cysteines provided direct information about their location. CDAP was successfully employed to a wide variety of labeling conditions. CDAP was reactive between pH 2.0 and 8.0 with a maximum labeling efficiency at pH 5.0. Its reactivity was not affected by excipients, salt or denaturant. Storing CDAP in an organic solvent increased its intrinsic stability. It was demonstrated that CDAP can be employed as a thiol-directed probe to investigate structural heterogeneity of proteins by examining the accessibility of unpaired cysteine residues. CDAP is a unique cysteine-labeling reagent because it is reactive under acidic conditions. This provides an advantage over other sulfhydryl labeling reagents as it avoids potential thiol-disulfide exchange. Optimization of the cyanylation reaction allowed the utilization of CDAP as a thiol-directed probe to investigate accessibility of sulfhydryl groups in proteins under various formulation conditions to monitor structural heterogeneity.

Compact Labeling Scheme for XML Ancestor Queries

XML documents are often viewed as trees (basically the parse tree of the document), and queries over such documents typically test for ancestor relationships among tree nodes. Search engines process such queries using an index structure summarizing the ancestor relations. In the index, each document item (tree node) is identified using some logical id (node label), such that, given two labels, the engine can determine the ancestor relationship between the corresponding nodes. The length of the labels is a main factor of the index size. Therefore, reducing this length, even by a constant factor, is a critical issue. In this work we consider the following problem. Given a rooted XML tree T, label the nodes of T in the most compact way such that given the labels of two nodes, one can determine in constant time, by looking at the labels only, whether one node is an ancestor of the other. Labelings currently being used are all variants of the following interval scheme. Number the leaves say from left to right and label each node with a pair consisting of the numbers of its smallest and largest leaf descendants. An ancestor query then amounts to an interval containment test on the labels. The maximum label length using this scheme is 2 log n, where n is the number of nodes in the tree. (All logarithms in this paper are to base 2.) The focus of this work is finding a scheme that works best in practice on real XML data. We suggest an orthogonal prefix-based approach, where the labeling is such that an ancestor query roughly amounts to testing whether one label is a prefix of the other. We present several new labeling schemes based on this approach and analyze their performance both theoretically and empirically.

Direct fluorescent staining and analysis of proteins on microspheres using CBQCA

General methods for accurate determination of microsphere surface protein loading are needed for applications from protein arrays to molecular assembly studies. Current methods include bulk absorption measurements of stained microspheres or use of known fluorescently tagged binding partners, which limit sensitivity and general applicability, respectively. Microspheres bearing covalently coupled proteins were stained with 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) using different incubation times and dye concentrations to determine optimal staining conditions. The CBQCA fluorescence of microspheres (measured by flow cytometry) bearing known amounts of protein were used to generate standard curves of CBQCA fluorescence response versus the amount of microsphere surface protein. CBQCA was also used to stain noncovalent protein interactions. Maximal labeling was attained within 1 h with 1 mM CBQCA. Linear fluorescence response occurred between 8 x 10(4) and 1 x 10(6) proteins/microsphere. CBQCA staining did not disturb noncovalent protein interactions. We have developed methods using CBQCA and flow cytometry to quickly and simply quantify the amount of protein on the surface of a microsphere. Importantly, this approach could be extended to other formats (e.g., chips). Further, because it does not disturb noncovalent protein-protein interactions, it may be possible to use this approach to detect protein interactions without the use of purified prestained probes.

Tolerance of kidney beans (Phaseolus vulgaris) to bentazon alone and when mixed with imazamox

Tolerance of Montcalm and Redhawk kidney beans to bentazon, imazamox plus fomesafen and imazamox plus bentazon applied postemergence at the maximum label rate in soybeans and twice that rate were studied at two Ontario locations (Exeter and Ridgetown) in 2001 and 2002. There were no differences between the two cultivars in their response to the herbicides tested. At Exeter in 2001, bentazon reduced plant height by 10 and 12% and yield by 33 and 22% at the label and twice that rate, respectively. Yield was also reduced by 11% at Exeter in 2002 at twice the label rate. Imazamox plus fomesafen reduced plant height equally by 14% and yield was reduced by 22 and 30% at the label rate and twice the label rate, respectively, at Exeter in 2001. Imazamox plus bentazon in Exeter in 2001 reduced plant height equally by 8% and yield by 20 and 14% at the label rate and twice the label rate, respectively. There were no negative effects on plant height, dry weight and yield at the other site-yrs. This research suggests that bentazon, imazamox plus fomesafen and imazamox plus bentazon applied postemergence can cause severe crop injury and yield reduction in kidney bean production under certain environmental conditions. Key words: Bentazon, crop tolerance, fomesafen; imazamox, Montcalm, Phaseolus vulgaris, Redhawk.

Function-dependent Conformational Changes of the ABCG2 Multidrug Transporter Modify Its Interaction with a Monoclonal Antibody on the Cell Surface

The human ABCG2 protein is an important primary active transporter for hydrophobic compounds in several cell types, and its overexpression causes multidrug resistance in tumors. A monoclonal antibody (5D3) recognizes this protein on the cell surface. In ABCG2-expressing cells 5D3 antibody showed a saturable labeling and inhibited ABCG2 transport and ATPase function. However, at low antibody concentrations 5D3 binding to intact cells depended on the actual conformation of the ABCG2 protein. ATP depletion or the addition of the ABCG2 inhibitor Ko143 significantly increased, whereas the vanadate-induced arrest of ABCG2 strongly decreased 5D3 binding. The binding of the 5D3 antibody to a non-functional ABCG2 catalytic center mutant (K86M) in intact cells was not affected by the addition of vanadate but still increased with the addition of Ko143. In isolated membrane fragments the ligand modulation of 5D3 binding to ABCG2 could be analyzed in detail. In this case 5D3 binding was maximum in the presence of ATP, ADP, or Ko143, whereas the non-hydrolysable ATP analog, adenosine 5'-(beta,gamma-imido)triphosphate (AMP-PNP), and nucleotide trapping by vanadate decreased antibody binding. In membranes expressing the ABCG2-K86M mutant, ATP, ADP, and AMP-PNP decreased, whereas Ko143 increased 5D3 binding. Based on these data we suggest that the 5D3 antibody can be used as a sensitive tool to reveal intramolecular changes, reflecting ATP binding, the formation of a catalytic intermediate, or substrate inhibition within the transport cycle of the ABCG2 protein.

Labeling Schemes for Small Distances in Trees

We consider labeling schemes for trees, supporting various relationships between nodes at small distance. For instance, we show that given a tree T and an integer k we can assign labels to each node of T such that given the label of two nodes we can decide, from these two labels alone, if the distance between v and w is at most k and, if so, compute it. For trees with n nodes and $k\geq 2$, we give a lower bound on the maximum label length of $\log n + \Omega(\log \log n)$ bits, and for constant k, we give an upper bound of log n + O(log log n). Bounds for ancestor, sibling, connectivity, and bi- and triconnectivity labeling schemes are also presented.

Strawberry Black Root Rot and Berry Yield Are Not Affected by Terbacil Herbicide

The effects of terbacil herbicide on strawberry (Fragaria ×ananassa Duch. `Honeoye') yield and black root rot disease were determined in field plots at two locations in Connecticut over 4 years. Terbacil treatments at up to four times the maximum label dosage caused some temporary foliar chlorosis but did not affect the health of structural or perennial roots and associated feeder roots. Development of secondary root growth (perennial roots) was not influenced by terbacil. Terbacil had no effect on the quantity of lesion nematodes [Pratylenchus penetrans (Cobb) Filip &amp; Schur. Stek.] extracted or the amount of the fungal pathogen Rhizoctonia fragariae Husain and McKeen isolated from strawberry roots. At both locations, R. fragariae was common on plant roots by the fourth year. Terbacil treatments did not affect strawberry yields in terms of number or weight of ripe berries per plot. Our results indicate that terbacil does not contribute to black root rot or decreased yields in `Honeoye' strawberry. Chemical name used: 5-chloro-3-(1,1-dimethylethyl)-6-methyl-2,4-(1H,3H)-pyrimidinedione (terbacil).

Comparative effects of pH and Vision herbicide on two life stages of four anuran amphibian species.

Vision, a glyphosate-based herbicide containing a 15% (weight:weight) polyethoxylated tallow amine surfactant blend, and the concurrent factor of pH were tested to determine their interactive effects on early life-stage anurans. Ninety-six-hour laboratory static renewal studies, using the embryonic and larval life stages (Gosner 25) of Rana clamitans, R. pipiens, Bufo americanus, and Xenopus laevis, were performed under a central composite rotatable design. Mortality and the prevalence of malformations were modeled using generalized linear models with a profile deviance approach for obtaining confidence intervals. There was a significant (p < 0.05) interaction of pH with Vision concentration in all eight models, such that the toxicity of Vision was amplified by elevated pH. The surfactant is the major toxic component of Vision and is hypothesized, in this study, to be the source of the pH interaction. Larvae of B. americanus and R. clamitans were 1.5 to 3.8 times more sensitive than their corresponding embryos, whereas X. laevis and R. pipiens larvae were 6.8 to 8.9 times more sensitive. At pH values above 7.5, the Vision concentrations expected to kill 50% of the test larvae in 96-h (96-h lethal concentration [LC50]) were predicted to be below the expected environmental concentration (EEC) as calculated by Canadian regulatory authorities. The EEC value represents a worst-case scenario for aerial Vision application and is calculated assuming an application of the maximum label rate (2.1 kg acid equivalents [a.e.]/ha) into a pond 15 cm in depth. The EEC of 1.4 mg a.e./L (4.5 mg/L Vision) was not exceeded by 96-h LC50 values for the embryo test. The larvae of the four species were comparable in sensitivity. Field studies should be completed using the more sensitive larval life stage to test for Vision toxicity at actual environmental concentrations.

First evidence of Dinophysistoxin-1 ester and carcinogenic polycyclic aromatic hydrocarbons in smoked bivalves collected in the Patagonia fjords

Diarrhetic shellfish poisoning (DSP) is a gastrointestinal disease caused by fat-soluble polyether toxins produced by dinoflagellates and accumulated in shellfish. Up to the present, only four fat-soluble polyethers have been known as diarrhetic shellfish toxins. Among them,*** Okadaic acid, Dinophysistoxin-1, Dinophysistoxin-2 and Dinophysistoxin-3. Outbreaks associated with DSP have occurred in the Chilean Patagonia fjords since 1970. Native people, who live in small communities close to the southern fjords, smoke fresh shellfish. During this popular smoking procedure, they impregnate the shellfish with polycyclic aromatic hydrocarbons, as incomplete combustion products, which are potent carcinogenic compounds, this product is sold in local markets without phycotoxins analysis or inclusion in any monitoring program. The present paper shows, DSP phycotoxins quantitation, using high performance liquid chromatography with fluorescent and mass spectrometric detection and the measurements of polycyclic aromatic hydrocarbons by gas chromatography with mass detection, in smoked shellfish samples. The presence of Dinophysistoxin-3, the Dinophysistoxin-1 ester (7- O-acyl-derivatives of dinophysistoxin-1), was assessed in all shellfish samples analyzed. The 7-OH in Dinophysistoxin-1 was esterified with palmitic fatty acid. The shellfish meat contains seven polycyclic aromatic hydrocarbons, among them fluoranthene, phenanthrene, anthracene, pyrene and benzo[a]pyrene. The gas chromatography-mass spectrometry analysis showed four of the six most frequent carcinogenic polycyclic aromatic hydrocarbons reported. The content of benzo[a]pyrene in the Razor Clam and Ribbed Mussel were 78.61 and 4.94 ng/g of shellfish dry weight, respectively. In both cases the benzo[a]pyrene amounts were greater than the acceptable tolerance limits of 1 μg/kg of sample. The Razor Clam samples also show amount further above the maximum label regulated by FAO/WHO (10 μg/kg). The presence of both type of compounds in the smoked shellfish samples analyzed, correspond to a dangerous combination, where the polycyclic aromatic hydrocarbons are carcinogenic compounds by themselves and DTX-1, is a potent tumor promoter.

256.Development of a label retaining cell method to identify stem cells in mouse endometrium

Epithelial and stromal stem/progenitor cells have been demonstrated in human endometrium by their clonogenic activity (1). To understand the regulation and growth kinetics of endometrial stem cells (SCs), an animal model is being established using the label retaining cell (LRC) approach. Tissue SCs are quiescent and will retain a DNA synthesis label (BrdU), while the label is diluted out in more mature dividing cells during a chase period. Those cells retaining the label over long period of time are LRCs and have been identified as SCs (2). An optimum time to initially label the majority of the cells is critical. We hypothesised that during mouse endometrial development, there is a growth window for maximum labelling of epithelial and stromal cells. The aim of this study was to determine the optimum age and length of time for maximum endometrial BrdU labelling. Two stages of endometrial growth were exploited: (1) postnatal growth before gland development (3 days old – P3); and (2) oestrogen induced growth in ovariectomised prepubertal mice (4–5 weeks old). C57/CBA P3 mice received subcutaneous BrdU injections twice daily for 3 consecutive days. Four to five-week-old ovariectomised mice received BrdU filled mini-osmotic pumps implanted subcutaneously for 7 and 14 days, with daily oestrogen injections. Mice were killed 4 h after the last treatment, uteri collected, and BrdU labelled cells were detected by immunohistochemistry. Results are shown in the table below (see PDF file). The data shows that maximum labelling for epithelial and stromal cells was achieved for P3 mice, possibly due to greater endometrial growth of the Müllerian duct compared to oestrogen induced endometrial growth following ovariectomy. Chase experiments are currently in progress on P3 labelled mice to identify the location of LRCs in mouse endometrial glands and stroma. (1) Chan RW, Schwab KE, Gargett CE (2004) Biol. Reprod. 70, in press. (2) Morris RJ, Potten CS (1994) Cell Prolif. 27, 279–289.

Augmented regeneration of partial liver allograft induced by nuclear factor-kappaB decoy oligodeoxynucleotides-modified dendritic cells.

To investigate the effect of NF-kappaB decoy oligodeoxynuleotides (ODNs) - modified dendritic cells (DCs) on regeneration of partial liver allograft. Bone marrow (BM)- derived DCs from SD rats were propagated in the presence of GM-CSF or GM-CSF+IL-4 to obtain immature DCs or mature DCs, respectively. GM-CSF-propagated DCs were treated with double-strand NF-kappaB decoy ODNs containing two NF-kappaB binding sites or scrambled ODNs. Allogeneic (SD rat to LEW rat) 50% partial liver transplantation was performed. Normal saline (group A), GM-CSF -propagated DCs (group B), GM-CSF+IL-4 - propagated DCs (group C), and GM-CSF+NF-kappaB decoy ODNs (group D) or scrambled ODNs -propagated DCs (group E) were injected intravenously into recipient LEW rats 7 days prior to liver transplantation and immediately after transplantation. DNA synthesis (BrdU labeling) and apoptosis of hepatocytes were detected with immunostaining and TUNEL staining postoperative 24 h, 48 h, 72 h and 84 h, respectively. Liver graft-resident NK cell activity, hepatic IFN-gamma mRNA expression and recipient serum IFN-gamma level at the time of the maximal liver allograft regeneration were measured with (51)Cr release assay, semiquantitative RT-PCR and ELISA, respectively. Regeneration of liver allograft was markedly promoted by NF-kappaB decoy ODNs-modified immature DCs but was significantly suppressed by mature DCs, the DNA synthesis of hepatocytes peaked at postoperative 72 h in group A, group B and group E rats, whereas the DNA synthesis of hepatocytes peaked at postoperative 84 h in group C rats and 48 h in group D rats, respectively. The maximal BrdU labeling index of hepatocytes in group D rats was significantly higher than that in the other groups rats. NF-kappaB decoy ODNs-modified immature DCs markedly suppressed but mature DCs markedly promoted apoptosis of hepatocytes, liver-resident NK cell activity, hepatic IFN-gamma mRNA expression and recipient serum IFN-gamma production. At the time of the maximal regeneration of liver allograft, the minimal apoptosis of hepatocytes, the minimal activity of liver-resident NK cells, the minimal hepatic IFN-gamma mRNA expression and serum IFN-gamma production were detected in group D rats. The apoptotic index of hepatocytes, the activity of liver- resident NK cells, the hepatic IFN-gamma mRNA expression level and the serum IFN-gamma level in group D rats were significantly lower than that in the other groups rats at the time of the maximal regeneration of liver allograft. The data suggest that the augmented regeneration of partial liver allograft induced by NF-kappaB decoy ODNs-modified DCs may be attributable to the reduced apoptotic hepatocytes, the suppressed activity of liver-resident NK cells and the reduced IFN-gamma production.