Sequestration of the abrin A chain to the nucleus by BASP1 increases the resistance of cells to abrin toxicity

Abrin, a type II ribosome-inactivating protein, comprises A and B subunits wherein the A subunit harbours toxin activity and the B subunit has a galactose-specific lectin activity. The entry of the protein inside the cell is through the binding of the B chain to cell surface glycoproteins followed by receptor-mediated endocytosis and retrograde transport. A previous study from our laboratory showed that different cell lines exhibited differences of as great as similar to 200-fold in abrin toxicity, prompting the present study to compare the trafficking of the toxin within cells. Observations made in this regard revealed that the abrin A chain, after being released into the cytosol, is sequestered into the nucleus through interaction with a cellular protein of similar to 25 kDa, BASP1 (brain acid-soluble protein 1). The nuclear localization of the A chain is seen predominantly in cells that are less sensitive to abrin toxicity and dependent on the levels of BASP1 in cells. The sequestration by BASP1 renders cells increasingly resistant to the inhibition of protein synthesis by abrin and the nucleus act as a sink to overcome cellular stress induced

Perturbation of nucleo-cytoplasmic transport affects size of nucleus and nucleolus in human cells

Size regulation of human cell nucleus and nucleolus are poorly understood subjects. 3D reconstruction of live image shows that the karyoplasmic ratio (KR) increases by 30-80% in transformed cell lines compared to their immortalized counterpart. The attenuation of nucleo-cytoplasmic transport causes the KR value to increase by 30-50% in immortalized cell lines. Nucleolus volumes are significantly increased in transformed cell lines and the attenuation of nucleo-cytoplasmic transport causes a significant increase in the nucleolus volume of immortalized cell lines. A cytosol and nuclear fraction swapping experiment emphasizes the potential role of unknown cytosolic factors in nuclear and nucleolar size regulation.

The Role of the Subthalamic Nucleus in L-DOPA Induced Dyskinesia in 6-Hydroxydopamine Lesioned Rats

14 p.

A study of the ^9 Be nucleus

The reaction ⁷Li(³He, p)⁹Be has been used to measure excitations and intrinsic widths of levels in ⁹Be below the ⁷Li + d threshold. Previously unreported levels have been found at excitations of (13.78 ± .03) MeV and (16.671 ± .008) MeV with widths of (590 ± 60) keV and (41 ± 4) keV respectively. Two overlapping levels have been found at (11.81 ± .02) MeV and (11.29 ± .03) MeV with widths of (400 ± 30) keV and (620 ± 70) keV respectively. Branching ratios from ⁹Be levels populated in this reaction to the ground and first excited states of ⁸Be have been measured by observing the associated protons in coincidence with the decay neutrons. Branching ratios were found to be:

Excitation in ⁹Be .... Branching Ratio.......... Final Nucleus.........

(MeV) .......................... (percent) .....................................

.. 2.43 ........................... 7.5 ± 1.5 .............. ⁸Be(g.s.)

.. 3.03 ........................... 87 ± 13......................................

.. 4.65 ........................... 13 ± 4.......................................

.. 6.76 .............................. ≤ 2 ......................................

.. 11.29 ...............................≤ 2 ......................................

.. 11.81 ...............................≤ 3 ......................................

.. 6.76 ........................... .41 ≤ B.R. ≤ .69 ....... ⁸Be(2⁺)

.. 11.29 ........................... 14 ± 4 .......................................

.. 11.81 ........................... 12 ± 4 .......................................

Corresponding reduced widths for neutron emission are calculated and a comparison of the results with the expectations of current nuclear models is made. In particular the measured branching ratio to ⁸Be(g.s.) from ⁹Be(2.43 MeV) corresponds to an f-wave reduced with θ²_f = 2.1 x 10^-2, in units of ħ²/mR², with R = 4.35 fm. A comparison of this value with that predicted by a Nilsson model calculation, in which ⁹Be is taken to be a deformed nucleus, is discussed. The measured value for θ²_f is found to be consistent with that expected on the basis of measured E2-transition rates between rotational levels in ⁹Be.

Cholinergic neurons within the nucleus basalis magnocellularis in bilaterally lesioned rats with 192IgG-saporin.

En el presente estudio se han estudiado las neuronas colinérgicas del núcleo basal magnocelular y sus áreas de proyección en ratas lesionadas bilateralmente con la inmunotoxina 192IgG-saporina.

The effect of temperature on the duration of spawning markers—migratory-nucleus and hydrated oocytes and postovulatory follicles—in the multiple-batch spawner Japanese flounder (Paralichthys olivaceus)

The duration of spawning markers (e.g. signs of previous or imminent spawnings) is essential information for estimating spawning frequency of fish. In this study, the effect of temperature on the duration of spawning markers (i.e., oocytes at early migratory nucleus, late migratory nucleus, and hydrated stages, as well as new postovulatory follicles) of an indeterminate multiple-batch spawner, Japanese f lounder (Paralichthys olivaceus), was evaluated. Cannulation was performed to remove samples of oocytes, eggs, and postovulatory follicles in individual females at 2–4 hour intervals over 27–48 hours. The duration of spawning markers was successfully evaluated in 14 trials ranging between 9.2° and 22.6°C for six females (total length 484–730 mm). The durations of spawning markers decreased exponentially with temperature and were seen to decrease by a factor of 0.16, 0.36, 0.30, and 0.31 as temperature increased by 10°C for oocytes at early migratory nucleus, late migratory nucleus, and hydrated stages, and new postovulatory follicles, respectively. Thus, temperature should be considered when estimating spawning frequency from these spawning markers, especially for those fish that do not spawn synchronously in the population.

Estimation of ploidy and hybridization status by erythrocyte and nucleus size analysis in carps

The present experiment was designed to observe whether the nuclear volume and area are affected by the ploidy and hybrid status of the individual. Polyploidy was induced by heat shock treatment given at 44 ± 0.5°C for 30 seconds and 45 seconds which was found to be most effective (64.7%) for induction of triploidy in Cyprinus carpio. Cell and nuclear volume and cell and nuclear area varied significantly in triploid fishes as compared to those of controls. Triploid fishes showed significantly higher growth compared to diploid counterparts. It was also observed that catla x rohu hybrid and its parents showed significant difference in the nuclear volume and area of their erythrocytes. Except nuclear volume, all the parameters were significantly different between catla and catla x rohu hybrid. The hybrids showed a closer relationship with catla as compared to rohu.

Establishment of satellite nucleus of genetically improved farmed tilapia (GIFT strain) in Bangladesh

The aim of this study was to evaluate growth performance of the GIFT (Genetically Improved Farmed Tilapia) strain after one generation of selection for increased body weight at Bangladesh Fisheries Research Institute. Founder stock comprised of 30 families having 300 individuals of the GIFT strain were introduced from Malaysia through \X!orldFish Center in March 2005. The founder stock was reared in 100 m2 hapa for three months and then individually tagged using Passive Integrated Transponder (PIT) at the weight between 30 and 40 g. After tagging, all the fish were communally grown out in pond until harvest. Breeding value for body weight was estimated using SAS and ASREML ranging from 4.17 to 9.70 g for males and 4.24 to 9.36 g for females. The best 40 females and 40 males from the founder stock were then selected to produce progeny of the first generation (F-1). From each family 25 female and 25 male fingerlings were sampled and tagged using PIT. A total of 2,000 tagged fish from 40 families were stocked in a pond (1000 m2) for a continuation of the selection program. In addition, surplus fish after tagging were also reared together with progeny of the founder stock in cistern ecology for growth evaluation. The mean weight of the F-1 generation of GIFT fish was 7.2% greater than that of the founder population (non selected population).

Distribution of rDNA in the nucleus of Giardia lamblia - Detection by Ag-I silver stain

OBJECTIVE: To determine whether rDNA of Giardia lamblia forms a nucleolus organizer region (NOR)-like structure and is in a very primitive state. STUDY DESIGN: G lamblia was used as the experimental animal, with Euglena gracilis as the control. The distribution was demonstrated indirectly by the modified Ag-I silver technique, which can specifically indicate the NOR under both light and electron microscopes. RESULTS: In the ultrathin sections of silver-stained Euglena cells, all the silver grains were concentrated in the fibrosa of the nucleolus, while no grains found in the cytoplasm, nucleoplasm, condensed chromosomes or pars granulosa of the nucleus. In the silver-stained Giardia cells, no nucleolus was found; a few silver grains were scattered in the nucleus but were not concentrated in any specific region. CONCLUSION: The distribution of silver grains in G lamblia showed that the transcription of rDNA occurs inside the nucleus, though no nucleolus is present. It is possible that chromosomes are in a very primitive state in diplomonad cells; as each chromosome has few prRNA genes, the transcription is independent of a nucleolus. These results imply that the rDNA of Giardia does not form a NOR-like structure and seems to represent a very primitive state in the evolution of the nucleolus.

The giant panda (Ailurapoda melanoleuca) somatic nucleus can dedifferentiate in rabbit ooplasm and support early development of the reconstructed egg

The giant panda skeletal muscle cells, uterus epithelial cells and mammary gland cells from an adult individual were cultured and used as nucleus donor for the construction of interspecies embryos by transferring them into enucleated rabbit eggs. All the three kinds of somatic cells were able to reprogram in rabbit ooplasm and support early embryo development, of which mammary gland cells were proven to be the Lest, followed by uterus epithelial cells and skeletal muscle cells. The experiments showed that direct injection of mammary gland cell into enucleated rabbit ooplasm, combined with in vivo development in ligated rabbit oviduct, achieved higher blastocyst development than in vitro culture after the somatic cell was injected into the perivitelline space and fused with the enucleated egg by electrical stimulation. The chromosome analysis demonstrated that the genetic materials in reconstructed blastocyst cells were the same as that in panda somatic cells. In addition, giant panda mitochondrial DNA (mtDNA) was shown to exist in the interspecies reconstructed blastocyst. The data suggest that (i) the ability of ooplasm to dedifferentiate somatic cells is not species-specific; (ii) there is compatibility between interspecies somatic nucleus and ooplasm during early development of the reconstructed egg.

Microsatellite DNA analysis proves nucleus of interspecies reconstructed blastocyst coming from that of donor giant panda

A method for DNA isolation from early development of blastocyst and further analysis of nuclear and mitochondrial DNA was developed in present study. Total DNA was prepared from interspecies reconstructed blastocyst and a giant panda specific microsatellite locus g(010) was successfully amplified. DNA sequencing of the PCR product showed that two sequences of reconstructed blastocysts are the same as that of positive control giant panda. Our results prove that the nucleus of interspecies reconstructed blastocyst comes from somatic nucleus of donor giant panda.