Anti-glioma effect of 7-H-pyrrolo [2,3-di]pyrimidine derivative via induction of cell death

Purpose: To investigate the effect of 7-H-pyrrolo[2,3-di]pyrimidine derivative (7-HPPD) on glioma cell growth.Methods: Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, while apoptosis was assessed in Hoechst 33342 stained cells using flow cytometry. Changes in cell morphology were assessed by scanning electron microscopy (SEM).Results: Glioma cell viability showed a concentration-dependent decrease on exposure to 7-HPPD for 72 h (p < 0.05). The viabilities of C6, U251 and U87 cells were reduced by 24, 28 and 31 %, respectively, following treatment with 20 μM 7-HPPD. Exposure to various concentrations of 7-HPPDresulted in a marked decrease in BrdU LI and cAMP levels in C6, U251 and U87 glioma cells (p < 0.05). Moreover, 7-HPPD induced apoptosis in U251 and U87 cell cultures, as was evident in the condensation of chromatin material, presence of apoptotic bodies and intense blue fluorescence.Treatment of U251 cells with 7-HPPD for 72 h led to a significant increase in the proportion of cells in G0/G1 phase, and significant decrease in percentage of cells in G2/M and S phases. The population of rounded cells showed a significant rise with increase in 7-HPPD concentration from 10 to 20 μM (p < 0.05).Conclusion: 7-HPPD inhibits growth and proliferation of glioma cells by inducing apoptosis. Therefore, it has a potential for application in glioma chemotherapy. Keywords: Glioma, Fluorescence, Metastasis, Apoptosis, Infiltration

Chromosome count, meiotic abnormalities and pollen sterility in Lahaul sweetvetch (Hedysarum astragaloides Benth. ex Baker, Fabaceae), an endemic and threatened species from India

Male meiotic studies were carried out on eight different accessions of Hedysarum astragaloides Benth. ex Baker (Fabaceae), an endemic and threatened species of northwest Himalaya, India. Although genetic factors such as meiosis, chromosome number, and ploidy level may be causative for the evolution, endemism, rare distribution or even extinction of the species, no detailed information exists. Keeping this in mind H. astragaloides has been studied cytologically. Male meiotic investigations revealed diploid level (2n=2x=14) for species and normal meiotic course in the accessions from the Manali Hills resulting in nearly 100% pollen fertility. However, the accessions scored from the Manimahesh Hills and Pangi Valley depicted inter-pollen mother cell transfer of chromatin material and structural heterozygosity for reciprocal translocations. Consequent upon these meiotic anomalies, some pollen sterility (21%) resulted. On account of this sweeping genetic outcome, the incidence of anomalies such as this in an endemic and threatened species warrants grave consideration. It is sensible to conclude that conservation measures should include the collection of germplasm from the localities where plants are meiotically stable with high gametic fertility, to ensure good germination and healthy plants for future use. Seeds from meiotically normal individuals should be given priority for inclusion in seed banks.

Nucleoli migration coupled with cytomixis

Nucleolus was regarded as a mere ribosome-producing factory for many decades however, in the past two decades; this has come to the light that it is a plurifunctional nuclear domain which plays vital role in many cellular processes. Despite many advances, much remains to be revealed about the nucleolus association with cellular processes. While exploring the chromosomal diversity in the high altitude plants of the Indian cold desert region in North West Himalayas we came across 140 wild plants, out of these, pollen mother cell (PMC) in 31 plants showed the occurrence of additional nucleoli. Coincidently, all these plants also depicted the phenomenon of cytomixis i.e. inter-PMC chromatin material extrusion/transfer among/between adjacent meiocytes. Interestingly, it was noticed that nucleolus accompanied the chromatin material during process of cytomixis. During, and after the migration of nucleolus, change in its shape, size and number has also been discussed. So the current paper here in correlated the inter-PMC migration of nucleolus to the occurrence of cytomixis.

Cytomictic Anomalous Male Meiosis and 2nPollen Grain Formation inMertensia echioidesBenth. (Boraginaceae) from Kashmir Himalaya

PresentlyMertensia echioidesBenth. (Boraginaceae) collected from Kashmir Himalaya, India, is cytologically analyzed for the first time revealing2n=2x=24(diploid). Interestingly we found 4.3–6.2% syncytic meiocytes/PMCs with2n=4x=48(tetraploid) in addition to normal meiocytes (2n=24) during male meiosis. These comparatively larger PMCs (pollen mother cells) lead to the formation of fertile giant2npollen grains. A frequency of 6.4–13.3% PMCs shows transfer of chromatin material at prophase-I and, therefore, results in aneuploid meiocytes. Whole chromatin transfer by the process of cytomixis could also have led to the formation of tetraploid cells. Translocation heterozygosity is also evident in the form of multivalents in 12–17% diploid (2x) meiocytes at diakinesis and metaphase-I and is reported for the first time in this species. The syncytes formed depict open chain hexavalent and quadrivalent formation in the three populations with different frequencies. Moreover chromatin stickiness at metaphase-I is observed in 45% of PMCs in population-1 (P-1). Syncyte or unreduced PMC formation leading to unreduced fertile gametes is here speculated to act as a possible way out for infraspecific polyploidization in the species.

Mesangial Cell-Specific Antibodies Are Central to the Pathogenesis of Lupus Nephritis

Not only is nephritis a common complaint in systemic lupus erythematosus, but it is also the most life-threatening complication of the disease. Anti-double-stranded DNA antibodies (Abs), which are found in up to 80% of these patients, might be nephritogenic per se. That is, they may cross-react with mesangial cell (MC) surface proteins, such as alpha-actinin and annexin A2, they may cross-react with mesangial matrix protein such as laminine and fibronectin, or they may recognize chromatin material previously deposited in the glomeruli. The consequence of the binding of anti-MC Abs may be their internalization, which results in activation and proliferation of these MCs. In turn, these activated MCs are suspected of promoting immune complex formation by sequestering and thereby protecting chromatin from degradation. The present paper will explain the mechanisms through which such autoAbs may initiate nephritis.

Electron microscopic study of sperm head differentiation in the lizard Agama stellio

Early differentiation of the spermatid of Agama stellio is demonstrated by two anterior nuclear depressions, occupied by two proacrosomal vesicles, which fuse to form one vesicle. Later, this vesicle exhibits an acrosomal granule in its midposterior portion. The space between the posterior acrosomal membrane and the nuclear envelope is occupied by a subacrosomal fibrous layer which later exhibits a subacrosomal granule posterior to the acrosomal granule. The acrosomal vesicle and the nuclear depression flatten and later elongate. The acrosomal granule spreads and assumes the inverted V shape of the acrosomal vesicle, and the subacrosomal material assumes a feathery shape capping the nuclear prolongation. The subacrosomal granule on top of this feathery material forms a long, cross-striated subacrosomal rod which extends towards the tip of the acrosome. The chromatin material undergoes condensation into spirally oriented fibers, which eventually become homogeneous and dense. This process is accompanied by a change in the orientation of the manchette microtubules, which initially occur as rings around the nucleus and are eventually found parallel to the longitudinal axis of the nucleus.

Alkaline proteinase localization in myoblasts.

Recent interest in elucidating the role of non-lysosomal proteases in intracellular protein catabolism in muscle has led to various investigations with three alkaline proteases: a trypsin-like, a chymotrypsin-like, and a high molecular weight cysteine proteinase. Although in vitro biochemical assays have revealed the catabolic potential of at least two of these proteases, confirmation of their presence in muscle cells has been difficult. In this study immunohistochemical techniques were employed to localize each of these proteases in rat myoblasts. Antisera against the trypsin-like and chymotrypsin-like proteinase (both serine proteinases) showed strong localization in the cytoplasm immediately around the nucleus. Both also stained chromatin material in the nucleus of these cells. Fluorescent localization of the high molecular weight cysteine proteinase (Proteinase I) also appeared to be cell-associated in the myoblasts. The use of myoblasts in cell culture sections of whole muscle was advantageous, since localization of the proteases could be assessed in the absence of other cell types.

Gametogenesis and fertilization in Dirofilaria immitis (Nematoda: Filarioidea)

SUMMARYGametogenesis in Dirofilaria immitis has been studied principally by means of the aceto-orcein chromosomal squash technique, but with additional ultrastructural observations. A terminal germinative zone, in which a continuous and rapid division of germ cells occurs, has been identified in the gonoduct of both male and female worms. Approximately 20% of cells within these germinative zones were in arrested mitotic division following the incubation in vitro of excised gonads in 0·01% colchicine for 4 h. All primary spermatocytes within a 1–2 cm length of the testis proximal to the germinative zone were at the prophase of the 1st meiotic division. In the corresponding region of the ovary, the primary oocytes were similarly at the prophase of the 1st meiotic division in 75% of female worms examined but in the remaining 25% all primary oocytes possessed markedly less condensed, probably interphase nuclei. A possible hormonal control of the cyclical development of primary oocytes, but not primary spermatocytes in D. immitis is suggested. In most of the remaining length of the gonoducts beyond this region of cells at meiotic prophase, the chromatin material of both primary spermatocytes and oocytes is decondensed. Recondensation of chromosomes in the spermatocytes is observed just prior to entry into the seminal vesicle, where meiosis I is completed and meiosis II takes place. In the primary oocyte, completion of meiosis only occurs after fertilization within the seminal receptacle by an entire male gamete. Following the 2 meiotic divisions in the oocyte and subsequent extrusion of the 2 polar bodies, the haploid chromosome complement of the female unites with that of the male, re-establishing the diploid number of the zygote (2n = 10). Male chromosomes within the oocyte remain visible throughout late oogenesis and fusion occurs without the formation of pronuclei.

An electron-microscope study of the intranucleolar chromatin during nucleologenesis in root meristematic cells of Allium cepa

The various states of condensation of the chromatin material contained inside the lacunar regions of the reforming nucleolus in Allium cepa, have been investigated by means of conventional electron-microscope techniques. The observations reveal that, in the emerging early to late telophase nucleoli, the intralacunar chromatin material in question appears both in an extended and a condensed condition; from late telophase th the mid G1 period of interphase, the intralacunar chromatin material of the rapidly growing and developing nucleoli is present in an extended state only. An attempt is made to interpret these morphological findings in the light of current knowledge concerning the structural relationship of the nucleolar organizing region of the nucleolar chromosome with the interphase nucleolus in plant cells. The relevant observational evidence would be consistent with the view that the chromatin-containing lacunar regions of the reforming nucleolus in Allium cepa correspond, in fact, to cross- or oblique sections of a meandering channel through which the nucleolar organizing segment of the nucleolar chromosome passes. Assuming the applicability to intranucleolar chromatin of the general concept of condensed-inactive versus extended-active chromatin, it is concluded that gradual uncoiling and subsequent decondensation of the chromatin of the nucleolar organizing region in the form of a convoluted loop structure are key morphological and functional events associated with the process of nucleologenesis in the species investigated.

THE INFLUENCE OF CHROMOSOME CONTENT ON THE SIZE AND SHAPE OF SPERM HEADS IN DROSOPHILA MELANOGASTER AND THE DEMONSTRATION OF CHROMOSOME LOSS DURING SPERMIOGENESIS

ABSTRACT The volumes of sperm heads were estimated from three-dimensional reconstructions of serially sectioned bundles of nearly mature spermatid nuclei. Cysts from males in which all sperm are expected to have comparable amounts of chromatin (X/Y and In(3LR)/+) show unimodal frequency distributions of nuclear volumes, whereas cysts from males in which meiotic segregation is expected to deliver unequal amounts of chromatin material to spermatid nuclei show two (XY/O and XY/Y) or more (T(2;3)/+ and C(2L);C(2R)) modes. The mean volumes of the subpopulations in these cases are related in the same proportions as the metaphase lengths of their chromosomal complements. Thus the volumes of sperm nuclei are proportional to their DNA content. Sperm head shape, on the other hand, does not appear to be very sensitive to chromosomal constitution, as heads of different size do not vary greatly in shape.—The numbers of sperm heads in the various size classes in a cyst depart from mendelian expectations; these departures are caused by the elimination, during individualization, of chromosomes contained within micronuclei that are formed in spermatids at the end of the second meiotic division. The effect of this chromosome loss is to increase the proportion of nullosomic gametes in the sperm pool.—The relative frequencies of XY-bearing and nullo-X, nullo-Y sperm in XY/O males were estimated from the volume measurements. Taking this estimate as a measure of the fertilizing population, it is possible to infer from the change in sex ratio over time following insemination, that XY-bearing sperm have an advantage of 1.5 over nullo-X, nullo-Y sperm in leaving the seminal receptacle of the female for fertilization of ova.