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Chromosomal anomalies and disturbance of transcriptional activity at the pachytene stage of meiosis: relationship to male sterility.

Morphological analysis of pachytene spermatocytes obtained from male mice carrying three chromosomal rearrangements--a Robertsonian translocation, Rb(X-2)2Ad; an autosomal reciprocal translocation, T(16;17)43H; and a tertiary trisomic, Ts(113)70H--demonstrated frequent association between the XY bivalent and the T43H and T70H translocation chromosomes. Quantitative autoradiographic data revealed that the normal transcriptional inactivity of the XY bivalent was not significantly disturbed, in contrast to that of the 16;17 quadrivalent and the extra 1(13) marker chromosome. These results are interpreted as an extension of the XY inactivation process to the associated autosomes and discussed in relation to male sterility.

Animals↗

Cytogenetics of collared lemmings (Dicrostonyx groenlandicus). I. Meiotic behavior and evolution of the neo-XY sex-chromosome system.

Electron-microscopic analysis of surface-spread synaptonemal complexes at pachynema and light-microscopic analysis of chromosomal configurations at diakinesis/metaphase I corroborate the hypothesized neo-XY derivation of the sex chromosomes of Dicrostonyx groenlandicus. Although an intact neo-XY pairing configuration was observed in a relatively small percentage of the pachytene cells in each individual, the high incidence of neo-XY bivalents at diakinesis/metaphase I suggests that the other observed pachytene configurations were artifacts of the physical stresses of the surface-spreading procedure. The very low frequency (0.6%) of univalent neo-X and neo-Y chromosomes at diakinesis and metaphase I is attributable to consistent synapsis and recombination between their homologous autosomally derived segments. The resultant stability of the sex bivalent through metaphase I may have increased the efficacy of sex-chromosome segregation, and thereby played a mechanistic role in the evolutionary incorporation of the neo-XY sex-chromosome constitution in D. groenlandicus.

Animals↗

The insulin-like growth factor-I receptor. Structure, ligand-binding mechanism and signal transduction.

The nonclassical binding kinetics of IGF-I and insulin to their respective receptors, suggestive of negative cooperativity, can be readily explained by our recently proposed novel binding mechanism whereby the bivalent ligand bridges the two receptor alpha-subunits alternatively at opposite sites in a symmetrical receptor structure. The bivalent binding mechanism also explains bell-shaped bioactivity curves. The possible role of different binding modes versus differences in downstream signaling by insulin and IGF-I in producing specific mitogenic or metabolic responses is discussed.

Animals↗

Dissociating task-set selection from task-set inhibition in the prefrontal cortex.

Patients with focal lesions in the left (n=7) and right (n=4) prefrontal cortex were compared with controls (n=16) in a task-switching experiment using four different, simple spatial tasks. Each of these tasks involved a left-right decision, either regarding an arrow, the word "left" or "right," a circle position, or the direction of a moving line. We compared performance on trials that required rule switches versus rule repetitions (local switch costs) and we compared performance between blocks with bivalent stimuli (two dimensions present) and blocks with univalent stimuli (only one dimension present) to assess global switch costs. Patients with left prefrontal lesions, but not patients with right prefrontal lesions, exhibited increased costs on trials in which the relevant dimension switched (local switch costs), but also on no-switch trials with bivalent stimuli (global costs). We also assessed task-set inhibition in the form of the backward-inhibition effect [increased response times to recently abandoned tasks; Mayr, U., & Keele, S. Changing internal constraints on action: The role of backward inhibition. Journal of Experimental Psychology: General, 129, 4-26, 2000]. Although left frontal patients showed normal inhibition, right frontal patients showed no evidence for inhibition. These results suggest a neurocognitive dissociation between task-set selection and inhibition.

Acoustic Stimulation↗

Impairment of isoproterenol, H2 histamine, and prostaglandin E1 response of human granulocytes after incubation in vitro with live influenza vaccines.

The release of the lysosomal enzyme beta-glucuronidase from granulocytes follows incubation in vitro with complement-activated zymosan particles. Release of beta-glucuronidase is inhibited by isoproterenol, histamine, and prostaglandin E1. This in vitro model was used to study the effect of incubating a live, bivalent (A + B) influenza vaccine on the granulocyte response to the agonists described. After incubation in vitro with the live, bivalent influenza vaccine, there was a significantly impaired granulocyte reponse to all 3 agonists. The change in the response of polymorphonuclear leukocytes to isoproterenol was similar to an impairment in beta-adrenergic response found during respiratory infections in vivo. The viral-induced changes in the granulocyte response to isoproterenol may reflect similar alteration in other tissues, such as variable control of the airways and provide one explanation for the occurrence of airway dysfunction during respiratory infections.

Adult↗

Survival improvement in patients with medullary thyroid carcinoma who undergo pretargeted anti-carcinoembryonic-antigen radioimmunotherapy: a collaborative study with the French Endocrine Tumor Group.

PURPOSE: No effective therapy is currently available for the management of patients with metastatic medullary thyroid carcinoma (MTC). The efficacy of pretargeted radioimmunotherapy (pRAIT) with bispecific monoclonal antibody (BsMAb) and a iodine-131 (131I) -labeled bivalent hapten is evaluated. PATIENTS AND METHODS: Twenty-nine patients with advanced, progressive MTC, as documented by short serum calcitonin doubling times (Ct DTs), received an anti-carcinoembryonic antigen (CEA)/anti-diethylenetriamine pentaacetic acid (DTPA) -indium BsMAb, followed 4 days later by a 131I-labeled bivalent hapten. Overall survival (OS) was compared with 39 contemporaneous untreated MTC patients with comparable prognostic indicators. RESULTS: OS was significantly longer in high-risk, treated patients (Ct DT < 2 years) than in high-risk, untreated patients (median OS, 110 v 61 months; P < .030). Forty-seven percent of patients, defined as biologic responders by a more than 100% increase in CtDT, experienced significantly longer survival than nonresponders (median OS, 159 v 109 months; P < .035) and untreated patients (median OS, 159 v 61 months; P < .010). Treated patients with bone/bone-marrow disease had a longer survival than patients without such involvement (10-year OS, 83% v 14%; P < .023). Toxicity was mainly hematologic and related to bone/bone-marrow tumor spread. CONCLUSION: pRAIT against CEA induced long-term disease stabilization and a significantly longer survival in high-risk patients with Ct DTs less than 2 years, compared with similarly high-risk, untreated patients. Ct DT and bone-marrow involvement appear to be prognostic indicators in MTC patients who undergo pRAIT.

Antibodies, Monoclonal↗

Identification and analysis of DYAD: a gene required for meiotic chromosome organisation and female meiotic progression in Arabidopsis.

The dyad mutant of Arabidopsis was previously identified as being defective in female meiosis. We report here the analysis of the DYAD gene. In ovules and anthers DYAD RNA is detected specifically in female and male meiocytes respectively, in premeiotic interphase/meiotic prophase. Analysis of chromosome spreads in female meiocytes showed that in the mutant, chromosomes did not undergo synapsis and formed ten univalents instead of five bivalents. Unlike mutations in AtDMC1 and AtSPO11 which also affect bivalent formation as the univalent chromosomes segregate randomly, the dyad univalents formed an ordered metaphase plate and underwent an equational division. This suggests a requirement for DYAD for chromosome synapsis and centromere configuration in female meiosis. The dyad mutant showed increased and persistent expression of a meiosis-specific marker, pAtDMC1::GUS during female meiosis, indicative of defective meiotic progression. The sequence of the putative protein encoded by DYAD did not reveal strong similarity to other proteins. DYAD is therefore likely to encode a novel protein required for meiotic chromosome organisation and female meiotic progression.

Amino Acid Sequence↗

Organization and pairing of meiotic chromosomes in the ciliate Tetrahymena thermophila.

During meiotic prophase in the ciliate Tetrahymena thermophila micronuclei dramatically elongate and form thread-like crescents. The arrangement of the chromosomes within the crescent as well as the timing of chromosome pairing and recombination with respect to the elongation process have been subjects of ongoing debate. Here, we addressed these issues by means of fluorescence in situ hybridization, labeling of individual chromosomes by BrdU (BrdU-painting) and by immunostaining of the recombination protein, Rad51. BrdU-painting indicated that chromosomes are arranged as parallel bundles within the crescent, and telomere-directed fluorescent in situ hybridization (FISH) revealed that most if not all telomeres are assembled near one end of the developing crescent. Prior to full crescent formation, Rad51 localizes to chromatin as numerous foci. Locus-specific FISH demonstrated that close pairing of homologues only occurs in the full crescent. Meiotic DNA double-strand break formation and the initiation of recombination thus seem to precede close pairing. A synaptonemal complex was not detected. We conclude that the chromosomes adopt a polarized arrangement within the crescent, probably resembling the classical bouquet arrangement. Furthermore, we propose that the elongated shape of meiotic micronuclei promotes the parallel arrangement of chromosomes and supports the juxtaposition of homologous regions in the absence of a synaptonemal complex. Several pieces of evidence indicate the presence of one to four chiasmata per bivalent, which would call for crossover interference to explain regular bivalent formation in spite of this low mean number. Tetrahymena might, therefore, pose a case of interference in the absence of a synaptonemal complex.

Animals↗

Assembly pathway of the anastral Drosophila oocyte meiosis I spindle.

Oocyte meiotic spindles of many species are anastral and lack centrosomes to nucleate microtubules. Assembly of anastral spindles occurs by a pathway that differs from that of most mitotic spindles. Here we analyze assembly of the Drosophila oocyte meiosis I spindle and the role of the Nonclaret disjunctional (Ncd) motor in spindle assembly using wild-type and mutant Ncd fused to GFP. Unexpectedly, we observe motor-associated asters at germinal vesicle breakdown that migrate towards the condensed chromosomes, where they nucleate microtubules at the chromosomes. Newly nucleated microtubules are randomly oriented, then become organized around the bivalent chromosomes. We show that the meiotic spindle forms by lateral associations of microtubule-coated chromosomes into a bipolar spindle. Lateral interactions between microtubule-associated bivalent chromosomes may be mediated by microtubule crosslinking by the Ncd motor, based on analysis of fixed oocytes. We report here that spindle assembly occurs in an ncd mutant defective for microtubule motility, but lateral interactions between microtubule-coated chromosomes are unstable, indicating that Ncd movement along microtubules is needed to stabilize interactions between chromosomes. A more severe ncd mutant that probably lacks ATPase activity prevents formation of lateral interactions between chromosomes and causes defective microtubule elongation. Anastral Drosophila oocyte meiosis I spindle assembly thus involves motor-associated asters to nucleate microtubules and Ncd motor activity to form and stabilize interactions between microtubule-associated chromosomes during the assembly process. This is the first complete account of assembly of an anastral spindle and the specific steps that require Ncd motor activity, revealing new and unexpected features of the process.

Animals↗

Localization of two mammalian cyclin dependent kinases during mammalian meiosis.

Mammalian meiotic progression, like mitotic cell cycle progression, is regulated by cyclins and cyclin dependent kinases (CDKs). However, the unique requirements of meiosis (homologous synapsis, reciprocal recombination and the dual divisions that segregate first homologues, then sister chromatids) have led to different patterns of CDK expression. Here we show that Cdk4 colocalizes with replication protein A (RPA) on the synaptonemal complexes (SCs) of newly synapsed axes of homologously pairing bivalents, but disappears from these axes by mid-pachynema. The switch from the mitotic pattern of expression occurs during the last two spermatogonial divisions. Cdk2 colocalizes with MLH1, a mismatch repair protein at sites of reciprocal recombination in mid-late pachynema. In addition Cdk2 localizes to the telomeres of chromosomal bivalents throughout meiotic prophase. The mitotic pattern of expression of Cdk2 remains unchanged throughout the spermatogonial divisions, but is altered in meiosis of the spermatocytes.

Animals↗

Inversion heterozygosity in females of the newt Notophthalmus viridescens and its influence on chiasma distribution.

In a wild population of the American newt Notophthalmus viridescens 15 females out of a total of 94 were found to be heterozygous for a paracentric inversion which includes almost the whole of the longer arm of the smallest chromosome (XI). The inversion was recognized in preparations of lampbrush chromosomes because it transfers the sequential loops, which normally lie close to the telomere, to a position neighbouring the centromere. Because of inversion the transcriptional polarity of the sequential loops is reversed vis-à--vis the chromosome as a whole. In normal bivalents XI (both in male and female meiosis) each arm pair generally forms a single chiasma close to the telomeres (proterminal localization). In bivalents XI heterozygous for the inversion no chiasmata are formed between the mutually inverted longer arm pairs, presumably because they fail to synapse, but chiasma frequency in the non-inverted shorter arm pairs is increased, and the normal restraint on chiasma distribution in this arm pair is lifted. An explanation is offered in terms of the availability of recombination nodules, and the time of their association with the synaptonemal complex.

Animals↗

Ultrastructural organization, sites of transcription and distribution of fibrillar centres in the nucleolus of the mouse oocyte.

The emergence of newly formed nucleoli and their development have been studied in mouse oocytes from pachytene to diplotene stages. At mid-pachytene, the nucleolus first appears as a fibrillar centre surrounded by a layer of electron-dense fibrils and penetrated by chromatin fibres emanating from the secondary constriction region of the nucleolar bivalent. Since this bivalent contains 2 paired nucleolar organizers, 2 nucleoli are formed in a symmetrical fashion. At advanced pachytene, the nucleoli are extended by strands of fibrillar component which become fibrillogranular distally. The 2 nucleoli fuse together at late pachytene. At diplotene, the nucleolus becomes large and reticulated. The development of the nucleolonema coincides with the appearance of numerous secondary fibrillar centres. Three-dimensional reconstruction of the reticulated nucleolus shows that the number of fibrillar centres largely exceeds that of nucleolar organizers. Radioautography after [3H]uridine incorporation demonstrates that during the first step of nucleologenesis the labelling is limited to the layer of electron-dense fibrils surrounding the fibrillar centre. Study of the time course of tritiated uridine incorporation from pachytene to diplotene shows that the labelling extends with the extending strands of fibrillar component. In the fully developed nucleolus, all fibrillar strands are labelled and contain, therefore, actively transcribed rDNA. These observations suggest that the rDNA, which is initially compacted in the primary fibrillar centre at the onset of nucleogenesis, progressively unravels and becomes distributed throughout the fibrillar parts of the nucleolonema. The lack of labelling of the secondary fibrillar centres suggests that they are zones of inactivity of the ribosomal genes where the rDNA remains locally compacted. A model of the ultrastructural organization of the nucleolus is proposed based on our observations.

Animals↗

Formation of water-insoluble gel in dry-coated tablets for the controlled release of theophylline.

Sodium alginate (ALNa) of a natural polysaccharide is known to form a water-insoluble gel when combined with a bivalent metal. In this study, we prepared tablets containing ALNa and calcium gluconate (GLCa) as a bivalent metal, and studied the application of the water-insoluble gel involving the controlled release of a test drug by permeation of water. Dry-coated tablets containing theophylline (TP) as a model drug, ALNa and GLCa were prepared by the dry power compression method. The controlled release of TP was evaluated by the dissolution test according to JP XIII. The release rate was extremely high for the tablets which contained only TP and GLCa. A zero order or sigmoidal release profile was observed for the tablets that contained only TP and ALNa. On the other hand, the lowest dissolution rate and a sigmoidal release profile were observed for the tablet containing TP and GLCa in its core and ALNa in its outer phase. These results suggest that dry-coated tablets containing ALNa and GLCa and prepared by the direct powder compression method would be useful for the controlled release of drugs.

Alginates↗

Transfer of small chromosome fragments of Agropyron elongatum to wheat chromosome via asymmetric somatic hybridization.

The chromosome constitution of hybrids and chromatin patterns of Agropyron elongatum (Host)Neviski in F5 somatic hybrid lines II -1-3 and I-1-9 between Triticum aestivum L. and A. elongatum were analyzed. Based on the statistic data of pollen mother cells, F5 I-1-9 and II-1-3 had 20-21 bivalents with a frequency of 84.66% and 85.28%, of which, 89.83% and 89.57% were ring bivalents. The result indicated that both hybrid lines were basically stable in the chromosome constitution and behavior. RAPD analysis showed that the two hybrids contained biparental and integrated DNA. GISH (Genome in situ hybridization) revealed that in the form of small chromosome segments, A. elongatum chromatin was scattered on 4-6 wheat chromosomes near by the region of centromere and telomere in the two hybrid lines. SSR analysis indicated that A. elongatum DNA segments were distributed on the 2A, 5B, 6B and 2D wheat chromosomes in the hybrids, which was in accordance with the GISH results that small-segments intercalated poly-site.

Agropyron↗

The synaptic process in Locusta migratoria spermatocytes by synaptonemal complex analysis.

We describe the synaptic process during meiotic prophase in spermatocytes from Locusta migratoria, using synaptonemal complex (SC) spreads analyzed by light (LM) and electron (EM) microscopy. At leptotene, a stage of short duration, unpaired axial elements begin to be assembled. Synapsis starts at zygotene, beginning usually at the terminal regions of the bivalents, either at the proximal, centromeric end or at the distal, non-centromeric end; interstitial initiation of synapsis was only occasionally observed in the longer chromosomes. Pairing is asynchronous, and shorter chromosomes are the first to complete synapsis. At pachytene all bivalents are fully synapsed. Diplotene is characterized by the progressive fragmentation of SCs; fragmentation is asynchronous, and affects mainly the longer chromosomes, while the shorter ones maintain their morphology up to late diplotene.

Animals↗

Targeting acetylcholinesterase to treat neurodegeneration.

Neurodegenerative disorders, such as Alzheimer's disease, are often characterised by the degeneration of the cholinergic system. Thus, the aim of many treatment regimens is to support this system either by means of muscarinic agonists or by inhibitors of acetylcholinesterase (AChE), the latter being able to increase the concentration of acetylcholine. However, both pharmacological groups of drugs can only help in the beginning of the progressive disease. The finding that the occupation of the peripheral anionic site of AChE is able to stop the formation of the amyloid plaque led to the development of bivalent ligands that occupy both the active and the peripheral site. This dual action might be more beneficial for treatment of Alzheimer s disease than simple inhibition of the acetylcholine hydrolysis. Thus, the new bivalent ligands are the focus of this review.

Acetylcholinesterase↗

Brain regions mediating flexible rule use during development.

During development, children improve at retrieving and using rules to guide their behavior and at flexibly switching between these rules. In this study, we used functional magnetic resonance imaging to examine the changes in brain function associated with developmental changes in flexible rule use. Three age groups (8-12, 13-17, and 18-25 years) performed a task in which they were cued to respond to target stimuli on the basis of simple task rules. Bivalent target stimuli were associated with different responses, depending on the rule, whereas univalent target stimuli were associated with fixed responses. The comparison of bivalent and univalent trials enabled the identification of regions modulated by demands on rule representation. The comparison of rule-switch and rule-repetition trials enabled the identification of regions involved in rule switching. We have used this task previously in adults and have shown that ventrolateral prefrontal cortex (VLPFC) and the (pre)-supplementary motor area (pre-SMA/SMA) have dissociable roles in task-switching, such that VLPFC is associated most closely with rule representation, and pre-SMA/SMA is associated with suppression of the previous task set (Crone et al., 2006a). Based on behavioral data in children (Crone et al., 2004), we had predicted that regions associated with task-set suppression would show mature patterns of activation earlier in development than regions associated with rule representation. Indeed, we found an adult-like pattern of activation in pre-SMA/SMA by adolescence, whereas the pattern of VLPFC activation differed among children, adolescents, and adults. These findings suggest that two components of task-switching--rule retrieval and task-set suppression--follow distinct neurodevelopmental trajectories.

Adolescent↗

Sex-dependent frequency and type of autosomal univalency at the first meiotic metaphase in mouse germ cells.

Univalents at the first meiotic metaphase in mouse spermatocytes occur mainly in the XY pair, making it difficult to compare the amounts of univalency in males and females. In this study, the amounts of autosomal univalency in male and female meiosis were compared using the model strain CBA-T6, in which univalency of the small marker autosome pair T6 has been shown to occur very frequently in spermatocytes. Mice from inbred CBA and DBA strains were also analysed. The total frequencies of univalency (sex chromosomes plus autosomes) in metaphase I spermatocytes were 45.6% in CBA, 36.9% in CBA-T6, and 37.3% in DBA males. The aneuploidy in metaphase II spermatocytes ranged from 1.4 to 3% in these strains, which was in agreement with previous findings that most primary spermatocytes with abnormal chromosome configurations are arrested in their development before metaphase II. In the CBA-T6 strain, autosomal univalency at metaphase I mostly involved chromosome pair T6; however, its frequency differed significantly between the sexes, amounting to 18.9% in spermatocytes and 4.3% in oocytes. In the CBA strain, autosomal univalents at metaphase I were seen in 7.7% of the spermatocytes and 1.4% of the oocytes and, in DBA mice, in 4.9% of the spermatocytes and 3.8% of the oocytes. However, in DBA oocytes, when univalency occurred it usually concerned a greater number of bivalents in one cell (range: 2-19 disjoined bivalents), a phenomenon very rare in males of this strain. This study shows that univalent formation differs between the male and female types of meiosis.

Animals↗