Disparate molecular evolution of two types of repetitive DNAs in the genome of the grasshopper Eyprepocnemis plorans.

Wide arrays of repetitive DNA sequences form an important part of eukaryotic genomes. These repeats appear to evolve as coherent families, where repeats within a family are more similar to each other than to other orthologous representatives in related species. The continuous homogenization of repeats, through selective and non-selective processes, is termed concerted evolution. Ascertaining the level of variation between repeats is crucial to determining which evolutionary model best explains the homogenization observed for these sequences. Here, for the grasshopper Eyprepocnemis plorans, we present the analysis of intragenomic diversity for two repetitive DNA sequences (a satellite DNA (satDNA) and the 45S rDNA) resulting from the independent microdissection of several chromosomes. Our results show different homogenization patterns for these two kinds of paralogous DNA sequences, with a high between-chromosome structure for rDNA but no structure at all for the satDNA. This difference is puzzling, considering the adjacent localization of the two repetitive DNAs on paracentromeric regions in most chromosomes. The disparate homogenization patterns detected for these two repetitive DNA sequences suggest that several processes participate in the concerted evolution in E. plorans, and that these mechanisms might not work as genome-wide processes but rather as sequence-specific ones.

Clinical application of the UroLift® prostatic urethral lift in Spain: consensus on the treatment of lower urinary tract symptoms associated with urinary flow obstruction and secondary to benign prostatic hyperplasia (BPH).

Benign prostatic hyperplasia (BPH) is an increasingly common pathology in the adult male. BPH increases after the age of 40-45 years, and its management consumes an enormous amount of resources. The UroLift® System is an approved technology designed to treat lower urinary tract symptoms (LUTS) secondary to BPH and is used to perform the prostatic urethral lift (PUL) procedure. Various urology specialists in Spain with experience in PUL have prepared this consensus document. Endorsed by the Spanish Urology Association, its information is based on the most recent findings. The main objective of this document is to disseminate the consensus recommendations among all professionals treating patients with LUTS/BPH. Both primary care physicians and urologists can assess and offer PUL as an effective, minimally invasive treatment.

Lack of association of ACP1 gene with inflammatory bowel disease: a case-control study.

The red cell acid phosphatease (ACP1) gene, which encodes a low molecular weight phosphotyrosine phosphatase (LMW-PTP), has been suggested as a common genetic factor of autoimmunity. In the present study, we aimed to investigate the possible influence of ACP1 polymorphisms in the susceptibility of inflammatory bowel disease (IBD). A total of 1271 IBD Spanish patients [720 Crohn's disease (CD) and 551 ulcerative colitis (UC)] and 1877 healthy subjects were included. Four single-nucleotide polymorphisms (SNPs), rs10167992, rs11553742, rs7576247 and rs3828329, were genotyped using TaqMan SNP genotyping assays. Common ACP1 alleles (i.e. ACP1*A, ACP1*B and ACP1*C) were determined by two of these SNPs. After the analysis, no evidence of association of the ACP1 genetic variants was found with CD or UC. Therefore, our results suggest that the ACP1 gene may not play a relevant role in the development of IBD.

Level of heat shock proteins decreases in individuals carrying B-chromosomes in the grasshopper Eyprepocnemis plorans.

We analyzed the effect of B-chromosome presence on expression level of heat shock protein 70 (Hsp70) in cerebral ganglion and gonad in both males and females of the grasshopper Eyprepocnemis plorans. Two natural Spanish populations, Salobreña (Granada) and Torrox (Málaga) were assayed, the former harbouring a neutralized (non-driving) B-chromosome (B(2)) and the latter a parasitic (driving) B-chromosome (B(24)). The analysis was performed by Western blotting, immunostaining and densitometric measuring expression level of the Hsp70 family in adult individuals. The results showed that Hsp70 levels of testis were significantly higher in Salobreña than Torrox, and were significantly lower in testes of B-carrying males from both populations. A similar effect was observed in the ovary of females from Torrox. No effect was, however, observed in cerebral ganglia in any sex or population. B-chromosome effects in Torrox showed a dose-dependent pattern. The results point to an interesting interaction between B-chromosome and stress protein expression in reproductive tissue.

Degradation of a series of fluorinated acrylates and methacrylates initiated by OH radicals at different temperatures.

Rate coefficients for the gas-phase reactions of OH radicals with a series of fluorinated acrylates and methacrylates: 2,2,2-trifluoroethylmethacrylate (k 1), 1,1,1,3,3,3-hexafluoroisopropylacrylate (k 2), 1,1,1,3,3,3-hexafluoroisopropylmethacrylate (k 3), and 2,2,2-trifluoroethylacrylate (k 4) have been measured for the first time as a function of temperature in the range 290-308 K. The kinetic data obtained were used to derive the following Arrhenius expressions (in units of cm3 per molecule per s): k 1 = (2.13 ± 0.68) × 10-18 exp[(4745 ± 206)/T], k 2 = (8.72 ± 0.68) × 10-15 exp[(2166 ± 205)/T], k 3 = (6.30 ± 0.51) × 10-17 exp[(3721 ± 153)/T] and k 4 = (3.93 ± 0.43) × 10-16 exp[(3140 ± 129)/T]. The experiments were performed at normal atmospheric pressure in synthetic air using a 1080 L photoreactor and coupled with FTIR analysis to monitor the decay of the substances of interest and the reference compounds. The obtained negative temperature dependencies are in agreement with a mechanism implying an initial addition of the OH radical to the double bond. Atmospheric implications are discussed with reference to the rate coefficients obtained as a function of the temperature.

Complications of transrectal prostate biopsy in our context. International multicenter study of 3350 patients. / Complicaciones de la biopsia transrectal de próstata en nuestro medio. Estudio multicéntrico internacional de 3.350 pacientes.

INTRODUCTION: Prostate cancer is the most common visceral neoplasm in men and the second one in the United States with the highest mortality behind lung cancer and ahead of colorectal cancer. While prostate cancer mortality rates have been reduced in the United States, Austria, United Kingdom and France, 5-year survival rates have been incremented in Sweden, probably due to a higher diagnostic activity and non-lethal tumor detection. TRPB usually has low rates of serious complications, with a not negligible number of minor complications. Mortality directly associated with this procedure is low and usually related to septic shock. The main complications derived from prostate biopsy can be infectious (mild or severe) and non-infectious (hematuria consistent with hemorrhage, urethral bleeding, rectal bleeding or hemospermia, acute urinary retention, pain or vasovagal reactions). MATERIAL AND METHOD: The objective of the study is to compare three usual TRPB protocols and their relationship with the incidence of complications. Retrospective multicenter observational study conducted in three countries (Spain, Italy and Portugal). We have reviewed the medical records of 3350 men who underwent TRPB to evaluate the existence of prostate cancer, with a minimum evolutionary control of 6months. RESULTS: The mean age was 65,50years, median 66, range 43-79. The subgroup analysis showed that younger patients had higher rates of acute urine retention (AUR) (P=.0000001). Likewise, our results revealed that younger patients presented more procedural pain (P=.0000001) than older patients. Regarding PSA, the mean value was 10.44, SD 7.73, median 8.15, range 0.98-68.09. A higher body mass index (BMI) was not associated with further infection (P=.000004). When performing the multivariate analysis, it was found that the significant variables in the general group were: age (P=.0013), PSA (P=.0402), local infiltration anesthesia (P=.0001) and prophylaxis with metronidazole +tobramycin +amoxicillin/clavulanic acid +gentamicin (P=.0001), presenting a normal distribution with high confidence interval (95%) and significant correlation. Prophylaxis is the most significant variable for no complications and pain (P=.0001), age (P=.0013) and prophylaxis (P=.0001) are for bleeding, age (P=.0013), prophylaxis and PSA (P=.0001) are for infection, and finally, age (P=.0013), anesthesia with local infiltration and prophylaxis (P=.0001) and PSA (P=.0402) are for AUR. CONCLUSIONS: Sedation has fewer side effects and complications related to the transrectal prostate biopsy procedure with respect to transrectal local anesthesia. The choice of the antibiotic prophylaxis scheme is decisive in the onset of complications arising from the performance of a transrectal prostate biopsy.

Microdissection and chromosome painting of X and B chromosomes in the grasshopper Eyprepocnemis plorans.

The relative location of 2 repetitive DNAs, i.e. ribosomal (rDNA) and a tandemly repeated satellite DNA (satDNA), with respect to the centromere, suggested that B chromosomes in the grasshopper Eyprepocnemis plorans derived intraspecifically from the X chromosome. To test this hypothesis, we microdissected X and B chromosomes and amplified the obtained DNA by 2 different procedures, the conventional DOP-PCR method and the single-cell whole-genome amplification GenomePlex method. We then generated DNA probes to perform chromosome painting. Our results have confirmed that X and B chromosomes share many DNA sequences between them and with most of the autosomes, especially at locations where the satDNA and rDNA reside, in consistency with previous information. This supports the hypothesis of an intraspecific origin of B chromosomes in E. plorans. Nevertheless, the present results did not help to clarify whether Bs were derived from the X chromosome or else from 1 or more autosomes.

Spatial sensing in fibroblasts mediated by 3' phosphoinositides.

The directed movement of fibroblasts towards locally released platelet-derived growth factor (PDGF) is a critical event in wound healing. Although recent studies have implicated polarized activation of phosphoinositide (PI) 3-kinase in G protein-mediated chemotaxis, the role of 3' PI lipids in tyrosine kinase-triggered chemotaxis is not well understood. Using evanescent wave microscopy and green fluorescent protein-tagged Akt pleckstrin homology domain (GFP-AktPH) as a molecular sensor, we show that application of a shallow PDGF gradient triggers a markedly steeper gradient in 3' PI lipids in the adhesion zone of fibroblasts. Polar GFP-AktPH gradients, as well as a new type of radial gradient, were measured from front to rear and from the periphery to the center of the adhesion zone, respectively. A strong spatial correlation between polarized 3' PI production and rapid membrane spreading implicates 3' PI lipids as a direct mediator of polarized migration. Analysis of the temporal changes of 3' PI gradients in the adhesion zone revealed a fast diffusion coefficient (0.5 microm(2)/s) and short lifetime of 3' PIs of <1 min. Together, this study suggests that the tyrosine kinase-coupled directional movement of fibroblasts and their radial membrane activity are controlled by local generation and rapid degradation of 3' PI second messengers.

Green fluorescent protein (GFP)-tagged cysteine-rich domains from protein kinase C as fluorescent indicators for diacylglycerol signaling in living cells.

Cysteine-rich domains (Cys-domains) are approximately 50-amino acid-long protein domains that complex two zinc ions and include a consensus sequence with six cysteine and two histidine residues. In vitro studies have shown that Cys-domains from several protein kinase C (PKC) isoforms and a number of other signaling proteins bind lipid membranes in the presence of diacylglycerol or phorbol ester. Here we examine the second messenger functions of diacylglycerol in living cells by monitoring the membrane translocation of the green fluorescent protein (GFP)-tagged first Cys-domain of PKC-gamma (Cys1-GFP). Strikingly, stimulation of G-protein or tyrosine kinase-coupled receptors induced a transient translocation of cytosolic Cys1-GFP to the plasma membrane. The plasma membrane translocation was mimicked by addition of the diacylglycerol analogue DiC8 or the phorbol ester, phorbol myristate acetate (PMA). Photobleaching recovery studies showed that PMA nearly immobilized Cys1-GFP in the membrane, whereas DiC8 left Cys1-GFP diffusible within the membrane. Addition of a smaller and more hydrophilic phorbol ester, phorbol dibuterate (PDBu), localized Cys1-GFP preferentially to the plasma and nuclear membranes. This selective membrane localization was lost in the presence of arachidonic acid. GFP-tagged Cys1Cys2-domains and full-length PKC-gamma also translocated from the cytosol to the plasma membrane in response to receptor or PMA stimuli, whereas significant plasma membrane translocation of Cys2-GFP was only observed in response to PMA addition. These studies introduce GFP-tagged Cys-domains as fluorescent diacylglycerol indicators and show that in living cells the individual Cys-domains can trigger a diacylglycerol or phorbol ester-mediated translocation of proteins to selective lipid membranes.

Localized biphasic changes in phosphatidylinositol-4,5-bisphosphate at sites of phagocytosis.

Phagocytosis requires localized and transient remodeling of actin filaments. Phosphoinositide signaling is believed to play an important role in cytoskeletal organization, but it is unclear whether lipids, which can diffuse along the membrane, can mediate the focal actin assembly required for phagocytosis. We used imaging of fluorescent chimeras of pleckstrin homology and C1 domains in live macrophages to monitor the distribution of phosphatidylinositol-4,5-bisphosphate (4,5-PIP(2)) and diacylglycerol, respectively, during phagocytosis. Our results reveal a sequence of exquisitely localized, coordinated steps in phospholipid metabolism: a focal, rapid accumulation of 4,5-PIP(2) accompanied by recruitment of type Ialpha phosphatidylinositol phosphate kinase to the phagosomal cup, followed by disappearance of the phosphoinositide as the phagosome seals. Loss of 4,5-PIP(2) correlated with mobilization of phospholipase Cgamma (PLCgamma) and with the localized formation of diacylglycerol. The presence of 4, 5-PIP(2) and active PLCgamma at the phagosome was shown to be essential for effective particle ingestion. The temporal sequence of phosphoinositide metabolism suggests that accumulation of 4,5-PIP(2) is involved in the initial recruitment of actin to the phagocytic cup, while its degradation contributes to the subsequent cytoskeletal remodeling.

CaMKIIbeta functions as an F-actin targeting module that localizes CaMKIIalpha/beta heterooligomers to dendritic spines.

Ca2+/calmodulin-dependent protein kinase II (CaMKII) is a serine/threonine protein kinase that regulates long-term potentiation and other forms of neuronal plasticity. Functional differences between the neuronal CaMKIIalpha and CaMKIIbeta isoforms are not yet known. Here, we use green fluorescent protein-tagged (GFP-tagged) CaMKII isoforms and show that CaMKIIbeta is bound to F-actin in dendritic spines and cell cortex while CaMKIIalpha is largely a cytosolic enzyme. When expressed together, the two isoforms form large heterooligomers, and a small fraction of CaMKIIbeta is sufficient to dock the predominant CaMKIIalpha to the actin cytoskeleton. Thus, CaMKIIbeta functions as a targeting module that localizes a much larger number of CaMKIIalpha isozymes to synaptic and cytoskeletal sites of action.

Effect of progesterone administration on the ovarian response to superovulatory treatments in cattle.

To evaluate ovarian response in Angus cows previously treated with progesterone (P4), animals were randomly assigned to two groups: T600 group (n=14), 600 mg of P4/day. P4 was injected from days 3 to 7 of the estrous cycle. On day 7, superovulatory treatments began. The control group (n=12) was given vehicle only. The superovulatory treatments in the control group began on days 7-9 of the estrous cycle. The superovulatory total treatment dose of 400mg NIH FSH P1 was given twice a day over a 4-day period. Ultrasonography of the ovaries was conducted 3 days preceding the initiation of superovulatory treatment, every 24h. In both groups, an additional ultrasonographic evaluation was made at 24h after the end of superovulatory treatment. Blood samples were collected 4 days preceding the initiation of superovulatory treatment, every 24h. Additional samples were taken from the P600 group for 12 day after of initiation of superovulatory treatment every 24h, except on the fifth day after the initiation of superovulatory treatment. In the P600 group, P4 concentrations were greater than in the control group (P<0.01) and remained over 1 ng/ml up to day 11 after beginning of superovulatory treatment. The diameter of the dominant follicle was larger in the animals of the control group (P<0.01). Cows of the P600 group had a greater number of Class I (3-4mm) follicles (P<0.01). A significant day and treatment effect (P<0.01) were observed in Class II (5-9 mm) follicles. Effects due to treatment on the number of Class III follicles (P<0.05) were observed. In the P600 group, no estrous post-superovulatory was observed and there were no ovulations that occurred. Conversely, 100% of the cows of the control group showed estrous. In the P600 group, there were a greater number of Class III follicles (P<0.01) and a lesser number of Class II follicles (P<0.05) at 24h after the end of superovulatory. In the control group, 66.7% of the cows responded to superovulatory treatments. In conclusion, the daily administration of 600 mg of P4, from days 3 to 7 of the estrous cycle, produces an increase of plasma concentrations of this hormone from day 4, resulting in changes in follicular dynamics (absence of follicles greater than 10mm of diameter and an increase of the population of Class I follicles). As to the ovarian stimulation using Folltropin V in animals receiving a daily injection of 600 mg of P4 from days 3 to 7 of the estrous cycle, a greater population of follicles>or=10mm developed by 24h after superovulatory treatments were completed.

Evaluation of a health promotion intervention for skin cancer prevention in Spain: the SolSano program.

The incidence of skin cancer is increasing worldwide, and Spain is no exception. SolSano is the first Spanish health education program for sun safety directed at elementary school children. The objective was to evaluate SolSano's effects on students' knowledge, attitudes and practices about sun safety. A non-randomized, before/after, community intervention without control group, with schools as the unit of intervention, was used for the study. Five thousand eight hundred and forty-five children from 215 Aragonese Primary Schools (Grades 1-2) participated in the program in their classes during the 2004-2005 academic year. The educational package contained an activity guide for teachers, a workbook for each pupil, a poster and an informative pamphlet for families. The pre-test and post-test surveys were similar and were composed of two parts: the first part uses the 'Draw and Write research strategy' and the second part was a questionnaire. One thousand five hundred and twenty-two students completed both questionnaires, 49.2% were boys, and the mean age was 6.6; 45.7% self-reported pale skin and easy sunburn and 48% dark skin and rarely sunburn; 72.3% of the children reported having dark hair and eyes, and 51.6% freckles or moles. The mean score for the complete survey significantly increased by 1.55 points (1.38-1.72) after the intervention (p < 0.001), and girls did better than boys. Sunscreens were the most-commonly employed sun protection strategy while strategies such as seeking shadow and wearing clothes exhibited the greatest increase after the SolSano program [percentage increase of 19.3% (16.4-22.3) and 26.8% (23.4-30.3), respectively]. At baseline, 35.8% of children reported sunburns during the previous summer compared with 23.5% after the program. SolSano also achieved a slight reduction in the percentage of students who desired to be tanned. Our study demonstrates that significant knowledge can be acquired, attitudes regarding the healthiness of a tan can be modified and intentions to change sun protection behaviour can be promoted by well-designed educational programs.

Molecular memory by reversible translocation of calcium/calmodulin-dependent protein kinase II.

Synaptic plasticity is thought to be a key process for learning, memory and other cognitive functions of the nervous system. The initial events of plasticity require the conversion of brief electrical signals into alterations of the biochemical properties of synapses that last for much longer than the initial stimuli. Here we show that a regulator of synaptic plasticity, calcium/calmodulin-dependent protein kinase IIalpha (CaMKII), sequentially translocates to postsynaptic sites, undergoes autophosphorylation and gets trapped for several minutes until its dissociation is induced by secondary autophosphorylation and phosphatase 1 action. Once dissociated, CaMKII shows facilitated translocation for several minutes. This suggests that trapping of CaMKII by its targets and priming of CaMKII translocation may function as biochemical memory mechanisms that change the signaling capacity of synapses.

Control of astrocyte Ca(2+) oscillations and waves by oscillating translocation and activation of protein kinase C.

BACKGROUND: Glutamate-induced Ca2+ oscillations and waves coordinate astrocyte signaling responses, which in turn regulate neuronal excitability. Recent studies have suggested that the generation of these Ca2+ oscillations requires a negative feedback that involves the activation of conventional protein kinase C (cPKC). Here, we use total internal reflection fluorescence (TIRF) microscopy to investigate if and how periodic plasma membrane translocation of cPKC is used to generate Ca2+ oscillations and waves. RESULTS: Glutamate stimulation of astrocytes triggered highly localized GFP-PKCgamma plasma membrane translocation events, induced rapid oscillations in GFP-PKCgamma translocation, and generated GFP-PKCgamma translocation waves that propagated across and between cells. These translocation responses were primarily mediated by the Ca2+-sensitive C2 domains of PKCgamma and were driven by localized Ca2+ spikes, by oscillations in Ca2+ concentration, and by propagating Ca(2+) waves, respectively. Interestingly, GFP-conjugated C1 domains from PKCgamma or PKCdelta that have been shown to bind diacylglycerol (DAG) also oscillated between the cytosol and the plasma membrane after glutamate stimulation, suggesting that PKC is repetitively activated by combined oscillating increases in Ca(2+) and DAG concentrations. The expression of C1 domains, which increases the DAG buffering capacity and thereby delays changes in DAG concentrations, led to a marked prolongation of Ca(2+) spikes, suggesting that PKC activation is involved in terminating individual Ca(2+) spikes and waves and in defining the time period between Ca(2+) spikes. CONCLUSIONS: Our study suggests that cPKCs have a negative feedback role on Ca(2+) oscillations and waves that is mediated by their repetitive activation by oscillating DAG and Ca(2+) concentrations. Periodic translocation and activation of cPKC can be a rapid and markedly localized signaling event that can limit the duration of individual Ca(2+) spikes and waves and can define the Ca(2+) spike and wave frequencies.

Differential codes for free Ca(2+)-calmodulin signals in nucleus and cytosol.

BACKGROUND: Many targets of calcium signaling pathways are activated or inhibited by binding the Ca(2+)-liganded form of calmodulin (Ca(2+)-CaM). Here, we test the hypothesis that local Ca(2+)-CaM-regulated signaling processes can be selectively activated by local intracellular differences in free Ca(2+)-CaM concentration. RESULTS: Energy-transfer confocal microscopy of a fluorescent biosensor was used to measure the difference in the concentration of free Ca(2+)-CaM between nucleus and cytoplasm. Strikingly, short receptor-induced calcium spikes produced transient increases in free Ca(2+)-CaM concentration that were of markedly higher amplitude in the cytosol than in the nucleus. In contrast, prolonged increases in calcium led to equalization of the nuclear and cytosolic free Ca(2+)-CaM concentrations over a period of minutes. Photobleaching recovery and translocation measurements with fluorescently labeled CaM showed that equalization is likely to be the result of a diffusion-mediated net translocation of CaM into the nucleus. The driving force for equalization is a higher Ca(2+)-CaM-buffering capacity in the nucleus compared with the cytosol, as the direction of the free Ca(2+)-CaM concentration gradient and of CaM translocation could be reversed by expressing a Ca(2+)-CaM-binding protein at high concentration in the cytosol. CONCLUSIONS: Subcellular differences in the distribution of Ca(2+)-CaM-binding proteins can produce gradients of free Ca(2+)-CaM concentration that result in a net translocation of CaM. This provides a mechanism for dynamically regulating local free Ca(2+)-CaM concentrations, and thus the local activity of Ca(2+)-CaM targets. Free Ca(2+)-CaM signals in the nucleus remain low during brief or low-frequency calcium spikes, whereas high-frequency spikes or persistent increases in calcium cause translocation of CaM from the cytoplasm to the nucleus, resulting in similar concentrations of nuclear and cytosolic free Ca(2+)-CaM.

Histone H3 lysine 9 acetylation pattern suggests that X and B chromosomes are silenced during entire male meiosis in a grasshopper.

The facultative heterochromatic X chromosome in leptotene spermatocytes of the grasshopper Eyprepocnemis plorans showed marked hypoacetylation for lysine 9 in the H3 histone (H3-K9) with no sign of histone H2AX phosphorylation. Since H3-K9 hypoacetylation precedes the meiotic appearance of phosphorylated H2AX (gamma-H2AX), which marks the beginning of recombinational DNA double-strand breaks (DSBs), it seems that meiotic sex-chromosome inactivation (MSCI) in this grasshopper occurs prior to the beginning of recombination and hence synapsis (which in this species begins later than recombination). In addition, all constitutively heterochromatic chromosome regions harbouring a 180-bp tandem-repeat DNA and rDNA (B chromosomes and pericentromeric regions of A chromosomes) were H3-K9 hypoacetylated at early leptotene even though they will synapse at subsequent stages. This also suggests that meiotic silencing in this grasshopper might be independent of synapsis. The H3-K9 hypoacetylated state of facultative and constitutive heterochromatin persisted during subsequent meiotic stages and was even apparent in round spermatids. Finally, the fact that B chromosomes are differentially hypoacetylated in testis and embryo interphase cells suggests that they might be silenced early in development and remain this way for most (or all) life-cycle stages.

Physical mapping of rDNA and satDNA in A and B chromosomes of the grasshopper Eyprepocnemis plorans from a Greek population.

Adult males and females of the grasshopper Eyprepocnemis plorans from a Greek population were analysed by C-banding, silver impregnation and double FISH for two DNA probes, i.e. ribosomal DNA (rDNA) and a 180-bp tandem repeat DNA (satDNA). This population shows characteristics of rDNA location in A chromosomes that are intermediate between those previously reported for eastern (Caucasus) and western (Spain and Morocco) populations. The four rDNA clusters revealed by FISH in chromosomes X, 9, 10 and 11 in Greek specimens imply two more than the two observed in chromosomes 9 and 11 in the Caucasus, but less than the 12 observed in all chromosomes in Morocco. Remarkably, the X chromosome bears one of the new rDNA locations in Greece with respect to the Caucasus, but it appears to be inactive, in contrast to X chromosomes in western populations, which are usually active. B chromosomes were very frequent in the Greek population, and three variants differing in size were observed, all of these being largely composed of rDNA, with the exception of a small pericentromeric satDNA cluster. The high B frequency suggests that B chromosomes in this population might behave parasitically, in resemblance to Bs in western populations.

Histone H2AX phosphorylation is associated with most meiotic events in grasshopper.

It is widely accepted that the H2AX histone in its phosphorylated form (gamma-H2AX) is related to the repair of DNA double-strand breaks (DSBs). In several organisms, gamma-H2AX presence has been demonstrated in meiotic processes such as recombination and sex chromosome inactivation during prophase I (from leptotene to pachytene). To test whether gamma-H2AX is present beyond pachytene, we have analysed the complete sequence of changes in H2AX phosphorylation during meiosis in grasshopper, a model organism for meiotic studies at the cytological level. We show the presence of phosphorylated H2AX during most of meiosis, with the exception only of diplotene and the end of each meiotic division. During the first meiotic division, gamma-H2AX is associated with i) recombination, as deduced from its presence in leptotene-zygotene over all chromosome length, ii) X chromosome inactivation, since at pachytene gamma-H2AX is present in the X chromosome only, and iii) chromosome segregation, as deduced from gamma-H2AX presence in centromere regions at first metaphase-anaphase. During second meiotic division, gamma-H2AX was very abundant at most chromosome lengths from metaphase to telophase, suggesting its possible association with the maintenance of chromosome condensation and segregation.

Influence of different doses of progesterone treatments on ovarian follicle status in beef cows.

To determine a dose of progesterone (P4) that allow ovarian follicular wave control, Aberdeen Angus cows were randomly assigned into four groups: T600 (n=5), 600 mg of P4/day; T400 (n=5), 400 mg of P4/day; T200 (n=4), 200mg of P4/day and Control (n=4) (excipient only). Progesterone was injected from day 3 to 9 of estrous cycle. Ultrasonographies and blood sample collections were performed daily from day 2 to 10 and on day 15 of the estrous cycle. Additionally, an ultrasonographic study was conducted on day 13. Progesterone concentrations were different among all groups (P<0.01). The diameter of the dominant follicle was greater for control than for T200, T400 and T600 groups (P<0.01); there was no difference between T200 and T400 (P>0.05), but they had a greater diameter follicle than the T600 group (P<0.01). The growth rate of the dominant follicle between day 3 and 7 of estrous cycle was greater for control group (1.63+/-0.3 mmday(-1)) than for T200 (0.56+/-0.19 mmday(-1), P<0.05), T400 (0.6+/-0.23 mmday(-1), P<0.05) and T600 (0.11+/-0.13 mmday(-1), P<0.01) groups. The mean number of class I follicles (3-4mm) per day for the entire experimental period was less for the control group than for T200 (P<0.05), T400 and T600 (P<0.01) groups (3.7+/-1.3; 5.3+/-1.3; 6.6+/-1.8 and 8.1+/-1.9, respectively). The mean number for the T200 group was less than for T600 (P<0.05) and similar for T400 and T600 groups (P>0.05). The number of class III follicles was greater for control group than for the other groups (P<0.01). T200 and T400 groups had similar numbers of class III follicles (P>0.05) and both had greater numbers of follicles than the T600 group (P<0.05). The diameter of the corpus luteum of the T600 group (15.8+/-1.6 mm) was less than for control (21.0+/-2.5 mm, P<0.01), T200 (19.3+/-2.7 mm, P<0.01) and T400 (20.0+/-2.2 mm) groups (P<0.05). The mean diameter of corpus luteum of T200 was similar to T400 (P>0.05), but different from the control group (P<0.05). In conclusion, the daily intramuscular administration of 200mg or more of progesterone from day 3 to 9 of the estrous cycle indicates that plasma concentrations of progesterone can be used to modify the pattern of follicular development during the follicular wave. From day 5 of the estrous cycle, progesterone concentrations greater than 15 ng/ml (T600 group: 600 mg/day of progesterone from day 3 to 9 of the estrous cycle) inhibit dominant follicle development, increase the class I follicle populations (3-4 mm) and diminish the development of the corpus luteum.