Search

This work presents retreat measurements obtained along a 400 m long cliff section located in the Podes Peninsula (43º 37‘ 35’ ‘N, 5º 54’ 4 ‘’ W; Peñas Cape, N Spain). In this cliff, 30 m high and developed on Devonian limestone, dolomite and marl,active coastal erosion and slope movements are observed (Jiménez-Sánchez et al., 2019). The methodology included i) the acquisition and processing of SAR data from Sentinel-1 satellite (from 2018 to 2020), ii) the generation of ground deforma-tion velocity maps and deformation time series by means of the PSIG software and the GEP service, iii) the comparison of DInSAR results with the displacement measurements from 9 topographic nails and 12 pheno survey markers installed in the area and obtained with a total station between 2018 and 2020. The results confirm the need to combine several monitoring techniques in the analysis of retreating cliffs.

The research reported focuses on the multi-causal landslide risk in Sandnes (Norway) in connection with the climate change, the use of the land and the geological/geotechnical properties of the urban subsoil. The municipality is giving special focus on the potential climate change challenges in the development plans of the city and the weather is a significant triggering factor of landslides. The average annual precipitation is 1290 mm and the Norwegian Environmental Agency (NEA) estimates a 10 % increase by 2100 and a 40 % increase for the short-term rainfall (3 h). In addition, the NEA expects sea level to rise between 59 and 89. The study includes records of land use, demographic growth (+1.5 % annually), historical maps and aerial imagery. Events occurred in the past have been inventoried by interviews and document retrieval. Additionally, geological, geotechnical and hydrogeological data from surveys performed in the area have been gathered. The data material is georeferenced in a GIS database for visualization and further treatment. We give special attention to the quality and density of the data to account for the uncertainty and lack of information for certain subareas within the geographical boundaries. This will provide guidance for future work such as soil sampling campaigns. A multi-layered model has been created combining the different aspects studied: elevation, slope, hydrology, historical information and geological data. We model the urban area theoretically affected by the sea level rise and the potential effects on different soils. Finally, the work includes preliminary landslide hazard and vulnerability maps of the present urban plot.

The main objective of this study was to determine differences in sperm recovered from the three segments of the epididymis, i.e., head, corpus and cauda, using routine measurements of sperm quality and DNA fragmentation assessment. There was a larger sperm concentration (P < 0.05) in the head (3.34 × 106 sperm/mL) compared with the corpus and cauda. The percentage of normal morphology was higher in the cauda but had lower motility (P < 0.05). The DNA fragmentation index was higher for sperm obtained from the head and body of the epididymis (P < 0.01) compared with the cauda. The DNA fragmentation index values were 12% higher (P < 0.01) using Wright stain compared with DAPI stain in sperm from the head and corpus of the epididymis, but similar in the cauda. Spermatobioscopy parameters match the maturation changes that occur in the epididymal tract with higher quality sperm being from the cauda, but with a lower concentration compared to sperm found in the head.

El objetivo de este trabajo fue analizar la curva de fragmentación del ADN espermático en semen criopreservado de 10 sementales de la raza de Lidia y observar su evolución a diferentes intervalos de tiempo preestablecidos. Se descongelaron las dosis a 37ºC y se incubaron durante 10 días a 38,6ºC, simulando la temperatura corporal de la hembra. Se analizó el porcentaje de fragmentación del ADN espermático de cada muestra a las 4, 24, 48 y 72 horas y a los 4, 5, 6, 7, 8, 9 y 10 días de incubación utilizando el kit Sperm-Halomax®-Bos-FF (ChromaCell SL, Madrid, Spain) para microscopía de fluorescencia. Los resultados obtenidos indican que: (i) Los niveles de fragmentación basal, así como la velocidad de degradación del ADN espermático a diferentes tiempos de incubación, presentan importantes diferencias interindividuales. (ii) A falta del estudio de un mayor número de muestras, se podría considerar que en esta raza, la integridad de la molécula de cromatina se rompe cuando el semen es incubado a una temperatura de 38,6ºC, pero aún así, puede permanecer estable durante un prolongado período de tiempo. Aquellos sementales con altos porcentajes de fragmentación y que acumulan daños en su ADN espermático rápidamente, tendrían escasas posibilidades de conseguir la fertilización. Por tanto, sería interesante conocer la curva de fragmentación del ADN de cada muestra seminal criopreservada, porque proporcionaría información muy valiosa para establecer la mejor estrategia de reproducción asistida y esclarecer problemas de infertilidad o de falta de desarrollo embrionario cuando se aplican estas técnicas.

This study examined the dynamic response of Spermatozoa DNA Fragmentation after sex selection in bulls using a MoFlo(®) SX (Beckman Coulter, Miami FL) spermatozoa sorter. The dynamic response of spermatozoa DNA fragmentation refers to the changing values of SDF, i.e., rate of SDF (rSDF), when analyzed periodically over a set incubation time at 37 °C. A dynamic assessment of SDF using non-sorted and sex-sorted spermatozoa samples during 72 h of incubation at 37 °C was performed. Results showed a reduced DNA longevity in sex-sorted frozen-thawed spermatozoa, with spermatozoa DNA damage appearing between 24 h and 48 h. The baseline SDF level was higher in conventional frozen-thawed than in sex-sorted frozen-thawed spermatozoa samples; while the reverse occurred for the rSDF. The afore-mentioned result produced a crossover point between both dynamic tendencies of SDF for sex-sorted versus conventional samples. We defined this crossover point as the Crossover Positioning Time (CPT) or the time (in hours) where both curves crossover after a period of spermatozoa incubation at 37 °C. The point at which the CPT occurs could be used as an indicator of the rSDF for individual bulls after X- and Y-chromosome bearing spermatozoa selection. CPT values produced a window of SDF ranging between 24 h and 48 h in the present experiment. It is proposed that higher values for CPT are indicative of bulls presenting chromatin that is more resistant to the external stressors affecting spermatozoa DNA after spermatozoa sorting.

Sperm DNA fragmentation was analysed in 113 semen samples obtained from different rams over a period of one year. Semen samples were collected from: unvaccinated rams between January and June (Control group 1; CG1); vaccinated rams at least 70 days after vaccination between October and December (Control group 2; CG2); vaccinated rams 20 days after vaccination (Vaccinated group 1; VG1); and vaccinated rams 40 days after vaccination (Vaccinated group 2; VG2). Results show Miloxan, the vaccine of interest in this study, increased the percentage of sperm cells with fragmented DNA by 10-fold on average (from 6.5±7.9 to 63.4±24.2). However, the negative impact of vaccination on sperm DNA fragmentation appeared to reversible; near normal levels of sperm DNA fragmentation had been restored 40 days after vaccination (21.7±10.6). On the basis of these data, the use of semen samples from vaccinated animals should be avoided until at least one month after vaccination.

In situ digestion with the restriction endonuclease (RE) Sau3A (Sau3A REISD) uncovers a polymorphism for the pericentromeric heterochromatin of human chromosome 3, which can be positively stained (3+) or not (3–), and has proven useful to differentiate donor and recipient cells after sex-matched bone marrow transplantation and to analyze the so-called hemopoietic chimerism. The aim of the present investigation was to obtain insight into the molecular basis of such polymorphism to optimize its use for chimerism quantification using methodological approaches other than REISD. To this end, fluorescence in situ hybridization (FISH) assays using probes for the satellite DNA sequences that mainly constitute chromosome 3 pericentromeric heterochromatin (α-satellite and satellite 1 DNA) were performed on control and Sau3A-digested chromosomes. The results obtained suggest that chromosome 3 α-satellite DNA is digested in all individuals studied, irrespective of the karyotype obtained by Sau3A REISD (3++, 3+–, 3--), and thus it does not seem to be involved in the polymorphism uncovered by Sau3A on this chromosome. Satellite 1 DNA is not digested in any case, and shows a polymorphism for its domain size, which correlates with the polymorphism uncovered by Sau3A in such a way that 3+ chromosomes show a large domain (3L) and 3– chromosomes show a small domain (3S). It seems, therefore, that the cause of the polymorphism uncovered by Sau3A on the pericentromeric region of chromosome 3 is a difference in the size of the satellite 1 DNA domain. Small satellite 1 DNA domains fall under the resolution level of REISD technique and are identified as 3–.Key words: heterochromatin, α-satellite DNA, classical satellite DNA, satellite 1 DNA, restriction endonucleases, FISH.

This paper aimed at investigating the potential use of sperm DNA fragmentation (SDF) to improve the routine screening of infertility of Holstein bulls. Cryopreserved sperm samples from 201 Holstein bulls provided by an AI center were used in the analyses of SDF at 0 (SDF_0) and 6 (SDF_6) h of incubation at 37°C. A refinement of the sperm chromatin dispersion test implemented in the Sperm-Halomax kit was employed to measure SDF. Records on routinely collected semen traits (volume, concentration, mass and individual motility evaluated in the fresh ejaculate, and individual motility in post-thawed semen straws) were provided by the AI center. Artificial insemination bull fertility was obtained from official field recording as successful or failed insemination. The results show that the average SDF was low (around 3.5%) at 0 and 6 h of incubation. A moderate effect of inbreeding depression was found. Estimated heritability for SDF traits were moderately high (0.41 and 0.29 for SDF_0 and SDF_6, respectively) and estimated repeatability of SDF measures in the same animal were high (0.73 and 0.70 for SDF_0 and SDF_6, respectively). An overall estimated service bull value (ESBV) obtained through statistical modeling that allowed for adjustment of systematic environmental effects not specific to a bull and of the female contribution to fertility, and the estimated genetic values (EGV) were obtained from field-recorded AI information. The ESBV and EGV were also obtained for all semen traits. Moderately large and negative Pearson correlation coefficients were observed between SDF traits and male fertility ranging from (-0.43 to -0.50; P0.001). Results of stepwise regression analyses showed that SDF_6 had the largest partial r(2) (0.15 to 0.26) among all semen characteristics. Overall, the selected semen traits explained 25% and 31% of the observed variability in bull fertility measured as EGV and ESBV, respectively. When looking at the predictive ability of bull fertility categories, the results of discriminant and logistic regression analyses showed that low-fertility bulls (those in the 10th or lower percentile in the fertility distribution) can be accurately identified by using measures of SDF alone or in combination with sperm motility. Values of SDF around 7% to 10% could be used as indicators of low AI success.

The rate of increase of sperm DNA fragmentation (rsDF) in fresh and frozen-thawed and processed sperm samples after a density gradient for sperm selection was analysed after 0, 0.5, 1.5, 4.5, 6, 24, 48 and 72 h of incubation at 37 °C, in five donors with proven fertility. The results showed that: (i) sperm DNA fragmentation (sDF) at baseline in fresh samples (14.3 ± 3.3) was lower than that obtained after freeze-thawing and selection (19.4 ± 4.1), significant differences; (ii) After 6 h of incubation the mean sDF in fresh samples (24.2 ± 10.2) was significantly lower than that in frozen-thawed samples (45.3 ± 7.1); (iii) Subsequently, the rsDF in fresh semen samples was 1.6% per h after 6 h of incubation, while after thawing and selection the rsDF was 4.3% per h; The tendency to increase in sDF showed high R(2) values (R(2) = 0.90) for exponential functions in case of fresh samples, whereas R(2) values for linear functions were higher after sperm selection (R(2) = 0.97). These results indicate that differences in sperm DNA fragmentation dynamics before and after storage are an important issue that must be considered for storage of sperm to be used for artificial reproduction techniques.

This study examined DNA damage and postthaw motility of white-tailed deer sperm (n = 28) before and after sex selection and conventional sorting using MoFlo XDP SX flow cytometry. Semen samples from the same individuals were treated in 4 different ways: 1) chilled-extended sperm samples (without glycerol); 2) cryopreserved conventional samples, samples directly cryopreserved after the addition of extenders; 3) cryopreserved conventionally sorted samples, sorted samples to remove the dead sperm subpopulation; and 4) cryopreserved sex-sorted samples; sorted samples to remove the dead sperm subpopulation and separation of X- and Y-chromosome-bearing sperm. In all the cases (n = 6), conventional samples showed decreased postthaw motilities (43 ± 26%) when compared with X-sorted samples (59 ± 20%; P0.05) and Y-sorted samples (54 ± 20%; P0.05). The DNA fragmentation baseline was5% for frozen-thawed conventional samples, but even less after sex sorting and conventional sorting: 2.4 and 1.7%, respectively. On the other hand, conventional samples showed greater (P0.05) DNA fragmentation than the sex-sorted sperm (n = 6) at 96 h (average of 4.8 ± 4.5% and 5.3 ± 4%, respectively). Conventionally sorted samples (n = 8) did not have greater (P0.05) DNA fragmentation when compared with the sex-sorted samples. Fragmentation of DNA on X-chromosome and Y-chromosome-bearing sorted sperm were not significantly different (n = 10, P0.05) after 96 h (2.6 ± 3.6% and 2.2 ± 0.5%, respectively). Future research should be implemented for examining the fertilizing potential of sex-sorted white-tailed deer sperm (e.g., AI fertility trials).

To update the ad hoc Committee of the Cerebrovascular Diseases Study Group of The Spanish Neurological Society guidelines on prevention of ischaemic stroke (IS) and transient ischaemic attack (TIA).We reviewed available evidence on risk factors and means of modifying them to prevent ischaemic stroke and TIA. Levels of evidence and recommendation grades are based on the classification of the Centre for Evidence-Based Medicine.This first section summarises the recommendations for action on the following factors: blood pressure, diabetes, lipids, tobacco and alcohol consumption, diet and physical activity, cardio-embolic diseases, asymptomatic carotid stenosis, hormone replacement therapy and contraceptives, hyperhomocysteinemia, prothrombotic states and sleep apnea syndrome.Changes in lifestyle and pharmacological treatment for hypertension, diabetes mellitus and dyslipidemia, according to criteria of primary and secondary prevention, are recommended for preventing ischemic stroke.

The Spanish Stroke Group published the "Plan for stroke healthcare delivery" in 2006 with the aim that all stroke patients could receive the same degree of specialised healthcare according to the stage of their disease, independently of where they live, their age, gender or ethnicity. This Plan needs to be updated in order to introduce new developments in acute stroke.A committee of 19 neurologists specialised in neurovascular diseases representing different regions of Spain evaluated previous experience with this Plan and the available scientific evidence according to published literature.The new organised healthcare system must place emphasis on the characteristics of the different care levels with promotion of Reference Stroke Hospitals, set up less restrictive Stroke Code activation criteria that include new therapeutic options, establish new standard measures for endovascular treatment and develop tele-medicine stroke networks.

The increase in the ageing population in the last decades has led to an increased frequency of cancer-associated complications. Among these, neurological disorders stand out, as they appear in 10-30% of patients with systemic neoplasia. Neoplastic meningitis accounts for 4-15% of patients with solid tumours and it has a poor prognosis. The objective of this paper is to describe the clinical, imaging and prognostic characteristics as well as cerebrospinal fluid findings in a series of neoplastic meningitis.We performed a retrospective review of all patients admitted to the Hospital Universitario of Gran Canaria Dr. Negrín with clinical suspicion of neoplastic meningitis between 1990 and 2008. We selected 37 patients with an average age ranging from 15 to 75 years old. A total of 81.8% of the cases in which a primary tumour was found were associated with solid tumours (24.2% were located in the breast, and 24.2% in the lung). The most frequent sign of cranial nerve dysfunction was dyplopia, which was observed in 32.4% of the cases. The average survival rate after diagnosis was 87.9 days (12.6 weeks). The cerebrospinal fluid cytology was positive in 46.4% of the cases.Neoplastic meningitis is a severe complication of both solid and haematological tumours. We stress the importance of maintaining a high level of suspicion to achieve early diagnosis, since the average survival probability for neoplastic meningitis patients is low.

Idiopathic intracranial hypertension (IICH) is characterized by a rising in intracranial hypertension without evidence of intracranial expansive disease. Diagnostic criteria of headache related to IICH are described in the International Classification of Headache Disorders. In clinical practice, however, headache related to IICH may be heterogeneous.Clinical charts of patients with IICH admitted to the Hospital Universitario de Gran Canaria Dr. Negrin between 1990 and 2007 were retrospectively reviewed.Fifty-five patients were included, with a ratio female/male of 8.1:1. Headache was present in 85.4%. The headache was continuous in 63.8% of patients, and diffuse in 51%.Headache related to IICH is heterogeneous, and may mimic primary headache, so a high level of suspicion is needed to avoid diagnostic delay.

The objective of this work is to describe the clinical, etiological and prognostic differences in diabetic and non-diabetic patients presenting ischemic stroke.We performed a retrospective review of all patients with ischemic stroke requiring admission to the Neurology Service at the University Hospital of Gran Canaria Dr. Negrín (Spain) during our study period (1 Jan, 2004 -31 Dec, 2006). We collected demographic, clinical and prognostic variables.A total of 633 patients were included, 256 (40.4%) were diabetic and 377 (59.6%) were non-diabetic. The group of diabetic patients presented a mean age of 69.2or = 9.3 years (65.4or = 13.9 years in non-diabetic). Mean made up 63.7% of the diabetic patients and women 36.3% (62.2% men and 37.8% women in non-diabetics). A total of 85.9% of diabetic patients presented arterial hypertension (62.9% in non-diabetics); 49.2% had previous hyperlipidemia (31% in non-diabetics); 22% had ischemic heart disease (12.7% in nondiabetics); and 8.6% had peripheral arteriopathy (4.8% in non-diabetics). In the group of diabetic patients, 25% presented atherothrombotic stroke (16.4% in non-diabetics); 20.3% were cardioembolic (24.4% in non-diabetics); 26.1% were lacunar (22% in non-diabetics); and 25.7% had undetermined origin (27.2% in non-diabetics). The modified Rankin Scale at discharge was +/- in 65.3% of diabetic patients and 62.5% in non-diabetics.In our series, diabetic patients frequently present other conventional vascular risk factors and show a tendency to more atherothrombotic strokes. We have found no significant differences in relation to prognosis.

Whole-comparative genomic hybridization (W-CGH) allows identification of chromosomal polymorphisms related to highly repetitive DNA sequences localized in constitutive heterochromatin. Such polymorphisms are detected establishing competition between genomic DNAs in an in situ hybridization environment without subtraction of highly repetitive DNA sequences, when comparing two species from closely related taxa (same species, sub-species, or breeds) or somewhat related taxa. This experimental approach was applied to investigating differences in highly repetitive sequences of three sheep breeds (Castellana, Ojalada, and Assaf). To this end, W-CGH was carried out using mouflon (sheep ancestor) chromosomes as a common target to co-hybridize equimolar quantities of two genomic DNAs obtained from either Castellana, Ojalada or Assaf sheep breeds. The results showed that the amount of constitutive heterochromatin is greater in all pericentromeric heterochromatin regions of acrocentric chromosomes than in metacentric or sex chromosomes. Additionally, when W-CGH was performed using DNAs from the Iberian breeds Castellana and Ojalada, chromosomal pericentromeric regions revealed quantitatively and qualitatively a presence of DNA families similar to that obtained from any of the above-cited breeds. On the contrary, when the DNA used in W-CGH experiments was obtained from Assaf, as compared to either Castellana or Ojalada, two different pericentromeric DNA families of highly repetitive sequences could be detected. Lastly, sex chromosomes were shown to be homogeneous among all breeds and thus revealed no detectable constitutive heterochromatin. W-CGH results were confirmed using DNA breakage detection-FISH experiments (DBD-FISH) carried out on lymphocytes. As a whole, the results showed that two different repetitive DNA families are present in the pericentromeric heterochromatin of the sheep breeds studied here. Additionally, they suggest a differential presence of these distinct repetitive DNA families in Castellana and Ojalada breeds as compared to the Assaf breed. Finally, the results of W-CGH after using mouflon as the targeted chromosomes also show that the two DNA families are present in the ancestor.

Cerebral venous sinus thrombosis (CVSTs) constitutes an infrequent location of the venous thrombotic disease. It is caused by diverse factors, although up to 35% of the CVSTs can remain without aetiologic diagnosis. Coagulation disorders have been involved in up to 75% of the CVSTs. AIM. To describe the prevalence of coagulation disorders in a consecutive series of CVSTs.We performed a retrospective revision of the clinical history of all patients with CVST admitted in our service between January, 1st 1995 and December, 31st 2005. The thrombophilic study was carried out a posteriori in whatever individual who lacked it.We studied twenty-one cases (7 men and 14 women), within an age range of 16-69 years old (corresponding to an average of 35,4 +/- 14,4 years old). The thrombophilic study was carried out in eighteen patients. Five (27,7%) presented some hereditary coagulation disorder; in four of them, a hormonal cause coexisted.Recent advances in the diagnostic procedures for the detection of thrombophilias and hereditary prothrombotic mutations have enabled the specialists to be more precise in the aetiologic diagnosis of suspected CVST. Even so, the apparent cause for a high percentage of patients with CVST has not been found yet.

Whole-comparative genomic hybridization (W-CGH) has been used to exemplify a simple methodology which allows identifying and mapping whole genome differences for highly repetitive DNA sequences between two related species of unknown genomic background. The use of this technique to the species binomy Arcyptera fusca/Arcyptera tornosi has allowed the identification of different DNA families mainly concentrated within the para-/peri-centromeric and distal heterochromatic regions of different chromosomes, which are differentially expanded in both genomes. Additionally, W-CGH allowed chromosome mapping of particular euchromatic regions immersed in the chromosome arms which have been affected by processes of DNA amplification and losses. A molecular approach was also conducted to analyse satellite DNA families in these species. We have found three different families showing an unequal representation in both species. Two of these families showed a centromeric location (EcoRV-390CEN and Sau3A-419CEN), whereas the last one was located at distal heterochromatic regions (Sau3A-197TEL). As A. fusca is a widely distributed species represented in most European high mountains, whereas A. tornosi is an endemic species represented in the Iberian Peninsula, the differences and resemblances reported here offer a good basis to support a close evolutionary relationship between both of the actually isolated species. Finally, W-CGH allowed identification of an asynchronic pattern of heterochromatin condensation through early prophase (characteristic in both species) which is uncommon or probably has been poorly analysed within classical early condensing chromosome domains through meiosis. The congruence of the obtained cytological and molecular results is analysed in light of the ancestral genome relationship between both species.

Infertility affects almost 20% of couples in reproductive age and the male factor being responsible of 50% of this infertility. Among the classic parameters that determine a good seminal quality such as sperm motility, sperm morphology or the quality of the of acrosomes and/or sperm membranes, the integrity of the DNA molecule is crucial to carry out a successful fertilization. Nevertheless, the study of this parameter has not been straightforward approached. This fact has shunned its incorporation, as a routine technique, within a standard seminogram. The aim of the present review is to summarize and update those technologies that are considered more successful to study sperm DNA fragmentation with special emphasis to: 1) the levels of technological complexity and the possibility of its use in laboratories of andrology, according with the equipment and the resources available, and 2) the effects and possible implications of high level of sperm DNA fragmentation for fecundation, embryo development and fertility.

The mixed success of equine artificial insemination programs using chilled and frozen-thawed semen is most likely associated with the variable response of the sperm cell to the preservation process and the fact that stallions are not selected on the basis of reproductive performance. We propose that the traditional indicators of sperm viability do not fully account for male factor infertility in the stallion and that knowledge of sperm DNA damage in the original semen sample and during semen processing may provide a more informed explanation of an individual stallion's reproductive potential. This study reports on the validation of a sperm DNA fragmentation test based on the sperm chromatin dispersion test (SCD) for stallion spermatozoa and on its application to semen that was chilled (4 degrees C; n=10) or frozen-thawed (n=13). Semen samples were collected by artificial vagina and the proportion of sperm with fragmented DNA determined. Seminal plasma was then removed by centrifugation and the sperm pellet re-suspended in commercial extenders prior to being chilled or cryopreserved using standard industry protocols. Chilled semen was cooled slowly to 4 degrees C and stored for 1h before commencing the analysis; cryopreserved semen was thawed and immediately analyzed. Following chilling or cryopreservation, the semen samples were incubated at 37 degrees C and analyzed for SCD after 0, 4, 6, 24 and 48 h storage. The results of this investigation revealed that there was no significant difference in the sperm DNA fragmentation index (sDFI) of sperm evaluated initially after collection compared to those tested immediately after chilling or cryopreservation. However, within 1h of incubation at 37 degrees C, both chilled and frozen-thawed spermatozoa showed a significant increase in the proportion of sDFI; after 6h the sDFI had increased to over 50% and by 48 h, almost 100% of the sperm showed DNA damage. While the sDFI of individual stallions at equivalent times of incubation was variable, an analysis of the rate of change of sDFI revealed no difference between stallions or the way in which the semen was preserved. In terms of sperm DNA fragmentation dynamics, the highest intensity of sperm DNA damage occurred in the first 6h of incubation. We suggest that the SCD test can be used as a routine assessment tool for the development and refinement of preservation protocols designed to reduce stallion sperm DNA damage.

In situ digestion with the restriction endonuclease (RE) Sau3A (Sau3A REISD) uncovers a polymorphism for the pericentromeric heterochromatin of human chromosome 3, which can be positively stained (3+) or not (3-), and has proven useful to differentiate donor and recipient cells after sex-matched bone marrow transplantation and to analyze the so-called hemopoietic chimerism. The aim of the present investigation was to obtain insight into the molecular basis of such polymorphism to optimize its use for chimerism quantification using methodological approaches other than REISD. To this end, fluorescence in situ hybridization (FISH) assays using probes for the satellite DNA sequences that mainly constitute chromosome 3 pericentromeric heterochromatin (alpha-satellite and satellite 1 DNA) were performed on control and Sau3A-digested chromosomes. The results obtained suggest that chromosome 3 alpha-satellite DNA is digested in all individuals studied, irrespective of the karyotype obtained by Sau3A REISD (3++, 3+-, 3--), and thus it does not seem to be involved in the polymorphism uncovered by Sau3A on this chromosome. Satellite 1 DNA is not digested in any case, and shows a polymorphism for its domain size, which correlates with the polymorphism uncovered by Sau3A in such a way that 3+ chromosomes show a large domain (3L) and 3- chromosomes show a small domain (3S). It seems, therefore, that the cause of the polymorphism uncovered by Sau3A on the pericentromeric region of chromosome 3 is a difference in the size of the satellite 1 DNA domain. Small satellite 1 DNA domains fall under the resolution level of REISD technique and are identified as 3-.

Chimerism quantification (CQ) after sex-matched bone marrow transplantation (BMT) is based on the identification of autosomal differences distinguishable at the chromosomal level, such as variations within constitutive heterochromatin between the recipient and the donor. The probability of finding distinguishable recipient/donor pairs at the karyotypic level depends on the frequency of the chromosome variants or morphs in the population, on whether recipient and donor are related, and if so, their kinship relation. We have developed a population genetics-based method that allows the estimation of the percentage of post-BMT CQ expected to be informative using any autosomal polymorphic marker. This method has been developed for the most common transplant situations, such as sibling-matched recipient/donor pairs, haploidentical related (parental/filial) pairs, and unrelated pairs. The method developed was applied to a polymorphism of the pericentromeric region of chromosome 3. This polymorphism becomes evident after in situ digestion with the restriction endonuclease Sau3A, and can be successfully used for CQ. It has been estimated that approximately 59% of the cases of BMT from unrelated donors, 36% of those from sibling donors, and 42% from parental/filial donors, are expected to be distinguishable for post-BMT CQ using this approach.

To study acute cerebrovascular ischemia in young persons in our environment.A retrospective study was made of 68 patients, aged between 15 and 45, with transient ischemic accidents (TIA) or cerebral infarcts. Etiological factors, clinical features, diagnostic groups according to the TOAST classification and prognosis were analyzed.In our series there was a ratio of 1.26 in favor of the women. TIAs made up 29.4% of the cases. The most commonly affected territory was that of the carotid artery. In persons over the age of 30 there was greater prevalence of this pathology and of the risk factors diabetes, arterial hypertension and dyslipaemia. There was a significant association with migraine in those aged under 30. The biggest diagnostic group was that of infarct of unknown origin (39.7%), probably because of the specificity of the classification. The functional condition on follow-up was good: only 4.4% of the patients had a score greater than 3 on the Rankin scale. There were no deaths either during the acute phase or subsequently. Recurrences were seen in 10.2% of the patients.The results obtained in our series are similar to those published in the literature. In acute cerebrovascular ischaemia in young persons, full diagnostic investigations should be carried out in order to give suitable treatment.

The era of the longitudinal differentiation of metaphase chromosomes (chromosome banding) dawned in 1968 when Caspersson and coworkers described the use of quinacrine mustard as a fluorescent compound which heterogeneously stained fixed human chromosomes, thus producing the so-called Q-bands. Around the same time, Pardue and Gall (1970) were experimenting with the in situ hybridization of nucleic acids to localize satellite DNAs in mouse chromosomes. Twenty five years later this technique is an indispensable tool in the approach to basic problems in various fields of Genetics and Biology. It was also in the early 1970’s that Sumner et al. (1971) and Sumner (1972) unveiled techniques of easy application to produce G- and C-bands by means of chemical pretreatment of metaphase chromosomes. The basis of chromosomal differentiation was established. Thus, the use of fluorochromes which are able to recognize GC- or AT-enriched DNA motifs, physicochemical modifications of the chromatin, and the incorporation of modern molecular techniques into cytogenetics would prove to be the first steps along new pathways in the production of longitudinal differentiation of metaphase chromosomes or interphase nuclei.

Fixed chromosomes from human amniotic fluid cells and peripheral blood lymphocytes were digested in situ with exonuclease III and the single stranded DNA obtained was used as template for an in situ random primer extension. Under these conditions an R banding pattern, more evident in lymphocytes than in amniocytes, was obtained. Nevertheless, constitutive heterochromatin of chromosomes 1, 16, Yq, and mainly the pericentromeric region of chromosome 9 was far more intensely labelled in amniocytes than in lymphocytes. Fluorescence in situ hybridisation with a specific classical satellite DNA probe, showed that this differential labelling was dependent on a greater sensitivity of chromosome 9 constitutive heterochromatin to exonuclease III digestion in amniocytes than in lymphocytes, thus indicating qualitative differences in this region between both human cellular materials.

interaction between stallion and extender with respect to freezability. To evaluate the differences between “poor” and “good” freezers, the stallions grouped into the same cluster with both the extenders (5 and 3, respectively) were considered.When datawere compared prior to freezing, no differences were reported for TM, DFI, MI, HF and mitochondrial membrane potential, while VAP showed higher values in “good” freezer (P < 0.01). After centrifugation, “poor” freezers presented a worsening in TM (P < 0.01). After thawing, “good” freezers had lower DFI (P< 0.01) and higher TM (P< 0.01) and VAP (P< 0.01) compared to “poor” freezers. Furthermore, lower MI (P < 0.001) and greater HF (P < 0.01) and percentage of sperm with inactive mitochondria (P< 0.05)were reported for “poor” freezers. These results show that the damage induced by cryopreservation in “poor” freezers affects several sperm compartments: DNA, cell membranes and mitochondria. The discordant clustering of some stallions associated with different extenders, confirms previous observations suggesting the use of freezing protocols specific for each individual stallion to improve results semen cryopreservation.

We used X-ray microanalysis to study the changes induced in mouse metaphase chromosomes as a result of digestion with the restriction endonuclease HaeIII. The phosphorus X-ray signal was used as a marker for DNA and the sulfur signal for protein. Calcium, iron, copper, and zinc were also detected. HaeIII induced a loss of phosphorus from both the centromeres and chromosome arms, but the losses in the arms were much greater. These changes were accompanied by an increase in the electron density of the centromeres and a reduction in that of the arms. No reduction in the sulfur signal in either arms or centromeres occurred as a result of HaeIII digestion. Except for calcium, which showed only a moderate reduction, the inorganic ions exhibited very large losses as a result of HaeIII digestion. The differentiation of chromosome arms and centromeres as a result of HaeIII digestion is therefore not simply due to differential loss of DNA but also involves structural reorganization of the chromatin, as shown by electron microscopy. This reorganization does not involve loss of proteins but may be correlated with changes in the amounts of inorganic ions known to be involved in chromatin condensation.

Two types of extra heterochromatin, in the form of either supernumerary segments or B-chromosomes, have been observed in natural populations of Gomphocerus sibiricus. The presence of a heterochromatic supernumerary segment affecting the M7 pair in a population from the Valle de Aran disturbs the normal separation of this bivalent at anaphase I of meiosis and non-disjunction products, respectively deficient or duplicate for the M7, are present at metaphase II.

The electrical resistivity tomography method is able to provide a high resolution section of subsoil where to interpret the faults structure in depth. However, the results with this technique can vary depending on the acquisition array used. In this work we compare three surveys with different array configurations (dipole-dipole, Schlumberger and Wenner) with the aim to optimize the electrical method for the study of fault zones. All of them were applied within a same line of investigation in order to characterize the Ventaniella fault. This is a significant structure in NW Iberia that shows a low but persistent seismicity in some of its segments. It runs for at least 450 km following a NW-SE direction from the continental platform offshore in the Bay of Biscay to the Duero Basin across the Cantabrian Mountains within the Iberian plate. The data obtained has allowed to constrain to what extent the various array configurations are useful in determining internal structure, position and range of depths of the main fault and associated structures.

The spinal abscess is an infrequent infectious complication whose diagnosis is often delayed, therefore bringing about significant morbidity and death rates. In the last years, its incidence has raised progressively. The objective of this work is to present the clinical features of a series of spinal abscesses.The clinical histories of all patients diagnosed of spinal cord abscess were evaluated retrospectively.We obtained seven cases, with a 5/2 male/female proportion, all subjects being over 50 years old. Incidence was 0.58/10,000 admissions per year. All patients debuted with local pains, but only 6/7 (90%) had fever. Focal neurological signs appeared in all patients within the first 0-5 day-period; they were weakness (86%) and sensitive alterations (43%). The most frequent germ was Staphylococcus aureus ( 5 0%). Diabetes mellitus existed in 5/7 cases (71.5%). Four patients were treated by surgery; three of them (75%) had flaccid motor deficit as sequel and the other died. Medical treatment alone was maintained in 3/7 (43%); two experienced partial improvement without recovery of the motor functions and another died.Spinal epidural abscess is a rare, although serious, disease. For this reason, we should maintain a high level of suspicion when a patient has spinal or radicular pain associated with systemic symptoms such as fever or deterioration in the general state of health, especially in patients50 years under weakening situations.

A homogeneous heterochromatic zone obtained with the direct application of the C-banding technique is unravelled into 2 sub-zones with the successive application of orcein staining and the C-banding method in the grasshopperArcyptera fusca.