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Between 2008 and 2010, 14 isolates of Avibacterium paragallinarum were identified as serovar C-1 in Mexico. All isolates were obtained from commercial laying hens suffering infectious coryza despite a history of vaccination. The enterobacterial repetitive intergenic consensus-based PCR genotyping showed that all isolates had a common pattern. Until recently, serovars A-1, A-2, B-1, and C-2 were the serovars prevalent in Mexico. Serovar C-1 has been identified in Japan and recently in the Americas in Ecuador. Our current study suggests that Av. paragallinarum serovar C-1 is an emerging serovar in Mexico. Our results also indicate that the Mexican isolates of Av. paragallinarum serovar C-1 may have a clonal relationship. Knowledge of the genetic diversity of Av. paragallinarum may be of value in understanding vaccine performance and identifying the best combination to achieve broader protection.

When genomic DNA from the European flat oyster Ostrea edulis L. was digested by BclI enzyme, a band of about 150 bp was observed in agarose gel. After cloning and sequencing this band and analysing their molecular characteristics and genomic organization by means of Southern blot, in situ hybridisation, and polymerase chain reaction (PCR) protocols, we concluded that this band is an interspersed highly repeated DNA element, which is related in sequence to the flanking regions of (CT)-microsatellite loci of the species O. edulis and Crassostrea gigas. Furthermore, we determined that this element forms part of a longer repetitive unit of 268 bp in length that, at least in some loci, is present in more than one copy. By Southern blot hybridisation and PCR amplifications-using primers designed for conserved regions of the 150-bp BclI clones of O. edulis-we determined that this repetitive DNA family is conserved in five other oyster species (O. stentina, C. angulata, C. gigas, C. ariakensis, and C. sikamea) while it is apparently absent in C. gasar. Finally, based on the analysis of the repetitive units in these oyster species, we discuss the slow degree of concerted evolution in this interspersed repetitive DNA family and its use for phylogenetic analysis.

Objective: To verify whether chromosomes 1, 4, and 6 have a role in determining oocyte viability., Design: Retrospective study., Setting: Reproductive Medicine Unit, Società Italiana Studi Medicina della Riproduzione, Bologna, Italy., Patient(s): Eighty-five patients with a normal karyotype who had undergone an assisted conception cycle with chromosomal analysis of first polar bodies for chromosomes 13, 15, 16, 18, 21, and 22 (first panel). A clinical pregnancy was obtained in 43 patients, whereas 42 patients were not pregnant., Intervention(s): After conclusion of clinical pregnancies to delivery or abortion, first polar bodies from 85 patients were reanalyzed for chromosomes 1, 4, and 6 (second panel)., Main Outcome Measure(s): Aneuploidy frequency, clinical pregnancy outcome., Result(s): The aneuploidy rate contributed by chromosome 1, 4, and 6 to the oocytes that were normal for the first panel was significantly higher in the nonpregnant patients (28%) versus the pregnant patients (11%), whereas no difference resulted between term pregnancies (11%) and abortions (10%). This trend was also observed when studying the first polar bodies from the oocytes that originated the transferred embryos. The frequency of aneuploidy for chromosomes 1 and 4 was comparable with that of chromosomes 15, 16, 21, and 22., Conclusion(s): Aneuploidy of chromosomes 1, 4, and 6 seems to be related to failed implantation and not to spontaneous abortions., (Copyright © 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Background: To estimate the incidence of aneuploidy in relation to patients' characteristics, the type of hormonal stimulation and their response to induction of multiple follicular growth, 4163 first polar bodies (PB1s) were analyzed., Methods: Five hundred and forty four infertile couples underwent 706 assisted conception cycles (640 with poor prognosis indications and 66 controls) in which chromosomal analysis of PB1 for the chromosomes 13, 15, 16, 18, 21 and 22 was performed. Results were evaluated in a multivariate analysis., Results: The proportion of normal oocytes was directly correlated (P < 0.01) with (i) the number of mature oocytes and (ii) the establishment of a clinical pregnancy; and inversely correlated (P < 0.01) with (i) female age, (ii) causes of female infertility (endometriosis, abortions, ovulatory factor), (iii) poor prognosis indications (female age, number of previous cycles, multiple poor prognosis indications), (iv) number of FSH units per oocyte and (v) number of FSH units per metaphase II oocyte. There was a weak significance of frequency (P < 0.05) between type of abnormality (originated by chromatid predivision, chromosome non-disjunction or combined mechanisms in the same oocyte) and groups of the studied variables, rather than to a specific abnormality or a specific chromosome., Conclusions: The type of infertility had a significant effect on errors derived from the first meiotic division, whose incidence was significantly higher in the presence of endometriosis or of an ovulatory factor, and in women that experienced repeated abortions. Each aneuploidy event was found to be dependent not on a specific variable, but on groups of variables. In addition, the tendency of chromosomal abnormalities to occur simultaneously implies that the deriving aneuploidies can be of any type.

Background: Some authors have indicated that the cognitive decline may be due to an inadequate nutritional status., Aim of Study: To determine the association between food and nutrient intakes and cognitive capacity score in a group of institutionalized elderly people., Methods: The study subjects were 178 elderly (> or = 65 years of age) institutionalized people from the Madrid region. The diets of these subjects were recorded using the precise weighing method over a 7-day period, and their cognitive capacity assessed using the Short Portable Mental Status Questionnaire (SPMSQ). Subjects were grouped into those who did not incur errors (SPMSQ = 0) and who incurred one or more errors (SPMSQ > 0). Since an association was seen between the SPMSQ test score and age (r = 0.2030; p < 0.01), the subjects were also grouped according to whether they were above or below the percentile 50 (P(50)) for this variable (83 years)., Results: The subjects with no errors in the SPMSQ test (32%) consumed greater quantities of cereals, eggs, oils, and fats. After adjusting for energy intake and educational level an inverse association was seen between fish and vegetable consumption and cognitive capacity score. In addition, these subjects had greater intakes of carbohydrates, polyunsaturated fatty acids, riboflavin, and vitamins C, D, and E. After adjusting for energy intake, a negative relationship was found between cognitive capacity score and the intake of fibre, vitamin B(6), and folic acid., Conclusion: In general, the subjects of our study showed an adequate mental capacity, but those who made no errors in the SPMSQ test had more satisfactory diets. This shows the importance of the diet in the maintenance of cognitive function.

Current methods for analysis of spectroscopic optical coherence tomography (SOCT) signals suffer from an inherent tradeoff between time (depth) and frequency (wavelength) resolution. Here, we present a dual window (DW) method for reconstructing time frequency distributions (TFDs) that applies two orthogonal Gaussian windows that independently determine the spectral and temporal resolution. The effectiveness of the method is demonstrated in simulations and in processing of measured OCT signals that contain fields which vary in time and frequency. The DW method yields TFDs that maintain high spectral and temporal resolution and are free from the artifacts and limitations commonly observed with other processing methods.

In the course of a histopathological survey performed to discover the cause of mass mortality of the striped clam Chamelea gallina in the Balearic Islands (Spain, Mediterranean Sea), we detected a Marteilia-like parasite in 3 clams. Molecular methods were applied to identify the parasite. DNA extracted from a paraffin block was used to carry out a PCR assay for Marteilia refringens detection based on a rDNA sequence of the parasite (the intergenic spacer of ribosomal genes, IGS). The nucleotide sequence of the IGS amplified fragment and the positive signal obtained by in situ hybridisation analysis with a M. refringens-specific probe allowed us to confirm the presence of this parasite in the digestive gland tissue of C. gallina.

Introduction: The 22q11.2 deletion syndrome is a contiguous gene deletion syndrome with an incidence rate of 1/4,000-6,000 live births. The most specific clinical features are: congenital conotruncal heart diseases, palate anomalies, hypocalcaemia, immunity and learning problems, and a characteristic facial phenotype. The objective of this work is to review the presenting phenotype and clinical features of children with 22q11.2 deletion syndrome as a guide for early diagnosis., Patients and Methods: Retrospective study of 22 patients with 22q11.2 deletion syndrome diagnosed at our hospital in the time period 2004-2007. Variables analyzed: incidence, sex, age at diagnosis, presenting phenotype, clinical features, positive family history, mortality and natural history., Results: From a total of 22 patients, 63 % were males, and the median age at diagnosis was of 4.5 years. Presenting pheno-type: congenital heart disease, milestones delay, velopharyngeal incompetence, hypocalcaemia, and mental retardation/psychiatric disturbances., Clinical Features: congenital heart disease (84 %), velopharyngeal incompetence (47 %), milestones delay and learning disabilities (79 %). All of the deletions were de novo, except in one case where the deletion was present as mosaicism in the father. Three patients died, due to congenital heart disease., Conclusions: Clinical expression is widely variable, although a characteristic phenotype exists. Patients with heart disease are diagnosed earlier than other patients with unusual presenting phenotype such as congenital dysphagia. It is important to recognize less common phenotypes at early ages in order to provide multidisciplinary monitoring and accurate genetic counselling.

Agro-food industrial processes produce a large amount of residues, most of which are organic. One of the possible solutions for the treatment of these residues is anaerobic digestion in bioreactors. A novel 18-L bioreactor for treating waste water was designed based on pneumatic agitation and semispherical baffles. Flow patterns were visualized using the particle tracer technique. Circulation times were measured with the particle tracer and the thermal technique, while mixing times were measured using the thermal technique. Newtonian fluid and two non-Newtonian fluids were used to simulate the operational conditions. The results showed that the change from Newtonian to non-Newtonian properties reduces mixed zones and increases circulation and mixing times. Circulation time was similar when evaluated with the thermal and the tracer particle methods. It was possible to predict dimensionless mixing time (theta (m)) using an equivalent Froude number (Fr (eq)).

A highly debated problem in Acipenseridae taxonomy is whether Acipenser oxyrinchus (North American Atlantic sturgeon) and A. sturio (European Atlantic sturgeon) are true species: a detailed comparison of their karyotypes could provide relevant information. Here we describe for the first time the karyotype of A. oxyrinchus (2n = 121 +/- 3), and its features, among which the constitutive heterochromatin, revealed by C-banding technique, the distribution of telomeric regions, and the 5S rRNA genes, detected by FISH. The results reveal that A. oxyrinchus and A. sturio karyotypes and features are quite similar. Moreover, comparing the results obtained through hybridization by FISH with HindIII and PstI satellite DNA in these and in other sturgeon species, no hybridization signals are detected in A. sturio and A. oxyrinchus, while A. stellatus and A. gueldenstaedtii show hybridization. Thus A. sturio and A. oxyrinchus appear very similar from a cytogenetic point of view: these and molecular data repeatedly point out that A. sturio and A. oxyrinchus represent a sister clade in comparison to all other sturgeon species up to now studied.

Objectives: To evaluate the validity of clinical diagnoses of attention deficit/hyperactivity disorder (AD/HD)., Design: Descriptive, cross-sectional study., Setting: Mental Health Care area 6, Murcia, Spain., Participants: The sample consists of those patients referred consecutively to the child psychiatry clinic between July and September 2005., Main Measurements: The presumptive diagnosis of AD/HD in the paediatric department referral was compared with the definitive one in psychiatry (gold standard). Sensitivity, specificity and predictive values and probability quotients were calculated., Results: Seventy-five patients were included; 12% were lost. Sixty-six cases were studied, 15 with AD/HD and 51 with other diagnoses (23 with differential diagnoses from AD/HD). Sensitivity was 86.7% (95% CI, 69.5-100); specificity, 54.9% (95% CI, 41.3-68.6); positive predictive value, 36.1% (95% CI, 20.4-51.8); and negative predictive value, 93.3% (95% CI, 84.4-100)., Conclusions: Validity of diagnosis was moderate with high sensitivity, low specificity and a low positive predictive value, the latter calculated for low AD/HD prevalence. The clinical picture of AD/HD tends to be oversized. The presumptive diagnosis in primary care behaved as a screening test. The clinical diagnosis in primary care should be complemented with other diagnostic tests that provide greater specificity.

Here, a new satellite-DNA family is isolated and characterized from wedge sole, Dicologoglossa cuneata Moreau, 1881 (Pleuronectiformes), a fish having a small genome. This satellite-DNA family of sequences was isolated by conventional cloning after digestion of genomic DNA with the DraI restriction enzyme. Repeat units are 171 bp in length with a high AT content (63%). Several runs of consecutive adenines and thymines were found, and concomitantly computer analyses revealed that these regions are prone to acquire stable sequence-directed curvature. Especially remarkable is that the DraI sequences are composed almost entirely of the repetition of up to fourteen 9-bp motifs (T/C)GTC(A/C)AAAA similar to other vertebrate centromeric satellite-DNA sequences. In fact, we demonstrate the origin of this satellite through duplication of this motif plus the addition of a stretch of cytosines. The centromeric location and the presence in this satellite-DNA sequence of not only different vertebrate motifs (CENP-B box, pJalpha) but also others such as the CDEIII motif of Saccharomyces cerevisiae reveal a possible role in centromere function. All these characteristics provide important information on the origin, function, and the evolution of the centromeric satellite DNAs in wedge sole., (Copyright 2008 S. Karger AG, Basel.)

Gallibacterium was recently established as a new genus including organisms previously reported as Pasteurella anatis, [Actinobacillus] salpingitidis and avian Pasteurella haemolytica-like organisms. The aim of the present study was to develop a PCR method allowing unambiguous identification of Gallibacterium. PCR primers positioned in the 16S rRNA (1133fgal) and 23S rRNA (114r) genes were defined and their specificity was subsequently tested on 122 strains. Twenty-five of the strains represented all of the presently available 15 phenotypic variants of Gallibacterium from different geographical locations, 22 other strains represented other poultry associated bacterial species or bacteria which could pose a differential diagnostic problem including members of the families Pasteurellaceae, Enterobacteriaceae and Flavobacteriaceae, and finally 75 Gallibacterium field strains isolated from Mexican chicken egg-layers. Specific amplicons were generated in all 100 Gallibacterium strains tested, whereas none of the non-Gallibacterium strains tested positive. Correct identification was confirmed by hybridization with the Gallibacterium specific probe GAN850. Two internal amplification control strategies were successfully incorporated into the PCR assay, one based on amplification of the house-keeping gene rpoB (sharing target DNA) and another based on addition of trout DNA (foreign target DNA) and amplification with beta-actin specific primers. In conclusion, the described PCR assay enables specific identification of Gallibacterium and will thus stand as a strong alternative to the present diagnostic methods.

Background: Health system delay (HSD) is an important issue in tuberculosis (TB) control. This report investigates HSD and associated factors in a cohort of Spanish culture-confirmed TB patients., Methods: Data were collected from clinical records. Using logistic regression with two different cut-off points to define HSD (median and 75th percentile), adjusted odds ratios were used to estimate the association between HSD and different variables., Results: A total of 5184 culture-confirmed TB cases were included. Median and 75th percentile HSD were 6 and 25 days respectively. HSD significantly greater than the median was associated with: age >44 years, past or present intravenous drug use, diagnosis at a primary-care centre, prior preventive therapy, positive histology, request for drug-sensitivity testing, presence of silicosis or neoplasia in addition to TB, presence of non-TB related symptoms, and gastrointestinal site. HSD greater than the 75th percentile was related to the same variables, with the exception of diagnosis at a primary-care centre, positive histology, silicosis, non-TB-related symptoms and gastrointestinal site, for which the association disappeared; in contrast, an association with female gender emerged., Conclusion: Despite free health care being universally available in Spain, there are some groups of TB patients whose treatment is unduly delayed.

Ribosomal DNA in sturgeon is informative when analyzed at the molecular level because it bears unique characteristics that are, to a certain extent, ancestral within vertebrates. In this paper, we examine the structure and the molecular evolution of the 5S ribosomal DNA (rDNA) region in 13 sturgeon species, comparing both the 5S ribosomal RNA (rRNA) genes and the non-transcribed spacer (NTS) sequences between the coding regions. We have found that different NTS and 5S gene variants are intermixed in the 5S rDNA arrays of the different sturgeon species and that all variants are ancestral, having been maintained over many millions of years. Using predictive models, we have found similar levels of sequence diversity in the coding regions, as well as in the non-coding region, but fixed interspecific differences are underrepresented for 5S genes. However, contrary to the expectations, we have not found fixed differences between NTS sequences when comparing many pairs of species. Specifically, when they belong to the same phylogeographic clade of the four into which the sturgeon is divided, but fixation of mutations and divergence is found between species belonging to different phylogeographic clades. Our results suggest that the evolution of the two parts of the 5S rDNA region cannot be explained exclusively as the outcome of a balance between mutational, homogenizing (i.e., gene conversion as a predominant force in sturgeon), and selective forces. Rather, they suggest that other factors (i.e., hybridization) might be superimposed over those forces and thus could to some extent be masking their effects.

This study characterizes a repetitive DNA family of sequences in sturgeon, the PstI satellite DNA. We have found a high degree of preservation for these sequences, which are present in all 13 species analyzed, including within the genera Acipenser, Huso, and Scaphirhynchus of the family Acipenseridae. This is one of the most ancient satellite DNAs found to date, because it has been estimated to be more than 100 million years old. Alternatively, to the current view that most satellite DNAs are species-specific or preserved in a few closely related species, the PstI family and other previously characterized sturgeon satellite DNA, the HindIII, represent the most fascinating exceptions to the rapid sequence change usually undergone by satellite DNAs. Here, we compare the evolutionary pattern of these two satellite DNA families, PstI and HindIII, which differ markedly in length, sequence, and nucleotide composition. We have found that, in contrast to the situation in most other living beings, a high degree of preservation, a slow sequence change rate and slowed concerted evolution, appears to be a general rule for sturgeon satellite DNAs. The possible causes for all these features are discussed in the light of the evolutionary specifics found within these ancient organisms.

Background: Patient delay was investigated in a cohort of TB patients identified from May 1996 until April 1997 in 13 Autonomous Regions in Spain. The study covered almost 67% of the total Spanish population., Methods: Data were collected from clinical records. Using unconditional logistic regression with two different cut-off points to define 'patient delay' (the median and 75th percentile), the association between patient delay and different factors was estimated., Results: A total of 7,037 cases were included. Median and 75th percentile delays were 22 and 57 days respectively. Factors associated with patient delay greater than the median (p<0.05) were: non-respiratory symptoms of TB and age over 14 years, although the effect of age was not linear. Furthermore, an interaction was observed between intravenous drug user (IDU) and HIV status, in that, whereas patient delay was greater in IDUs than in non-IDUs among cases whose HIV status was either negative or unknown, among HIV-positive patients no such IDU-related differences were in evidence. Factors associated with extreme patient delay (greater than the 75th percentile) were essentially the same, but the above-described interaction disappeared, with IDU status showing no direct effect. In addition, likelihood of extreme patient delay increased in the case of alcoholism and female gender and decreased in the case of chronic renal failure, corticoid treatment, prison inmates and residents of old age homes., Conclusion: Although there is a universally enjoyed right to health care in Spain, some groups of TB patients could nevertheless be experiencing problems in seeking medical attention.

Gibberellic acid production was studied in different fermentation systems. Free and immobilized cells of Gibberella fujikuroi cultures in shake-flask, stirred and fixed-bed reactors were evaluated for the production of gibberellic acid (GA3). Gibberellic acid production with free cells cultured in a stirred reactor reached 0.206 g/L and a yield of 0.078 g of GA3/g biomass.

Objective: To know the clinical features of nursing home residents with pneumonia comparing with patients with Community-acquired pneumonia and identify the main prognostic index of mortality., Material and Methods: Longitudinal prospective study including all the elderly patients hospitalized in Cantoblanco Hospital of Madrid during the year 2001 for pneumonia and classified according to the Fine prognosis index and the SEPAR criteria., Results: Of the 78 patients with pneumonia, 27 came from Residence, with an average of age of 86.85(+/- 6.43) years old, opposite to 83.11 (+/- 5.87) years in patients with Community acquired pneumonia ( p<0.05). Of all of them, 33,3% belonged to class IV and 66.7% to class V of Fine. Of all the variables studied, only the age (p= 0.03) and the hypoxemia (p= 0.03) were statistical significant., Conclusions: Nursing home residents with pneumonia are older and have more prevalence of morbi-mortality than those with Community acquired pneumonia. In our study, the age and the hypoxemia were the two independent prognosis factors associate to more mortality.

Objective: To study the differences in cognitive status with respect to food habits and energy and nutrient intake in a group of non institutionalised, elderly people., Design: Prospective study., Setting: The study subjects were independently living, elderly people who spent some of their time at day centres in the Comunidad de Madrid (the Madrid region). The study centres were selected by the Madrid City Hall., Subjects: The study included 168 elderly people aged 65-90 y. All accepted the invitation to participate, met all inclusion criteria, and were free of significant cognitive impairment., Interventions: Dietary intake was monitored using a 'food record' for 7 consecutive days including a Sunday. In addition, the 'precise individual weighing' method was used for 5 days in order to monitor the meals taken by the subjects at the centres' canteens. Cognitive capacity was measured using Folstein's mini-mental state examination (MMSE), validated for the Spanish population., Results: Subjects with an adequate cognitive capacity (MMSE>/=28) showed a greater intake of total foods, fish, and alcoholic drinks, but took less foods from the 'various' group (chocolates, cakes, etc). These subjects had a more adequate intake of fatty acids and cholesterol, and a greater intake of vitamins implicated in correct brain function (thiamine, folic acid, vitamin C)., Conclusions: Subjects with satisfactory intellectual function generally had a better diet. This shows the importance of correct nutrition in its maintenance.

The aim of this study was to examine associations between vitamin E status and cognitive performance in elderly people. The study subjects were a group of 34 men and 86 women, aged 65-91 y, who were free of significant cognitive impairment. Dietary intake was monitored using a "weighed food record" for 5 consecutive days including a Sunday. Serum levels of alpha-tocopherol and cholesterol were determined by HPLC and colorimetric methods, respectively. The cognitive capacity of subjects was tested using the Pfeiffer's Mental Status Questionnaire (PMSQ). Subjects with vitamin E intakes lower than 50% of those recommended had higher PMSQ scores, demonstrating a greater number of errors in comparison to participants with a greater intake of the vitamin (0.91 +/- 1.22 vs. 0.47 +/- 0.60, respectively, P < 0.05). Subjects who made no errors in the PMSQ test had significantly higher serum alpha-tocopherol concentrations (19.7 +/- 8.6 micromol/L in men and 20.0 +/- 8.4 micromol/L in women) and alpha-tocopherol/cholesterol ratios (3.5 +/- 2.0 micromol/mmol in men 2.9 +/- 1.4 micromol/mmol in women) compared with those who made errors (alpha-tocopherol 15.1 +/- 5.6 micromol/L in men and 14.9 +/- 6.1 micromol/L in women; alpha-tocopherol/cholesterol ratio 2.4 +/- 0.8 and 2.3 +/- 1.3 micromol/mmol in men and women, respectively). This study shows there to be a relationship between vitamin E status and cognitive function, and that vitamin E status could be improved in this population of elderly individuals.

Setting: Thirteen Autonomous Regions in Spain., Objective: To study the incidence of all forms of tuberculosis (TB) and investigate clinical practice in TB., Design: Cases of all forms of tuberculosis diagnosed in the study setting from May 1986 to April 1997 were identified though active search of different databases. Clinical and epidemiological information on cases was collected from clinical records or by interview with physicians., Results: The overall incidence of all forms of tuberculosis was 38.5/100,000 and the incidence of smear-positive disease was 13.83/100,000. Most cases (97.1%) were Spanish nationals, with rates higher in men than in women (52.7/100,000 vs. 24.87/100,000) and in groups aged 25-34 and 75 years and over (61.35/100,000 and 59.35/100,000, respectively). Disseminated forms were frequent (6.6%), and the most common risk factor was human immunodeficiency virus (HIV) infection (17.7% of cases). Hospitalisation was common (71.6%). Microbiological confirmation of diagnosis was sought for 87.7% of the cases (91.8% of pulmonary vs. 75.5% of extra-pulmonary cases), and 65.2% were culture-positive (73.8% of pulmonary vs. 39.7% of extra-pulmonary cases). HIV-infected patients were treated in almost equal proportions with three or four drugs (49.7% and 48.2%, respectively), while HIV-negative cases or those whose HIV status was unknown were usually treated with three drugs., Conclusion: The epidemiological pattern of TB in Spain is different to other industrialised countries in the age distribution of cases and the proportions of foreigners and cases with HIV infection. Microbiological confirmation of diagnosis is more common in pulmonary than in extra-pulmonary disease, and treatment with four drugs more frequent in HIV-positive cases.

We examined the composition and evolution of a large heterochromatic region present in the genomes of certain species of the genus Muscari (Hyacinthaceae). We found that in Muscari comosum this heterochromatic region is composed mainly of a satellite DNA family, which we named MCSAT. Molecular analyses and in situ hybridization revealed that, through the evolution of Muscari species, the MCSAT sequences have been progressively amplified in several species of the genus, such as M. matritensis and M. dionysicum, attaining enormous amplification in the genome of M. comosum. We discuss the characteristics of this satellite DNA family, which, being exclusively amplified in one chromosome pair of M. comosum, constitute the major exception to the equilocal model of satellite DNA and heterochromatin distribution. Also, we discuss the possibility that the amplification of these sequences in a single chromosome could have contributed to a progressive increase in the asymmetry of the karyotypes in Muscari species.

A simple procedure for the simultaneous determination of modafinil, its acid and sulfone metabolites in plasma is described. The assay involved an extraction of the drug, metabolites and internal standard from plasma with a solid-phase extraction using C18 cartridges. These compounds were eluted by methanol. The extract was evaporated to dryness at 40 degrees C under a gentle stream of nitrogen. The residue was redissolved in 250 microl of mobile-phase and a 30 microl aliquot was injected via an automatic sampler into the liquid chromatograph and eluted with the mobile-phase (26%, v/v acetonitrile in 0.05 M orthophosphoric acid buffer adjusted to pH 2.6) at a flow-rate of 1.1 ml/min on a C8 Symmetry cartridge column (5 microm, 150 mm x 3.9 mm, Waters) at 25 degrees C. The eluate was detected at 225 nm. Intra-day coefficients of variation ranged from 1.0 to 2.9% and inter-day coefficients from 0.9 to 6.1%. The limits of detection and quantitation of the assay were 0.01 microg/ml and 0.10 microg/ml respectively.