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1.  Draft Genome Sequences of Two Burkholderia multivorans Sequential Isolates from a Chronic Lung Infection of a Cystic Fibrosis Patient 
Genome Announcements  2015;3(1):e01531-14.
Burkholderia multivorans belongs to the Burkholderia cepacia complex, which comprises opportunistic pathogens infecting cystic fibrosis (CF) patients. Here, we report the genome sequences and annotations of two sequential B. multivorans clinical isolates (D2095 and D2214) displaying different traits. The differences in the genomic contents of these isolates may provide clues regarding the evolution of B. multivorans within the airways of a CF patient.
PMCID: PMC4333657  PMID: 25676757
2.  Males and Females Contribute Unequally to Offspring Genetic Diversity in the Polygynandrous Mating System of Wild Boar 
PLoS ONE  2014;9(12):e115394.
The maintenance of genetic diversity across generations depends on both the number of reproducing males and females. Variance in reproductive success, multiple paternity and litter size can all affect the relative contributions of male and female parents to genetic variation of progeny. The mating system of the wild boar (Sus scrofa) has been described as polygynous, although evidence of multiple paternity in litters has been found. Using 14 microsatellite markers, we evaluated the contribution of males and females to genetic variation in the next generation in independent wild boar populations from the Iberian Peninsula and Hungary. Genetic contributions of males and females were obtained by distinguishing the paternal and maternal genetic component inherited by the progeny. We found that the paternally inherited genetic component of progeny was more diverse than the maternally inherited component. Simulations showed that this finding might be due to a sampling bias. However, after controlling for the bias by fitting both the genetic diversity in the adult population and the number of reproductive individuals in the models, paternally inherited genotypes remained more diverse than those inherited maternally. Our results suggest new insights into how promiscuous mating systems can help maintain genetic variation.
PMCID: PMC4277350  PMID: 25541986
3.  Deciphering the Genome Repertoire of Pseudomonas sp. M1 toward β-Myrcene Biotransformation 
Pseudomonas sp. M1 is able to mineralize several unusual substrates of natural and xenobiotic origin, contributing to its competence to thrive in different ecological niches. In this work, the genome of M1 strain was resequenced by Illumina MiSeq to refine the quality of a published draft by resolving the majority of repeat-rich regions. In silico genome analysis led to the prediction of metabolic pathways involved in biotransformation of several unusual substrates (e.g., plant-derived volatiles), providing clues on the genomic complement required for such biodegrading/biotransformation functionalities. Pseudomonas sp. M1 exhibits a particular sensory and biotransformation/biocatalysis potential toward β-myrcene, a terpene vastly used in industries worldwide. Therefore, the genomic responsiveness of M1 strain toward β-myrcene was investigated, using an RNA sequencing approach. M1 cells challenged with β-myrcene(compared with cells grown in lactate) undergo an extensive alteration of the transcriptome expression profile, including 1,873 genes evidencing at least 1.5-fold of altered expression (627 upregulated and 1,246 downregulated), toward β-myrcene-imposed molecular adaptation and cellular specialization. A thorough data analysis identified a novel 28-kb genomic island, whose expression was strongly stimulated in β-myrcene-supplemented medium, that is essential for β-myrcene catabolism. This island includes β-myrcene-induced genes whose products are putatively involved in 1) substrate sensing, 2) gene expression regulation, and 3) β-myrcene oxidation and bioconversion of β-myrcene derivatives into central metabolism intermediates. In general, this locus does not show high homology with sequences available in databases and seems to have evolved through the assembly of several functional blocks acquired from different bacteria, probably, at different evolutionary stages.
PMCID: PMC4316614  PMID: 25503374
genome sequencing; biocatalysis; terpenes; Pseudomonas; RNA-seq; genomic island
4.  Correction of Angle Class II division 1 malocclusion with a mandibular protraction appliances and multiloop edgewise archwire technique 
Korean Journal of Orthodontics  2014;44(5):268-277.
A Brazilian girl aged 14 years and 9 months presented with a chief complaint of protrusive teeth. She had a convex facial profile, extreme overjet, deep bite, lack of passive lip seal, acute nasolabial angle, and retrognathic mandible. Intraorally, she showed maxillary diastemas, slight mandibular incisor crowding, a small maxillary arch, 13-mm overjet, and 4-mm overbite. After the diagnosis of severe Angle Class II division 1 malocclusion, a mandibular protraction appliance was placed to correct the Class II relationships and multiloop edgewise archwires were used for finishing. Follow-up examinations revealed an improved facial profile, normal overjet and overbite, and good intercuspation. The patient was satisfied with her occlusion, smile, and facial appearance. The excellent results suggest that orthodontic camouflage by using a mandibular protraction appliance in combination with the multiloop edgewise archwire technique is an effective option for correcting Class II malocclusions in patients who refuse orthognathic surgery.
PMCID: PMC4192529  PMID: 25309867
Class II malocclusion; Mandibular advancement; Orthodontic treatment
5.  Arthroscopic Double-Locked Stitch: A New Technique for Suturing Rotator Cuff Tears 
Arthroscopy Techniques  2014;3(2):e223-e225.
There are a number of reasons for failed rotator cuff tear repair. In such cases the suture-tendon interface seems to be the most vulnerable area, especially when tendon degeneration is present. We describe a new technique, the arthroscopic double-locked suture, that increases the tendon fixation and has the added benefit of being placed parallel to the blood vessels, therefore avoiding damage to the tendon vascularization. The suture may be achieved by use of knots or knotless anchors and suture passers, without the need for any additional instrumentation. The new technique is especially helpful in cases in which the tendon is retracted and degeneration is present, impeding the use of the double-row technique or its transosseous equivalents.
PMCID: PMC4044500  PMID: 24904764
6.  Proteomic Profiling of Burkholderia cenocepacia Clonal Isolates with Different Virulence Potential Retrieved from a Cystic Fibrosis Patient during Chronic Lung Infection 
PLoS ONE  2013;8(12):e83065.
Respiratory infections with Burkholderia cepacia complex (Bcc) bacteria in cystic fibrosis (CF) are associated with a worse prognosis and increased risk of death. In this work, we assessed the virulence potential of three B. cenocepacia clonal isolates obtained from a CF patient between the onset of infection (isolate IST439) and before death with cepacia syndrome 3.5 years later (isolate IST4113 followed by IST4134), based on their ability to invade epithelial cells and compromise epithelial monolayer integrity. The two clonal isolates retrieved during late-stage disease were significantly more virulent than IST439. Proteomic profiling by 2-D DIGE of the last isolate recovered before the patient’s death, IST4134, and clonal isolate IST439, was performed and compared with a prior analysis of IST4113 vs. IST439. The cytoplasmic and membrane-associated enriched fractions were examined and 52 proteins were found to be similarly altered in the two last isolates compared with IST439. These proteins are involved in metabolic functions, nucleotide synthesis, translation and protein folding, cell envelope biogenesis and iron homeostasis. Results are suggestive of the important role played by metabolic reprogramming in the virulence potential and persistence of B. cenocepacia, in particular regarding bacterial adaptation to microaerophilic conditions. Also, the content of the virulence determinant AidA was higher in the last 2 isolates. Significant levels of siderophores were found to be secreted by the three clonal isolates in an iron-depleted environment, but the two late isolates were more tolerant to low iron concentrations than IST439, consistent with the relative abundance of proteins involved in iron uptake.
PMCID: PMC3862766  PMID: 24349432
7.  Machine Learning Techniques for Arterial Pressure Waveform Analysis 
The Arterial Pressure Waveform (APW) can provide essential information about arterial wall integrity and arterial stiffness. Most of APW analysis frameworks individually process each hemodynamic parameter and do not evaluate inter-dependencies in the overall pulse morphology. The key contribution of this work is the use of machine learning algorithms to deal with vectorized features extracted from APW. With this purpose, we follow a five-step evaluation methodology: (1) a custom-designed, non-invasive, electromechanical device was used in the data collection from 50 subjects; (2) the acquired position and amplitude of onset, Systolic Peak (SP), Point of Inflection (Pi) and Dicrotic Wave (DW) were used for the computation of some morphological attributes; (3) pre-processing work on the datasets was performed in order to reduce the number of input features and increase the model accuracy by selecting the most relevant ones; (4) classification of the dataset was carried out using four different machine learning algorithms: Random Forest, BayesNet (probabilistic), J48 (decision tree) and RIPPER (rule-based induction); and (5) we evaluate the trained models, using the majority-voting system, comparatively to the respective calculated Augmentation Index (AIx). Classification algorithms have been proved to be efficient, in particular Random Forest has shown good accuracy (96.95%) and high area under the curve (AUC) of a Receiver Operating Characteristic (ROC) curve (0.961). Finally, during validation tests, a correlation between high risk labels, retrieved from the multi-parametric approach, and positive AIx values was verified. This approach gives allowance for designing new hemodynamic morphology vectors and techniques for multiple APW analysis, thus improving the arterial pulse understanding, especially when compared to traditional single-parameter analysis, where the failure in one parameter measurement component, such as Pi, can jeopardize the whole evaluation.
PMCID: PMC4251397  PMID: 25562520
pulse wave analysis; arterial pressure waveform; machine learning; arterial stiffness; augmentation index
8.  Burkitt's Lymphoma in a Pregnant Woman: Case Report and Review of the Literature 
Burkitt's lymphoma (BL) is an aggressive B-cell malignancy with very high proliferation rate, more common in males than females. Here, we describe a case of Burkitt's lymphoma in a 24-week pregnant woman with cervical and abdominal involvement. The common genetic event of virtually all BL is a reciprocal chromosomal translocation involving the proto-oncogene MYC and one of the Ig gene heavy or light chain loci. Supportive treatment was administered until early delivery, after which the patient was treated according to protocol LMB96. Pregnancy and tumorogenesis share some important events such as immunologic tolerance, angiogenesis, and editing the host immune response. Little is known about the relationship between these events in pregnancy and in tumorogenesis.
PMCID: PMC3666439  PMID: 23762688
9.  Towards the Description of the Genome Catalogue of Pseudomonas sp. Strain M1 
Genome Announcements  2013;1(1):e00146-12.
Pseudomonas sp. strain M1 is a soil isolate with remarkable biotechnological potential. The genome of Pseudomonas sp. M1 was sequenced using both 454 and Illumina technologies. A customized genome assembly pipeline was used to reconstruct its genome sequence to a single scaffold.
PMCID: PMC3569285  PMID: 23405299
10.  Microsatellite markers for identification and parentage analysis in the European wild boar (Sus scrofa) 
BMC Research Notes  2012;5:479.
The wild boar (Sus scrofa) is among the most widespread mammal species throughout the old world. Presently, studies concerning microsatellites in domestic pigs and wild boars have been carried out in order to investigate domestication, social behavior and general diversity patterns among either populations or breeds. The purpose of the current study is to develop a robust set of microsatellites markers for parentage analyses and individual identification.
A set of 14 previously reported microsatellites markers have been optimized and tested in three populations from Hungary, Portugal and Spain, in a total of 167 samples. The results indicate high probabilities of exclusion (0.99999), low probability of identity (2.0E-13 – 2.5E-9) and a parentage assignment of 100%.
Our results demonstrate that this set of markers is a useful and efficient tool for the individual identification and parentage assignment in wild boars.
PMCID: PMC3475110  PMID: 22943565
Sus scrofa; Parentage assignment; Individual identification; Microsatellite markers; Wild boar
11.  Is prnt a Pseudogene? Identification of Ram Prt in Testis and Ejaculated Spermatozoa 
PLoS ONE  2012;7(8):e42957.
A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrPC), expressed by the prion gene (prnp), into an abnormally folded isoform (PrPSc) with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel), and prnt (that encodes the prion protein testis specific - Prt) with sprn (shadow of prion protein gene, that encodes Shadoo or Sho) genes, constitute the “prion gene complex”. Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA) immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated with its phenylalanine residue.
PMCID: PMC3427297  PMID: 22937002
12.  Surgical Treatment of Iatrogenic Rectourinary Fistula—York-Mason Technique—a Case Report 
ISRN Urology  2011;2011:292517.
Introduction. Recto-urinary fistulas resulting from trauma or surgery are a serious and debilitating complication. They represent a challenge not only because of the difficulty on choosing the best technique to solve them but also because of the risk of recurrence. Spontaneous cure is rare. Materials and Methods. We describe the case of a 61-years-old man that on the 9th postoperative day of a laparoscopic radical prostatectomy (LRP) started with fecaluria and liquid faeces. Recto-urinary fistula was confirmed at the 10th postoperative day by CT scan and contrast enema. Discussion. We chose the York-Mason technique, because it is simple to perform, effective and has minimal morbidity. This is a posterior, transrectal, and transsphincteric approach, carried out on healthy tissues without previous scarring phenomena. Results. The postoperative period progressed without complications, and the patient discharged on the 4th day. The closure of the fistula was confirmed radiologically by retrograde cystography after 4 weeks allowing the removal of drainage catheter. The reconstruction of intestinal transit was carried out 2 months later. Conclusion. The York-Mason technique, a transrectal and transsphincteric approach with minimal morbidity, proved to be effective on the resolution of the recto-urinary fistula, a rare complication of the radical prostatectomy.
PMCID: PMC3196981  PMID: 22084795
13.  Hematological abnormalities and 22q11.2 deletion syndrome 
The 22q11.2 deletion syndrome (22q11DS) is a common genetic disease characterized by broad phenotypic variability. Despite the small number of studies describing hematological alterations in individuals with 22q11DS, it appears that these abnormalities are more frequent than previously imagined. Thus, the objective of our study was to report on a patient with 22q11DS presenting thrombocytopenia and large platelets and to review the literature. The patient, a 13-year-old boy, was originally evaluated due to craniofacial dysmorphia and speech delay. He also had a history of behavioral changes, neuropsychomotor delay and recurrent otitis/sinusitis. The identification of a 22q11.2 microdeletion by fluorescent in situ hybridization diagnosed the syndrome. Despite his hematological alterations, he only had a history of epistaxis and bruising of the upper and lower limbs. Assessments of the prothrombin time, thrombin time, partial thromboplastin time, bleeding time, fibrinogen levels and platelet aggregation (including the ristocetin induced platelet aggregation test) were all normal. Hematological alterations observed in 22q11DS are directly related to the genetic disorder itself (especially in respect to deletion of the GPIb gene) and secondary to some clinical findings, such as immunodeficiency. Macrothrombocytopenia is increasingly being considered a feature of the broad spectrum of 22q11DS and may potentially be a clinical marker for the syndrome.
PMCID: PMC3520641  PMID: 23284264
Blood platelets; Thrombocytopenia; Bernard-Soulier syndrome; DiGeorge syndrome; In situ hybridization; Chromosomes, Human, Pair 22
14.  Impact of positive surgical margins on biochemical relapse after radical retropubic prostatectomy (RRP) 
RP (radical prostatectomy) technique continues the major treatment option for men with potential cure and life expectancy exceeding 10 years. The aim of the study is to assess the impact of PSM on BR (biochemical relapse), to identify PSM risk factors, to clarify the factors involved in BR in the absence of PSM.
Material and methods
Consultation of 171 medical-records from patients submitted to RRP (radical retropubic prostatectomy) between January/2000-December/2005. Mean-age: 64 yr. Mean – PSA (positive surgical margin): 11.88 ng/ml. Clinical staging: 67.8% cT1, 32.2% cT2. GS: ≤6 (66.1%), =7 (21.1%), 8-10 (12.3%). PS: pT0 1.2%, pT2 50.3%, pT3a 36.3%, pT3b 12.9%, pT4 0.6%. pathological Gleason score: ≤6 39.2%, =7 40.9%, 8-10 19.3%. RB definition was PSA ≥0.2 ng/ml. Adjusted Odds-Ratios with 95% confidence intervals (CI) were estimated through univariate logistic regression.
There were PSM in 46 specimens, 28 had single PSM and 18 multiple PSM (≥2). BR occurred in 57 patients (33.3%), with an average time after surgery of 23.5 months – 26 patients had PSM and 31 had not. Statistical significant results for BR in variables PSA, PS and PSM. Quadruples if PSM (p <0.0001), triples in single PSM (p = 0.01) and is 6x higher in multiple PSM (p = 0.001). Regarding factors that influence the presence of PSM, only PS ≥pT3a reach statistical significance (p <0.0001). Patients with BR but without PSM (54.38%), variables statistically significant were: initial PSA >10, (p = 0.029) and pathological Gleason score ≥8 with a risk nearly 4x higher than pathological Gleason score ≤6 (p = 0.027).
Statistical risk analysis concluded that the presence of PSM in RRP is strongly influenced by PS ≥pT3a. The presence of PSM and their number increase significantly the risk of BR compared to other factors. In the absence of PSM, the factors that seem to be crucial and with greater impact on BR are initial PSA>10 and pathological Gleason score ≥8.
PMCID: PMC3921750  PMID: 24578898
biochemical relapse; positive surgical margins; radical retropubic prostatectomy
15.  Additive effects of LPL, APOA5 and APOE variant combinations on triglyceride levels and hypertriglyceridemia: results of the ICARIA genetic sub-study 
BMC Medical Genetics  2010;11:66.
Hypertriglyceridemia (HTG) is a well-established independent risk factor for cardiovascular disease and the influence of several genetic variants in genes related with triglyceride (TG) metabolism has been described, including LPL, APOA5 and APOE. The combined analysis of these polymorphisms could produce clinically meaningful complementary information.
A subgroup of the ICARIA study comprising 1825 Spanish subjects (80% men, mean age 36 years) was genotyped for the LPL-HindIII (rs320), S447X (rs328), D9N (rs1801177) and N291S (rs268) polymorphisms, the APOA5-S19W (rs3135506) and -1131T/C (rs662799) variants, and the APOE polymorphism (rs429358; rs7412) using PCR and restriction analysis and TaqMan assays. We used regression analyses to examine their combined effects on TG levels (with the log-transformed variable) and the association of variant combinations with TG levels and hypertriglyceridemia (TG ≥ 1.69 mmol/L), including the covariates: gender, age, waist circumference, blood glucose, blood pressure, smoking and alcohol consumption.
We found a significant lowering effect of the LPL-HindIII and S447X polymorphisms (p < 0.0001). In addition, the D9N, N291S, S19W and -1131T/C variants and the APOE-ε4 allele were significantly associated with an independent additive TG-raising effect (p < 0.05, p < 0.01, p < 0.001, p < 0.0001 and p < 0.001, respectively). Grouping individuals according to the presence of TG-lowering or TG-raising polymorphisms showed significant differences in TG levels (p < 0.0001), with the lowest levels exhibited by carriers of two lowering variants (10.2% reduction in TG geometric mean with respect to individuals who were homozygous for the frequent alleles of all the variants), and the highest levels in carriers of raising combinations (25.1% mean TG increase). Thus, carrying two lowering variants was protective against HTG (OR = 0.62; 95% CI, 0.39-0.98; p = 0.042) and having one single raising polymorphism (OR = 1.20; 95% CI, 1.39-2.87; p < 0.001) or more (2 or 3 raising variants; OR = 2.90; 95% CI, 1.56-5.41; p < 0.001) were associated with HTG.
Our results showed a significant independent additive effect on TG levels of the LPL polymorphisms HindIII, S447X, D9N and N291S; the S19W and -1131T/C variants of APOA5, and the ε4 allele of APOE in our study population. Moreover, some of the variant combinations studied were significantly associated with the absence or the presence of hypertriglyceridemia.
PMCID: PMC2877669  PMID: 20429872
16.  Lipoprotein ratios: Physiological significance and clinical usefulness in cardiovascular prevention 
Low-density lipoprotein (LDL) cholesterol concentration has been the prime index of cardiovascular disease risk and the main target for therapy. However, several lipoprotein ratios or “atherogenic indices” have been defined in an attempt to optimize the predictive capacity of the lipid profile. In this review, we summarize their pathophysiological aspects, and highlight the rationale for using these lipoprotein ratios as cardiovascular risk factors in clinical practice, specifying their cut-off risk levels and a target for lipid-lowering therapy. Total/high-density lipoprotein (HDL) cholesterol and LDL/HDL cholesterol ratios are risk indicators with greater predictive value than isolated parameters used independently, particularly LDL. Future recommendations regarding the diagnosis and treatment of dyslipidemia, including instruments for calculating cardiovascular risk or action guidelines, should include the lipoprotein ratios with greater predictive power which, in view of the evidence-based results, are none other than those which include HDL cholesterol.
PMCID: PMC2747394  PMID: 19774217
apolipoproteins; cholesterol ratios; predictive power; cardiovascular disorders
17.  Lipoprotein lipase activity and mass, apolipoprotein C-II mass and polymorphisms of apolipoproteins E and A5 in subjects with prior acute hypertriglyceridaemic pancreatitis 
BMC Gastroenterology  2009;9:46.
Severe hypertriglyceridaemia due to chylomicronemia may trigger an acute pancreatitis. However, the basic underlying mechanism is usually not well understood. We decided to analyze some proteins involved in the catabolism of triglyceride-rich lipoproteins in patients with severe hypertriglyceridaemia.
Twenty-four survivors of acute hypertriglyceridaemic pancreatitis (cases) and 31 patients with severe hypertriglyceridaemia (controls) were included. Clinical and anthropometrical data, chylomicronaemia, lipoprotein profile, postheparin lipoprotein lipase mass and activity, hepatic lipase activity, apolipoprotein C II and CIII mass, apo E and A5 polymorphisms were assessed.
Only five cases were found to have LPL mass and activity deficiency, all of them thin and having the first episode in childhood. No cases had apolipoprotein CII deficiency. No significant differences were found between the non-deficient LPL cases and the controls in terms of obesity, diabetes, alcohol consumption, drug therapy, gender distribution, evidence of fasting chylomicronaemia, lipid levels, LPL activity and mass, hepatic lipase activity, CII and CIII mass or apo E polymorphisms. However, the SNP S19W of apo A5 tended to be more prevalent in cases than controls (40% vs. 23%, NS).
Primary defects in LPL and C-II are rare in survivors of acute hypertriglyceridaemic pancreatitis; lipase activity measurements should be restricted to those having their first episode during chilhood.
PMCID: PMC2705373  PMID: 19534808
18.  Omega 3 fatty acids induce a marked reduction of apolipoprotein B48 when added to fluvastatin in patients with type 2 diabetes and mixed hyperlipidemia: a preliminary report 
Mixed hyperlipidemia is common in patients with diabetes. Statins, the choice drugs, are effective at reducing lipoproteins that contain apolipoprotein B100, but they fail to exert good control over intestinal lipoproteins, which have an atherogenic potential. We describe the effect of prescription omega 3 fatty acids on the intestinal lipoproteins in patients with type 2 diabetes who were already receiving fluvastatin 80 mg per day.
Patients with type 2 diabetes and mixed hyperlipidemia were recruited. Fasting lipid profile was taken when patients were treated with diet, diet plus 80 mg of fluvastatin and diet plus fluvastatin 80 mg and 4 g of prescription omega 3 fatty acids. The intestinal lipoproteins were quantified by the fasting concentration of apolipoprotein B48 using a commercial ELISA.
The addition of 4 g of prescription omega 3 was followed by significant reductions in the levels of triglycerides, VLDL triglycerides and the triglyceride/HDL cholesterol ratio, and an increase in HDL cholesterol (P < 0.05). Fluvastatin induced a reduction of 26% in B100 (P < 0.05) and 14% in B48 (NS). However, the addition of omega 3 fatty acids enhanced this reduction to 32% in B100 (NS) and up to 36% in B48 (P < 0.05).
Our preliminary findings therefore suggest an additional benefit on postprandial atherogenic particles when omega 3 fatty acids are added to standard treatment with fluvastatin.
PMCID: PMC2631503  PMID: 19133114
20.  Physiological Analysis of the Expression of the Styrene Degradation Gene Cluster in Pseudomonas fluorescens ST 
The effects of different carbon sources on expression of the styrene catabolism genes in Pseudomonas fluorescens ST were analyzed by using a promoter probe vector, pPR9TT, which contains transcription terminators upstream and downstream of the β-galactosidase reporter system. Expression of the promoter of the stySR operon, which codes for the styrene two-component regulatory system, was found to be constitutive and not subject to catabolite repression. This was confirmed by the results of an analysis of the stySR transcript in P. fluorescens ST cells grown on different carbon sources. The promoter of the operon of the upper pathway, designated PstyA, was induced by styrene and repressed to different extents by organic acids or carbohydrates. In particular, cells grown on succinate or lactate in the presence of styrene started to exhibit β-galactosidase activity during the mid-exponential growth phase, before the preferred carbon sources were depleted, indicating that there is a threshold succinate and lactate concentration which allows induction of styrene catabolic genes. In contrast, cells grown on glucose, acetate, or glutamate and styrene exhibited a diauxic growth curve, and β-galactosidase activity was detected only after the end of the exponential growth phase. In each experiment the reliability of the reporter system constructed was verified by comparing the β-galactosidase activity and the activity of the styrene monooxygenase encoded by the first gene of the styrene catabolic operon.
PMCID: PMC91985  PMID: 10742204

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