Bone development and fracture healing is normal in mice that have a defect in the development of the lymphatic system.

Ectopic lymphatics form in bone and promote bone destruction in diseases such as Gorham-Stout disease, generalized lymphatic anomaly, and kaposiform lymphangiomatosis. However, the role lymphatics serve in normal bone development and repair is poorly understood. The objective of this study was to characterize bone development and fracture healing in mice that have a defect in the development of the lymphatic vasculature. We found that bones in wild-type adult mice and mouse embryos did not have lymphatics. We also found that bone development was normal in Vegfr3 (Chy/Chy) embryos. These mice do not have lymphatics and die shortly after birth. To determine whether lymphatics serve a role in postnatal bone development and fracture healing, we analyzed bones from Vegfr3 (wt/Chy) mice. These mice are viable and have fewer lymphatics than wild-type mice. We found that postnatal bone development and fracture healing was normal in Vegfr3 (wt/Chy) mice. Taken together, our results suggest that lymphatics do not play a major role in normal bone development or repair.

Nanobiofertilization as a novel technology for highly efficient foliar application of Fe and B in almond trees.

Nanofertilization is postulated as a new technology to deal with the environmental problems caused by the intensive use of traditional fertilizers. One of the aims of this new technology is to improve foliar fertilization, which has many environmental advantages, but currently there are numerous factors that limit its efficiency. In this research, the objective was to study the potential of membrane vesicles derived from plant material as nanofertilizers of iron (Fe) and boron (B) for foliar application in almond trees (Prunus dulcis L.). The results show that the application of vesicles caused invaginations in the plasma membrane of the leaf cells. Also, the increase in leaf B and Fe was greater when these elements were applied in an encapsulated form rather than in a non-encapsulated form. The distribution of these elements in leaf tissues indicated the existence of an intracellular element transport pathway and accumulation areas, enabling greater element entry and mobility.

MS tools for a systematic approach in survey for carotenoids and their common metabolites.

Metabolism of the carotenoids in humans is often reserved to bioconversion of provitamin A carotenoids in retinoids and oxidative/degradative products arising from enzymatic eccentric cleavage, yielding apo-carotenoids. Nevertheless, additional metabolic routes should be available, and some of the structures of those metabolites have only been anticipated. The identification of carotenoids and their metabolites is commonly performed by HPLC coupled to MS. The acquisition of MS data in different instrumental modes and hardware configurations allows the detailed structural description of the target compound. Indeed, the MS data of carotenoids and their metabolites available in literature could be collected in a database to automatically perform a screening of the theoretical MS data included in the database with the MS experimental values. This review describes the required fundamentals for a systematic pursuit of carotenoids and their metabolites. Highlights on the use of appropriate hyphenated HPLC-MS systems and the requirements for the identification are discussed, while the application of software tools to apply filtering rules and the implementation of post-processing workflow, which uses two orthogonal criteria for the identification from the mass spectra data, are described.

The reduced osteogenic potential of Nf1-deficient osteoprogenitors is EGFR-independent.

Neurofibromatosis type 1 (NF1) is a common genetic disorder caused by mutations in the NF1 gene. Recalcitrant bone healing following fracture (i.e. pseudarthrosis) is one of the most problematic skeletal complications associated with NF1. The etiology of this condition is still unclear; thus, pharmacological options for clinical management are limited. Multiple studies have shown the reduced osteogenic potential of Nf1-deficient osteoprogenitors. A recent transcriptome profiling investigation revealed that EREG and EGFR, encoding epiregulin and its receptor Epidermal Growth Factor Receptor 1, respectively, were among the top over-expressed genes in cells of the NF1 pseudarthrosis site. Because EGFR stimulation is known to inhibit osteogenic differentiation, we hypothesized that increased EREG and EGFR expression in NF1-deficient skeletal progenitors may contribute to their reduced osteogenic differentiation potential. In this study, we first confirmed via single-cell mRNA sequencing that EREG over-expression was associated with NF1 second hit somatic mutations in human bone cells, whereas Transforming Growth Factor beta 1 (TGFß1) expression was unchanged. Second, using ex-vivo recombined Nf1-deficient mouse bone marrow stromal cells (mBMSCs), we show that this molecular signature is conserved between mice and humans, and that epiregulin generated by these cells is overexpressed and active, whereas soluble TGFß1 expression and activity are not affected. However, blocking either epiregulin function or EGFR signaling by EGFR1 or pan EGFR inhibition (using AG-1478 and Poziotinib respectively) did not correct the differentiation defect of Nf1-deficient mBMSCs, as measured by the expression of Alpl, Ibsp and alkaline phosphatase activity. These results suggest that clinically available drugs aimed at inhibiting EGFR signaling are unlikely to have a significant benefit for the management of bone non-union in children with NF1 PA.

Second nonmelanoma skin cancer in Spain: frequency and chronology.

BACKGROUND: Follow-up of patients with nonmelanoma skin cancer (NMSC) is mainly aimed at the early diagnosis of local recurrences and early detection of a subsequent or second NMSC (sNMSC). However, the frequency and chronology of sNMSC have not been widely studied in the literature. OBJECTIVES: To assess the frequency and chronology of developing an sNMSC. METHODS: This was a retrospective longitudinal study of patients with a first NMSC excised and diagnosed at the Hospital Universitario Virgen Macarena (Seville, Spain) between 1 January 1998 and 31 December 2000 and followed up until 31 December 2009. Age, sex, histological type of NMSC (basal cell carcinoma, squamous cell carcinoma), anatomical location, and the diagnosis date of both the first NMSC and the sNMSC were recorded. Main outcomes assessed were the frequency of developing an sNMSC, the annual probability density and the median sNMSC-free survival through the Kaplan-Meier model. RESULTS: A total of 926 patients with a first single NMSC were enrolled and followed up for a median time of 81.9 months. Two hundred and nine patients (22.6%) developed an sNMSC in a median time of 28.3 months. Probability density of a sNMSC ranged from 0.003 to 0.004 for the first 5 years of follow-up. Age ≥ 40 years (P = 0.02), and first NMSC excised from the limbs (P = 0.03) were predictors of developing an sNMSC. CONCLUSIONS: The development of an sNMSC is a common event in patients previously diagnosed with an NMSC especially during the first 5 years of follow-up. This finding should be taken into consideration when designing follow-up guidelines for patients with NMSC.

Hipertensión arterial pulmonar en paciente con neumonía intersticial no específica y con test de vasorreactividad positivo / Pulmonary arterial hypertension in patients with non-specific interstitial pneumonia and with positive vasoreactivity test

La hipertensión pulmonar (HP) es una enfermedad que se caracteriza por un aumento progresivo de la resistencia vascular pulmonar, que da lugar a insuficiencia ventricular derecha y a una muerte prematura. Esta puede ser idiopática o asociarse a diferentes entidades, entre las que destacan las colagenopatías, los tóxicos y las enfermedades pulmonares, como son las neumopatías intersticiales. Presentamos el caso de una paciente con diagnóstico de HP e imágenes indicativas de enfermedad intersticial en la tomografía computarizada de alta resolución y que posteriormente fue diagnosticada de una neumonía intersticial no específica. Es importante destacar la prueba de vasorreactividad positiva en el cateterismo cardíaco, que posiblemente indicaría la presencia de dos entidades independientes en el presente caso (AU)

Pulmonary hypertension (PH) is a disease characterized by a progressive increase of pulmonary vascular resistance, which gives rise to right ventricular failure and premature death. This may be idiopathic or be associated to different conditions, standing out among them collagen, toxic diseases and lung conditions such as interstitial lung diseases. We present the case of a female patient diagnosed of PH with images suggestive of interstitial disease in the high resolution computed tomography and who was subsequently diagnosed of a non-specific interstitial pneumonia. It is important to stress the positive vaso-reactivity test in cardiac catheterization that may indicate the presence of two independent conditions in the present case (AU)

Headspace solid-phase microextraction of oil matrices heated at high temperature and phthalate esters determination by gas chromatography multistage mass spectrometry.

A solvent-free analytical approach based on headspace solid-phase microextraction (SPME) of oil matrices heated at high temperatures coupled to gas chromatography with mass spectrometry detector (GC-ion trap) has been developed for the determination of phthalic acid esters (PAEs) in oil matrices without sample manipulation. For this study, three fibers, i.e., 85 microm-polyacrylate (PA), 50/30 microm-divinylbenzene-carboxen-polydimethylsiloxane (DVB/CAR/PDMS) and 100 microm-polydimethylsiloxane (PDMS) were tested. Variables affecting the SPME headspace composition such as incubation sample temperature, sample incubation time and fiber exposition time were optimized. The optimal values found were 250 degrees C for sample incubation temperature and 30 min for incubation and extraction time. PA fiber was not suitable for the lightest polar phthalates which showed poor extraction and repeatability values. PDMS fiber had very poor response for some of the heavier and non-polar phthalates, whereas DVB/CAR/PDMS fiber showed the best response and repeatability values for the majority of the phthalates studied. The main benefit of the analytical method proposed is the absence of sample manipulation and hence avoidance of possible contamination coming from glassware, environment, solvents and samples.

Response of nitrogen metabolism to boron toxicity in tomato plants.

Boron (B) toxicity has become important in areas close to the Mediterranean Sea where intensive agriculture has been developed. The objective of this research was to study the effects of B toxicity (0.5 mM and 2.0 mM B) on nitrogen (N) assimilation of two tomato cultivars that are often used in these areas. Leaf biomass, relative leaf growth rate (RGR(L)), concentration of B, nitrate (NO(3) (-)), ammonium (NH(4) (+)), organic N, amino acids and soluble proteins, as well as nitrate reductase (NR), nitrite reductase (NiR), glutamine synthase (GS), glutamate synthetase (GOGAT) and glutamate dehydrogenase (GDH) activities were analysed in leaves. Boron toxicity significantly decreased leaf biomass, RGR(L), organic N, soluble proteins, and NR and NiR activities. The lowest NO(3) (-) and NH(4) (+) concentration in leaves was recorded when plants were supplied with 2.0 mM B in the root medium. Total B, amino acids, activities of GS, GOGAT and GDH increased under B toxicity. Data from the present study prove that B toxicity causes inhibition of NO(3) (-) reduction and increases NH(4) (+) assimilation in tomato plants.

Un encéfalo funcional integrado, no un encéfalo parcelado / An integrated functional brain, not a partitioned brain

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Involvement of lignification and membrane permeability in the tomato root response to boron toxicity.

To gain an insight into the role of lignification and membrane permeability in the root response to boron (B) toxicity, lignification-related enzymes and a number of physiological and oxidative stress parameters were analyzed in two tomato (Solanum lycopersicum L.) cultivars (Kosaco and Josefina) subjected to 0.05 (control), 0.5 and 2mM B during 16 days. 2mM B supply inhibited root growth and increased the root B concentration in both tomato cultivars. Although excess B increased the hydrogen peroxide (H2O2) concentration in Kosaco, no major changes were observed in other oxidative-stress-related parameters. High levels of B supply also induced higher lignin deposition in Kosaco roots but did not in Josefina ones. The latter result was associated with an increase of the polyphenol oxidase (PPO), guaiacol peroxidase (GPOX) and soluble syringaldazine peroxidase (SPOX) activity in Kosaco roots. Boron toxicity did not induce lipid peroxidation but increased the leakage of K(+) and the passive efflux of B in tomato roots. We conclude that high concentrations of B do not cause major oxidative or membrane damage in tomato roots. The data also indicate that high levels of B supply induce a higher lignin deposition in Kosaco roots but not in Josefina ones. This phenomenon suggests that lignification is not an essential factor reducing root growth in tomato plants, however, it proves that exist a high genotypic variation in response to excess B at root level.

Characterization and quantification of the hydrocarbons fraction of the subcutaneous fresh fat of Iberian pig by off-line combination of high performance liquid chromatography and gas chromatography.

Hydrocarbons in fresh subcutaneous fat of Iberian pig have been analyzed by GC-MS after fractionation of the unsaponifiable fraction with a new off-line combination of HPLC and GC method. The new method proposed improves the recovery and simultaneous quantification of terpenic hydrocarbons in comparison to the traditional LC method. When necessary and for identification purposes, selective ion monitoring (SIM) was used as acquisition mode in GC-MS. To determine the position of the double bonds in the unsaturated hydrocarbon chain the dimethyl disulfide derivatives (DMDS) were obtained. To elucidate the structure of the branched 1-alkenes the hydrocarbon fraction was submitted to hydrogenation. Thirty-five compounds have been identified, including n-alkanes, n-alkenes, branched (n-1,n-2-dimethyl-1-alkenes) and terpenic hydrocarbons, being the most abundant n-alkenes and n-alkanes of even chain of n-C12-n-C26. Besides the hydrocarbons already described in bibliography, a new diterpenic hydrocarbon, ent-kaurene, have been identified for the first time. The compound reported as Neophytadiene by other authors, has been identified as a 20 atoms hydrocarbon with two double bonds, the 7,11,15-trimethyl-heptadeca-1,4-diene.

Solid-phase extraction gas chromatography-ion trap-mass spectrometry qualitative method for evaluation of phenolic compounds in virgin olive oil and structural confirmation of oleuropein and ligstroside aglycons and their oxidation products.

Phenolic compounds in Spanish virgin olive oil were analyzed by GC-MS after an SPE diol cartridge extraction and clean-up procedure. Posterior derivatization to trimethylsilyl (TMS) ethers using a mixture of hexamethyldisilazane:dimethylclorosilane (HMDS:DMCS) in pyridine (3:1:9) was performed. Several compounds were detected and 21 of them were identified. Free phenols such as hydroxytyrosol, tyrosol, tyrosyl and hydroxytyrosyl acetate, and aldehydic and dialdehydic forms of elenolic acid linked to tyrosol and hidroxytyrosol were the most abundant compounds. Likewise, oxidation products coming from the aldehydic and dialdehydic forms of elenolic acid, and of ligstroside and oleuropein aglycons, were detected, and their structure confirmed by other mass spectrometry technique, i.e., HPLC-APCI-MS. Individual oxidation products were isolated from an oxidized sample by preparative HPLC, converted to TMS ethers and re-analyzed by GC-MS. When necessary and for identification purposes, selective ion monitoring, namely, GC-MS-SIM, was employed. This is the first time that structures of oxidized forms are determined by GC-MS.

Main polyphenols in the bitter taste of virgin olive oil. Structural confirmation by on-line high-performance liquid chromatography electrospray ionization mass spectrometry.

Twenty virgin olive oils of extra quality and different bitter intensity were submitted to sensory evaluation and to the determination of polyphenols. A linear regression analysis was carried out assuming, as an independent variable, bitter intensity perceived by tasters, as an independent variable, the concentration (mmol/kg) of dialdehydic and aldehydic forms oleuropein aglycon, and dialdehydic and aldehydic forms ligstroside aglycon. Structural confirmation of these compounds was done by online high-performance liquid chromatography-electrospray ionization-collison-induced dissociation-mass spectrometry. The results obtained demonstrate the essential role played by this compound in the bitter taste of virgin olive oil.

Achromobacter xylosoxidans bacteremia: a 10-year analysis of 54 cases.

Fifty-four cases of Achromobacter xylosoxidans bacteremia diagnosed over a 10-year period in patients from 2 months to 87 years of age were reviewed. Fifty-two episodes were nosocomial. The most frequent underlying condition was neoplasm (solid or hematological). The source of infection was a contaminated intravenous catheter in 35 patients (60%) and pneumonia in 6 patients. Eight (15%) patients died. The only risk factors significantly associated with mortality were age over 65 years and neutropenia. The results of in vitro susceptibility studies of the isolates showed that antibiotic therapy with antipseudomonal penicillins or carbapenems would be a reasonable choice. An epidemiological study conducted in the hemodialysis unit showed Achromobacter xylosoxidans in tap water and on the hands of two healthcare workers but not in the hemodialysis systems. Patients were probably contaminated when healthcare workers manipulated the intravenous catheters without wearing gloves.

Pancreatic ascites: study of therapeutic options by analysis of case reports and case series between the years 1975 and 2000.

OBJECTIVE: Because of the low incidence of pancreatic ascites, only case reports and case series have been published, and no randomized controlled trials have been performed to find out which is the best therapeutic approach. The aim of this study was to evaluate the utility of the different treatments for pancreatic ascites by a thorough review of all case reports described in the literature in which an independent analysis of the efficacy of the treatment administered to each patient is possible. METHODS: We conducted an analysis of all case reports and case series of pancreatic ascites published between 1975 and 2000 in which clinical data of every patient could be identified individually. A total of 139 cases were studied. Clinical characteristics, treatments administered, and response to therapy of every patient were registered. Conservative therapy included drainage of ascitic fluid, total parenteral nutrition and diet, and somatostatin analogues. Interventional therapy was either endoscopic or surgical. RESULTS: After multivariate analysis, the only treatments related to success were surgery (adjusted OR = 8.2, 95% CI = 3.0-22.9) and transpapillary stent (adjusted OR = 7.3, 95% CI = 0.8-62.9). No significant relationship was found between failure or death and the use of other treatments, age, sex, year of publication, underlying disease, site of leakage, or serum amylase levels. The apparent lack of effect of somatostatin analogues could be attributed to the small number of cases and the heterogeneity of the dosages. CONCLUSIONS: Conservative therapy is not advisable for pancreatic ascites because of the high proportion of failures. Interventional therapy with surgery or transpapillary stent has a positive effect in the clinical outcome.

Justificación de la denominación de nuestraespecialidad: algiología / Rationale for the name of our speciality: algiology

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