Major lipophilic pigments in Atlantic seaweeds as valuable food ingredients: Analysis and assessment of quantification methods.

Current trends towards the use of ingredients from natural origin in food, cosmetic and pharmaceutical industry, place macroalgae as a good reservoir of novel compounds. Among them, lipophilic major pigments such as chlorophylls and fucoxanthin, are of great interest because of their multiple applications as bioactive compounds and dyes. In this work, a mid-polarity medium was used to extract pigments from twenty-four species from North coast of Spain, including brown (Phaeophyceae) and red macroalgae (Rhodophyta). The fucoxanthin and chlorophyll a content was assessed by means of two different methods, spectrophotometric and high-performance liquid chromatography coupled to diode array detection (HPLC-DAD). The effect of dried processing on the pigment content of selected species was also evaluated. A linear relationship between the extractability of fucoxanthin and chlorophyll a was observed, being the highest content recorded among members belonging to the order Fucales and Undaria pinnatifida. This work provides good insights about the content on pigments in Spanish North Atlantic macroalgae with future commercial value in different industrial fields, as well as a critical overview of the suitability of the quantification methods and challenges related to their effect in results evaluation.

Approach to the correction of drooping tip: common problems and solutions.

The drooping tip deformity is both a bothersome aesthetic feature and functional impairment of the nose. Both static and dynamic factors may affect tip appearance and it seems logical to take into account these factors when planning correction of drooping tip. Many studies have examined this topic, but its treatment remains controversial. In order to make nasal tip surgery successful, it is useful to identify the keystone anatomical characteristics of the tip itself. Naso-labial angle, nostril axis, tip rotation angle according to Frankfort plane and columellar-facial angle may be measured to assess nasal tip position. The present study focuses on the authors' personal experience on the key anatomic changes of the nose that deserve correction and on the main surgical steps needed to achieve consistent results when dealing with a drooping tip. Pre- and post-operative nasal tip rotation and projection were studied. Correction of the drooping tip was accomplished by an open or closed septorhinoplasty approach according to patient's needs. The surgical techniques mostly employed for tip repositioning was septum straightening (41/41) and tongue-in-groove (36/41 cases) (87.8%). A columellar strut was used in 8/41 (19.51%) cases. LLC cephalic resection was applied in 29/41 patients (70.73%), LLC re-orienting sutures were made in 18/41 cases (43.9%) and lateral crural overlay was needed in 2/41 (4.8%). The key anatomic changes of the nose that deserve correction and the surgical steps needed to ease the often intriguing pre-operative decision-making process are reviewed.

Draft Genome Sequence of the Host-Restricted Salmonella enterica Serovar Abortusovis Strain SS44.

Salmonella enterica serovar Abortusovis is a pathogen strictly adapted to ovines, in which it causes abortion. To enhance our understanding of this pathogen, we assembled the first draft sequence of an S. Abortusovis genome (strain SS44). The obtained genomic data might facilitate the study of S. enterica evolution and host adaptation.

Detection and characterization of Mycoplasma spp. and Salmonella spp. in free-living European tortoises (Testudo hermanni, Testudo graeca, and Testudo marginata).

Free-living and captive chelonians might suffer from upper respiratory tract disease (URTD), a pathology primarily caused by Mycoplasma agassizii. Wild tortoises can also be an important reservoir of Salmonella spp., which are commensal in the host reptile but are potential zoonotic agents. Between July 2009 and June 2010, we screened free-living European tortoises (spur-thighed tortoises Testudo graeca, Hermann's tortoises Testudo hermanni, marginated tortoises Testudo marginata) temporarily housed in a wildlife center in Italy. We molecularly characterized 13 Mycoplasma isolates detected in all Testudo spp. studied, and three PCR-positive animals showed typical URTD clinical signs at the time of sampling. Three Salmonella enterica serotypes (Abony, Potsdam, Granlo), already related to reptile-associated human infections, were also identified. These results highlight the potential role played by wildlife recovery centers in the spread and transmission of pathogens among wild chelonians and to humans.

Bacteraemia with pleural effusions complicating typhoid fever caused by high-level ciprofloxacin-resistant Salmonella enterica serotype Typhi.

An unusual case of bacteraemia with bilateral pleural effusion caused by Salmonella enterica serotype Typhi in a 10-year-old previously healthy girl is reported. The organism was isolated from pleural fluid aspirate and from blood, and exhibited high-level ciprofloxacin resistance (MIC 16 µg/ml) associated with triple mutations in the QRDRs of the gyrA and parC genes leading to the amino-acid changes Ser83→Phe and Asp87→Asn in gyrA and Ser80→Ile in parC. The patient was successfully treated with parenteral ceftriaxone and intercostal chest tube drainage. The case is notable because of the important issue of antimicrobial resistance in S. Typhi and the therapeutic dilemma faced by clinicians regarding the empirical use of ciprofloxacin and newer fluoroquinolones.

Asymmetry of the two-beam geometry in EMCD experiments.

We analyse theoretically the influence of the asymmetry of the two-beam geometry on quantitative measurements of the energy-loss magnetic chiral dichroism. Our simulations indicate that this asymmetry is not very strong inside or close to the Thales circle, but in other regions of the diffraction plane it can hinder an accurate extraction of the orbital to spin moment ratio.

Synthetic, structural and biochemical studies of polynuclear platinum(II) complexes with heterocyclic ligands.

"Non-classical" di- and trinuclear Pt(II) complexes with polydentate nitrogen ligands; ionic [(PtCl(2))(2)(tptz)(2)(mu-PtClNCPh)]Cl (1) [tptz =2,4,6-tris(2-pyridyl)-1,3,5-triazine], [(PtCl(2))(2)(bptz)(2)(mu-Pt)]Cl(2) (2) [bptz = 3,6-bis(2-pyridyl)-1,2,4,5-tetrazine] and neutral [(PtCl(2))(2)(tptz)(2)(mu-PtCl(2))](H(2)O)(4) (3), [(PtCl(2))(2)(mu-tppz)](CHCl(3)) (4) [tppz = 2,3,5,6-tetra(2-pyridyl)pyrazine] complexes, have been prepared and structurally characterized. The neutral tptz and tppz complexes present three and two separate PtCl(2) moieties, respectively, in a cis position, presumably acting in a bifunctional mode towards DNA; the cationic tptz and bptz complexes contain monofunctional and bifunctional bridging Pt(II) moieties, respectively, (other Pt(II) moieties in the complexes are bifunctional). All complexes were tested for their biological activity. Both tptz complexes, neutral and ionic, show a potent cytotoxic activity and reduced cell viability in a concentration-dependent manner that was evaluated in a panel of different cancer cell lines: human HT29 colon-rectal carcinoma, HepG2 hepatoma, MDA-MB-231 breast cancer and MG63 osteosarcoma cells; their activity was higher than cisplatin, IC50 values have been calculated for the active compounds and flow cytometric analysis for the tptz complexes performed. Therefore, these new platinum drugs warrant further investigation into their antitumor activity against different types of tumors.

Prevalence and Antibiotic-Resistance of Salmonella Isolated from Food in Morocco

Background: Salmonellosis remains one of the most frequent food-borne diseases worldwide; especially in developing countries. The emergence of antimicrobial resistance in Salmonella isolates from food can potentially compromise the treatment of these infections. This investigation was conducted for the first time in Morocco both to detect the occurrence of Salmonella in foods as well as to determine the antibiotic resistance profile of the Salmonella isolates. Methodology: In total; 11;516 food samples collected from 2002 to 2005 were investigated. Isolated Salmonella were characterized by serotyping and susceptibilities were determined for 15 antimicrobial drugs using the disc diffusion assay. Results: The overall percentage of Salmonella prevalence (n=105) was 0.91with rates of 71for slaughterhouses and 9for seafood. Sixteen different serotypes were identified among 104 Salmonella enterica isolates including serotypes Infantis (n=25); Bredeney (n=13); Blokley (n=11); Typhimurium (n=9); Mbandaka (n=8); Branderup II (n=7); and Kiambu (n=6); 1 isolate of Salmonella enterica belonged to subspecies II salamae. Twenty-nine percent of isolates (n=30/105) were resistant to at least one antimicrobial. Resistance to tetracycline was the most common finding (21); followed by resistance to ampicillin (13); amoxicillin+clavulanic acid (9); streptomycin (7); chloramphenicol (4) and nalidixic acid (3;8). None of the isolates was resistant to 3rd-cephalosporin and fluoroquinolones (i.e. ciprofloxacin). Multidrug resistance (MDR) was seen in 9.5of the isolates; mainly in S. Typhimurium DT104 with R-type ACSSuT and S. Hadar. Conclusions: Despite a low frequency of Salmonella isolation; S. Typhimurium DT104 was identified in the first step of the food chain. The study points out the need control antibiotic resistance in Salmonella isolated from food in Morocco to avoid the spread of MDR

Prevalence and Antibiotic-Resistance of Salmonella Isolated from Food in Morocco

Background: Salmonellosis remains one of the most frequent food-borne diseases worldwide; especially in developing countries. The emergence of antimicrobial resistance in Salmonella isolates from food can potentially compromise the treatment of these infections. This investigation was conducted for the first time in Morocco both to detect the occurrence of Salmonella in foods as well as to determine the antibiotic resistance profile of the Salmonella isolates. Methodology: In total; 11;516 food samples collected from 2002 to 2005 were investigated. Isolated Salmonella were characterized by serotyping and susceptibilities were determined for 15 antimicrobial drugs using the disc diffusion assay. Results: The overall percentage of Salmonella prevalence (n=105) was 0.91with rates of 71for slaughterhouses and 9for seafood. Sixteen different serotypes were identified among 104 Salmonella enterica isolates including serotypes Infantis (n=25); Bredeney (n=13); Blokley (n=11); Typhimurium (n=9); Mbandaka (n=8); Branderup II (n=7); and Kiambu (n=6); 1 isolate of Salmonella enterica belonged to subspecies II salamae. Twenty-nine percent of isolates (n=30/105) were resistant to at least one antimicrobial. Resistance to tetracycline was the most common finding (21); followed by resistance to ampicillin (13); amoxicillin+clavulanic acid (9); streptomycin (7); chloramphenicol (4) and nalidixic acid (3;8). None of the isolates was resistant to 3rd-cephalosporin and fluoroquinolones (i.e. ciprofloxacin). Multidrug resistance (MDR) was seen in 9.5of the isolates; mainly in S.. Typhimurium DT104 with R-type ACSSuT and S. Hadar. Conclusions: Despite a low frequency of Salmonella isolation; S. Typhimurium DT104 was identified in the first step of the food chain. The study points out the need control antibiotic resistance in Salmonella isolated from food in Morocco to avoid the spread of MDR

Optimal aperture sizes and positions for EMCD experiments.

The signal-to-noise ratio (SNR) in energy-loss magnetic chiral dichroism (EMCD)--the equivalent of X-ray magnetic circular dichroism (XMCD) in the electron microscope--is optimized with respect to the detector shape, size and position. We show that an important increase in SNR over previous experiments can be obtained when taking much larger detector sizes. We determine the ideal shape of the detector but also show that round apertures are a good compromise if placed in their optimal position. We develop the theory for a simple analytical description of the EMCD experiment and then apply it to dynamical multibeam Bloch wave calculations and to an experimental data set. In all cases it is shown that a significant and welcome improvement of the SNR is possible.

Magnetic circular dichroism in electron energy loss spectrometry.

The measurement of circular dichroism in the electron microscope is a new, emerging method and, as such, it is subject to constant refinement and improvement. Different ways can be envisaged to record the signal. We present an overview of the key steps in the energy-loss magnetic chiral dichroism (EMCD) experiment as well as a detailed review of the methods used in the intrinsic way where the specimen is used as a beam splitter. Lateral resolution up to 20-30 nm can be achieved, and the use of convergent beam techniques leads to an improved S/N ratio. Dichroic effects are shown for Ni and Co single crystal; as a counterexample, measurements were carried also for a non-magnetic (Ti) sample, where no dichroic effect was found.

Mono- and polynuclear complexes of Pt(II) with polypyridyl ligands. Synthesis, spectroscopic and structural characterization and cytotoxic activity.

An array of poly- and mononuclear complexes of Pt(II) with polypyridyl ligands is reported. The framework complexes [(PtCl(2))(2)(bpp)(2)(micro-PtCl(2))](H(2)O)(2) [bpp=2,3-bis(2-pyridyl)pyrazine], [PtCl(2)(micro-tptz)PtClNCPh]Cl [tptz=2,4,6-tris(2-pyridyl)-1,3,5-triazine], and mononuclear PtCl(2)(NH(2)dpt) [NH(2)dpt=4-amino-3,5-bis(2-pyridyl)-1,2,4-triazole] have been prepared and structurally characterized. Both neutral and ionic complexes are present, with bifunctional and monofunctional Pt(II) moieties, whose size and shape enable them to behave as novel scaffolds for DNA binding. Pt(II) complexes were tested for their biological activity. Cell viability assay and flow cytometric analysis demonstrated that these complexes, particularly [PtCl(2)(micro-tptz)PtClNCPh]Cl, were effective death inducers in human colon rectal carcinoma HT29 cells and their cytotoxic activity was higher than that exerted by cisplatin. Morphological analysis of treated HT29 cells, performed by fluorescence microscopy after Hoechst 33258 staining, showed the appearance of the typical features of apoptosis. Moreover, our results suggested that mitochondria are involved in apoptosis induced by Pt(II) complexes in HT29 cells as demonstrated by dissipation of mitochondrial transmembrane potential.

SipA, SopA, SopB, SopD and SopE2 effector proteins of Salmonella enterica serovar Typhimurium are synthesized at late stages of infection in mice.

Salmonella pathogenicity island (SPI)-1 is essential for invasion of non-phagocytic cells, whereas SPI-2 is required for intracellular survival and proliferation in phagocytes. Some SPI-1 effectors, however, are induced upon invasion of both phagocytic and non-phagocytic cells, suggesting that they may also be required post-invasion. In the present work, the presence was analysed of SipA, SopA, SopB, SopD and SopE2 effector proteins of Salmonella enterica serovar Typhimurium in vitro and in vivo during murine salmonellosis. Tagged (3xFLAG) strains of S. enterica serovar Typhimurium were inoculated intraperitoneally or intragastrically to BALB/c mice and recovered from the spleen and mesenteric lymph nodes of moribund mice. Tagged proteins were detected by SDS-PAGE and immunoblotting with anti-FLAG antibodies. In vitro experiments showed that SPI-1 effector proteins SipA, SopA, SopB, SopD and SopE2 were secreted under SPI-1 conditions. Interestingly, it was found that S. enterica serovar Typhimurium continued to synthesize SipA, SopB, SopD and SopE2 in colonized organs for several days, regardless of the route of inoculation. Together, these results indicate that SPI-1 effector proteins may participate in the late stages of Salmonella infection in mice.

Molecular characterization of ciprofloxacin-resistant Salmonella enterica serovar Typhi and Paratyphi A causing enteric fever in India.

OBJECTIVES: To define the genetic characteristics and resistance mechanisms of clinical isolates of Salmonella enterica serovar Typhi (S. Typhi) and S. enterica serovar Paratyphi A (S. Paratyphi A) exhibiting high-level fluoroquinolones resistance. METHODS: Three S. Typhi and two S. Paratyphi A ciprofloxacin-resistant isolates (MICs > 4 mg/L) were compared with isolates with reduced susceptibility to ciprofloxacin (MICs 0.125-1 mg/L) by PFGE, plasmid analysis, presence of integrons and nucleotide changes in topoisomerase genes. RESULTS: In S. Typhi and Paratyphi A, a single gyrA mutation (Ser-83-->Phe or Ser-83-->Tyr) was associated with reduced susceptibility to ciprofloxacin (MICs 0.125-1 mg/L); an additional mutation in parC (Ser-80-->Ile, Ser-80-->Arg, Asp-69-->Glu or Gly-78-->Asp) was accompanied by an increase in ciprofloxacin MIC (> or = 0.5 mg/L). Three mutations conferred ciprofloxacin resistance: two in gyrA (Ser-83-->Phe and Asp-87-->Asn or Asp-87-->Gly) and one in parC. This is the first report of parC mutations in S. Typhi. Ciprofloxacin-resistant S. Typhi and S. Paratyphi A differed in their MICs and mutations in gyrA and parC. Moreover S. Typhi harboured a 50 kb transferable plasmid carrying a class 1 integron (dfrA15/aadA1) that confers resistance to co-trimoxazole and tetracycline but not to ciprofloxacin. PFGE revealed undistinguishable XbaI fragment patterns in ciprofloxacin-resistant S. Typhi as well as in S. Paratyphi A isolates and showed that ciprofloxacin-resistant S. Typhi have emerged from a clonally related isolate with reduced susceptibility to ciprofloxacin after sequential acquisition of a second mutation in gyrA. CONCLUSIONS: To our knowledge this is the first report of molecular characterization of S. Typhi with full resistance to ciprofloxacin. Notably, the presence of a plasmid-borne integron in ciprofloxacin-resistant S. Typhi may lead to a situation of untreatable enteric fever.

Detection of magnetic circular dichroism using a transmission electron microscope.

A material is said to exhibit dichroism if its photon absorption spectrum depends on the polarization of the incident radiation. In the case of X-ray magnetic circular dichroism (XMCD), the absorption cross-section of a ferromagnet or a paramagnet in a magnetic field changes when the helicity of a circularly polarized photon is reversed relative to the magnetization direction. Although similarities between X-ray absorption and electron energy-loss spectroscopy in a transmission electron microscope (TEM) have long been recognized, it has been assumed that extending such equivalence to circular dichroism would require the electron beam in the TEM to be spin-polarized. Recently, it was argued on theoretical grounds that this assumption is probably wrong. Here we report the direct experimental detection of magnetic circular dichroism in a TEM. We compare our measurements of electron energy-loss magnetic chiral dichroism (EMCD) with XMCD spectra obtained from the same specimen that, together with theoretical calculations, show that chiral atomic transitions in a specimen are accessible with inelastic electron scattering under particular scattering conditions. This finding could have important consequences for the study of magnetism on the nanometre and subnanometre scales, as EMCD offers the potential for such spatial resolution down to the nanometre scale while providing depth information--in contrast to X-ray methods, which are mainly surface-sensitive.

Characterization of virulence factors in the newly described Salmonella enterica serotype Keurmassar emerging in Senegal (sub-Saharan Africa).

From 2000 to 2001, nine strains of Salmonella enterica belonging to the new serotype Keurmassar have been isolated from human and poultry samples at the Senegalese National Salmonella and Shigella Reference Laboratory at the Pasteur Institute, in Dakar. All strains carried virulence factors including Salmonella Pathogenicity Islands (SPI)-1, -2, -3 and -5 encoded genes. Strains did not harbour virulence plasmid. Ribotyping analysis revealed a single clone identical to Salmonella Decatur isolated in Zimbabwe. These data suggest that strains are closely related, and may have been spread clonally. In this new serotype, insertion sequence IS200 is not present.

Synthesis, characterization, and cytotoxic activity of copper(II) and platinum(II) complexes of 2-benzoylpyrrole and X-ray structure of bis[2-benzoylpyrrolato(N,O)]copper(II).

Copper(II) and platinum(II) complexes of 2-benzoylpyrrole (2-BZPH) were synthesized and characterized with IR, 1H and 13C NMR spectroscopies and coordination geometry with ligands arranged in transoid fashion. The crystal structure of [Cu(II)(2-BZP)2] was determined by X-ray diffraction. Death of complex treated Jurkat cells was measured by flow cytometry. The bis-chelate complexes [Cu(II)(2-BZP)2] and [Pt(II)(2-BZP)2] adopt square-planar coordination geometry with ligands, arranged in transoid fashion. Concentrations of 1-10 microM Platinum(II) complexes reduced cell survival from 100% to 20%, in contrast to the copper(II) complex which caused no cell death at a concentration of 10 microM. While the Pt(II) complexes may have damaged DNA to induce cell death, treatment with the Cu(II) complex did not induce Jurkat cell death.

IS200 fingerprinting of Salmonella enterica serotype Abortusovis strains isolated in Iran.

Salmonella enterica serovar Abortusovis is one of the most common pathogens responsible for abortion in sheep. In Iran, the spread of Abortusovis is highly dependent on the nomadic life style. In this study we performed IS200 fingerprinting to identify the clonal lines circulating in Iran. All the isolates contained 4 or 5 copies of the transposon and could be classified in 4 genotypes. A single genotype was highly prevalent and very likely it has circulated in Iran since 1970. All the isolates showed a high degree of relatedness.

Stroke related to carotid artery dissection in a young patient with Takayasu arteritis, systemic lupus erythematosus and antiphospholipid antibody syndrome.

Autoimmune diseases are rarely the cause of stroke even in the young age group in association with cervical artery dissection and collagen vascular diseases. Takayasu arteritis is a chronic, idiopathic, inflammatory disease that primarily affects large vessels, such as the aorta and its main branches. Takayasu arteritis rarely coexists with systemic lupus erythematosus, and only few cases have been reported in association with the presence of antiphospholipid antibodies. We describe a young patient with right internal carotid artery dissection and subsequent stroke who presented with all three syndromes. Although this patient met the diagnostic criteria for each syndrome, systemic lupus erythematosus, Takayasu arteritis and the antiphospholipid antibody syndrome, it remains unlikely that the three disorders are not related. We suggest a single disimmune disorder may have led to carotid artery dissection.