Impact assessment of multiple pressures on ecosystem services with a state and transition model: Application to Posidonia oceanica seagrass meadows.

A significant challenge in the integration of ecosystem services into decision-making processes lies in effectively capturing the dynamics of marine socio-ecological systems, including their evolutionary pathways, equilibrium states, and tipping points. This paper explores the evolutionary trajectories of a vital marine ecosystem endemic to the Mediterranean Sea: the Posidonia oceanica seagrass meadows, in response to various drivers of change. A state-and-transition model is employed to assess the ecosystem services provided by P. oceanica across different states defined by selected transitions, such as overfishing, fragmentation, pollution, and invasion by non-native species. To apply this model, scientific expertise is combined with field data generated using the Ecosystem-Based Quality Index to evaluate the conservation status of P. oceanica. This integrated approach allows for the representation of the ecosystem services offered by the meadows across different states, leveraging ecological data. The findings highlight the disproportionate impact on provisioning services, particularly sea urchins and commercial fish production, which suffer the most under various stressors. Notably, when these services decline to critical levels, the meadows cease to provide significant benefits. Finally, a synthesized representation is presented, merging ecological insights with monitoring data, offering a framework that is more accessible to stakeholders and decision-makers.

Site-selective-induced isomerization of formamide.

The new capacity of X-ray free-electron laser (XFEL) facilities to produce multi-color X-ray femtosecond pulses paves the way to explore ultrafast phenomena in matter induced by X-ray photons. In the present study, we exploit the site-selectivity and the high temporal resolution of a two-color X-ray femtosecond pump-probe sequence to investigate the isomerization of formamide. The pump pulse excites a particular atomic site in the molecule, while the probe pulse captures changes in the chemical environment at a remote atomic site. The response of the system is found to strongly depend on the nature of the excited site, and the core excitation provides selective control over chemical bond breaking. In particular, we show that the N1s → π* transition can induce the isomerization reaction in formamide and demonstrate the possibility to observe in real time hydrogen migration by measuring the chemical shifts with the X-ray probe pulse. This work opens a unique prospect for X-ray-induced photochemistry at XFEL facilities.

Proteolysis and biogenic amine buildup in high-pressure treated ovine milk blue-veined cheese.

Penicillium roqueforti plays an important role in the ripening of blue-veined cheeses, mostly due to lactic acid consumption and to its extracellular enzymes. The strong activity of P. roqueforti proteinases may bring about cheese over-ripening. Also, free amino acids at high concentrations serve as substrates for biogenic amine formation. Both facts result in shorter product shelf-life. To prevent over-ripening and buildup of biogenic amines, blue-veined cheeses made from pasteurized ovine milk were high-pressure treated at 400 or 600 MPa after 3, 6, or 9 wk of ripening. Primary and secondary proteolysis, biogenic amines, and sensory characteristics of pressurized and control cheeses were monitored for a 90-d ripening period, followed by a 270-d refrigerated storage period. On d 90, treatments at 400 MPa had lowered counts of lactic acid bacteria and P. roqueforti by less than 2 log units, whereas treatments at 600 MPa had reduced lactic acid bacteria counts by more than 4 log units and P. roqueforti counts by more than 6 log units. No residual α-casein (CN) or κ-CN were detected in control cheese on d 90. Concentrations of ß-CN, para-κ-CN, and γ-CN were generally higher in 600 MPa cheeses than in the rest. From d 90 onwards, hydrophilic peptides were at similar levels in pressurized and control cheeses, but hydrophobic peptides and the hydrophobic-to-hydrophilic peptide ratio were at higher levels in pressurized cheeses than in control cheese. Aminopeptidase activity, overall proteolysis, and free amino acid contents were generally higher in control cheese than in pressurized cheeses, particularly if treated at 600 MPa. Tyramine concentration was lower in pressurized cheeses, but tryptamine, phenylethylamine, and putrescine contents were higher in some of the pressurized cheeses than in control cheese. Differences in sensory characteristics between pressurized and control cheeses were generally negligible, with the only exception of treatment at high pressure level (600 MPa) at an early ripening stage (3 wk), which affected biochemical changes and sensory characteristics.

High-pressure processing decelerates lipolysis and formation of volatile compounds in ovine milk blue-veined cheese.

Enzyme-rich cheeses are prone to over-ripening during refrigerated storage. Blue-veined cheeses fall within this category because of the profuse growth of Penicillium roqueforti in their interior, which results in the production of highly active proteinases, lipases, and other enzymes responsible for the formation of a great number of flavor compounds. To control the excessive formation of free fatty acids (FFA) and volatile compounds, blue-veined cheeses were submitted to high-pressure processing (HPP) at 400 or 600 MPa on d 21, 42, or 63 after manufacture. Cheeses were ripened for 30d at 10°C and 93% relative humidity, followed by 60 d at 5°C, and then held at 3°C until d 360. High-pressure processing influenced the concentrations of acetic acid and short-chain, medium-chain, and long-chain FFA. The effect was dependent on treatment conditions (pressure level and cheese age at the time of treatment). The lowest concentrations of acetic acid and FFA were recorded for cheeses treated at 600 MPa on d 21; these cheeses showed the lowest esterase activity values. Acetic acid and all FFA groups increased during ripening and refrigerated storage. The 102 volatile compounds detected in cheese belonged to 10 chemical groups (5 aldehydes, 12 ketones, 17 alcohols, 12 acids, 35 esters, 9 hydrocarbons, 5 aromatic compounds, 3 nitrogen compounds, 3 terpenes, and 1 sulfur compound). High-pressure processing influenced the levels of 97 individual compounds, whereas 68 individual compounds varied during refrigerated storage. Total concentrations of all groups of volatile compounds were influenced by HPP, but only ketones, acids, esters, and sulfur compounds varied during refrigerated storage. The lowest total concentrations for most groups of volatile compounds were recorded for the cheese pressurized at 600 MPa on d 21. A principal component analysis combining total concentrations of groups of FFA and volatile compounds discriminated cheeses by age and by the pressure level applied to HPP cheeses.

Enhancement of vibrational excitation and dissociation of H2(+) in infrared laser pulses.

We study vibrational excitations, dissociation, and ionization of H(2)(+) in few-cycle laser pulses over a broad wavelength regime. Our results of numerical simulations supported by model calculations show a many orders-of-magnitude enhancement of vibrational excitation and dissociation (over ionization) of the molecular ion at infrared wavelengths. The enhancement occurs without any chirping of the pulse, which was previously applied to take account of the anharmonicity of the molecular vibrations. The effect is related to strong-field two- and higher-order photon transitions between different vibrational states.

Effect of high-pressure treatment of ewe raw milk curd at 200 and 300 MPa on characteristics of Hispánico cheese.

Hispánico cheese is a semihard variety made from a mixture of cow and ewe milks. Production of ewe milk declines in summer and autumn. To surmount the seasonal shortage of ewe milk and prevent the inactivation of milk enzymes by pasteurization, curd made in spring from ewe raw milk was pressurized at 200 and 300 MPa and stored frozen for 4 mo. Thawed ewe milk curds were added to fresh curd made from pasteurized cow milk for the manufacture of experimental Hispánico cheeses. Control cheese was made from a mixture of pasteurized cow and ewe milk in the same proportions as those used for experimental cheeses. Experimental cheeses exhibited lower dry matter content, higher aminopeptidase activity and total free amino acid concentration, and higher levels of acetic and propionic acids, aldehydes, alcohols, and esters compared with control cheese. In contrast, the concentration of total free fatty acids and ketones and the levels of textural parameters were significantly higher in control cheese. The use of ewe raw milk curd pressurized at 200 and 300 MPa, stored frozen and thawed for Hispánico cheese manufacture, was generally beneficial for cheese characteristics and increased cheese yield because of the lower dry matter content of experimental cheeses.

Microbial dynamics during the ripening of a mixed cow and goat milk cheese manufactured using frozen goat milk curd.

To overcome the seasonal shortage of goat milk in mixed milk cheese manufacture, pasteurized goat milk curd and high-pressure-treated raw goat milk curd manufactured in the spring were held at -24 °C for 4 mo, thawed, and mixed with fresh cow milk curd for the manufacture of experimental cheeses. Control cheeses were made from a mixture of pasteurized cow and goat milk. The microbiota of experimental and control cheeses was studied using culture-dependent and culture-independent techniques. Bacterial enumeration by classical methods showed lactic acid bacteria to be the dominant population in both control and experimental cheeses. In total, 681 isolates were grouped by partial amplified rDNA restriction analysis (ARDRA) into 4 groups and identified by 16S rRNA gene sequencing as Lactococcus lactis ssp. lactis (563 isolates), Leuconostoc pseudomesenteroides (72 isolates), Lactobacillus spp. (34 isolates), and Lc. lactis ssp. cremoris (12 isolates). Temporal temperature gradient gel electrophoresis (TTGE) analysis of cheese showed (1) the predominance of Lc. lactis in all cheeses; (2) the presence of Leu. pseudomesenteroides population in all cheeses from d 15 onward; (3) the presence of a Lactobacillus plantarum population in control cheese until d 15 and in experimental cheeses throughout the ripening period. Due to the most diverse and complete set of peptidases present in the genus Lactobacillus, the prevalence of this population in experimental cheeses could give rise to differences in cheese flavor between experimental and control cheeses.

Photoionization with orbital angular momentum beams.

Intense laser ionization expands Einstein's photoelectric effect rules giving a wealth of phenomena widely studied over the last decades. In all cases, so far, photons were assumed to carry one unit of angular momentum. However it is now clear that photons can possess extra angular momentum, the orbital angular momentum (OAM), related to their spatial profile. We show a complete description of photoionization by OAM photons, including new selection rules involving more than one unit of angular momentum. We explore theoretically the interaction of a single electron atom located at the center of an intense ultraviolet beam bearing OAM, envisaging new scenarios for quantum optics.

Proteolysis, lipolysis, volatile compounds, texture, and flavor of Hispánico cheese made using frozen ewe milk curds pressed for different times.

Hispánico cheese is manufactured in Spain from a mixture of cow and ewe milk. Production of ewe milk varies throughout the year, with a peak in spring and a valley in summer and autumn. To overcome this seasonal shortage, curd from spring ewe milk may be frozen and used for cheese manufacture some months later. In the present work, ewe milk curds pressed for 15, 60, or 120 min were held at -24 degrees C for 4 mo, thawed, cut to 1-mm pieces, and mixed with fresh cow milk curd for the manufacture of experimental Hispánico cheeses. Control cheese was made from a mixture of pasteurized cow and ewe milk in the same (80:20) proportion. Cheeses, made in duplicate experiments, were analyzed throughout a 60-d ripening period. No significant differences between cheeses were found for lactic acid bacteria counts, dry matter content, hydrophilic peptides, 47 out of 68 vol.tile compounds, texture, and flavor characteristics. On the other hand, differences of minor practical significance between experimental and control cheeses, unrelated to the use of frozen ewe milk curd or the pressing time of ewe milk curd, were found for pH value, aminopeptidase activity, proteolysis, hydrophobic peptides, free amino acids, free fatty acids, and the remaining 21 vol.tile compounds. It may be concluded that the use of frozen ewe milk curd in the manufacture of Hispánico cheese does not alter its main characteristics.

Contribution of autochthonous lactic acid bacteria to the typical flavour of raw goat milk cheeses.

In this study, a collection of 298 wild strains of lactic acid bacteria (LAB) isolated from raw goat milk cheeses, belonging to 8 genera and 24 species, was investigated for their ability to generate volatile compounds. Sensory evaluation showed that goat milk cultures of Leuconostoc and Lactococcus strains reached the highest scores for dairy odour attributes. Fifty six LAB strains with the highest sensory evaluation scores (one per each ten, or fraction, of those belonging to the same species) were selected for gas chromatography-mass spectrometry (GC-MS) analysis. A total of 34 volatile compounds (five carboxylic acids, eleven alcohols, six aldehydes, six ketones, one ester, and five miscellaneous compounds) were detected in lactic curds made with each one of 56 selected LAB strains. The number of volatile compounds in individual curds varied from 15 to 33 while the total abundance of volatile compounds in individual curds ranged from 1.98- to 17.50-fold the total abundance in uninoculated curd. Major differences in volatile generation were related to amino acid catabolism.

Melanoma primario maligno variedad rabdoide. A Propósitode un caso / Malignant primary melanoma rhabdoid variety. about a case

Resumen El melanoma primariovariedad rabdoide es unapresentación pocofrecuente.Reconocido como un subtipo histopatológico distinto de melanoma maligno generalmente observado en tumores metastásicos o recurrentes.El diagnóstico definitivo requiere el estudio de inmunomarcación y la identificación de células neoplásicas con marcadores melanocíticos. Clínicamente se han reportado mayormente de tipo nodular y amelanótico.

Summary Rhabdoid melanoma has been recognized as a histopathological subtype of malignant melanoma. It generally presents as a recurrent tumor, so its presentation as a primary lesion is infrecuent.Definitive diagnosis requires the study of immunostaining and the identification of neoplastic cells with melanocytic markers. Clinically, mostly nodular and amelanotic types have been reported.

Growth stimulation of a proteinase positive Lactococcus lactis strain by a proteinase negative Lactococcus lactis strain.

Lactococcus lactis AMP15/pAMP31(D471R) is a proteinase negative, lactose negative strain with a modified oligopeptide transport system, and potential as a debittering agent due to its efficient utilization of hydrophobic peptides. Five wild L. lactis strains of dairy origin, which produced cheeses of high flavour quality, were cocultured with L. lactis AMP15/pAMP31(D471R) in an attempt to select adequate combinations of strains for use as defined cheese starters with potential debittering ability. Four of these strains, L. lactis B6, K16, M21 and P21, inhibited growth of L. lactis AMP15/pAMP31(D471R) at a level of 10(6) to 10(7) cfu mL(-1) after 24 h of incubation, even though production of bacteriocin-like compounds could only be proven for L. lactis M21. When L. lactis AMP15/pAMP31(D471R) was cocultured with the fifth strain, L. lactis N22, its growth was significantly (P<0.001) inhibited whereas growth of L. lactis N22 was significantly stimulated. The nature of the interaction was studied and it was established that L. lactis N22 is auxotrophic for folate, a compound produced and excreted by L. lactis AMP15/pAMP31(D471R).

Volatile compounds and odour characteristics of seven species of dehydrated edible seaweeds.

The volatile fraction of dehydrated edible seaweeds belonging to seven species (Himanthalia elongata, Laminaria ochroleuca, Palmaria palmata, Porphyra umbilicalis, Saccharina latissima, Ulva lactuca and Undaria pinnatifida) was analyzed by gas chromatography-mass spectrometry, after solid-phase microextraction of samples. Thirty-six hydrocarbons, 34 ketones, 28 aldehydes, 23 alcohols, 8 carboxylic acids, 6 halogenated compounds, 4 furans, 3 esters, 2 sulphur compounds, 2 pyrazines, 1 pyridine and 1 amine were detected among the 151 volatile compounds found in seaweeds. There were significant differences between seaweed species for all the volatile compounds. Hydrocarbons reached their highest levels in U. pinnatifida, ketones in P. umbilicalis, aldehydes in P. palmata and P. umbilicalis, alcohols in P. umbilicalis, carboxylic acids in S. latissima, and halogenated compounds in L. ochroleuca and S. latissima. Sensory analysis revealed that P. palmata, U. lactuca and H. elongata were the seaweeds showing the most potent seafood odour and seaweed odour characteristics.

Microbiota of high-pressure-processed Serrano ham investigated by culture-dependent and culture-independent methods.

The microbiota of Serrano dry-cured ham of different chemical composition, subjected or not to high-pressure processing (HPP), was investigated using culture-dependent and culture-independent methods. Microbial counts were submitted to analysis of variance with physicochemical parameters (aw, NaCl concentration, salt-in-lean ratio and intramuscular fat content) or HPP as main effects. In untreated hams, physicochemical parameters significantly affected counts of aerobic mesophiles, psychrotrophs, and moulds and yeasts. NaCl concentration and fat content influenced the levels of four and three of the five studied microbial groups, respectively, whereas no influence of aw was stated. The HPP treatment had a significant effect on counts of all investigated microbial groups. Culture-independent methods showed the presence of bacteria such as Staphylococcus equorum, Staphylococcus succinus, Bacillus subtilis and Cellulosimicrobium sp., moulds like Penicillium commune, Aspergillus fumigatus, Sclerotinia sclerotiorum, Eurotium athecium and Moniliella mellis, and yeasts like Debaryomyces hansenii and Candida glucosophila. Absence of B. subtilis bands and weaker bands of E. athecium were recorded for HPP-treated hams. The higher microbial levels found in lean ham might result in a quicker deterioration. HPP treatment confirmed its suitability as a procedure to control spoilage microorganisms. DGGE did not seem to be sensitive enough to highlight changes caused by HPP treatment in the microbiota of ham, but contributed to the detection of microbial species not previously found in ham.

Influence of physicochemical parameters and high pressure processing on the volatile compounds of Serrano dry-cured ham after prolonged refrigerated storage.

One hundred and three volatile compounds were detected by solid-phase microextraction followed by gas chromatography-mass spectrometry in 30 ripened Serrano dry-cured hams, submitted or not to high pressure processing (HPP) and afterwards held for 5months at 4°C. The effect of ham physicochemical parameters and HPP (600MPa for 6min) on volatile compounds was assessed. Physicochemical parameters primarily affected the levels of acids, alcohols, alkanes, esters, benzene compounds, sulfur compounds and some miscellaneous compounds. Intramuscular fat content was the physicochemical parameter with the most pronounced effect on the volatile fraction of untreated Serrano ham after refrigerated storage, influencing the levels of 38 volatile compounds while aw, salt content and salt-in-lean ratio respectively influenced the levels of 4, 4 and 5 volatile compounds. HPP treatment affected 21 volatile compounds, resulting in higher levels of alkanes and ketones and lower levels of esters and secondary alcohols, what might affect Serrano ham odor and aroma after 5months of refrigerated storage.

Hetero-site-specific X-ray pump-probe spectroscopy for femtosecond intramolecular dynamics.

New capabilities at X-ray free-electron laser facilities allow the generation of two-colour femtosecond X-ray pulses, opening the possibility of performing ultrafast studies of X-ray-induced phenomena. Particularly, the experimental realization of hetero-site-specific X-ray-pump/X-ray-probe spectroscopy is of special interest, in which an X-ray pump pulse is absorbed at one site within a molecule and an X-ray probe pulse follows the X-ray-induced dynamics at another site within the same molecule. Here we show experimental evidence of a hetero-site pump-probe signal. By using two-colour 10-fs X-ray pulses, we are able to observe the femtosecond time dependence for the formation of F ions during the fragmentation of XeF2 molecules following X-ray absorption at the Xe site.

Analysis of the human proopiomelanocortin gene promoter in a small cell lung carcinoma cell line reveals an unusual role for E2F transcription factors.

The small cell lung carcinoma (SCLC) cell line DMS-79 has been used as a model for studying the molecular mechanism underlying the ectopic ACTH syndrome. We previously showed that two domains of the human Proopiomelanocortin (POMC) gene promoter were specifically active in DMS-79 cells. The present study focuses on the more distal one, Domain IV (-376/-417). DNaseI footprinting experiments identified a single binding site for DMS-79 cell proteins in this domain. Gel-shift and sequence analysis indicated that E2F proteins might bind this site. Indeed, proteins from DMS-79 cells which bind this site (i) have in vitro DNA binding properties indistinguishable from those of E2F proteins (ii) form, like E2F proteins, multiprotein complexes which can be dissociated by sodium deoxycholate and (iii) are recognized by antibodies directed against E2F proteins. Further, we show that the rat POMC distal promoter domain contains a homologous sequence which constitutes a natural mutant of the human POMC E2F binding site, since it does not bind E2F. We show by transient transfection that this natural mutant, in the context of the rat POMC promoter, is not active in DMS-79 cells by contrast to the human POMC E2F binding site. We conclude that E2F binding is required for the activity of Domain IV in DMS-79 cells and contributes to the expression of the POMC gene in SCLC. Further studies are required to elucidate the role of E2F factors in POMC gene transcription in SCLC cells, but our results have identified mechanisms different from those in pituitary corticotroph cells that are used by these SCLC tumor cells.

Multiplex PCR/LDR for detection of K-ras mutations in primary colon tumors.

Point mutations in codons 12, 13, and 61 of the K-ras gene occur early in the development of colorectal cancer and are preserved throughout the course of tumor progression. These mutations can serve as biomarkers for shed or circulating tumor cells and may be useful for diagnosis of early, curable tumors and for staging of advanced cancers. We have developed a multiplex polymerase chain reaction/ligase detection reaction (PCR/LDR) method which identifies all 19 possible single-base mutations in K-ras codons 12, 13, and 61, with a sensitivity of 1 in 500 wild-type sequences. In a blinded study, 144 paraffin-embedded archival colon carcinomas were microdissected and K-ras mutations determined by both dideoxy-sequencing and multiplex PCR/LDR. Results were concordant for 134 samples. The ten discordant samples were re-evaluated using higher sensitivity uniplex PCR/LDR, and the original multiplex PCR/LDR result was confirmed in nine of these ten cases. Multiplex PCR/LDR was able to identify mutations in solid tumors or paraffin-embedded tissues containing a majority of wild-type stromal cells, with or without microdissection. The technique is well suited for large scale studies and for analysis of clinical samples containing a minority population of mutated cells.

Cheesemaking with a Lactococcus lactis strain expressing a mutant oligopeptide binding protein as starter results in a different peptide profile.

Lactic starters used for cheese manufacture play an important role in the production of bitter peptides and their degradation to non-bitter products. The oligopeptide transport system (Opp) of lactococci is essential for milk peptide utilization. The periplasmic substrate binding protein serves to capture the substrate with high affinity and to deliver it to a membrane-bound complex that translocates it inside the cell. Prt(+)- and Lac(+)-derivatives of MG1363 DeltaoppA strains expressing a wild-type MG1363 OppA or a mutant OppA with a single point mutation at residue 471 (OppA(D471R)) from a plasmid were constructed. These strains were used as lactic starters in cheese manufacture to improve flavour quality by removing hydrophobic peptides from the cheese matrix, through their preferential transport by OppA(D471R). Cheeses made with these strains were not significantly different from control cheeses after 1 day of ripening with respect to bacterial counts, pH and proteolysis, and only slight differences were recorded after 9 and 20 days of ripening. HPLC chromatograms of the hydrophilic and hydrophobic peptides present in the water-soluble fraction of experimental cheeses showed significant differences in peptide content as well as in peak profiles. These results suggest a different peptide utilization in the strain expressing OppA(D471R) and make it suitable for use as starter to improve cheese quality.

High expression of the POU factor Brn3a in aggressive neuroendocrine tumors.

A new family of POU transcription factors called Brn plays a role in development of the brain and some neuroendocrine structure. Because a member of this family, Brn3a, is present in the ACTH-producing mouse pituitary tumor AtT-20, binds to POMC promoter, and stimulates its activity, we studied its human homolog in ACTH-secreting or nonsecreting tumors of pituitary and bronchial origins. A specific and quantitative reverse transcription-PCR assay was developed to assess Brn3a transcripts in tumor ribonucleic acid. Brn3a transcript levels were invariably low (< 5 x 10(-6) arbitrary units) in four GH-, two PRL-, three gonadotropin-, and seven of eight ACTH-producing pituitary adenomas. A single highly invasive ACTH-secreting pituitary adenoma in a patient who ultimately died with liver metastases, and the mouse corticotroph tumor cell line AtT-20 had high Brn3a transcripts levels at 3 x 10(-5) and 4 x 10(-4) arbitrary units, respectively. Five typical bronchial carcinoids had barely detectable levels (< 5 x 10(-6) arbitrary units), whereas seven of eight small cell carcinomas of the lung (SCCLs) had extremely high levels (between 10(-3)-10(-1) arbitrary units); six of seven atypical bronchial carcinoids had intermediate values, between 10(-6) and 5 x 10(-3) arbitrary units. Although nine bronchial tumors produced POMC, there was no association between Brn3a levels and POMC gene expression; the two tumors with the highest POMC messenger ribonucleic acid contents were two bronchial carcinoids with barely detectable Brn3a levels. A gel mobility shift assay was performed with a rat CRH promoter probe that binds Brn3a; extracts of the POMC-producing human SCCL line DMS-79, which contained high levels of Brn3a transcripts, generated the same specific complex as did AtT-20 cell extracts. These data show that Brn3a gene expression in neuroendocrine tumors is not correlated with POMC gene expression; rather, it is strikingly elevated in the highly aggressive tumors, independently of their POMC status and their pituitary or nonpituitary origin.