Clinical insights gained by refining the 2016 WHO classification of diffuse gliomas with: EGFR amplification, TERT mutations, PTEN deletion and MGMT methylation.

BACKGROUND: Significant advances in the molecular profiling of gliomas, led the 2016 World Health Organization (WHO) Classification to include, for the first-time, molecular biomarkers in glioma diagnosis: IDH mutations and 1p/19q codeletion. Here, we evaluated the effect of this new classification in the stratification of gliomas previously diagnosed according to 2007 WHO classification. Then, we also analyzed the impact of TERT promoter mutations, PTEN deletion, EGFR amplification and MGMT promoter methylation in diagnosis, prognosis and response to therapy in glioma molecular subgroup. METHODS: A cohort of 444 adult gliomas was analyzed and reclassified according to the 2016 WHO. Mutational analysis of IDH1 and TERT promoter mutations was performed by Sanger sequencing. Statistical analysis was done using SPSS Statistics 21.0. RESULTS: The reclassification of this cohort using 2016 WHO criteria led to a decrease of the number of oligodendrogliomas (from 82 to 49) and an increase of astrocytomas (from 49 to 98), while glioblastomas (GBM) remained the same (n = 256). GBM was the most common diagnosis (57.7%), of which 55.2% were IDH-wildtype. 1p/19q codeleted gliomas were the subgroup associated with longer median overall survival (198 months), while GBM IDH-wildtype had the worst outcome (10 months). Interestingly, PTEN deletion had poor prognostic value in astrocytomas IDH-wildtype (p = 0.015), while in GBM IDH-wildtype was associated with better overall survival (p = 0.042) as well as MGMT promoter methylation (p = 0.009). EGFR amplification and TERT mutations had no impact in prognosis. Notably, EGFR amplification predicted a better response to radiotherapy (p = 0.011) and MGMT methylation to chemo-radiotherapy (p = 0.003). CONCLUSION: In this study we observed that the 2016 WHO classification improved the accuracy of diagnosis and prognosis of diffuse gliomas, although the available biomarkers are not enough. Therefore, we suggest MGMT promoter methylation should be added to glioma classification. Moreover, we found two genetic/clinical correlations that must be evaluated to understand their impact in the clinical setting: i) how is PTEN deletion a favorable prognostic factor in GBM IDH wildtype and an unfavorable prognostic factor in astrocytoma IDH wildtype and ii) how EGFR amplification is an independent and strong factor of response to radiotherapy.

Preparative scale purification of fucosyl-N-acetylglucosamine disaccharides and their evaluation as potential prebiotics and antiadhesins.

Fucosyl-N-acetylglucosamine disaccharides are important core structures that form part of human mucosal and milk glyco-complexes. We have previously shown that AlfB and AlfC α-L-fucosidases from Lactobacillus casei are able to synthesize fucosyl-α-1,3--N-acetylglucosamine (Fuc-α1,3-GlcNAc) and fucosyl-α-1,6-N-acetylglucosamine (Fuc-α1,6-GlcNAc), respectively, in transglycosylation reactions. Here, these reactions were performed in a semipreparative scale, and the produced disaccharides were purified. The maximum yields obtained of Fuc-α1,3-GlcNAc and Fuc-α1,6-GlcNAc were 4.2 and 9.3 g/l, respectively. The purified fucosyl-disaccharides were then analyzed for their prebiotic effect in vitro using strains from the Lactobacillus casei/paracasei/rhamnosus group and from Bifidobacterium species. The results revealed that 6 out of 11 L. casei strains and 2 out of 6 L. rhamnosus strains tested were able to ferment Fuc-α1,3-GlcNAc, and L. casei BL87 and L. rhamnosus BL327 strains were also able to ferment Fuc-α1,6-GlcNAc. DNA hybridization experiments suggested that the metabolism of Fuc-α1,3-GlcNAc in those strains relies in an α-L-fucosidase homologous to AlfB. Bifidobacterium breve and Bibidobacterium pseudocatenolatum species also metabolized Fuc-α1,3-GlcNAc. Notably, L-fucose was excreted from all the Lactobacillus and Bifidobacterium strains fermenting fucosyl-disaccharides, except from strains L. rhamnosus BL358 and BL377, indicating that in these latest strains, L-fucose was catabolized. The fucosyl-disaccharides were also tested for their inhibitory potential of pathogen adhesion to human colon adenocarcinoma epithelial (HT29) cell line. Enteropathogenic Escherichia coli (EPEC) strains isolated from infantile gastroenteritis were used, and the results showed that both fucosyl-disaccharides inhibited adhesion to different extents of certain EPEC strains to HT29 cells in tissue culture.

Collective deviance: Scaling up subjective group dynamics to superordinate categories reveals a deviant ingroup protection effect.

Six experiments examined responses to groups whose attitudes deviated from wider social norms about asylum and immigration (in the United Kingdom), or about taxation levels (in the U.S.). Subjective group dynamics (SGD) theory states that people derogate in-group individuals who deviate from prescriptive in-group norms. This enables members to sustain the subjective validity of those norms and, hence, a positive social identity. Research also shows that in-group deviants who accentuate the difference between the in-group and out-group norm (e.g., extremists) are derogated less than deviants who attenuate that difference (e.g., a member who veers toward the outgroup's norm; Abrams et al., 2000). We hypothesize that these effects and the associated group dynamics should scale up when people evaluate deviant groups that are part of larger in-categories. Consistent with SGD theory, participants in Experiments 1, 2, and 3 derogated an in-category attenuating deviant group and upgraded an out-category attenuating deviant group relative to groups that consolidated or accentuated the respective norms of those categories-thereby reinforcing in-category norms relative to out-category norms. Across all experiments, this pattern of differential evaluation was associated with greater subjective validity of the in-category norm. We also hypothesized a novel Deviant Ingroup Protection (DIP) effect, wherein people should curtail derogation of an in-category deviant group when that group is their own. Consistent with this hypothesis, participants in Experiments 4, 5, and 6 evaluated an accentuating in-group, or an attenuating in-group, equally to or more positively than other in-category groups. Implications for political and organizational entrenchment are discussed. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Infant Gut Microbial Metagenome Mining of α-l-Fucosidases with Activity on Fucosylated Human Milk Oligosaccharides and Glycoconjugates.

The gastrointestinal microbiota members produce α-l-fucosidases that play key roles in mucosal, human milk, and dietary oligosaccharide assimilation. Here, 36 open reading frames (ORFs) coding for putative α-l-fucosidases belonging to glycosyl hydrolase family 29 (GH29) were identified through metagenome analysis of breast-fed infant fecal microbiome. Twenty-two of those ORFs showed a complete coding sequence with deduced amino acid sequences displaying the highest degree of identity with α-l-fucosidases from Bacteroides thetaiotaomicron, Bacteroides caccae, Phocaeicola vulgatus, Phocaeicola dorei, Ruminococcus gnavus, and Streptococcus parasanguinis. Based on sequence homology, 10 α-l-fucosidase genes were selected for substrate specificity characterization. The α-l-fucosidases Fuc18, Fuc19A, Fuc35B, Fuc39, and Fuc1584 showed hydrolytic activity on α1,3/4-linked fucose present in Lewis blood antigens and the human milk oligosaccharide (HMO) 3-fucosyllactose. In addition, Fuc1584 also hydrolyzed fucosyl-α-1,6-N-acetylglucosamine (6FN), a component of the core fucosylation of N-glycans. Fuc35A and Fuc193 showed activity on α1,2/3/4/6 linkages from H type-2, Lewis blood antigens, HMOs and 6FN. Fuc30 displayed activity only on α1,6-linked l-fucose, and Fuc5372 showed a preference for α1,2 linkages. Fuc2358 exhibited a broad substrate specificity releasing l-fucose from all the tested free histo-blood group antigens, HMOs, and 6FN. This latest enzyme also displayed activity in glycoconjugates carrying lacto-N-fucopentaose II (Lea) and lacto-N-fucopentaose III (Lex) and in the glycoprotein mucin. Fuc18, Fuc19A, and Fuc39 also removed l-fucose from neoglycoproteins and human α-1 acid glycoprotein. These results give insight into the great diversity of α-l-fucosidases from the infant gut microbiota, thus supporting the hypothesis that fucosylated glycans are crucial for shaping the newborn microbiota composition. IMPORTANCE α-l-Fucosyl residues are frequently present in many relevant glycans, such as human milk oligosaccharides (HMOs), histo-blood group antigens (HBGAs), and epitopes on cell surface glycoconjugate receptors. These fucosylated glycans are involved in a number of mammalian physiological processes, including adhesion of pathogens and immune responses. The modulation of l-fucose content in such processes may provide new insights and knowledge regarding molecular interactions and may help to devise new therapeutic strategies. Microbial α-l-fucosidases are exoglycosidases that remove α-l-fucosyl residues from free oligosaccharides and glycoconjugates and can be also used in transglycosylation reactions to synthesize oligosaccharides. In this work, α-l-fucosidases from the GH29 family were identified and characterized from the metagenome of fecal samples of breastfed infants. These enzymes showed different substrate specificities toward HMOs, HBGAs, naturally occurring glycoproteins, and neoglycoproteins. These novel glycosidase enzymes from the breast-fed infant gut microbiota, which resulted in a good source of α-l-fucosidases, have great biotechnological potential.

Mixed testicular germ cell tumour in a patient with previous pineal germinoma.

Germ cell tumours (GCT) are a relatively common malignancy in men aged 15-35 years. They occur most frequently in the gonads, but 3-5% have extragonadal origin, mainly in the pineal gland, neurohypophysis, mediastinum and retroperitoneum. Although intracranial germinomas may present with synchronous midline lesions, development of metachronous testicular germ cell primaries seems to be extremely rare, and confirmed dissemination of intracranial GCT to the testes has never been reported. We report the case of a 32-year-old man, with previously treated pineal germinoma at age 16 years, who later developed mixed GCT of the left testis.

The future in our hands: How citizenship efficacy ensures commitment to the national group facing institutional inefficacy.

Literature has shown that citizens' mistrust in national institutions has a negative impact on their involvement with, and commitment to, their national group. We examine the idea that citizenship efficacy beliefs may revert this process. We propose that facing institutional inefficacy to exert social control, beliefs that civic participation is effective, strengthens individuals' commitment to the national group. Participants (N = 176) were informed that national institutions were effective (vs. ineffective) in reacting to white-collar crime, and that citizens' civic/political participation had an effective (vs. ineffective) impact on government's decisions, the political system and their nation's future. Results suggest that citizenship efficacy beliefs are crucial to counteract or even revert citizens' disinvestment in the national group caused by the perception that the social control system is ineffective. We discuss the results in light of the theoretical and empirical framework of social psychology of citizenship and subjective group dynamics theory.

Social Dominance Orientation Boosts Collective Action Among Low-Status Groups.

We propose that low-status group members' support for group-based hierarchy and inequality (i.e., social dominance orientation; SDO) may represent an ideological strategy to guarantee the legitimacy of future ingroup status-enhancement. Specifically, we argue that, under unstable social structure conditions, SDO serves as an ideological justification for collective action tendencies aimed at competing for a higher status. In such context, SDO should be positively related with actions aimed to favor the ingroup (i.e., collective actions) by increasing group members' motivation to engage in direct competition with a relevant higher-status outgroup. We conducted two studies under highly competitive and unstable social structure contexts using real life groups. In Study 1 (N = 77), we induced Low vs. High Ingroup (University) Status and in Study 2 (N = 220) we used competing sports groups. Overall, results showed that, among members of low-status groups, SDO consistently increased individuals' motivation to get involved in actions favoring the ingroup, by boosting their motivation to compete with the opposing high-status outgroup. We discuss the results in light of the social dominance and collective action framework.

A Path Toward Inclusive Social Cohesion: The Role of European and National Identity on Contesting vs. Accepting European Migration Policies in Portugal.

The Western hemisphere has witnessed recent increased immigration flows generating social and political debate across Europe. In one view, migration flows represent an opportunity to construct a diverse social cohesion. In another view, migration flows are perceived as a threat to existent national cultures. This view is held by political nationalisms and right-wing populist forces installed in the majority of EU countries' parliaments, accentuating discrimination against immigrants and residents in Europe. We theorize that European identity predicts positive attitudes toward immigrants (prosocial behavior and support for inclusive policies), whereas national identity's predictions of attitudes toward immigrants' inclusion depends on participants' political tendency. Moreover, we test the mediation effect of positive (humanitarian concerns and economic benefit) and negative (jobs scarcity, cultural deterioration, and invasion) arguments used in political discourses regarding immigrants' inclusion on the relation between national and European identities and attitudes toward immigrants' inclusion. Results (Portuguese sample, N = 176) show that national identity predicts negative attitudes toward immigrants' inclusion, but only among right-wing individuals. Among left-wing individuals, national identity predicts less contestation to immigrant's inclusion sustained by humanitarian concerns. Interestingly, European identification weakened right-wing individuals' adherence to discriminatory arguments and increased perceived economic contribution that immigrants bring to society, increasing agreement with prosocial behavior and immigrants' inclusion. We discuss that European identity, sustained in humanitarian values and economic benefit, may stimulate a stronger multicultural social cohesion, intergroup trust, and social well-being based on democratic values, social justice, and equality, and on the respect for human dignity.

Lactobacillus casei extracellular vesicles stimulate EGFR pathway likely due to the presence of proteins P40 and P75 bound to their surface.

In the complex interplay of beneficial bacteria with the host, there are few examples of bacterial metabolites and effector molecules that have been consistently identified. Protective effects on the intestinal epithelium have been ascribed to P40 and P75, two well characterized cell wall muramidases, present in the culture supernatant of strains belonging to the taxon Lactobacillus casei/paracasei/rhamnosus. This work reports that Lactobacillus casei BL23 extracellular vesicles (BL23 EVs) have a small size (17-20 nm or 24-32 nm, depending on the method used) and contain lipoteichoic acid (LTA). Interestingly, all detected P40 and most of P75 were associated to EVs and possibly located at their external surface, as shown by proteinase K digestion. Biosensor assays showed that both proteins bind LTA and vesicles, suggesting that they could bind to ligands like LTA present on BL23 EVs. Native BL23 EVs have a moderate proinflammatory effect and they were able to induce phosphorylation of the epidermal growth factor receptor (EGFR), showing an effect similar to purified P40 and P75 and leading to the conclusion that the activity described in the supernatant (postbiotic) of these bacteria would be mainly due to P40 and P75 bound to EVs.

Effect of Computer-Simulated Leaders' Compromise on Members' Emotional State and Protest Behavior.

Participants (N = 119) played the "Dictator Game" (computer mediated) with two bogus computer-simulated players, one of whom, the Dictator, distributed money across ten trials, either as extremely unfair (Inflexible Dictator) or being less unfair (Flexible Dictator). The other player either protested against (Protest condition) or did not react (Apathy condition) to the dictator's decision, after each trial. We measured participants' self-reported anger and disinterest, physiological skin conductance (SCL), heart rate (HR), and number and type of comments directed to the Dictator. Anger and number of comments were lower in the Apathy than in the Protest condition. Participants' SCL, HR, and protest comments decreased in the Apathy condition, and increased in the Protest condition. Protest assumed a more punitive tone in the Inflexible than in the Flexible Dictator condition. We discuss these results' contribution to understand individuals' motivation to engage in protest and apathy, and the role of emotions in that process.

P40 and P75 Are Singular Functional Muramidases Present in the Lactobacillus casei /paracasei/rhamnosus Taxon.

Lactobacillus casei and Lactobacillus rhamnosus proteins P40 and P75 belong to a large family of secreted cell wall proteins that contain a carboxy(C)-terminal CHAP or NlpC/P60 superfamily domains. In addition to their peptidoglycan hydrolases activity, proteins in this family are specific antigens of pathogens, frequently responsible of interactions with the host. L. rhamnosus GG and L. casei BL23 purified P40 and P75 proteins have antiapoptotic activity by inducing the EGF/Akt pathway. The aim of this work was to study the genetics, phylogeny and dissemination of this family of proteins in the genus Lactobacillus as well as their characteristics and likely function. The scrutiny of their DNA encoding sequences revealed the presence of minisatellite DNA in the P75 encoding gene of L. casei/paracasei strains (cmuB) with intraspecific indels that gave raise to four different alleles (cmuB1-4), which are exclusive of this species. Phylogenic analyses suggest that both proteins are present mainly in the L. casei and Lactobacillus sakei phylogenomic groups. A P40 ancestral gene was possibly present in the common ancestor of Enterococcaceae, Lactobacillaceae and Streptococcaceae. P75 is also present in L. casei and L. sakei groups, but its evolution is difficult to explain only by vertical transmission. Antibodies raised against the N-terminal regions of P40 and P75 improved their immunological detection in culture supernatants as they recognized almost exclusively proteins of L. casei/paracasei/rhamnosus strains, highlighting their structural similarity, that allowed to detect them in different fermented dairy products that contained probiotic L. casei strains. Purified P40 and P75 proteins showed no evident lytic activity but they complemented L. casei BL23 cmuA and cmuB defective mutants, respectively, thus proving that they actively participate in cell division.

Innovation credit: when can leaders oppose their group's norms?

Two preliminary studies and 5 experiments examined judgments of leaders who challenge their group's norms. Participants viewed information about group members whose attitudes were normative or deviated in a pronormative or antinormative direction. The antinorm member was identified as (a) either a nonleader or an established leader (Study 1), (b) an ex-leader (Studies 2 and 5), or (c) a future leader (Studies 3, 4, and 5). Antinorm future leaders were judged more positively and were granted greater innovation credit (license to innovate and remuneration) relative to antinorm members, ex-leaders, and established leaders. Results are discussed in terms of the idea that leadership can accrue from prototypicality and can also confer the right to define prescriptive norms. However, innovation credit is only granted in the case of future leaders.

Polyphosphate in Lactobacillus and Its Link to Stress Tolerance and Probiotic Properties.

The synthesis of the inorganic polymer polyphosphate (poly-P) in bacteria has been linked to stress survival and to the capacity of some strains to sequester heavy metals. In addition, synthesis of poly-P by certain strains of probiotic lactobacilli has been evidenced as a probiotic mechanism due to the homeostatic properties of this compound at the intestinal epithelium. We analyzed the link between poly-P synthesis, stress response, and mercury toxicity/accumulation by comparing wild-type strains of Lactobacillus and their corresponding mutants devoid of poly-P synthesis capacity (defective in the poly-P kinase, ppk, gene). Results showed that resistance to salt (NaCl) and acidic (pH 4) stresses upon ppk mutation was affected in Lactobacillus casei, while no effect was observed in two different Lactobacillus plantarum strains. Inorganic [Hg(II)] and organic (CH3Hg) mercury toxicity was generally increased upon ppk mutation, but no influence was seen on the capacity to retain both mercurial forms by the bacteria. Notwithstanding, the culture supernatants of ppk-defective L. plantarum strains possessed a diminished capacity to induce HSP27 expression, a marker for cell protection, in cultured Caco-2 cells compared to wild-type strains. In summary, our results illustrate that the role of poly-P in stress tolerance can vary between strains and they reinforce the idea of probiotic-derived poly-P as a molecule that modulates host-signaling pathways. They also question the relevance of this polymer to the capacity to retain mercury of probiotics.

Changes in bone Pb accumulation: cause and effect of altered bone turnover.

This paper assesses the magnitude of Pb uptake in cortical and trabecular bones in healthy animals and animals with altered balance in bone turnover, and the impact of exposure to Pb on serum markers of bone formation and resorption. The results reported herein provide physiological evidence that Pb distributes differently in central compartments in Pb metabolism, such as cortical and trabecular bones, in healthy animals and animals with altered balance in bone turnover, and that exposure to Pb does have an impact on bone resorption resulting in OC-dependent osteopenia. These findings show that Pb may play a role in the etiology of osteoporosis and that its concentration in bones varies as a result of altered bone turnover characteristic of this disease, a long standing question in the field. In addition, data collected in this study are consistent with previous observations of increased half-life of Pb in bone at higher exposures. This evidence is relevant for the necessary revision of current physiologically based kinetic models for Pb in humans.

Noroviral p-particles as an in vitro model to assess the interactions of noroviruses with probiotics.

Noroviruses (NoVs) are the main etiologic agents of acute epidemic gastroenteritis and probiotic bacteria have been reported to exert a positive effect on viral diarrhea. The protruding (P) domain from NoVs VP1 capsid protein has the ability to assemble into the so-called P-particles, which retain the binding ability to host receptors. We purified the P-domains from NoVs genotypes GI.1 and GII.4 as 6X(His)-tagged proteins and determined that, similar to native domains, they were structured into P-particles that were functional in the recognition of the specific glycoconjugated receptors, as established by surface plasmon resonance experiments. We showed that several lactic acid bacteria (probiotic and non-probiotic) and a Gram-negative probiotic strain have the ability to bind P-particles on their surfaces irrespective of their probiotic status. The binding of P-particles (GI.1) to HT-29 cells in the presence of selected strains showed that bacteria can inhibit P-particle attachment in competitive exclusion experiments. However, pre-treatment of cells with bacteria or adding bacteria to cells with already attached P-particles enhanced the retention of the particles. Although direct viral binding and blocking of viral receptors have been postulated as mechanisms of protection against viral infection by probiotic bacteria, these results highlight the need for a careful evaluation of this hypothesis. The work presented here investigates for the first time the probiotic-NoVs-host interactions and points up the NoVs P-particles as useful tools to overcome the absence of in vitro cellular models to propagate these viruses.

Bone regeneration in osteoporotic conditions: healing of subcritical-size calvarial defects in the ovariectomized rat.

PURPOSE: Osteoporosis is a pathologic condition characterized by low bone mass and changes in the microarchitecture of the bone tissue. Although compromised bone strength and increased susceptibility to fracture have been established, little is known regarding the process of bone regeneration in osteoporotic conditions. Accordingly, this study sought to evaluate the intramembranous bone regeneration process in an ovariectomized rat model following the establishment of calvarial subcritical-size defects (sCSDs). MATERIALS AND METHODS: Calvarial sCSDs were established in rats that had been ovariectomized (Ovx) or sham-operated 2 months previously and left to heal, unfilled, for 6 months. Bone regeneration was assessed by radiographic, densitometric, histologic, and histometric analyses. RESULTS: Radiologic and histologic analyses showed reduced new bone formation in calvarial sCSDs in Ovx animals in comparison to sham animals. Densitometric analysis of radiologic images and histometric analysis showed significant quantitative differences between groups that converged to substantiate reduced bone regeneration in Ovx animals. CONCLUSIONS: The intramembranous ossification process is impaired in the Ovx rat model. This may suggest an impairment of the bone regeneration process in clinical conditions of postmenopausal osteoporosis and highlight the requirement for selective bone regenerative strategies in affected patients.

Membership status and subjective group dynamics: who triggers the black sheep effect?

In 3 experiments, participants (Ns = 50, 95, and 75, respectively) judged 2 ingroup or outgroup members who occupied 1 of 3 statuses--new members, full members, or marginal members. In each case, 1 of these members adopted a normative position and another supported a deviant position regarding a relevant issue. Participants upgraded normative ingroup full members and derogated deviant ingroup full members compared with all other members. In addition, derogation of deviant ingroup members was associated with a socializing and a punishing intention toward new members and full members, respectively. These results are discussed in terms of the group socialization model (e.g., Levine & Moreland, 1994) and the subjective group dynamics model (e.g., Marques, Paez, & Abrams, 1998).

Nitrogen isotopes determination in natural gas: analytical method and first results on magmatic, hydrothermal and soil gas samples.

A continuous-flow GC/IRMS technique has been developed to analyse delta(15)N values for molecular nitrogen in gas samples. This method provides reliable results with accuracy better than 0.15 per thousand and reproducibility (1sigma) within+/-0.1 per thousand for volumes of N(2) between 1.35 (about 56 nmol) and 48.9 muL (about 2 mumol). The method was tested on magmatic and hydrothermal gases as well as on natural gas samples collected from various sites. Since the analysis of nitrogen isotope composition may be prone to atmospheric contamination mainly in samples with low N(2) concentration, we set the instrument to determine also N(2) and (36)Ar contents in a single run. In fact, based on the simultaneously determined N(2)/(36)Ar ratios and assuming that (36)Ar content in crustal and mantle-derived fluids is negligible with respect to (36)Ar concentration in the atmosphere, for each sample, the degree of atmospheric contamination can be accurately evaluated. Therefore, the measured delta(15)N values can be properly corrected for air contamination.

Assessment of recharge and flowpaths in a limestone thermomineral aquifer system using environmental isotope tracers (Central Portugal).

We conducted chemical and isotopic analyses to develop and test a hydrogeological model of thermomineral water circulation in a limestone aquifer system at Caldas da Rainha (Central Portugal), contributing to future borehole-drilling and development strategies, with the aim of extracting waters with the best possible flow and/or temperature. The thermomineral waters (T approximately 33 degrees C) discharge from springs and boreholes located close to a locally N-S-oriented oblique fault (60 degrees E) that places loamy and detritic Upper Jurassic rocks and Hettangian-Rhaetian marls (and evaporitic deposits) in contact. (14)C determinations indicate a pmC content between 29.33+/-0.14 and 44.39+/-0.20 pmC. The presence of HCO[image omitted] , Ca(2+) (and Mg(2+)) are ascribed to water-limestone interactions, while Na(+), Cl(-) and SO[image omitted] concentrations are mainly associated with the dissolution of halite and gypsum lenses found along the regional syncline structure. The delta(18)O values of Caldas da Rainha thermomineral water were slightly lower than those of shallow groundwater from the Upper Jurassic rocks, suggesting the existence of two distinct aquifer systems. The different isotopic composition of water also indicates that the main recharge of the thermomineral waters likely occurs in the Middle and Lower Jurassic limestone formations of the Candeeiros Mountains. The presence of (3)H (from 1.1 to 2.8 TU) in some thermomineral borehole waters (showing rather similar geochemical signatures) suggests mixing of small amounts of shallow groundwater with thermomineral waters, as a result of leaking borehole casing construction or a recharge when the (3)H content in the atmosphere was higher than that at present. Caldas da Rainha thermomineral waters having delta(34)S(sulphate) and delta(18)O(sulphate) values between+14.9 and+19.1 per thousand and+11.1 and+16.2 per thousand, respectively, indicate that the sulphate is the result of water-rock interaction with evaporitic formations. The obtained chemical and isotopic data have informed the further development of a hydrogeological model that will be used by decision-makers, in order to contribute to the socio-economic development of the spa region.