Perfluoroalkyl acids (PFASs) are toxic pollutants ubiquitously found in the ecosystem. Recent investigations have focused on describing their environmental behavior and spreading. This includes transportation by water, where groundwater can continue to spread the contaminants a long time after the use has ended. This thesis surveys the existing literature on the reactions in soil that decides the presence and composition of PFASs in groundwater. A chemical groundwater investigation was recently done by Stockholm’s Environmental and Health Administration. The thesis presents a quantitative analysis of the PFASs’ content in Stockholm’s groundwater to verify if it conforms to the literature. A statistical analysis of the ratio between perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) is included to test if the PFAS composition in groundwater is influenced by soil reactions. The literature states that due to the high water solubility, low volatility and moderate adsorption, PFASs are likely to be encountered in groundwater. The adsorption depends on both PFASs’ and soil’s properties. The quantitative analysis confirms the influence of the PFASs’ properties in Stockholm’s groundwater, where both more short-chained PFASs and carboxylates are present. However, the analysis could not confirm the influence of the soil properties in adsorption at normal environmental conditions. The statistical analysis shows that the ratio of PFOS to PFOA is increased in groundwater in comparison to stormwater (p < 0.15). The increase cannot be explained by the hypothesis that the adsorption of PFAS governs this behavior, since PFOS is a sulfonate and longer than PFOA. An explanation is found in the degradation of precursors, which seems to contribute to PFOS concentrations in groundwater. This shows the important contribution of degradation of precursors to PFOS concentrations and motivates further investigations on the matter.

This study compared whether activation of muscle ergoreceptor afferents caused by isometric muscle contraction, activation of baroreceptor afferents induced by i.v. infusion of phenylephrine, or baroreceptor afferent inactivation, caused by carotid artery occlusion, elicit similar patterns of c-Fos induction in brainstem areas. Adult cats were anesthetized with α-chloralose, and in each case, the experimental intervention caused an increase in the arterial blood pressure. There were two sets of control experiments: in both, animals underwent the same surgical procedures but then either remained at rest for the entire study, or the tibial nerve was stimulated, as in the contraction group, following muscle paralysis with tubocurarine. Following the procedures, animals rested for 90 min to allow neuronal expression of c-Fos. Control cats showed very little c-Fos immunoreactivity (c-Fos-ir) in the brainstem. Muscle contraction induced c-Fos-ir expression mainly in the nucleus tractus solitarius, lateral reticular nucleus, lateral tegmental field, vestibular nucleus, subretrofacial nucleus, spinal trigeminal tract and in a lateral region of the periaqueductal grey (P 0.5-1.0). The majority of the c-Fos-ir was found in brainstem areas contralateral to the contracted muscle. In addition, muscle contraction induced c-Fos-ir in the dorsal horns of spinal segments L6-S1 on the ipsilateral side of the spinal cord. Phenylephrine infusion caused c-Fos-ir expression in the nucleus tractus solitarius, spinal trigeminal tract, solitary tract, and dorsal motor nucleus of the vagus. No c-Fos-ir was apparent in the periaqueductal grey. Carotid occlusions induced c-Fos-ir expression in the area postrema, nucleus tractus solitarius, solitary tract, and spinal trigeminal tract. Expression was bilateral. Areas that exhibited c-Fos-ir correspond to sites previously reported to release various neuropeptides in response to muscle contraction or carotid occlusions. These results indicate that the exercise pressor reflex and baroreflex activate similar, but not completely identical, sites in the brainstem.

Recent investigations of hypoxia physiology in the naked mole rat have opened up an interesting line of research into the basic physiological and genomic alterations that accompany hypoxia survival. The extent to which such findings connect the effect of hypoxia to metabolic rate (O2 consumption), core body temperature (Tb), and transcripts encoding the immediate early gene product (such as c-fos) under a constant ambient temperature (Ta) is not well known. We investigated this issue in the current study. Our first sets of experiments measured Tb and metabolic rates during exposure of naked mole rats to hypoxia over a constant Ta. Hypoxia significantly decreased metabolic rates in the naked mole rat. Although core Tb also decreased during hypoxia, the effect of hypoxia in suppressing core Tb was not significant. The second series of experiments revealed that c-fos protein and mRNA expression in the hippocampus neurons (CA1) increased in naked mole rats that were repeatedly exposed to 3% O2 for 60min per day for 5 days when compared to normoxia. Our findings provide evidence for the up-regulation of c-fos and suppression of metabolic rate in hypoxia tolerating naked mole rats under constant ambient temperature. Metabolic suppression and c-fos upregulation constitute part of the physiological complex associated with adaptation to hypoxia.

The upper cervical spinal region functions as an intraspinal controller of thoracic spinal reflexes and contributes to neuronal regulation of the ischemic myocardium. Our objective was to determine whether stimulation of the C2 cervical spinal cord (SCS) of rats modified the input signal at the thoracic spinal cord when cardiac ischemia-sensitive (sympathetic) afferents were activated by transient occlusion of the left anterior descending coronary artery (CoAO). Changes in c-Fos expression were used as an index of neuronal activation within the spinal cord and brain stem. The pattern of substance P (SP) release, a putative nociceptive transmitter, was measured using antibody-coated microprobes. Two SCS protocols were used: reactive SCS, applied concurrently with intermittent CoAO and preemptive, sustained SCS starting 15 min before and continuing during the repeated intermittent CoAO. CoAO increased SP release from laminae I and II in the T4 spinal cord above resting levels. Intermittent SCS with CoAO resulted in greater levels of SP release from deeper laminae IV-VII in T4 than CoAO alone. In contrast, SP release from laminae I and II was inhibited when CoAO was applied during preemptive, sustained SCS. Preemptive SCS likewise reduced c-Fos expression in the T4 spinal cord (laminae I-V) and nucleus tractus solitarius but increased expression in the intermediolateral cell column of T4 compared with CoAO alone. These results suggest that preemptive SCS from the high cervical region modulates sensory afferent signaling from the ischemic myocardium.

The Multichannel Analysis of Surface Waves (MASW) method traditionally uses an array of collinear vertical geophones to measure seismic wave propagation velocity at discrete points along the ground surface. Distributed fiber optic sensors (FOS) measure the average longitudinal strain over discrete lengths (i.e., zones) of a buried fiber optic cable. Such strain measurements can be used to assess ground motion and thus analyzed with the MASW method. To evaluate the feasibility of using FOS strain measurements in the MASW method, field experiments were conducted with both FOS and surface vertical geophones. Synthetic seismograms were also used to compare FOS to vertical and horizontal geophones and investigate the effect of installation depth and sensor type. Through the MASW method, shear wave (Vs) profiles from the FOS showed comparable results to those obtained with the geophones and achieved the same degree of uncertainty from the non-uniqueness of the MASW inversion process.

Our aim was to analyse (i) the presence of single nucleotide polymorphisms (SNPs) in the JUN and FOS core promoters in patients with rheumatoid arthritis (RA), knee-osteoarthritis (OA), and normal controls (NC); (ii) their functional influence on JUN/FOS transcription levels; and (iii) their associations with the occurrence of RA or knee-OA. JUN and FOS promoter SNPs were identified in an initial screening population using the Non-Isotopic RNase Cleavage Assay (NIRCA); their functional influence was analysed using reporter gene assays. Genotyping was done in RA (n = 298), knee-OA (n = 277), and NC (n = 484) samples. For replication, significant associations were validated in a Finnish cohort (OA: n = 72, NC: n = 548). Initially, two SNPs were detected in the JUN promoter and two additional SNPs in the FOS promoter in perfect linkage disequilibrium (LD). JUN promoter SNP rs4647009 caused significant downregulation of reporter gene expression, whereas reporter gene expression was significantly upregulated in the presence of the FOS promoter SNPs. The homozygous genotype of FOS promoter SNPs showed an association with the susceptibility for knee-OA (odds ratio (OR) 2.12, 95% confidence interval (CI) 1.2–3.7, p = 0.0086). This association was successfully replicated in the Finnish Health 2000 study cohort (allelic OR 1.72, 95% CI 1.2–2.5, p = 0.006). FOS Promoter variants may represent relevant susceptibility markers for knee-OA.

No description available.

Chronic stress has a cumulative physiological impact, causing dysregulation of multiple systems due to allostatic overload. There is growing evidence that one such system, the microbiota, is engaged in persistent bidirectional interplay with the brain—a phenomenon that influences neural function and behaviour. However, the functional role of the microbiota in stress-associated changes and the underlying pathways of communication are unknown. Using a murine model of depression, we demonstrate that chronic stress has top-down effects on the structure of the microbiota community, reducing its richness and diversity, altering its profile, and causing differential abundance of various bacterial genera. These structural changes have functional consequences, including in metabolic pathways responsible for the synthesis of short chain fatty acids, tryptophan, and tyrosine. Using a physiologically active bacteria, Lactobacillus rhamnosus (JB-1), we probed for bottom-up signalling in chronic stress. JB-1 attenuated deficits in anxiety-like and social behaviours, and induced systemic immunoregulatory effects, independent of affecting stress-induced changes in the microbiota. In examining possible mechanisms of gut-brain brain signalling, we observed that in unstressed mice, a single dose of JB-1 causes rapid expression of c-Fos—a marker of neuronal activation—in distributed areas of the brain within 165 minutes, absent behavioural changes. No such effects were observed with heat-killed JB-1, despite that both live and heat-killed preparations facilitated vagal activity. Sub-diaphragmatic vagotomy prevented neuronal activation in most but not all brain regions, suggesting that vagal signalling is critical but indicating the presence of additional independent pathways. Finally, only chronic JB-1 treatment increased ΔFosB expression in the brain, which is indicative of long-term neuronal adaptations, in association with behavioural changes. These studies demonstrate a role for bidirectional gut-brain signalling in chronic stress, and highlight the signalling pathways and brain regions through which gut bacteria exert their influence on host behaviour. / Thesis / Candidate in Philosophy / Stress, which is a leading risk factor for mental illnesses such as depression, drastically affects the microbiota—the community of intestinal bacteria. However, this influence is bidirectional as gut bacteria can also influence the brain. Thus, we sought to understand the role of the microbiota in the negative effects of stress and how these microorganisms interact with the brain. We observed that behavioural changes in mice after chronic stress were associated with inflammation and community-wide changes in the microbiota. Treatment with a bacterial strain, Lactobacillus rhamnosus (JB-1), attenuated changes in behaviour and inflammation, but had no effect on the microbiota composition. We observed that the brain rapidly responded to JB-1 via the vagus nerve, and that chronic treatment caused long-term changes in brain regions. This work will allow us to discover novel pathways that can be targeted with greater specificity in clinical settings, providing an innovative approach to treatment of psychiatric conditions.

Bifidobacteria are the predominant members of the infant gut, colonize adults to a lesser extent, and are recognized as beneficial microbes. Various bifidobacterial species produce ��-aminobutryic acid (GABA), the chief inhibitory neurotransmitter in the mammalian central nervous system. It is postulated that in order to produce GABA, the bifidobacterial genome must contain the gadB and gadC genes which encode a glutamate decarboxylase and a glutamate/GABA antiporter, respectively. Once exported by GadC, GABA is absorbed and transported systemically throughout the host. We hypothesize that specific dietary oligosaccharides will modulate bifidobacterial production of GABA due to varying intracellular concentrations of glutamate. To test this, 33 bifidobacterial strains were screened for GABA production via reverse phase HPLC. Interestingly, 10 strains contained both gadB and gadC genes, but only 8 strains produced detectable GABA in vitro. To further elucidate the extrinsic factors influencing GABA production, strains were subjected to different dietary components. Specifically, lactose and the dietary oligosaccharide FOS were evaluated for the ability to promote biosynthesis of intracellular glutamate and thus potentially GABA. Understanding the relationship between diet, bifidobacterial physiology, and GABA production may inform dietary interventions to modulate this neurotransmitter in vivo.

O hormônio adrenocorticotrópico, ACTH, regula função (esteroidogênese) e proliferação das células da córtex das glândulas adrenais através de um único receptor específico, ACTHR, que pertence à superfamília GPCR (G-protein coupled receptors). Embora o ACTHR tenha sido clonado há 8 anos, os mecanismos moleculares das ações mitogênica e anti-mitogênica de ACTH permanecem obscuros, cuja elucidação é objeto de estudo deste trabalho. A abordagem experimental consistiu na clonagem do ACTHR de células adrenocorticais Y-1 de camundongo e expressão funcional em fibroblastos 3T3 e células AR-1. Clones transfectantes, expressando estavelmente ACTHR, mostraram-se responsivos a ACTH através de: a) ativação de adenilato ciclase e b) indução de genes das famílias fos e jun. Por outro lado, medidas de síntese de DNA e proliferação celular indicam que ACTH não tem nenhum efeito mitogênico ou anti-mitogênico nos transfectantes ACTHR. O gene c-fos foi usado como alvo para testar vias de transdução de sinal ativadas por ACTH nos transfectantes 3T3 ACTHR. Estes testes mostraram que PKA, PKC e MAPK tem pouca ou nenhuma participação na indução de c-fos por ACTH nos clones 3T3 ACTHR, sugerindo que ACTHR pode ativar vias ainda não identificadas e motivando a busca de novas vias ativadas por ACTH nas células Y-1. Verificou-se que células Y-1 apresentam níveis constitutivamente elevados de AKT/PKB ativada (fosforilada), dependentes de PI3K e SRC, e que ACTH promove rápida desfosforilação de AKT via Gs/Adenilato Ciclase/cAMP/PKA. Ao promover a inativação de AKT, ACTH promove simultaneamente a indução da proteína p27'IND.Kip1', um mecanismo que contribui para a atividade anti-mitogênica de ACTH. / The adrenocorticotropic hormone, ACTH, regulates both function and proliferation of adrenocortical cells binding to a specific receptor, ACTHR, which belongs to superfamily of GPCR (G-protein coupled receptors). ACTHR was cloned a few years ago, but the molecular mechanisms underlying the mitogenic and anti-mitogenic actions of ACTH remain unknown, whose elucidation is aim of this project. The experimental approach was to clone the ACTHR from mouse Y-1 adrenocortical cells and to express the functional receptor in Balb 3T3 fibroblasts and AR-1 cells. Transfectant clones stably expressing the ACTHR respond to ACTH treatments by: a) adenylate cyclase activation and b) induction of fos and jun genes. On the other hand, experiments of DNA synthesis and cellular proliferation showed that ACTH has not mitogenic or anti-mitogenic effects in ACTHR transfectants. c-fos gene was used as a target to test signal transduction pathways activated by ACTH into 3T3 ACTHR transfectants. The results showed that PKA, PKC and MAPK have no relevant contribution on the ACTH c-fos induction in 3T3 ACTHR transfectants suggesting that ACTHR can activate signal transduction pathways still not identified. This conclusion prompted us to search for another signal transduction pathways triggered by ACTHR in Y-1 adrenocortical cells. This search led to the detection of high constitutive levels of activated AKT/PKB in Y-1 adrenocortical cells, which are dependent on PI3K and SRC. ACTH causes a strong and rapid downregulation of activated AKT in a Gs/Adenylate Cyclase/cAMP/PKA dependent manner. Apparently, dephosphorylation of AKT promoted by ACTH releases transcription factors that induce p27'IND.Kip1', a likely mechanism underlying ACTH anti-mitogenic effects in adrenocortical cells.