Doctor of Philosophy (PhD) / The investigation into the relationship between neural plasticity in the rat forebrain associated with opiate-induced behaviours yielded two major results. The major finding of the functional neuroanatomy of acute morphine dependence was that doses of naloxone that induced hyperalgesia following a brief exposure to morphine, in previously drug-naïve rats, caused a specific induction of the inducible transcription factor (itf) proteins c-Fos and zif268 in the extended amygdala. Moreover, doses of naloxone that caused a simple reversal in morphine analgesia failed to induce itf proteins in these same brain regions. This increase in itf proteins was specific to regions of the extended amygdala that receive and process nociceptive information relayed via the spino-parabrachio-amygdaloid pathway and was not observed in other regions that are involved in supraspinal pain modulation such as the rostral ventromedial medulla and the periaqueductal gray. We also found that acute morphine increased c-Fos protein in the basolateral amygdala and the major output nucleus of the central amygdala the medial subdivision. Acute morphine also up-regulated c-Fos protein in striatal, midbrain, and hypothalamic nuclei. A unique finding of the current study was that prolonged exposure to morphine was required to induce c-Fos in these brain regions, as the subsequent administration of naloxone 30-minutes after morphine either reversed or blocked this induction. These results indicate the potential role of the amygdala in analgesia following systemic morphine and in pain facilitation during acute morphine abstinence. Investigation into the neurons and circuitry that undergo long-term neuroplasticity in response to repeated morphine exposure revealed that network-level changes in the distribution of Fos protein in the nucleus accumbens and striatum predicted both tolerance to catalepsy and psychomotor sensitisation. Drug-naïve rats became profoundly cataleptic following morphine, an effect that rats with a drug-history became tolerant. Rats with a history of morphine exposure showed an increase in stereotyped behaviours compared to drug-naïve rats. The major finding of this study was that a shift in the induction of c-Fos protein from a matrix predominance in drug-naïve rats toward a patch predominance in drug-sensitised rats in the accumbens core predicted both tolerance to catalepsy and sensitisation of oral stereotyped behaviours. Acute injection of morphine in a drug-naïve rat induced catalepsy and increased the number of c-Fos-positive neurons in matrix striatopallidal projection neurons of the rostral accumbens core. An increase in activity of striatopallidal projection neurons, which give rise to the indirect pathway, could potentially increase inhibitory drive to the pedunculopontine nucleus (PPN). The PPN, long known as a site of termination for basal ganglia output, is thought to direct the outflow of incentive-motivational and sensorimotor information from the nucleus accumbens to pons, medullary, and spinal cord nuclei translating the incentive impact of the stimuli into appropriate motor, autonomic and emotive responses (Winn et al., 1997). Inhibition of this nucleus would cause the animal to be unable to initiate a movement and in effect lock up, which is precisely what cataleptic postures look like. In contrast c-Fos-positive neurons were decreased in the rostral matrix and increased in patch striatonigral projection neurons along the rostro-caudal extent of the accumbens core when morphine was administered to drug-sensitised rats. Striatonigral neurons located in the patch give rise to the direct pathway innervating the dopaminergic neurons in both substantia pars compacta and the dopamine rich islands in the substantia nigra pars reticulata (Berendse et al., 1992; Gerfen, 1992; Furuta et al., 2002). Activity of this pathway is thought to be involved in the initiation of movement (Gerfen, 1992; Gerfen and Wilson, 1996), however, when this pathway is overstimulated as is the case when morphine is injected in drug-sensitised rats this could potentially cause increased activity of PPN neurons leading to repetitive psychomotor behaviours or stereotypy. This data adds to the growing body of evidence that suggests that long-term neuroadaptations induced by drugs of abuse including morphine that lead to behavioural sensitisation involves the circuitry that includes the nucleus accumbens.

<p>Den vanligaste smittvägen för vårdrelaterade infektioner är genom direkt eller indirekt kontaktsmitta via personalens händer, kläder och tekniska utrustning. Röntgensjuksköterskor möter dagligen ett stort antal patienter och blir därför en potentiell smittväg för mikroorganismer, som skulle kunna spridas vidare på kort tid till patienter om den basala hygienen inte efterföljs. Syftet med detta arbete är att undersöka i vilken omfattning röntgensjuksköterskor utför den basala hygienen vid patientkontakt enligt Socialstyrelsens föreskrifter. Kvantitativ metod med enkät valdes där urvalet bestod av röntgensjuksköterskor som arbetade vid två röntgenavdelningar från två sjukhus i södra Sverige. Resultatet visade att majoriteten av röntgensjuksköterskorna följer Socialstyrelsens föreskrifter inom basal hygien och att röntgensjuksköterskorna upplever att det fanns fler faktorer på arbetsplatserna som främjade men även faktorer som hindrade utförandet av den basala hygienen.</p>

<p>The human skin provides us with an excellent protective barrier and possesses a remarkable ability of constant renewal. Basal cell carcinoma is the most common type of skin cancer. The aim of this project was to verify results from an earlier study investigating the molecular differences between basal cell carcinoma (BCC) and basal cells of normal human epidermis. In that study microdissection of cell populations from BCC and basal cells of normal epidermis respectively was performed in five cases of confirmed BCC. Following RNA extraction and amplification, a gene expression analysis was performed using a 46 k human cDNA microarray. Comparison of expression profiles showed a differential expression of approximately 300 genes in BCC. An upregulation of signaling pathways previously known to be of importance in BCC development could be observed, as well as a downregulation of differentiation markers, MHC class II molecules, and proteins active in scavenging of oxygen radicals. We wanted to confirm these findings for a number of selected genes, using real time PCR. The focal point of this project was microdissection of cells from BCC and subsequent isolation of RNA. Microdissection based methods offer a possibility of selecting well defined cell populations for further analysis by using a focused laser beam. Initially tests in order to optimize the method were also performed, concerning the dehydration process and choice of slides used in microdissection. Isolation of RNA may, as we experienced, be associated with problems due to destruction of RNA by degrading enzymes.</p>

Parkinson’s disease (PD) and dystonia are movement disorders related to dysfunctions of basal ganglia (BG). Deep brain stimulation (DBS) of the subthalamic nucleus (STN) and internal globus pallidus (GPi) are treatments for PD and dystonia. Previous research indicated that abnormally elevated oscillatory activities at the theta (3-10 Hz) beta frequency bands (11-30 Hz) may be related to parkinsonian and dystonic motor symptoms but their precise roles are not well understood. Recently, DBS of the pedunculopontine nucleus region (PPNR) has been used to treat PD patients with postural and gait dysfunctions, but movement-related PPNR activities had not been explored. We aimed to investigate movement-related local field potentials (LFP) recorded from the BG and PPNR in PD and dystonia patients. We recorded STN LFP from PD patients and subsequently applied the intrinsic STN theta, beta, and gamma (31-100 Hz) frequencies through DBS to study their effects on PD motor signs. We also recorded movement-related PPNR LFP in PD patients and movement-related GPi activities in patients with primary dystonia. Finally, we simultaneously recorded movement-related activities from the GPi and the motor thalamus in a patient with secondary dystonia. We found that DBS at the dopamine-dependent and movement-related intrinsic STN gamma frequencies, were as effective as traditionally used high frequencies (130-185 Hz) in reducing PD motor signs, but theta and beta frequencies did not worsen motor symptoms. Voluntary movements modulated two discrete movement-related frequencies in the theta and beta bands in the PPNR and these two frequencies interacted with the sensorimotor and frontal cortices during movements. We showed that voluntary movements modulated beta and gamma frequencies in the GPi. A resting ~5-18 Hz coherence between the GPi bilaterally was attenuated during movements in patients, which may be related to dystonia because this 5-18Hz coherence was also present between the GPi and motor thalamus in the patient with secondary dystonia. Our findings indicated that intrinsic STN gamma frequency oscillations were likely prokinetic rhythms but theta and beta frequencies may not contribute to PD motor symptoms. Voluntary movements modulated theta and beta frequencies in the PPNR, which may explain why PPNR DBS uses lower frequencies than those of the BG. The 5-18 Hz oscillatory activities in the BG-thalamic circuit may be a feature of dystonia.

Lymphocytic infiltration is a feature of basal breast cancer tumors. CXCL10 is a chemokine that was found expressed higher in basal tumor RNA compared to estrogen receptor positive tumor RNA. Both CXCL10 and its receptor CXCR3 were expressed in familial breast cancer tissues, a proportion of which have a basal phenotype. CXCL10 expression was associated with lymphocytic infiltration, and with CXCR3 expression. CXCL10 ligand and receptor were overexpressed individually or together in the human MCF7 cell line. Recombinant human CXCL10 was found to dose dependently decrease cell proliferation. CXCR3 and CXCL10-CXCR3 expressing cells had the potential for increased migration independent of CXCL10 concentration. Co-expression of both genes increased proMMP-2 levels, and conditioned media from one of these clones chemoattracted more of the CXCR3 clones, CXCL10-CXCR3 clones, and CD4+ T-lymphocytes. CXCL10 neutralization suggested that CXCL10 could play a role in this chemoattraction, though it is likely not the only factor involved.

Sox2 is the most frequently amplified oncogene in lung squamous cell carcinoma (SCC). Lung SCC arises in the proximal to central airways and is thought to originate from the p63-positive basal progenitor cells. Since Sox2 amplification occurs early in SCC pathogenesis, we investigated the oncogenic role of Sox2 using normal primary human lung basal progenitor cells. Although Sox2 is highly expressed in normal basal progenitors in a quiescent tracheal epithelium in vivo, we found that Sox2 expression decreases substantially during in vitro proliferation. When Sox2 expression is elevated in the proliferating basal cells in vitro to a level clinically observed in lung SCCs, Sox2 causes hyperplasia and promotes both squamous and Mucin16-positive glandular lineages at the expense of ciliated cell differentiation. Furthermore, our data suggest that the squamous and glandular-differentiating activity of Sox2 is differentially modulated by Receptor tyrosine kinase (RTK) and/or PI3-kinase signaling to promote squamous metaplasia of basal progenitor cells during SCC development.

Lymphocytic infiltration is a feature of basal breast cancer tumors. CXCL10 is a chemokine that was found expressed higher in basal tumor RNA compared to estrogen receptor positive tumor RNA. Both CXCL10 and its receptor CXCR3 were expressed in familial breast cancer tissues, a proportion of which have a basal phenotype. CXCL10 expression was associated with lymphocytic infiltration, and with CXCR3 expression. CXCL10 ligand and receptor were overexpressed individually or together in the human MCF7 cell line. Recombinant human CXCL10 was found to dose dependently decrease cell proliferation. CXCR3 and CXCL10-CXCR3 expressing cells had the potential for increased migration independent of CXCL10 concentration. Co-expression of both genes increased proMMP-2 levels, and conditioned media from one of these clones chemoattracted more of the CXCR3 clones, CXCL10-CXCR3 clones, and CD4+ T-lymphocytes. CXCL10 neutralization suggested that CXCL10 could play a role in this chemoattraction, though it is likely not the only factor involved.

Sox2 is the most frequently amplified oncogene in lung squamous cell carcinoma (SCC). Lung SCC arises in the proximal to central airways and is thought to originate from the p63-positive basal progenitor cells. Since Sox2 amplification occurs early in SCC pathogenesis, we investigated the oncogenic role of Sox2 using normal primary human lung basal progenitor cells. Although Sox2 is highly expressed in normal basal progenitors in a quiescent tracheal epithelium in vivo, we found that Sox2 expression decreases substantially during in vitro proliferation. When Sox2 expression is elevated in the proliferating basal cells in vitro to a level clinically observed in lung SCCs, Sox2 causes hyperplasia and promotes both squamous and Mucin16-positive glandular lineages at the expense of ciliated cell differentiation. Furthermore, our data suggest that the squamous and glandular-differentiating activity of Sox2 is differentially modulated by Receptor tyrosine kinase (RTK) and/or PI3-kinase signaling to promote squamous metaplasia of basal progenitor cells during SCC development.

Parkinson’s disease (PD) and dystonia are movement disorders related to dysfunctions of basal ganglia (BG). Deep brain stimulation (DBS) of the subthalamic nucleus (STN) and internal globus pallidus (GPi) are treatments for PD and dystonia. Previous research indicated that abnormally elevated oscillatory activities at the theta (3-10 Hz) beta frequency bands (11-30 Hz) may be related to parkinsonian and dystonic motor symptoms but their precise roles are not well understood. Recently, DBS of the pedunculopontine nucleus region (PPNR) has been used to treat PD patients with postural and gait dysfunctions, but movement-related PPNR activities had not been explored. We aimed to investigate movement-related local field potentials (LFP) recorded from the BG and PPNR in PD and dystonia patients. We recorded STN LFP from PD patients and subsequently applied the intrinsic STN theta, beta, and gamma (31-100 Hz) frequencies through DBS to study their effects on PD motor signs. We also recorded movement-related PPNR LFP in PD patients and movement-related GPi activities in patients with primary dystonia. Finally, we simultaneously recorded movement-related activities from the GPi and the motor thalamus in a patient with secondary dystonia. We found that DBS at the dopamine-dependent and movement-related intrinsic STN gamma frequencies, were as effective as traditionally used high frequencies (130-185 Hz) in reducing PD motor signs, but theta and beta frequencies did not worsen motor symptoms. Voluntary movements modulated two discrete movement-related frequencies in the theta and beta bands in the PPNR and these two frequencies interacted with the sensorimotor and frontal cortices during movements. We showed that voluntary movements modulated beta and gamma frequencies in the GPi. A resting ~5-18 Hz coherence between the GPi bilaterally was attenuated during movements in patients, which may be related to dystonia because this 5-18Hz coherence was also present between the GPi and motor thalamus in the patient with secondary dystonia. Our findings indicated that intrinsic STN gamma frequency oscillations were likely prokinetic rhythms but theta and beta frequencies may not contribute to PD motor symptoms. Voluntary movements modulated theta and beta frequencies in the PPNR, which may explain why PPNR DBS uses lower frequencies than those of the BG. The 5-18 Hz oscillatory activities in the BG-thalamic circuit may be a feature of dystonia.

Objetivos: El objeto de estudio es la membrana basal glomerular (M.B.G.) y la distribución de dos de las moléculas estructurales de las matrices extracelulares del corpúsculo renal, la fibronectina y la laminina. el estudio se ha realizado con cuatro especies aparentemente próximas de un mismo grupo zoológico (Rodentia) dado que experimentalmente, se incorporan nuevas especies como animales de laboratorio, comparando un animal estándar de laboratorio, con especies silvestres, con distintas estrategias tróficas.Material y métodos: Se han utilizado individuos adultos de Mus musculus (cepa isogénica BALB/c), y Mus spretus, Apodemus sylvaticus y Clethrionomys glareolus precedentes de capturas "in vivo" realizadas en el medio natural. El material histológico ha sido procesado mediante M.O. (tinción del PAS-Azul de Alcián/He. y dos técnicas de localización inmunohistoquímica, inmunoperoxidasa e inmunofluorescencia para anticuerpos anti-fibronectina y anti-laminina) y mediante M.E.T. (estándar y de inmunolocalización de anticuerpos anti-fibronectina y anti-laminina mediante oro coloidal).Resultados: La M.E.T. no muestra diferencias morfológicas entre los glomérulos de las diferentes especies utilizadas, excepto para la M.B.G. ya que en todas las especies silvestres utilizadas, el material que conforma toda la M.B.G. aparece con densidad uniforme. Con la inmunolocalización mediante oro coloidal los resultados para la distribución de fibronectina en M. musculus muestran marcaje positivo en la matriz mesangial, y negativo en la M.B.G. y en la membrana basal peritubular. Los resultados para M. spretus y C. glareolus son semejantes a los obtenidos para M. musculus; si bien con un valor de reacción menor al observado para M. musculus.Con la inmunolocalización mediante oro coloidal los resultados para la distribución de laminina, en M. musculus muestran marcaje positivo en la matriz mesangial, en la membrana basal tubular y en la M.B.G., en ésta última con tendencia a localizarse en las dos láminas raras. Los resultados para M. spretus y C. glareolus son semejantes a los obtenidos para M. musculus aunque con un valor de reacción menor al observado para el ratón de laboratorio.Las muestras procedentes de A. sylvaticus, no mostraron reactividad positiva frente a ninguno de los dos anticuerpos, anti-fibronectina y anti-laminina, utilizados.Conclusiones: La ultraestructura trilaminar de la M.B.G. presente en M. musculus no se observa en las otras especies utilizadas; en éstas existe densidad prácticamente uniforme en todo su espesor: Tampoco existe diferencia en el límite entre la M.B.G. pericapilar, la M.B.G. perimesangial y la matriz mesangial.En las especies M. musculus, M. spretus y C. glareolus la fibronectina se localiza única y exclusivamente en la matriz mesangial y en ningún caso en las M.B.G. mientras que la laminina se localiza tanto en las M.B.G. como en la cápsula de Bowman, la membrana basal tubular y la matriz mesangial. Mostrando en la M.B.G. pericapilar una ligera tendencia a localizarse en las láminas raras externa e interna.El marcaje menos intenso en M. spretus y C. glareolus y su ausencia en A. sylvaticus para la fibronectina y la laminina en la M.B.G. y la matriz mesangial puede indicar un carácter de divergencia filogenética.ENGLISH / DISTRIBUTION OF FIBRONECTIN AND LAMININ IN RENAL CORPUSCLE OF DIVERSE SPECIES OF RODENTSPropose: The aim of this study is the glomerular basement membrane (GBM) and the distribution of two of structural molecules of the extracellular matrices of the renal corpuscle: fibronectin and laminin. The study has been performed with four apparently closed species of a same zoological group (Rodentia) since experimentally, new species like laboratory animals are starting being breeded, comparing a standard laboratory animal with wild species with different trophic strategies.Material and methods: Adults exemplars of Mus musculus (isogenic strain BALB/c), Mus spretus, Apodemus sylvaticus and Clethrionomys glareolus proceeding of captures "in vivo" made in natural fields have been used. The histological material has been processed by means of OM (stain PAS-Alcian Blue/H) and two immunohistochemical techniques, immunoperoxidase and immunofluorescence for antibodies anti-fibronectin and anti-laminin and by TEM (standard and immunolocalization of antibodies anti-fibronectin and anti-laminin by colloidal gold).Results: TEM does not show morphological differences between the glomeruli of the different species used, except for the GBM , showing the material that conforms all the GBM a uniform density in all the wild species. With the immunolocalization by colloidal gold, the results for the distribution of fibronectin in M. musculus is positive in the mesangial matrix, and negative in the GBM and the peritubular basement membrane. The results for M. spretus and C. Glareolus are similar to obtained for M. musculus; although with a smaller reaction to that observed for M. musculus. With the immunolocalization by colloidal gold the results for the distribution of laminin, in M. musculus is positive in the mesangial matrix, the tubular basement membrane and the GBM; the latter with tendency to be located in two laminae rara. The results for M. spretus and C. glareolus are similar to obtained for M. musculus although with a smaller reaction to the observed for the laboratory mouse.The samples from A. sylvaticus, did not show positive reactivity again the anti-fibronectin and anti-laminin antibodies used.Conclusions: The trilaminar ultrastructure of the GBM presents in M. musculus is not observed in the other species used; in those a uniform density in all its thickness exists. Differences in the limit among the pericapillary GBM, the perimesangial GBM and the mesangial matrix does not exist either. In M. musculus, M. spretus and C. glareolus species fibronectin are exclusively located in the mesangial matrix and never in the GBM, whereas laminin is located mainly in the GBM and also in the Bowman's capsule, the tubular basement membrane and the mesangial matrix; showing in the pericapillary GBM a slight tendency to be located in external and internal laminae rara. The less intense labelling in M. spretus and C. glareolus and its absence in A. sylvaticus for fibronectin and laminin in the GBM and the mesangial matrix may suggest a philogenetic divergence.