To unite or disconnect : AmotL2 in tubulogenesis and tumor invasion

For an epithelial- or endothelial cell sheet or ‐tissue to form, individual cells must come together unite and connect. Contacts to adjacent cells and underlying matrix must be initiated, and the junctional proteins mediating the contacts must be further linked to intracellular cytoskeletal networks. By connecting neighboring cells into one unit, contractile actomyosin filaments allow for a closely interlinked, but yet dynamic tissue. Eventhough essential for developmental processes such as organ formation, the exact mechanism of how the cell‐cell junctions connect to the contractile actomyosin network has not yet been completely revealed. In the papers of this thesis, we identify the protein Angiomotin Like 2 (AmotL2 p100) as a linker between the contractile radial actin filaments and VE‐cadherin at the adherens junctions. Furthermore, AmotL2 p100 enables controlled‐ and synchronized morphological alterations of individual cells, which can further result in the creation of new tissue level structures, for example through tubulogenesis. We show the radial AmotL2‐mediated actin filaments to be crucial for force‐generation during morphological transformation and further aortic lumen expansion. Morphological transformations usually require organization and collaboration of several processes, such as formation/disassembly of cell‐cell and cell‐ECM contacts, establishment/disruption of apical-basal polarity and polymerization/disassembly of cytoskeletal filaments. Just like proper regulation of those intra‐ and inter cellular processes and –signals can result in complex structures with diverse morphologies and functionalities, deregulation of the same signals might cause devastating consequences for the function of an organ and the entire organism. In Paper IV of this thesis, we could identify a shorter-, hypoxia regulated AmotL2 p60 isoform. We show the actions of AmotL2 p60 to cause retainment of apical polarity proteins in cytoplasmic vesicles, hence preventing the establishment of apical-basal polarity. Furthermore, we could show AmotL2 p60 to weaken cell‐cell junctions and sensitize cells to growth factor stimuli. The combined actions of AmotL2 p60 cause tumor cell invasion into the surrounding extracellular matrix (ECM). In conclusion, we here provide data showing the two AmotL2 isoforms to possess entirely distinct functions, uniting cells into a multicellular structure, and disconnecting cells during tumor invasion, respectively

The E-cadherin/AmotL2 complex organizes actin filaments required for epithelial hexagonal packing and blastocyst hatching

Epithelial cells connect via cell-cell junctions to form sheets of cells with separate cellular compartments. These cellular connections are essential for the generation of cellular forms and shapes consistent with organ function. Tissue modulation is dependent on the fine-tuning of mechanical forces that are transmitted in part through the actin connection to E-cadherin as well as other components in the adherens junctions. In this report we show that p100 amotL2 forms a complex with E-cadherin that associates with radial actin filaments connecting cells over multiple layers. Genetic inactivation or depletion of amotL2 in epithelial cells in vitro or zebrafish and mouse in vivo, resulted in the loss of contractile actin filaments and perturbed epithelial packing geometry. We further showed that AMOTL2 mRNA and protein was expressed in the trophectoderm of human and mouse blastocysts. Genetic inactivation of amotL2 did not affect cellular differentiation but blocked hatching of the blastocysts from the zona pellucida. These results were mimicked by treatment with the myosin II inhibitor blebbistatin. We propose that the tension generated by the E-cadherin/AmotL2/actin filaments plays a crucial role in developmental processes such as epithelial geometrical packing as well as generation of forces required for blastocyst hatching.Peer reviewe

Genetic and clinical basis for two distinct subtypes of primary Sjögren's syndrome

Objectives Clinical presentation of primary Sjögren’s syndrome (pSS) varies considerably. A shortage of evidence-based objective markers hinders efficient drug development and most clinical trials have failed to reach primary endpoints. Methods We performed a multicentre study to identify patient subgroups based on clinical, immunological and genetic features. Targeted DNA sequencing of 1853 autoimmune-related loci was performed. After quality control, 918 patients with pSS, 1264 controls and 107 045 single nucleotide variants remained for analysis. Replication was performed in 177 patients with pSS and 7672 controls. Results We found strong signals of association with pSS in the HLA region. Principal component analysis of clinical data distinguished two patient subgroups defined by the presence of SSA/SSB antibodies. We observed an unprecedented high risk of pSS for an association in the HLA-DQA1 locus of odds ratio 6.10 (95% CI: 4.93, 7.54, P=2.2×10−62) in the SSA/SSB-positive subgroup, while absent in the antibody negative group. Three independent signals within the MHC were observed. The two most significant variants in MHC class I and II respectively, identified patients with a higher risk of hypergammaglobulinaemia, leukopenia, anaemia, purpura, major salivary gland swelling and lymphadenopathy. Replication confirmed the association with both MHC class I and II signals confined to SSA/SSB antibody positive pSS. Conclusion Two subgroups of patients with pSS with distinct clinical manifestations can be defined by the presence or absence of SSA/SSB antibodies and genetic markers in the HLA locus. These subgroups should be considered in clinical follow-up, drug development and trial outcomes, for the benefit of both subgroups.publishedVersio

RESPONSIBILITIES TO FOLLOW UP AND AFTERCARE WHEN PLACEMENTS BY SOCIALSERVICES ARE TERMINATED

Studiens syfte var att undersöka hur unga vuxna (18–35 år) som tidigare har varit placerade inom familjehem, stödboende eller HVB, har erfarit sin övergång mot vuxenlivet, när placeringen upphört. Studien är kvalitativ och sex semistrukturerade intervjuer utfördes. Tidigare forskning inom området har använts som kunskapsgrund för att stärka studiens tillförlitlighet. Denna studie tyder på liknande resultat som forskningen, att tidigare placerade unga saknade delvis stöd och delaktighet i sin utslussningsprocess och i övergången mot vuxenlivet. Empowerment och KASAM ligger som teoretisk grund i studien.En tematisk analys ledde fram till två centrala teman: Vikten av sociala hållbara relationer och, känsla av maktlöshet och brist på stöd. Hållbara sociala relationer var av vikt utifrån teorierna då kontinuitet och trygga relationer kunde stärka ungdomarnas egenmakt och förmåga att ta egna beslut samt göra övergången begriplig, hanterbar och meningsfull. Brister som identifierades var att en del av ungdomarna uppgav att de blev helt ”släppta” när de lämnade sin placering och inte fick något stöd efter det att de fyllt 18 år, vilket gjorde dem sårbara. Slutsatsen visar att det krävs en förändring i form av skärpta riktlinjer kring eftervården och uppföljningsansvar efter ungdomar har avslutat sin placering. The purpose of the study was to investigate how young adults (18–35 years) that hadpreviously been placed in foster care, supported housing or HVB, have experienced their transition towards adulthood, when the placement has ended. The study is qualitative and six semi-structured interviews were conducted. Previous research has been used to strengthen the study and indicates that previously placed youths lacked support and participation in their aftercare and transition towards adulthood. Empowerment and KASAM form the theoretical basis of the study. A Thematic Analysis led the study to identify two themes: The importance of sustainable social relationships and, Feeling of powerlessness and deficiencies in support. Sustainable social relationships were important as continuity and secure relationships could strengthen young people's empowerment, ability to make their own decisions and make the transition understandable, manageable, and meaningful. Deficienciesthat were identified were that some of the informants were "let go" when they left their placement and did not receive any support after they turned 18, which made them vulnerable. Furthermore, the results indicate that a change is required in the form of stricter guidelines regarding aftercare and follow-up responsibility

Overweight : A sensitivetopic for the school nurse to consider

Bakgrund: Barn med övervikt är något som ökar i Sverige och i världen. Ett barn med övervikt har större risker att senare i livet drabbas av ett flertal sjukdomar, som diabetes och hjärt- och kärlsjukdomar. Det är därför viktigt att sätta in resurser tidigt för att motverka övervikt. Skolsköterskan innehar en position som möjliggör att uppmuntra till goda levnadsvanor och på så vis förebygga övervikt hos barn.  Syfte: Belysa skolsköterskans erfarenhet av arbetet med överviktiga barn i grundskolan. Metod: En kvalitativ intervjustudie med semistrukturerade intervjuer har utförts medsjuksköterskor som har en specialistutbildning som skolsköterska, distriktssköterska eller barnsjuksköterska. Den insamlade datan analyserades med en kvalitativ innehållsanalys. Resultat: Skolsköterskorna beskriver att arbetet med övervikt hos elever samt det preventiva arbetet är utmanande. Övervikt anses vara ett känsligt ämne att prata omoch tidsbristen som skolsköterskan upplever underlättar inte. De lagstadgade hälsobesöken har en central roll för identifiering av övervikt. Resultatet visar att skolsköterskorna önskar mer utbildning, riktlinjer och samarbete både inom och utanför skolan kring övervikt. Konklusion: Resultatet i studien visar på att övervikt är ett känsligt ämne att samtala om med elever och föräldrar. För att kunna vända den ökande trenden behövs mera tid för skolsköterskorna att arbeta preventivt.Background: Overweight among children increases in Sweden and the world. A child with overweight has a higher risk for suffering from a number of diseases later in life, such as diabetes and cardiovascular-diseases. It is therefore important to counteract overweight. The school nurse can encourage good living habits and thus prevent overweight in children.  Aim: Illustrate the school nurse´s experience of working with overweight children in primary school. Method: A qualitative interview study with semi-structured interviews was conducted with nurses who have a specialist education as a school nurse, district nurse or pediatric nurse. The data was analyzed with a qualitative content analysis. Results: The school nurses work with overweight among children and the preventive work is challenging. Overweight is considered a sensitive topic to talk about and the lack of time that the school nurses’ experiences increases this challenge. The statutory health-visits helps in identifying overweight. The school nurses want more education, guidelines and cooperation both within and outside school around obesity.  Conclusion: The results of the study show that overweight is a sensitive topic to talk about with students and parents. To be able to reverse the increasing trend, more time is needed for school nurses to work preventively

Overweight : A sensitivetopic for the school nurse to consider

Bakgrund: Barn med övervikt är något som ökar i Sverige och i världen. Ett barn med övervikt har större risker att senare i livet drabbas av ett flertal sjukdomar, som diabetes och hjärt- och kärlsjukdomar. Det är därför viktigt att sätta in resurser tidigt för att motverka övervikt. Skolsköterskan innehar en position som möjliggör att uppmuntra till goda levnadsvanor och på så vis förebygga övervikt hos barn.  Syfte: Belysa skolsköterskans erfarenhet av arbetet med överviktiga barn i grundskolan. Metod: En kvalitativ intervjustudie med semistrukturerade intervjuer har utförts medsjuksköterskor som har en specialistutbildning som skolsköterska, distriktssköterska eller barnsjuksköterska. Den insamlade datan analyserades med en kvalitativ innehållsanalys. Resultat: Skolsköterskorna beskriver att arbetet med övervikt hos elever samt det preventiva arbetet är utmanande. Övervikt anses vara ett känsligt ämne att prata omoch tidsbristen som skolsköterskan upplever underlättar inte. De lagstadgade hälsobesöken har en central roll för identifiering av övervikt. Resultatet visar att skolsköterskorna önskar mer utbildning, riktlinjer och samarbete både inom och utanför skolan kring övervikt. Konklusion: Resultatet i studien visar på att övervikt är ett känsligt ämne att samtala om med elever och föräldrar. För att kunna vända den ökande trenden behövs mera tid för skolsköterskorna att arbeta preventivt.Background: Overweight among children increases in Sweden and the world. A child with overweight has a higher risk for suffering from a number of diseases later in life, such as diabetes and cardiovascular-diseases. It is therefore important to counteract overweight. The school nurse can encourage good living habits and thus prevent overweight in children.  Aim: Illustrate the school nurse´s experience of working with overweight children in primary school. Method: A qualitative interview study with semi-structured interviews was conducted with nurses who have a specialist education as a school nurse, district nurse or pediatric nurse. The data was analyzed with a qualitative content analysis. Results: The school nurses work with overweight among children and the preventive work is challenging. Overweight is considered a sensitive topic to talk about and the lack of time that the school nurses’ experiences increases this challenge. The statutory health-visits helps in identifying overweight. The school nurses want more education, guidelines and cooperation both within and outside school around obesity.  Conclusion: The results of the study show that overweight is a sensitive topic to talk about with students and parents. To be able to reverse the increasing trend, more time is needed for school nurses to work preventively

RESPONSIBILITIES TO FOLLOW UP AND AFTERCARE WHEN PLACEMENTS BY SOCIALSERVICES ARE TERMINATED

Studiens syfte var att undersöka hur unga vuxna (18–35 år) som tidigare har varit placerade inom familjehem, stödboende eller HVB, har erfarit sin övergång mot vuxenlivet, när placeringen upphört. Studien är kvalitativ och sex semistrukturerade intervjuer utfördes. Tidigare forskning inom området har använts som kunskapsgrund för att stärka studiens tillförlitlighet. Denna studie tyder på liknande resultat som forskningen, att tidigare placerade unga saknade delvis stöd och delaktighet i sin utslussningsprocess och i övergången mot vuxenlivet. Empowerment och KASAM ligger som teoretisk grund i studien.En tematisk analys ledde fram till två centrala teman: Vikten av sociala hållbara relationer och, känsla av maktlöshet och brist på stöd. Hållbara sociala relationer var av vikt utifrån teorierna då kontinuitet och trygga relationer kunde stärka ungdomarnas egenmakt och förmåga att ta egna beslut samt göra övergången begriplig, hanterbar och meningsfull. Brister som identifierades var att en del av ungdomarna uppgav att de blev helt ”släppta” när de lämnade sin placering och inte fick något stöd efter det att de fyllt 18 år, vilket gjorde dem sårbara. Slutsatsen visar att det krävs en förändring i form av skärpta riktlinjer kring eftervården och uppföljningsansvar efter ungdomar har avslutat sin placering. The purpose of the study was to investigate how young adults (18–35 years) that hadpreviously been placed in foster care, supported housing or HVB, have experienced their transition towards adulthood, when the placement has ended. The study is qualitative and six semi-structured interviews were conducted. Previous research has been used to strengthen the study and indicates that previously placed youths lacked support and participation in their aftercare and transition towards adulthood. Empowerment and KASAM form the theoretical basis of the study. A Thematic Analysis led the study to identify two themes: The importance of sustainable social relationships and, Feeling of powerlessness and deficiencies in support. Sustainable social relationships were important as continuity and secure relationships could strengthen young people's empowerment, ability to make their own decisions and make the transition understandable, manageable, and meaningful. Deficienciesthat were identified were that some of the informants were "let go" when they left their placement and did not receive any support after they turned 18, which made them vulnerable. Furthermore, the results indicate that a change is required in the form of stricter guidelines regarding aftercare and follow-up responsibility

HPV transcription in skin tumors.

BackgroundAlthough more than 95% of viral sequences found in skin tumors typically belong to human papillomaviruses (HPVs), HPV transcription has so far not been detected. As current technology allows very deep transcriptome sequencing, we examined skin tumors and precursor lesions for HPV transcription.MethodFresh frozen biopsies from 12 skin specimens (11/12 were positive for HPV DNA) were subjected to total RNA sequencing. The cervical cancer cell line CaSki was included as a positive control for HPV transcription.ResultsHPV RNA was detected and confirmed in 1/12 skin lesions at a median depth of 66 million reads per sample. One specimen was positive for HPV 110 transcripts mapping to E6, E7, E2/E4 and L2 open reading frames, as well as to a spliced E1^E4 transcript.ConclusionIn conclusion, the study revealed that a minority of skin lesions contains HPV transcripts and that HPV DNA detection does not predict HPV transcriptional activity