Functional Analysis of saxophone, the Drosophila Gene Encoding the BMP Type I Receptor Ortholog of Human ALK1/ACVRL1 and ACVR1/ALK2

In metazoans, bone morphogenetic proteins (BMPS) direct a myriad of developmental and adult homeostatic evens through their heterotetrameric type I and type II receptor complexes. We examined 3 existing and 12 newly generated mutations in the Drosophila type I receptor gene, saxophone (sax), the ortholog of the human Activin Receptor-Like. Kinasel and -2 (ALK1/ACVR1 and ALK2/ACVR1) genes. Our genetic analyses identified two distinct classes of sax alleles. The first class consists of homozygous viable gain-of-function (GOF) alleles that exhibit (1) synthetic lethality in combination with mutations in BMP pathway components, and (2) significant maternal effect lethality that can be rescued by an increased dosage of the BMP encoding gene, dpp(+). In contrast, the second class consists of alleles that are recessive lethal and do not exhibit lethality in combination with mutations in other BMP pathway components. The alleles in this second class are clearly loss-of-function (LOF) with both complete and partial loss-of-function mutations represented. We find that one allele in the second class of recessive lethals exhibits dominant-negative behavior, albeit distinct from the GOF activity of the first class of viable alleles. On the basis of the fact that the first class of viable alleles can be reverted to lethality and on our ability to independently generate recessive lethal sat mutations, our analysis demonstrates that sax is an essential gene. Consistent with this conclusion, we find that a normal sax transcript is produced by sax(P), a viable allele previously reported to be mill, and that this allele can be reverted to lethality. Interestingly, we determine that two mutations in the first: class of sax alleles show the same amino acid substitutions as mutations in the human receptors ALK1/ACVR1-1 and ACVR1/ALK2, responsible for cases of hereditary hemorrhagic telangiectasia type 2 (HHT2) and fibrodysplasia ossificans progressiva (FOP), respectively. Finally, the data presented here identify different functional requirements for the Sax receptor, support the proposal that Sax participates in a heteromeric receptor complex, and provide a mechanistic framework for future investigations into disease states that arise from defects in BMP/TGF-beta signaling.

Modell för att prediktera när tertiära glödskalsproblem uppstår

SSAB Tunnplåt har i Borlänge en produktionsanläggning innehållande varmvalsverk, kallvalsverk, normaliseringsanläggning, betsträckor, spaltsträckor, klippsträckor och ytbeläggningslinjer m.m. Genom hela varmvalsningsprocessen bildas oxid på den varma stålytan s.k. glödskal. Under pågående processer avlägsnas glödskalet för att ej påverka slutproduktens ytfinish och materialegenskaper. Fastän ytan spolas ren från glödskal före varje valsning händer det att det finns glödskalsproblem i form av grov yta på de färdiga banden. Anledningen till detta är att det hinner bildas glödskal i början av färdigverket efter att den sista glödskalsrensningen gjorts. Denna glödskalsbildning beror på en rad olika faktorer och troligen kombinationer av olika faktorer. En nyligen installerad ytsyningsanläggning detekterar återbildat (tertiärt) glödskal, detta presenteras som area fraktion av defekter per band. Detta mått används som resultat parameter när man med hjälp av ett verktyg för multivariatdataanalys analyserar ett stort antal valsningsparametrar som kan ha påverkan på det återbildade glödskalet. Analysen utförs med ett verktyg för multivariatdataanalys som heter Simca. Resultatet från analysen skall ses som ett hjälpmedel för att tala om inom vilka gränser valsverket skall köras för att undvika problemen med återbildat glödskal.

Implementera en guide för allmänna transporter med buss på mobiltelefoner med Java 2 Micro Edition

Detta examensarbete går ut på att ta fram en applikation, avsedd för mobiltelefoner, där man på ett enkelt sätt kan få reda på avgångarna för Dalarnas kollektivtrafik. Det stigande bensinpriset och bilåkningens negativa miljöpåverkan ligger som grund till att Etex AB och Balanz AB vill undersöka om det går att få människor att välja bussen framför personbilen. Om man genom denna applikation kan underlätta planeringen av människors dagliga resor, hur påverkas då valet av fortskaffningsmedel?Resultatet blev en applikation där man, via sin mobiltelefon, bland annat kan ta reda på avgångar för utvalda linjer av Dalatrafiks bussar, hantera linjekartor och synkronisera avgångar med telefonens kalender. Alla tidtabeller sparas i telefonen för att minska nätverkstrafiken och för att informationen enkelt ska kunna återanvändas.Dalatrafik är det företag som ansvarar för kollektivtrafiken i Dalarna. Det finns i dagsläget ingen möjlighet att få reda på avgångarna för deras bussar om man inte har tillgång till en tidtabell eller ringer till deras kundservice. Vissa kommuner har redan lösningar för tidtabellshantering i mobiltelefonen och nu vill Etex AB och Balanz AB, som är två företag som bl.a. arbetar med trådlösa applikationer, att även Dalarna ska få ta del av detta praktiska hjälpmedel.De flesta mobiltelefoner som säljs idag har stöd för att köra applikationer, eller MIDlets som de också kallas, gjorda i J2ME (Java 2 Platform, Micro Edition). Denna teknik har tagits fram av Sun för att göra applikationer som kan köras på mindre mobila enheter såsom mobiltelefoner och handdatorer. Kombinerar man J2ME med den ökande prestandan i dagens mobiltelefoner kan avancerade applikationer t.ex. spel med 3D-grafik, webbläsare och andra nyttiga program tillverkas.

Utveckling av System för att Kartlägga Cykelbanor

Cykeln har på senare år fått en allt större roll inom transportpolitiska sammanhang. Det finns ettintresse att öka användningen av cykel, framförallt av miljö- och hälsoskäl. Därmed finns detockså ett intresse för att kartlägga cykelvägar på samma sätt bilvägar är kartlagda. Om Sverigescykelvägar fanns samlade i en databas skulle utveckling och underhåll förenklas. Det skulle ävenvara möjligt att ta fram informationstjänster som t.ex. ruttplanering för cykelvägnätet.Målet med det här arbetet var att utveckla en applikation för att kartlägga cykelbanor.Applikationen skulle kunna köras på en PC under Windows XP. Koordinater skulle hämtas frånen GPS-enhet ansluten till datorn och plottas på en karta. Insamlade koordinater skulle ävenkunna omvandlas till noder och länkar för att beskriva vägnätets utsträckning och logiskakopplingar. Projektet genomfördes i samarbete med WM-data.WM-data tillhandahöll ett antal färdiga moduler för uppritning av karta samt hantering avdatabas och GPS-enhet. Problemet kunde grovt delas i tre sektioner; utveckla användargränssnittför applikationen, integrera tillhandahållna moduler i applikationen och utveckla den internalogiken för hantering av länkar och noder. Uppgifterna löstes mer parallellt än sekventiellt.Resultat blev ett program där de tre huvudfunktionerna från användarperspektivet är starta/stoppakartläggning samt upprätta kopplingspunkt (nod). Den applikation som utvecklades under det härprojektets gång har möjliggjort kartläggning av cykelbanor, och det finns ett flertal alternativ förvidareutveckling.

Germ line Cysts and the Formation of the Germinal Epithelium During the Female Gonadal Morphogenesis in Cyprinus Carpio (Teleostei: Ostariophysi: Cypriniformes)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ultrastructural aspects of the intercellular bridges between female bee germ cells

No ovário das abelhas as células germinativas e as células foliculares são interconectadas por pontes intercelulares mantidas abertas por reforços do citoesqueleto na membrana plasmática. As pontes entre as células germinativas têm comportamento dinâmico e provavelmente atuam na determinação do ovócito entre as células do clone formado pelas mitoses pré meióticas formando posteriormente uma via de transporte para que os produtos sintetizados pelas células nutridoras atinjam o ovócito durante sua maturação. Os elementos do citoesqueleto presentes nas pontes intercelulares das gônadas das abelhas são basicamente microfilamentos e microtúbulos, mas nas pontes entre os cistócitos pré-meióticos outro tipo de filamento (espesso de natureza não definida, associado a elementos do retículo endoplasmático) está presente, atravessando a ponte e prendendo-se através dos microfilamentos à membrana plasmática. Estes filamentos aparentemente controlam o vão da ponte. Terminada a fase de proliferação os cistócitos tomam a forma de uma roseta e um fusoma, formado pela convergência das pontes, aparece no centro desta. Nesta conformação os filamentos grossos não estão presentes. Nova mudança ocorre com a diferenciação do ovócito e das células nutridoras, com a reorientação de todas as pontes de maneira a canalizar o conteúdo das futuras células nutridoras para o ovócito.

Molecular analysis of the Von Hippel-Lindau (VHL) gene in a family with non-syndromic pheochromocytoma: the importance of genetic testing

Dois pacientes índices da família analisada neste estudo foram submetidos a adrenalectomia bilateral devido a feocromocitoma. Foi, então, realizado o estudo genético dos pacientes e de sete parentes de primeiro grau. Os dois pacientes com feocromocitoma e dois outros membros assintomáticos da família apresentaram a mutação c496G>T no exon 3 do gene VHL. A família perdeu seguimento médico. Três anos após a realização da avaliação genética, a irmã dos pacientes, portadora da mutação, foi encaminhada para o nosso serviço após uma gestação complicada por pré-eclampsia. Ela referia paroxismos sugestivos de feocromocitoma, mas as metanefrinas urinárias eram negativas. Entretanto, a tomografia computadorizada de abdômen evidenciou uma massa adrenal que também se contrastou na cintilografia com metaiodobenzilguanidina (MIBG). Esse estudo mostra que a análise molecular do paciente índice pode levar à identificação de parentes assintomáticos portadores da mutação. Além disso, mesmo com as metanefrinas urinárias negativas, a identificação de uma mutação específica levou a um aumento da suspeita e detecção de feocromocitoma na irmã dos afetados pela doença.

The generation and utilization of a cancer-oriented representation of the human transcriptome by using expressed sequence tags

Whereas genome sequencing defines the genetic potential of an organism, transcript sequencing defines the utilization of this potential and links the genome with most areas of biology. To exploit the information within the human genome in the fight against cancer, we have deposited some two million expressed sequence tags (ESTs) from human tumors and their corresponding normal tissues in the public databases. The data currently define approximate to23,500 genes, of which only approximate to1,250 are still represented only by ESTs. Examination of the EST coverage of known cancer-related (CR) genes reveals that <1% do not have corresponding ESTs, indicating that the representation of genes associated with commonly studied tumors is high. The careful recording of the origin of all ESTs we have produced has enabled detailed definition of where the genes they represent are expressed in the human body. More than 100,000 ESTs are available for seven tissues, indicating a surprising variability of gene usage that has led to the discovery of a significant number of genes with restricted expression, and that may thus be therapeutically useful. The ESTs also reveal novel nonsynonymous germline variants (although the one-pass nature of the data necessitates careful validation) and many alternatively spliced transcripts. Although widely exploited by the scientific community, vindicating our totally open source policy, the EST data generated still provide extensive information that remains to be systematically explored, and that may further facilitate progress toward both the understanding and treatment of human cancers.

Canonical P elements are transcriptionally active in the saltans group of Drosophila

Up to now, investigations of expression and regulation of P transposable element have been almost exclusively carried out with the Drosophila melanogaster canonical P element. Analyzing eight species of the saltans group, we detected transposase mRNA in germline tissues of D. saltans and D. prosaltans and repressor mRNA in somatic tissues of D. saltans and D. sturtevanti. Sequencing analysis suggested that these transcripts might belong to the canonical subfamily and that they can be transpositionally active only in D. saltans. dN and dS values of Adh and the P element suggested that the sequences found in D. saltans and D. prosaltans might have been present in the ancestor of the saltans subgroup and that the sequence found in D. sturtevanti might have been horizontally transferred from D. saltans.

Expressão de genes relacionados ao ciclo celular e proteção da mucosa gástrica em metaplasia intestinal e ulcera gástrica em comparação com câncer gástrico

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytogenetic characterization and evaluation of c-MYC gene amplification in PG100, a new Brazilian gastric cancer cell line

Gastric cancer is the fourth most frequent type of cancer and the second cause of cancer mortality worldwide. The genetic alterations described so far for gastric carcinomas include amplifications and mutations of the c-ERBB2, KRAS, MET, TP53, and c-MYC genes. Chromosomal instability described for gastric cancer includes gains and losses of whole chromosomes or parts of them and these events might lead to oncogene overexpression, showing the need for a better understanding of the cytogenetic aspects of this neoplasia. Very few gastric carcinoma cell lines have been isolated. The establishment and characterization of the biological properties of gastric cancer cell lines is a powerful tool to gather information about the evolution of this malignancy, and also to test new therapeutic approaches. The present study characterized cytogenetically PG100, the first commercially available gastric cancer cell line derived from a Brazilian patient who had a gastric adenocarcinoma, using GTG banding and fluorescent in situ hybridization to determine MYC amplification. Twenty metaphases were karyotyped; 19 (95%) of them presented chromosome 8 trisomy, where the MYC gene is located, and 17 (85%) presented a deletion in the 17p region, where the TP53 is located. These are common findings for gastric carcinomas, validating PG100 as an experimental model for this neoplasia. Eighty-six percent of 200 cells analyzed by fluorescent in situ hybridization presented MYC overexpression. Less frequent findings, such as 5p deletions and trisomy 16, open new perspectives for the study of this tumor.

Specific Activation of an I-Like Element in Drosophila Interspecific Hybrids

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genomic screening of testicular germ cell tumors from monozygotic twins

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genomic profile of a Li-Fraumeni-like syndrome patient with a 45,X/46,XX karyotype, presenting neither mutations in TP53 nor clinical stigmata of Turner syndrome

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Predictive models for mutations in mismatch repair genes: implication for genetic counseling in developing countries

Background: Lynch syndrome (LS) is the most common form of inherited predisposition to colorectal cancer (CRC), accounting for 2-5% of all CRC. LS is an autosomal dominant disease characterized by mutations in the mismatch repair genes mutL homolog 1 (MLH1), mutS homolog 2 (MSH2), postmeiotic segregation increased 1 (PMS1), post-meiotic segregation increased 2 (PMS2) and mutS homolog 6 (MSH6). Mutation risk prediction models can be incorporated into clinical practice, facilitating the decision-making process and identifying individuals for molecular investigation. This is extremely important in countries with limited economic resources. This study aims to evaluate sensitivity and specificity of five predictive models for germline mutations in repair genes in a sample of individuals with suspected Lynch syndrome. Methods: Blood samples from 88 patients were analyzed through sequencing MLH1, MSH2 and MSH6 genes. The probability of detecting a mutation was calculated using the PREMM, Barnetson, MMRpro, Wijnen and Myriad models. To evaluate the sensitivity and specificity of the models, receiver operating characteristic curves were constructed. Results: Of the 88 patients included in this analysis, 31 mutations were identified: 16 were found in the MSH2 gene, 15 in the MLH1 gene and no pathogenic mutations were identified in the MSH6 gene. It was observed that the AUC for the PREMM (0.846), Barnetson (0.850), MMRpro (0.821) and Wijnen (0.807) models did not present significant statistical difference. The Myriad model presented lower AUC (0.704) than the four other models evaluated. Considering thresholds of >= 5%, the models sensitivity varied between 1 (Myriad) and 0.87 (Wijnen) and specificity ranged from 0 (Myriad) to 0.38 (Barnetson). Conclusions: The Barnetson, PREMM, MMRpro and Wijnen models present similar AUC. The AUC of the Myriad model is statistically inferior to the four other models.