Untersuchung der Reaktion kultivierter humaner Keratinozyten auf das Binden von Autoantikörpern beim bullösen Pemphigoid. Festgestellt wurde eine erhöhte Freisetzung von t-Pa, sowie eine Umverteilung und Reduktion des Zielantigens unter Calciumeinfluß. / Experiments to the reactions of cultivated keratinocytes after binding of autoantibodies against BP180. We recognizzed elevated levels of t-Pa and an alterated distribution of the target antigen under calcium influence.

Meningokokken sind nach wie vor eine wichtige Ursache für Gehirnhautentzündungen und Sepsen weltweit, vor allem bei Kindern und Jugendlichen. Weil viele Pathomechanismen dieses Erregers bislang noch unvollständig verstanden sind, wurden im Rahmen der vorliegenden Doktorarbeit populationsbiologische und pathogenetische Aspekte von Neisseria meningitidis untersucht. Die Kapsel ist der hauptsächliche Pathogenitätsfaktor von Meningokokken und wichtig für die Besiedelung von neuen Wirten. Isolate von symptomfreien Trägern sind allerdings häufig unbekapselt. Um Ursachen oder Mechanismen für den Verlust der Kapselexpression aufzudecken, wurden insgesamt 166 Isolate der Bayerischen Meningokokkenträgerstudie untersucht. Alle Isolate besaßen sämtliche zur Kapselsynthese notwendigen Gene, exprimierten aber keine Kapsel. Bei 39 Isolaten fanden sich Längenvariationen in homopolymeren Sequenzen (slipped strand mispairing, SSM) in den Genen siaA und siaD. 46 Isolate enthielten Insertionselemente (IS1301, IS1016 und IS1106) in den Genen der Kapselsynthese. Irreversible Mutationen (Deletionen, Insertionen, Basensubstitutionen) wurden bei 47 Isolaten gefunden. Veränderungen der Promotorregion schienen keine Rolle zu spielen. Es wurden bei insgesamt sechs Isolaten zwei nicht-synonyme Mutationen in unmittelbarer Nähe zum putativen aktiven Zentrum der UDP-N- Acetylglukosamin-2-Epimerase entdeckt, die einen Verlust der Kapselsynthese erklären könnten. Insgesamt wurden keine Akkumulationen von Mutationen in defekten Genen gefunden und es gab auch keine Korrelationen zwischen den verschieden Ursachen und bestimmten klonalen Linien. Die erhaltenen Ergebnisse legen nahe, dass die meisten der zur Blockierung der Kapselexpression führenden Ereignisse erst im aktuellen Wirt aufgetreten sind und dass zumindest bei bestimmten klonalen Linien die Verbreitung von der Expression einer Kapsel abhängig ist. Viele pathogene Bakterien nutzen zur Infektion des Menschen die ubiquitär im Körper vorkommende Protease Plasmin. Dazu binden diese Plasmin oder das Proenzym Plasminogen. Auch Meningokokken interagieren mit Plasmin und Plasminogen. Im Rahmen der vorliegenden Arbeit konnten drei Rezeptormoleküle für Plasminogen identifiziert werden. Die drei Proteine Enolase, DnaK und Peroxiredoxin konnten mit verschiedenen Methoden auf der Oberfläche der Erreger nachgewiesen werden. Die Bindung des Plasminogens ist bei Meningokokken ausschließlich über Lysinreste der Rezeptoren vermittelt, die C-terminalen Lysinreste der hier identifizierten Rezeptormoleküle spielen aber, wenn überhaupt, nur eine untergeordnete Rolle. Die Bindung von Plasminogen war durch rekombinante Rezeptorproteine konzentrationsabhängig inhibierbar. Plasminogen konnte von Meningokokken auch aus dem Serum rekrutiert werden. Gebundenes Plasminogen war mit uPA (Urokinase Plasminogen Aktivator) aktivierbar und physiologisch aktiv, was durch die Degradation von Fibrinogen nachgewiesen wurde. Das gebundene Plasmin wurde durch die Bakterien vor der Desaktivierung durch α2- Antiplasmin geschützt. Des Weiteren konnte gezeigt werden, dass Meningokokken auch mit weiteren Faktoren des Fibrinolyse-Systems (uPA) interagieren. Sie rekrutierten uPA an ihre Oberfläche und gebundenes uPA war physiologisch aktiv. Die erhaltenen Ergebnisse bestärken die These, dass Meningokokken die Faktoren des Fibrinolyse-Systems für ihre Pathogenese nutzen. / Meningococci remain to be one of the major causes for meningitis and septicaemia worldwide, especially in children and young adults. Because many of the pathogen’s pathomechanisms still remain unresolved, in the present doctoral thesis population biology and pathogenetic aspects of Neisseria meningitidis were investigated. The capsule is the major virulence-factor of meningococci and important for the dispersal to new hosts. However, isolates extracted from healthy carriers are frequently unencapsulated. To reveal the causes or mechanisms underlying this loss of encapsulation 166 strains of the Bavarian Meningococci Carriage Study were analysed. All these strains possessed all the genes responsible for capsule expression but were acapsulate. Slipped strand mispairing was demonstrated for 39 isolates in the genes siaA and siaD. The insertion elements IS1016, IS1106 and IS1301 were responsible for the loss of encapsulation in other 46 strains and 47 isolates showed irreversible mutations (deletions, insertions and base exchanges) in capsule genes. Sequence alterations in the promoter region seemed not to be responsible for the loss of encapsulation. In close vicinity to the putative active site of the UDP-N-acetylglucosamine-2-epimerase two non-synonymous mutations were detected in altogether six strains. Altogether there was no accumulation of mutations in the uncovered defective genes and no correlation between state of encapsulation and specific clonal lineages could be revealed. The obtained results portray a scenario, were the loss of encapsulation happens in the recent host and where at least some clonal lineages are dependent on the capsule for dissemination. Many bacteria use the protease plasmin for their pathogenesis. For this they recruit plasmin or its precursor plasminogen to their surface. Meningococci were shown to interact with plasminogen, too. In the present thesis three meningococcal receptors for plasminogen were identified. The three receptor proteins enolase, DnaK and peroxiredoxin were shown to be localised extracellularly on the bacterial cell surface by diverse techniques. Binding of plasminogen was demonstrated to occur exclusively to lysine residues of the receptor molecules, but the C-terminal lysine residues of the newly identified receptors were not involved in this process. Recruitment of plasminogen to the bacterial surface could be inhibited by soluble, recombinant receptor proteins in a concentration dependent manner. Furthermore, meningococci were shown to be able to recruit plasminogen from human serum. Receptor bound plasminogen could be activated by uPA (urokinase plasminogen activator) and was physiologically active as demonstrated by degradation of fibrinogen. The bound plasmin was also protected from deactivation by α2-antiplasmin. In addition, Neisseria meningitidis was shown to interact with other components of the fibrinolytic system. The bacteria attached uPA to their surface and the bound uPA was physiologically active. These data provide further evidence for recruitment and usage of factors of the fibrinolytic system in pathogenesis of meningococci.

The urokinase receptor (uPAR) plays a central role in metastatic process. It???s evident uPAR is overexpressed across a variety of tumour cells and leads to the increased aggressiveness and poor prognosis of cancer. Inhibition of uPAR expression can block metastatic potential in many tumours. In addition, besides uPA, there are several other proteins which have been confirmed to interact with uPAR, such as vitronectin and integrins. These interactions also contribute to signal transduction and the functions of uPAR complex. Therefore, downregulation of uPAR expression by targeting uPAR mRNA or protein, or by regulating the uPAR partners would be potential therapeutic strategies for prevention of cancer metastasis. There are two main aspects contained in this thesis. Firstly, three specific DNAzymes targeting uPAR mRNA were designed to downregulate uPAR expression in vitro and their effects to decrease cancer cell invasion studied in a human osteosarcoma cell line Saos-2. The results showed that two of them (Dz483 and Dz720) cleaved uPAR transcript in vitro with high efficacy and specificity and the Dz720 inhibited uPAR protein levels by 55% in Saos-2 cells. Besides, the Dz720 significantly suppressed Saos-2 cell invasion using an in vitro matrigel assay. Secondly, two potential uPAR partners from yeast two-hybrid screening, a heat shock protein MRJ and an anti-apoptosis protein HAX-1, were characterised and their functions binding with uPAR investigated. The interactions were confirmed by co-immunoprecipitation, GST-pull down assay and confocal microscopy in cancer cells. In addition, there was a 50% increase in cell adhesion after transfection with MRJ. This increase in adhesion is dependent on the uPAR/full length MRJ interaction as cells transfected with the mutant construct containing only N-terminal region or C-terminal region of MRJ had no increase in cell adhesion. The observed increase in adhesion to vitronectin by MRJ was also blocked by an anti-uPAR domain I antibody suggesting that the induced adhesion is at least in part contributed by uPAR on the cell surface. Together, the identification of both MRJ and HAX-1 as uPAR interactors provides further insight into the intricate relationship between uPAR and other proteins which may develop potential approaches for cancer therapy.

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Motivation Soluble urokinase plasminogen activator receptor (suPAR) is a known inflammatory marker, which is found in various body fluids. SuPAR reflects the immune and pro–inflammatory status of patients caused by HIV and tuberculosis, amongst others. However, recent studies have shown that suPAR is related to cardiovascular function. The cardiovascular health of the black South African population is a major health concern as this group suffers mostly from hypertension and stroke, leading to an alarming increase in cardiovascular morbidity and mortality. SuPAR may be able to contribute to early detection and prevention of cardiovascular diseases. No studies regarding the associations of suPAR with cardiovascular function have been investigated on black South Africans. Objectives To investigate suPAR as a possible marker of cardiovascular function in African and Caucasian men and women, by determining possible gender and ethnic–specific associations of suPAR with cardiovascular function. Methodology There were 207 African and 314 Caucasian men and women (aged 20–79 yrs.) included in this study. High–sensitivity C–reactive protein, glucose, lipids and creatinine were determined in fasting serum and suPAR was analyzed in plasma samples. Blood pressure was measured using the OMRON apparatus (HEM–747), with a 5–min rest interval between measurements. The Finometer device was used to determine the Windkessel compliance and the carotid dorsalis–pedis pulse wave velocity (PWV) was measured with the Complior (SP acquisition system) on the left side of each subject in the supine position. The means, adjusted means and proportions were compared between the groups by using independent t–tests, analysis of co–variance and the chi–square test, respectively. Associations were investigated between cardiovascular variables and suPAR using single and multiple regression analyses with either pulse wave velocity, systolic blood pressure, diastolic blood pressure or Windkessel compliance as dependent variable. Covariates included were age, body mass index, smoking, alcohol use, physical activity, glucose and high–density lipoprotein cholesterol. Results and conclusion SuPAR levels were significantly higher in Africans (P<0.001) compared to Caucasians. After adjusting for body mass index, suPAR increased significantly with age in all groups, except for African women. Moreover, the suPAR levels of African men and women were significantly higher than the Caucasians within each age quartile. While adjusting for age and body mass index, the cardiovascular profiles of the African and Caucasian men were less favourable compared to women, but suPAR levels were significantly higher in Caucasian women compared to men. In single regression, various measures of cardiovascular function correlated with suPAR in African men and Caucasian men and women. After adjusting for confounders the associations disappeared in Caucasian women, and remained nonsignificant in the African women. However, the association between PWV and suPAR remained significant in African men (B=0.19; P=0.030), while the association of systolic blood pressure (B=0.20; P=0.017), diastolic blood pressure (B=0.17; P=0.020) and Windkessel compliance (B=–0.14; P=0.004) with suPAR remained significant in Caucasian men. In conclusion, Africans presented higher suPAR levels compared to Caucasians, even when stratified by age. Gender specific associations indicated that suPAR was associated with arterial stiffness in African and Caucasian men only, therefore, indicating that suPAR could be a possible biomarker for predicting cardiovascular dysfunction. / Thesis (M.Sc. (Physiology))--North-West University, Potchefstroom Campus, 2011.

Motivation Soluble urokinase plasminogen activator receptor (suPAR) is a known inflammatory marker, which is found in various body fluids. SuPAR reflects the immune and pro–inflammatory status of patients caused by HIV and tuberculosis, amongst others. However, recent studies have shown that suPAR is related to cardiovascular function. The cardiovascular health of the black South African population is a major health concern as this group suffers mostly from hypertension and stroke, leading to an alarming increase in cardiovascular morbidity and mortality. SuPAR may be able to contribute to early detection and prevention of cardiovascular diseases. No studies regarding the associations of suPAR with cardiovascular function have been investigated on black South Africans. Objectives To investigate suPAR as a possible marker of cardiovascular function in African and Caucasian men and women, by determining possible gender and ethnic–specific associations of suPAR with cardiovascular function. Methodology There were 207 African and 314 Caucasian men and women (aged 20–79 yrs.) included in this study. High–sensitivity C–reactive protein, glucose, lipids and creatinine were determined in fasting serum and suPAR was analyzed in plasma samples. Blood pressure was measured using the OMRON apparatus (HEM–747), with a 5–min rest interval between measurements. The Finometer device was used to determine the Windkessel compliance and the carotid dorsalis–pedis pulse wave velocity (PWV) was measured with the Complior (SP acquisition system) on the left side of each subject in the supine position. The means, adjusted means and proportions were compared between the groups by using independent t–tests, analysis of co–variance and the chi–square test, respectively. Associations were investigated between cardiovascular variables and suPAR using single and multiple regression analyses with either pulse wave velocity, systolic blood pressure, diastolic blood pressure or Windkessel compliance as dependent variable. Covariates included were age, body mass index, smoking, alcohol use, physical activity, glucose and high–density lipoprotein cholesterol. Results and conclusion SuPAR levels were significantly higher in Africans (P<0.001) compared to Caucasians. After adjusting for body mass index, suPAR increased significantly with age in all groups, except for African women. Moreover, the suPAR levels of African men and women were significantly higher than the Caucasians within each age quartile. While adjusting for age and body mass index, the cardiovascular profiles of the African and Caucasian men were less favourable compared to women, but suPAR levels were significantly higher in Caucasian women compared to men. In single regression, various measures of cardiovascular function correlated with suPAR in African men and Caucasian men and women. After adjusting for confounders the associations disappeared in Caucasian women, and remained nonsignificant in the African women. However, the association between PWV and suPAR remained significant in African men (B=0.19; P=0.030), while the association of systolic blood pressure (B=0.20; P=0.017), diastolic blood pressure (B=0.17; P=0.020) and Windkessel compliance (B=–0.14; P=0.004) with suPAR remained significant in Caucasian men. In conclusion, Africans presented higher suPAR levels compared to Caucasians, even when stratified by age. Gender specific associations indicated that suPAR was associated with arterial stiffness in African and Caucasian men only, therefore, indicating that suPAR could be a possible biomarker for predicting cardiovascular dysfunction. / Thesis (M.Sc. (Physiology))--North-West University, Potchefstroom Campus, 2011.

The urokinase receptor (uPAR) plays a central role in metastatic process. It???s evident uPAR is overexpressed across a variety of tumour cells and leads to the increased aggressiveness and poor prognosis of cancer. Inhibition of uPAR expression can block metastatic potential in many tumours. In addition, besides uPA, there are several other proteins which have been confirmed to interact with uPAR, such as vitronectin and integrins. These interactions also contribute to signal transduction and the functions of uPAR complex. Therefore, downregulation of uPAR expression by targeting uPAR mRNA or protein, or by regulating the uPAR partners would be potential therapeutic strategies for prevention of cancer metastasis. There are two main aspects contained in this thesis. Firstly, three specific DNAzymes targeting uPAR mRNA were designed to downregulate uPAR expression in vitro and their effects to decrease cancer cell invasion studied in a human osteosarcoma cell line Saos-2. The results showed that two of them (Dz483 and Dz720) cleaved uPAR transcript in vitro with high efficacy and specificity and the Dz720 inhibited uPAR protein levels by 55% in Saos-2 cells. Besides, the Dz720 significantly suppressed Saos-2 cell invasion using an in vitro matrigel assay. Secondly, two potential uPAR partners from yeast two-hybrid screening, a heat shock protein MRJ and an anti-apoptosis protein HAX-1, were characterised and their functions binding with uPAR investigated. The interactions were confirmed by co-immunoprecipitation, GST-pull down assay and confocal microscopy in cancer cells. In addition, there was a 50% increase in cell adhesion after transfection with MRJ. This increase in adhesion is dependent on the uPAR/full length MRJ interaction as cells transfected with the mutant construct containing only N-terminal region or C-terminal region of MRJ had no increase in cell adhesion. The observed increase in adhesion to vitronectin by MRJ was also blocked by an anti-uPAR domain I antibody suggesting that the induced adhesion is at least in part contributed by uPAR on the cell surface. Together, the identification of both MRJ and HAX-1 as uPAR interactors provides further insight into the intricate relationship between uPAR and other proteins which may develop potential approaches for cancer therapy.

Syfte: Att utreda sambandet mellan den lösliga formen av urokinas-receptorn (suPAR) i saliv hos patienter med artrit i käkleden (A-TMJ) och friska kontroller, för att skapa en grund för vidare forskning av suPAR som prediktor för inflammationsgraden i käkleden hos patienter med A-TMJ.Material och metod: En fall-kontrollstudie utfördes med 6 kontroller (medelåldern 31±11år) och 5 patienter med A-TMJ (medelåldern 24±5år). Undersökningen bestod av salivprov, registrering av blödning vid sondering (BoP), blodprovstagning, och undersökning av tuggsystemet där antalet smärtsamma käkledsrörelser (PM) mättes. Sist samlades käkledvätska in. Halten suPAR analyserades i saliv, plasma och käkledsvätska. Resultat: En signifikant skillnad mellan suPAR i saliv kunde ej påvisas (A-TMJ 4,4±3,91ng/ml, kontroller 4,96±4,80ng/ml), emellertid hade patienter en signifikant högre halt av suPAR i plasma (A-TMJ 2,71±0,62ng/ml, kontroller: 1,86±0,35ng/ml, P=0,017). Halten av suPAR i käkledsvätska mättes till 1,57±1,50ng/ml hos patienter men kunde inte detekteras hos kontroller. BoP mättes till 16±9% hos patienter och 14±7% hos kontroller, och median(IQR) för PM var 3(1) i höger käkled och 0(3) i vänster käkled hos patienter. Slutsatser: (i) Ingen slutsats kan dras gällande sambandet mellan suPAR i saliv och A-TMJ, men (ii) patienter med A-TMJ har till viss mån en högre smärta i käkleden vid käkledsrörelse medan deras koncentration av suPAR i plasma är högre jämfört med friska kontroller. Det verkar som (iii) BoP skulle kunna vara kopplat till suPAR i saliv. Resultat från denna studie bör tolkas med försiktighet på grund av litet stickprov, fortsatt forskning behövs för att klargöra sambandet mellan suPAR i saliv och A-TMJ. / Aims: To investigate the levels of soluble urokinase plasminogen activator receptor (suPAR) in saliva between patients with arthritis in the temporomandibular joint (A-TMJ) and healthy controls to create a foundation for further research of the potential predictive value of suPAR in patients with A-TMJ.Materials and method: A case- control study was conducted, 6 controls (mean age 31±11years) and 5 patients with A-TMJ (mean age 24±5years) enrolled in the study. Saliva, blood, synovial fluid (SF) were sampled, and the masticatory system was examined according to DC/TMD, and bleeding on probing (BoP) was assessed, as was painful mandibular movement (PM). The level of suPAR was analyzed in saliva, plasma and SF.Results: Level of salivary suPAR did not differ significantly between A-TMJ patients and healthy controls (P > 0.05). Patients had a significantly higher level of suPAR in plasma than controls (A-TMJ 2.71±0.62ng/mL, controls: 1.86±0.35ng/mL, P=0.017). suPAR level in SF was measured to 1.57±1.50ng/mL in A-TMJ patients and not detected in controls. BoP was 16±9% in patients and 14±7% in controls, and median(IQR) of PM was 3(1) in the right TMJ and 0(3) in the left in patients.Conclusions: (i) No conclusion can be drawn regarding suPAR in saliva and A-TMJ, but (ii) to some degree A-TMJ patients have higher PM meanwhile their plasma concentration of suPAR is higher than controls. A trend that (iii) higher BoP is connected with higher suPAR in saliva could be distinguished. Results must be interpreted with caution due to small study sample, more research is required to further elucidate the association between suPAR in saliva and A-TMJ.

Quebec platelet disorder (QPD) is a bleeding disorder characterized by a gain of function defect in fibrinolysis. The hallmark feature of QPD is the marked overexpression of urokinase plasminogen activator (uPA) in megakaryocytes (MK) and platelets. The genetic cause of QPD is a tandem duplication of a ~78 kb region that encompasses the uPA gene, PLAU. As the mechanism of PLAU overexpression is unknown, gene regulatory mechanisms specifically epigenetics were evaluated at the PLAU locus in QPD MK and granulocytes, a QPD unaffected lineage. The aims of the thesis were to assess if QPD is associated with 1) genome wide methylation changes of promoter CpG islands, particularly at PLAU and 2) genome wide changes of active histone modifications H3K27Ac, H3K36me3 and H3K4me2, particularly at the region of PLAU duplication. Methylation and active histone enrichment analysis revealed that in QPD and control subjects, PLAU promoter CpG island was characterized by unaltered hypo-methylation and changes in active histone peak enrichments that were within the realm of having one extra copy of PLAU in both MK and granulocytes. The findings imply that the PLAU CNV mutation does not induce altered promoter methylation status and/or significantly alter active histone markers as the reason for the marked PLAU overexpression in QPD MK. Instead, the rearrangement of an active enhancer element, particularly an H3K27Ac enhancer expressed in MK but not granulocytes, that is upstream of the second copy of PLAU might underlie the marked PLAU expression by differentiated QPD MK. The thesis provides novel insights into the epigenetic regulation of PLAU that will be crucial to identifying the mechanism underlying the aberrant PLAU expression in QPD. / Thesis / Master of Science (MSc)

Le cancer du sein est la principale cause de mortalité par cancer chez la femme. Deux des facteurs de transcription de la famille Fos, Fra-1 et Fra-2, sont surexprimés dans les cancers du sein agressifs et contribuent au phénotype tumoral en favorisant entre autres, la prolifération, la motilité et l'invasivité. De façon surprenante, les mécanismes moléculaires via lesquels Fra-1 et Fra-2 (et plus généralement le complexe transcriptionnel AP-1 dont ils sont des constituants) gouvernent la transcription de leurs gènes cibles sont quasi-inconnus. Dans ce contexte, en combinant diverses approches (immunoprécipitation de chromatine, interférence à l'ARN…), j'ai étudié les mécanismes moléculaires par lesquels Fra-1 et Fra-2 contrôlent la transcription dérégulée du gène de l'urokinase ou uPA (sérine protéase cruciale dans la progression tumorale et l'établissement de métastases) qui est l'un des nouveaux marqueurs utilisés en clinique pour la mise en place des choix thérapeutiques. Mes travaux montrent de façon originale que (i) Fra-1 et Fra-2 agissent de façon non redondante et coopèrent pour réguler l'expression d'uPA via leur fixation sur un enhancer AP-1 localisé à -1,9 kb du site d'initiation de la transcription (TSS), (ii) Fra-2 est nécessaire au recrutement de RNA Pol II au niveau de l'enhancer, tandis que Fra-1 stimule le passage de RNA Pol II de sa forme initiatrice à sa forme élongatrice et (iii) que la polymérase recrutée à l'enhancer rejoint le TSS par un mécanisme de « tracking », très rarement décrit dans la littérature, en produisant de petits ARNs non codants, bidirectionnels et instables. / Breast cancer is the most frequent malignant disease among women. Two transcription factors, Fra-1 and Fra-2, belonging to the Fos family members, are overexpressed in aggressive breast cancers and contribute to the tumorigenic phenotype by favoring proliferation, motility and invasion. Surprisingly, the molecular mechanisms governed by Fra-1 and Fra-2 (and more generally by the AP-1 transcriptional complex, which they are components of) for the transcription of their target genes are still largely unknown. In this context, by combining different approaches (chromatin immunoprecipitation, RNA interference…), I studied the molecular mechanisms orchestrated by Fra-1 and Fra-2 for the expression of the urokinase (or uPA) gene (encoding a serine protease crucial for tumor progression and metastasis), which is one of the new diagnostic markers now taken into consideration for deciding therapeutic strategies. Interestingly, my results show that (i) Fra-1 and Fra-2 have non redundant functions and cooperate for the transcriptional regulation of uPA through their binding to AP-1 enhancer located 1.9 kb upstream of the transcriptional start site (TSS), (ii) Fra-2 is required for the recruitment of RNA Pol II on this enhancer while Fra-1 allows the conversion of RNA Pol II initiating form into its elongating form and (iii) enhancer-recruited RNA Pol II reaches the TSS by a tracking mechanism, mechanism very rarely described in the literature, during which it synthetizes small, unstable bidirectional, non coding RNAs.