Alexandru Constantinescu - Academia.edu (original) (raw)

Papers by Alexandru Constantinescu

Research paper thumbnail of Versality in toric geometry

Research paper thumbnail of Polyhedra, lattice structures, and extensions of semigroups

Journal of the London Mathematical Society, Apr 15, 2020

For an arbitrary rational polyhedron we consider its decompositions into Minkowski summands and, ... more For an arbitrary rational polyhedron we consider its decompositions into Minkowski summands and, dual to this, the free extensions of the associated pair of semigroups. Being free for the pair of semigroups is equivalent to flatness for the corresponding algebras. Our main result is phrased in this dual setup: the category of free extensions always contains an initial object, which we describe explicitly. These objects seem to be related to unique liftings in log geometry. Further motivation comes from the deformation theory of the associated toric singularity. CONTENTS 1. Introduction 1 2. Free pairs of semigroups 3 3. Extending free pairs 7 4. The cone setup 13 5. The discrete setup 17 6. Lifting the η to T * 23 7. The universal co-Cartesian extension 28 8. The initial object property 31 9. Minkowski decompositions revisited 43 References 50 2. FREE PAIRS OF SEMIGROUPS 2.1. Relative boundaries of semigroups within two different setups. Let T ⊆ S be two commutative and cancellative (a + c = b + c ⇒ a = b) semigroups with identity (0 + a = a + 0 = a), satisfying S ∩ (−S) = {0}, i.e. S (and hence T) is pointed. This situation gives rise to the following notion of a relative boundary.

Research paper thumbnail of Krull Dimension and Other Properties of Graph-Related Algebra

The algebra of basic covers of a graph G, denoted byĀ(G), was introduced by Jürgen Herzog as a su... more The algebra of basic covers of a graph G, denoted byĀ(G), was introduced by Jürgen Herzog as a suitable quotient of the vertex cover algebra. In this paper we show that if the graph is bipartite thenĀ(G) is a homogeneous algebra with straightening laws and thus is Koszul. Furthermore, we compute the Krull dimension ofĀ(G) in terms of the combinatorics of G. As a consequence we get new upper bounds on the arithmetical rank of monomial ideals of pure codimension 2. Finally, we characterize the Cohen-Macaulay property and the Castelnuovo-Mumford regularity of the edge ideal of a certain class of graphs.

Research paper thumbnail of Integrating Adaptronics in Intelligent Measurement Technique and in Mechatronics and Integronics Systems for Integrated Control

Proceedings of the 23rd International DAAAM Symposium 2012, 2012

The paper presents hard and software architecture of adaptonics in intelligent measurement engine... more The paper presents hard and software architecture of adaptonics in intelligent measurement engineering and in mechatronics and integronics systems for integrated automatic control, through cooperation between several scientific and technical disciplines, to meet growing and changing demands about modern systems and new products simultaneously improved with the lowest development time and with a higher flexibility and more efficient.

Research paper thumbnail of Adaptronics – New Concept for the Future of Advanced Engineering and Intelligent Automatised Manufacturing

Proceedings of the 23rd International DAAAM Symposium 2012, 2012

The paper treats new concept of adaptronics for the future of advanced engineering and intelligen... more The paper treats new concept of adaptronics for the future of advanced engineering and intelligent automation, by bringing news and discoveries in the field of basic and applied science of mechatronics and integronics and as development vector of know-how in high-tech advanced domain by adapting scientific, complex, structural, functional and decisional of structures, products and systems, to change the operating mode of environment requirements, to control in active mode the vibration, the noise and the deformation, to recover energy from mechanics and inertial vibrating systems and to influence and monitories all structures of any kind.

Research paper thumbnail of Renal Survival in Children with Glomerulonephritis with Crescents: A Pediatric Nephrology Research Consortium Cohort Study

Journal of Clinical Medicine, 2020

There is no evidence-based definition for diagnosing crescentic glomerulonephritis. The prognosti... more There is no evidence-based definition for diagnosing crescentic glomerulonephritis. The prognostic implications of crescentic lesions on kidney biopsy have not been quantified. Our objective was to determine risk factors for end-stage kidney disease (ESKD) in patients with glomerulonephritis and crescents on kidney biopsy. A query of the Pediatric Nephrology Research Consortium’s Pediatric Glomerulonephritis with Crescents registry identified 305 patients from 15 centers. A retrospective cohort study was performed with ESKD as the primary outcome. Median age at biopsy was 11 years (range 1–21). The percentage of crescents was 3–100% (median 20%). Etiologies included IgA nephropathy (23%), lupus (21%), IgA vasculitis (19%) and ANCA-associated GN (13%), post-infectious GN (5%), and anti-glomerular basement membrane disease (3%). The prevalence of ESKD was 12% at one year and 16% at last follow-up (median = 3 years, range 1–11). Median time to ESKD was 100 days. Risk factors for ESKD i...

Research paper thumbnail of EUS-Guided Transgastric Drainage of Intraabdominal Fluid Collections

Chirurgia, 2018

Rezultate: Vârsta medie a subiecţilor incluşi în studiu a fost 51 de ani. Diametrul mediu al cole... more Rezultate: Vârsta medie a subiecţilor incluşi în studiu a fost 51 de ani. Diametrul mediu al colecţiilor intraabdominale a fost de 109 mm (34-250 mm), acesta corelându-se cu metoda de tratament aleasă (p<0.005): drenaj ghidat eco-endoscopic, drenaj percutan, intervenţie chirurgicală, ca monoterapie sau combinaţii între acestea. Mortalitatea peri-procedurală a fost de 9,3%, asociată în principal cu severitatea patologiei ori cu asocierea chirurgiei în tratament. Concluzii: În conluzie, prin studiul efectuat putem afirma că ecoendoscopia poate fi metoda de primă intenţie în a drena colecţiile intra abdominale perigastrice, pentru că reprezintă o metodă sigură şi eficientă, cu un success tehnic de 100% în timp ce succesul clinic de peste 80% asigură o mai bună calitate a vieţii. Pentru colecţiile ce au un diametru mai mare de 127 mm, ne putem aştepta totuşi la necesitate unui tratament combinat, ecoendoscopic şi chirurgical. Cuvinte cheie: colecţii intraabdominale, drenaj transgastric, percutan, chirurgie, ecoendoscopie

Research paper thumbnail of Efficacy of Endobiliary Radiofrequency Ablation using a Novel Endoluminal Radiofrequency Ablation Catheter in a Swine Model

Journal of Investigative Surgery, 2018

Purpose: Endobiliary radiofrequency ablation (RFA) is a new endoscopic ablative technique. Howeve... more Purpose: Endobiliary radiofrequency ablation (RFA) is a new endoscopic ablative technique. However, the ideal power setting for RFA has not yet been clarified. Therefore, we intended to evaluate the effects of endobiliary RFA according to time variations using novel RFA. Materials and methods: Nine female pigs were divided into three groups according to ablation time (60, 90, and 120 seconds) with the same setting (10 watts, 80 C). All pigs underwent endoscopic retrograde cholangiography (ERC) and endobiliary RFA in the common bile duct. Gross and histologic examinations were performed after 24 hours. Results: The ERC and application of the endobiliary RFA were 100% successful, and the post-RFA cholangiogram did not show contrast leakage. The median depth of microscopic ablation was significantly different among the three groups (60 vs. 90 vs. 120 seconds ¼ 1.90 (1.17-2.23) vs. 2.44 (2.31-2.60) vs. 2.52 (2.47-2.64) mm, p ¼ 0.018). There was also a linear relationship between ablation time and microscopic ablation depth (r 2 ¼ 0.552, p ¼ 0.002). However, no significant differences in macroscopic or microscopic ablation length were observed. In addition, there were focal ablation injuries in adjacent liver tissue in five of the nine pigs (2/3 in 60, 1/3 in 90, and 2/3 in 120 seconds). Conclusion: Endobiliary RFA using a novel RFA catheter resulted in controlled ablation with a linear relationship between microscopic ablation depth and ablation time in a swine model. Clinical studies are needed to validate the safe energy condition of endobiliary RFA in malignant biliary obstruction.

Research paper thumbnail of Predicting First-Year Relapses in Children With Nephrotic Syndrome

Pediatrics, 2000

Objective. More than half of the children diagnosed with nephrotic syndrome will have relapses. T... more Objective. More than half of the children diagnosed with nephrotic syndrome will have relapses. These can be infrequent relapses (IRs: <2 in 6 months or <3 in a year) or frequent relaspses (FRs: >2 in 6 months or >3 in a year). Patients who relapse while on alternate day steroids or within 1 month of discontinuation of steroid therapy are considered steroid-dependent (SD; J Pediatr. 1982; 101:514–518). Patients with an IR course have a better long-term prognosis, and many of them have minimal-change disease without mesangial hypercellularity or sclerosis. The purpose of our study was to identify factors at initial presentation that could predict the relapse pattern in the first year after diagnosis, without taking into consideration the histopathology found on renal biopsy. Design. We analyzed the medical records of children who were seen by us before March 1997 and followed for at least 1 year. Variables selected in the study were age, sex, race, presence or absence of ...

Research paper thumbnail of Abiotic Synthesis of Phosphoric Esters of Monosaccharides According to the “Cold Model”

Springer Netherlands eBooks, 1981

It is investigated, according to the “cold model”, the abiotic synthesis of some phosphoric ester... more It is investigated, according to the “cold model”, the abiotic synthesis of some phosphoric esters of monosaccharides by cold plasma decomposition of methane- water mixtures in the presence of apatite.This high frequence electric discharge induced the formation of a complex reaction product, containing alcohols, aldehydes, ketones and acids.

Research paper thumbnail of Expression of Mammalian Geranylgeranyltransferase Type-II in Escherichia coli and Its Application for in Vitro Prenylation of Rab Proteins

Protein Expression and Purification, 2001

protein trafficking in eukaryotic cells. In mammalian Mammalian geranylgeranyltransferase type II... more protein trafficking in eukaryotic cells. In mammalian Mammalian geranylgeranyltransferase type II cells more than 40 Rabs have been cloned and this (GGTase-II) is a 100-kDa heterodimer that catalyzes the number is increasing (1, 2). Individual Rab proteins transfer of two 20-carbon geranylgeranyl groups from have been implicated as essential mediators of vesicle geranylgeranyl pyrophosphate onto C-terminal cystargeting and fusion events (3, 4). For their function, teine residues of Rab GTPases. This modification is Rab proteins require modification by geranylgeranyl essential for the biological activity of Rab proteins. isoprenoids which allows them to reversibly associate Geranylgeranylation can be performed in vitro using with membrane in a tightly controlled fashion. Covalent recombinant GGTase-II but so far large-scale producattachment of geranylgeranyl groups to two C-terminal tion of the enzyme was challenging. We report here the cysteines is catalyzed by geranylgeranyltransferase design of a two plasmid expression system that will type II (GGTase-II). GGTase-II is a heterodimer comproduce GGTase-II at levels as high as 15 mg/L in Escheposed of tightly associated 68-kDa ␣ and 45-kDa ␤ subrichia coli. The protein was produced as a heterodimer units and belongs to the family of protein prenyltransfwith the ␣ subunit bearing a cleavable tandem 6Hiserases together with farnesyl transferase (FTase) and glutathione S-transferase (GST) tag that was used for geranylgeranyl transferase I (GGTase-I) (for review 5). two-step purification of the enzyme. Purified enzyme In contrast to other known prenyltransferases, was functionally active as determined by in vitro pre-GGTase-II does not recognize its protein substrate dinylation and phosphoisoprenoid binding assay. Furrectly, but functions in a concert with a protein chaperthermore, the GST-tagged GGTase-II was used for one termed REP (Rab escort protein) (6). preparative in vitro prenylation of the Rab7:REP-1 Posttranslational geranylgeranylation is crucial for complex. Using this procedure, 10 mg of doubly prenylated Rab7:REP-1 complex were obtained. ᭧ 2001 Aca-biological activity of Rab proteins since it ensures not demic Press only membrane association but also modulates the interaction of Rab proteins with their effectors and regulatory molecules. Traditionally, prenylated Rab proteins were either purified from tissue sources or from The low molecular mass GTP 2-binding proteins encoded by the Rab gene family play important roles in

Research paper thumbnail of cAMP-dependent Protein Kinase Type I Regulates Ethanol-induced cAMP Response Element-mediated Gene Expression via Activation of CREB-binding Protein and Inhibition of MAPK

Journal of Biological Chemistry, 2004

Research paper thumbnail of cAMP-dependent Protein Kinase Types I and II Differentially Regulate cAMP Response Element-mediated Gene Expression

Journal of Biological Chemistry, 2002

We have shown that ethanol induces translocation of cAMP-dependent protein kinase (PKA) to the nu... more We have shown that ethanol induces translocation of cAMP-dependent protein kinase (PKA) to the nucleus, cAMP response element-binding protein (CREB) phosphorylation, and cAMP response element-mediated gene transcription in NG108-15 cells. However, little is known about which PKA types regulate this process. We show here that under basal conditions NG108-15 cells contain type I PKA (C␤RI␤) primarily in cytosol and type II PKA (C␣RII␤) in the particulate and nuclear fractions. Antagonists of both type I and type II PKA inhibit forskolin-and ethanol-induced cAMP response element-mediated gene transcription. However, only the type II PKA antagonist inhibits forskolin-induced C␣ and ethanol-induced C␣ and RII␤ translocation to the nucleus and CREB phosphorylation; the type I antagonist is without effect. Our data suggest that forskolin-and ethanol-induced CREB phosphorylation and gene activation are differentially mediated by the two types of PKA. We propose that type II PKA is translocated and activated in the nucleus and induces CREB phosphorylation that is necessary but not sufficient for gene transcription. By contrast, type I PKA is activated in the cytoplasm, turning on a downstream pathway that activates other transcription cofactors that interact with phosphorylated CREB to induce gene transcription. cAMP signaling is a multiple component system that requires production of cAMP and induces activation of PKA, 1 phosphorylation of CREB, and cAMP-dependent gene expression. This pathway has been implicated in learning and memory (1, 2) as well as addictive behaviors (3) and responses to ethanol (4). Ethanol activates the cAMP pathway to its full extent by stimulating adenylyl cyclase activity (4), C␣ translocation to the nucleus (5), CREB phosphorylation (6), and CREmediated gene transcription (7).

Research paper thumbnail of Mechanochemical reactions of some polyurethane elastomers. 3. Prediction of long-term uniaxial tensile creep using linear compliance as a basis

Colloid & Polymer Science, 1986

Based on isochronous tests the stress-strain curve for thermoplastic polyurethane elastomers (Mol... more Based on isochronous tests the stress-strain curve for thermoplastic polyurethane elastomers (Moldotan D) is plotted and used for correcting the tensile creep data.

Research paper thumbnail of Versality in toric geometry

Research paper thumbnail of Polyhedra, lattice structures, and extensions of semigroups

Journal of the London Mathematical Society, Apr 15, 2020

For an arbitrary rational polyhedron we consider its decompositions into Minkowski summands and, ... more For an arbitrary rational polyhedron we consider its decompositions into Minkowski summands and, dual to this, the free extensions of the associated pair of semigroups. Being free for the pair of semigroups is equivalent to flatness for the corresponding algebras. Our main result is phrased in this dual setup: the category of free extensions always contains an initial object, which we describe explicitly. These objects seem to be related to unique liftings in log geometry. Further motivation comes from the deformation theory of the associated toric singularity. CONTENTS 1. Introduction 1 2. Free pairs of semigroups 3 3. Extending free pairs 7 4. The cone setup 13 5. The discrete setup 17 6. Lifting the η to T * 23 7. The universal co-Cartesian extension 28 8. The initial object property 31 9. Minkowski decompositions revisited 43 References 50 2. FREE PAIRS OF SEMIGROUPS 2.1. Relative boundaries of semigroups within two different setups. Let T ⊆ S be two commutative and cancellative (a + c = b + c ⇒ a = b) semigroups with identity (0 + a = a + 0 = a), satisfying S ∩ (−S) = {0}, i.e. S (and hence T) is pointed. This situation gives rise to the following notion of a relative boundary.

Research paper thumbnail of Krull Dimension and Other Properties of Graph-Related Algebra

The algebra of basic covers of a graph G, denoted byĀ(G), was introduced by Jürgen Herzog as a su... more The algebra of basic covers of a graph G, denoted byĀ(G), was introduced by Jürgen Herzog as a suitable quotient of the vertex cover algebra. In this paper we show that if the graph is bipartite thenĀ(G) is a homogeneous algebra with straightening laws and thus is Koszul. Furthermore, we compute the Krull dimension ofĀ(G) in terms of the combinatorics of G. As a consequence we get new upper bounds on the arithmetical rank of monomial ideals of pure codimension 2. Finally, we characterize the Cohen-Macaulay property and the Castelnuovo-Mumford regularity of the edge ideal of a certain class of graphs.

Research paper thumbnail of Integrating Adaptronics in Intelligent Measurement Technique and in Mechatronics and Integronics Systems for Integrated Control

Proceedings of the 23rd International DAAAM Symposium 2012, 2012

The paper presents hard and software architecture of adaptonics in intelligent measurement engine... more The paper presents hard and software architecture of adaptonics in intelligent measurement engineering and in mechatronics and integronics systems for integrated automatic control, through cooperation between several scientific and technical disciplines, to meet growing and changing demands about modern systems and new products simultaneously improved with the lowest development time and with a higher flexibility and more efficient.

Research paper thumbnail of Adaptronics – New Concept for the Future of Advanced Engineering and Intelligent Automatised Manufacturing

Proceedings of the 23rd International DAAAM Symposium 2012, 2012

The paper treats new concept of adaptronics for the future of advanced engineering and intelligen... more The paper treats new concept of adaptronics for the future of advanced engineering and intelligent automation, by bringing news and discoveries in the field of basic and applied science of mechatronics and integronics and as development vector of know-how in high-tech advanced domain by adapting scientific, complex, structural, functional and decisional of structures, products and systems, to change the operating mode of environment requirements, to control in active mode the vibration, the noise and the deformation, to recover energy from mechanics and inertial vibrating systems and to influence and monitories all structures of any kind.

Research paper thumbnail of Renal Survival in Children with Glomerulonephritis with Crescents: A Pediatric Nephrology Research Consortium Cohort Study

Journal of Clinical Medicine, 2020

There is no evidence-based definition for diagnosing crescentic glomerulonephritis. The prognosti... more There is no evidence-based definition for diagnosing crescentic glomerulonephritis. The prognostic implications of crescentic lesions on kidney biopsy have not been quantified. Our objective was to determine risk factors for end-stage kidney disease (ESKD) in patients with glomerulonephritis and crescents on kidney biopsy. A query of the Pediatric Nephrology Research Consortium’s Pediatric Glomerulonephritis with Crescents registry identified 305 patients from 15 centers. A retrospective cohort study was performed with ESKD as the primary outcome. Median age at biopsy was 11 years (range 1–21). The percentage of crescents was 3–100% (median 20%). Etiologies included IgA nephropathy (23%), lupus (21%), IgA vasculitis (19%) and ANCA-associated GN (13%), post-infectious GN (5%), and anti-glomerular basement membrane disease (3%). The prevalence of ESKD was 12% at one year and 16% at last follow-up (median = 3 years, range 1–11). Median time to ESKD was 100 days. Risk factors for ESKD i...

Research paper thumbnail of EUS-Guided Transgastric Drainage of Intraabdominal Fluid Collections

Chirurgia, 2018

Rezultate: Vârsta medie a subiecţilor incluşi în studiu a fost 51 de ani. Diametrul mediu al cole... more Rezultate: Vârsta medie a subiecţilor incluşi în studiu a fost 51 de ani. Diametrul mediu al colecţiilor intraabdominale a fost de 109 mm (34-250 mm), acesta corelându-se cu metoda de tratament aleasă (p<0.005): drenaj ghidat eco-endoscopic, drenaj percutan, intervenţie chirurgicală, ca monoterapie sau combinaţii între acestea. Mortalitatea peri-procedurală a fost de 9,3%, asociată în principal cu severitatea patologiei ori cu asocierea chirurgiei în tratament. Concluzii: În conluzie, prin studiul efectuat putem afirma că ecoendoscopia poate fi metoda de primă intenţie în a drena colecţiile intra abdominale perigastrice, pentru că reprezintă o metodă sigură şi eficientă, cu un success tehnic de 100% în timp ce succesul clinic de peste 80% asigură o mai bună calitate a vieţii. Pentru colecţiile ce au un diametru mai mare de 127 mm, ne putem aştepta totuşi la necesitate unui tratament combinat, ecoendoscopic şi chirurgical. Cuvinte cheie: colecţii intraabdominale, drenaj transgastric, percutan, chirurgie, ecoendoscopie

Research paper thumbnail of Efficacy of Endobiliary Radiofrequency Ablation using a Novel Endoluminal Radiofrequency Ablation Catheter in a Swine Model

Journal of Investigative Surgery, 2018

Purpose: Endobiliary radiofrequency ablation (RFA) is a new endoscopic ablative technique. Howeve... more Purpose: Endobiliary radiofrequency ablation (RFA) is a new endoscopic ablative technique. However, the ideal power setting for RFA has not yet been clarified. Therefore, we intended to evaluate the effects of endobiliary RFA according to time variations using novel RFA. Materials and methods: Nine female pigs were divided into three groups according to ablation time (60, 90, and 120 seconds) with the same setting (10 watts, 80 C). All pigs underwent endoscopic retrograde cholangiography (ERC) and endobiliary RFA in the common bile duct. Gross and histologic examinations were performed after 24 hours. Results: The ERC and application of the endobiliary RFA were 100% successful, and the post-RFA cholangiogram did not show contrast leakage. The median depth of microscopic ablation was significantly different among the three groups (60 vs. 90 vs. 120 seconds ¼ 1.90 (1.17-2.23) vs. 2.44 (2.31-2.60) vs. 2.52 (2.47-2.64) mm, p ¼ 0.018). There was also a linear relationship between ablation time and microscopic ablation depth (r 2 ¼ 0.552, p ¼ 0.002). However, no significant differences in macroscopic or microscopic ablation length were observed. In addition, there were focal ablation injuries in adjacent liver tissue in five of the nine pigs (2/3 in 60, 1/3 in 90, and 2/3 in 120 seconds). Conclusion: Endobiliary RFA using a novel RFA catheter resulted in controlled ablation with a linear relationship between microscopic ablation depth and ablation time in a swine model. Clinical studies are needed to validate the safe energy condition of endobiliary RFA in malignant biliary obstruction.

Research paper thumbnail of Predicting First-Year Relapses in Children With Nephrotic Syndrome

Pediatrics, 2000

Objective. More than half of the children diagnosed with nephrotic syndrome will have relapses. T... more Objective. More than half of the children diagnosed with nephrotic syndrome will have relapses. These can be infrequent relapses (IRs: <2 in 6 months or <3 in a year) or frequent relaspses (FRs: >2 in 6 months or >3 in a year). Patients who relapse while on alternate day steroids or within 1 month of discontinuation of steroid therapy are considered steroid-dependent (SD; J Pediatr. 1982; 101:514–518). Patients with an IR course have a better long-term prognosis, and many of them have minimal-change disease without mesangial hypercellularity or sclerosis. The purpose of our study was to identify factors at initial presentation that could predict the relapse pattern in the first year after diagnosis, without taking into consideration the histopathology found on renal biopsy. Design. We analyzed the medical records of children who were seen by us before March 1997 and followed for at least 1 year. Variables selected in the study were age, sex, race, presence or absence of ...

Research paper thumbnail of Abiotic Synthesis of Phosphoric Esters of Monosaccharides According to the “Cold Model”

Springer Netherlands eBooks, 1981

It is investigated, according to the “cold model”, the abiotic synthesis of some phosphoric ester... more It is investigated, according to the “cold model”, the abiotic synthesis of some phosphoric esters of monosaccharides by cold plasma decomposition of methane- water mixtures in the presence of apatite.This high frequence electric discharge induced the formation of a complex reaction product, containing alcohols, aldehydes, ketones and acids.

Research paper thumbnail of Expression of Mammalian Geranylgeranyltransferase Type-II in Escherichia coli and Its Application for in Vitro Prenylation of Rab Proteins

Protein Expression and Purification, 2001

protein trafficking in eukaryotic cells. In mammalian Mammalian geranylgeranyltransferase type II... more protein trafficking in eukaryotic cells. In mammalian Mammalian geranylgeranyltransferase type II cells more than 40 Rabs have been cloned and this (GGTase-II) is a 100-kDa heterodimer that catalyzes the number is increasing (1, 2). Individual Rab proteins transfer of two 20-carbon geranylgeranyl groups from have been implicated as essential mediators of vesicle geranylgeranyl pyrophosphate onto C-terminal cystargeting and fusion events (3, 4). For their function, teine residues of Rab GTPases. This modification is Rab proteins require modification by geranylgeranyl essential for the biological activity of Rab proteins. isoprenoids which allows them to reversibly associate Geranylgeranylation can be performed in vitro using with membrane in a tightly controlled fashion. Covalent recombinant GGTase-II but so far large-scale producattachment of geranylgeranyl groups to two C-terminal tion of the enzyme was challenging. We report here the cysteines is catalyzed by geranylgeranyltransferase design of a two plasmid expression system that will type II (GGTase-II). GGTase-II is a heterodimer comproduce GGTase-II at levels as high as 15 mg/L in Escheposed of tightly associated 68-kDa ␣ and 45-kDa ␤ subrichia coli. The protein was produced as a heterodimer units and belongs to the family of protein prenyltransfwith the ␣ subunit bearing a cleavable tandem 6Hiserases together with farnesyl transferase (FTase) and glutathione S-transferase (GST) tag that was used for geranylgeranyl transferase I (GGTase-I) (for review 5). two-step purification of the enzyme. Purified enzyme In contrast to other known prenyltransferases, was functionally active as determined by in vitro pre-GGTase-II does not recognize its protein substrate dinylation and phosphoisoprenoid binding assay. Furrectly, but functions in a concert with a protein chaperthermore, the GST-tagged GGTase-II was used for one termed REP (Rab escort protein) (6). preparative in vitro prenylation of the Rab7:REP-1 Posttranslational geranylgeranylation is crucial for complex. Using this procedure, 10 mg of doubly prenylated Rab7:REP-1 complex were obtained. ᭧ 2001 Aca-biological activity of Rab proteins since it ensures not demic Press only membrane association but also modulates the interaction of Rab proteins with their effectors and regulatory molecules. Traditionally, prenylated Rab proteins were either purified from tissue sources or from The low molecular mass GTP 2-binding proteins encoded by the Rab gene family play important roles in

Research paper thumbnail of cAMP-dependent Protein Kinase Type I Regulates Ethanol-induced cAMP Response Element-mediated Gene Expression via Activation of CREB-binding Protein and Inhibition of MAPK

Journal of Biological Chemistry, 2004

Research paper thumbnail of cAMP-dependent Protein Kinase Types I and II Differentially Regulate cAMP Response Element-mediated Gene Expression

Journal of Biological Chemistry, 2002

We have shown that ethanol induces translocation of cAMP-dependent protein kinase (PKA) to the nu... more We have shown that ethanol induces translocation of cAMP-dependent protein kinase (PKA) to the nucleus, cAMP response element-binding protein (CREB) phosphorylation, and cAMP response element-mediated gene transcription in NG108-15 cells. However, little is known about which PKA types regulate this process. We show here that under basal conditions NG108-15 cells contain type I PKA (C␤RI␤) primarily in cytosol and type II PKA (C␣RII␤) in the particulate and nuclear fractions. Antagonists of both type I and type II PKA inhibit forskolin-and ethanol-induced cAMP response element-mediated gene transcription. However, only the type II PKA antagonist inhibits forskolin-induced C␣ and ethanol-induced C␣ and RII␤ translocation to the nucleus and CREB phosphorylation; the type I antagonist is without effect. Our data suggest that forskolin-and ethanol-induced CREB phosphorylation and gene activation are differentially mediated by the two types of PKA. We propose that type II PKA is translocated and activated in the nucleus and induces CREB phosphorylation that is necessary but not sufficient for gene transcription. By contrast, type I PKA is activated in the cytoplasm, turning on a downstream pathway that activates other transcription cofactors that interact with phosphorylated CREB to induce gene transcription. cAMP signaling is a multiple component system that requires production of cAMP and induces activation of PKA, 1 phosphorylation of CREB, and cAMP-dependent gene expression. This pathway has been implicated in learning and memory (1, 2) as well as addictive behaviors (3) and responses to ethanol (4). Ethanol activates the cAMP pathway to its full extent by stimulating adenylyl cyclase activity (4), C␣ translocation to the nucleus (5), CREB phosphorylation (6), and CREmediated gene transcription (7).

Research paper thumbnail of Mechanochemical reactions of some polyurethane elastomers. 3. Prediction of long-term uniaxial tensile creep using linear compliance as a basis

Colloid & Polymer Science, 1986

Based on isochronous tests the stress-strain curve for thermoplastic polyurethane elastomers (Mol... more Based on isochronous tests the stress-strain curve for thermoplastic polyurethane elastomers (Moldotan D) is plotted and used for correcting the tensile creep data.