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PURPOSE: Both acne keloidalis nuchae (AKN) and cutis verticis gyrata (CVG) are scalp conditions predominantly affecting men. Both are characterized by dermal thickening and fibroblast hyperactivity. AKN typically occurs in the nuchal area, often involving the naturally occurring folds in the occipital region. The aim of this study was to determine the relationship between excessive scalp folding (CVG) and AKN. PATIENTS AND METHODS: A total of 108 patients with AKN seen over 11 years from July 2009 and November 2020 were retrospectively evaluated. Patients with AKN concomitant with CVG were selected for analysis. RESULTS: Seven of the 108 AKN patients had scalp-wide (widespread) AKN lesions, including 4 with CVG. In 3 of the 4 patients with concomitant AKN and CVG, the AKN was widespread, and its onset had preceded CVG by 1-2 years. In the fourth CVG patient, AKN lesions were confined to the nuchal area, and the CVG preceded AKN onset by several years. All patients were male, with a mean age of 35.8 years (overall) and 38.0 years (CVG group). CONCLUSION: We describe a previously unreported relationship between widespread AKN and CVG, with the development of AKN preceding CVG formation.

The utilization of poly(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) with entrapped fish oil (FO) loaded in collagen-based scaffolds for cutaneous wound healing using a porcine model is unique for the present study. Full-depth cutaneous excisions (5 x 5 cm) on the pig dorsa were treated with pure collagen scaffold (control, C), empty PLGA NPs (NP), FO, mupirocin (MUP), PLGA NPs with entrapped FO (NP/FO) and PLGA NPs with entrapped MUP (NP/MUP). The following markers were evaluated on days 0, 3, 7, 14 and 21 post-excision: collagen, hydroxyproline (HP), angiogenesis and expressions of the COX2, EGF, COL1A1, COL1A3, TGFB1, VEGFA, CCL5 and CCR5 genes. The hypothesis that NP/FO treatment is superior to FO alone and that it is comparable to NP/MUP was tested. NP/FO treatment increased HP in comparison with both FO alone and NP/MUP (day 14) but decreased (p < 0.05) angiogenesis in comparison with FO alone (day 3). NP/FO increased (p < 0.05) the expression of the CCR5 gene (day 3) and tended (p > 0.05) to increase the expressions of the EGF (day 7, day 14), TGFB1 (day 21) and CCL5 (day 7, day 21) genes as compared with NP/MUP. NP/FO can be suggested as a suitable alternative to NP/MUP in cutaneous wound treatment., info:eu-repo/semantics/openAccess

The utilization of poly(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) with entrapped fish oil (FO) loaded in collagen-based scaffolds for cutaneous wound healing using a porcine model is unique for the present study. Full-depth cutaneous excisions (5 x 5 cm) on the pig dorsa were treated with pure collagen scaffold (control, C), empty PLGA NPs (NP), FO, mupirocin (MUP), PLGA NPs with entrapped FO (NP/FO) and PLGA NPs with entrapped MUP (NP/MUP). The following markers were evaluated on days 0, 3, 7, 14 and 21 post-excision: collagen, hydroxyproline (HP), angiogenesis and expressions of the COX2, EGF, COL1A1, COL1A3, TGFB1, VEGFA, CCL5 and CCR5 genes. The hypothesis that NP/FO treatment is superior to FO alone and that it is comparable to NP/MUP was tested. NP/FO treatment increased HP in comparison with both FO alone and NP/MUP (day 14) but decreased (p < 0.05) angiogenesis in comparison with FO alone (day 3). NP/FO increased (p < 0.05) the expression of the CCR5 gene (day 3) and tended (p > 0.05) to increase the expressions of the EGF (day 7, day 14), TGFB1 (day 21) and CCL5 (day 7, day 21) genes as compared with NP/MUP. NP/FO can be suggested as a suitable alternative to NP/MUP in cutaneous wound treatment., info:eu-repo/semantics/openAccess

The utilization of poly(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) with entrapped fish oil (FO) loaded in collagen-based scaffolds for cutaneous wound healing using a porcine model is unique for the present study. Full-depth cutaneous excisions (5 x 5 cm) on the pig dorsa were treated with pure collagen scaffold (control, C), empty PLGA NPs (NP), FO, mupirocin (MUP), PLGA NPs with entrapped FO (NP/FO) and PLGA NPs with entrapped MUP (NP/MUP). The following markers were evaluated on days 0, 3, 7, 14 and 21 post-excision: collagen, hydroxyproline (HP), angiogenesis and expressions of the COX2, EGF, COL1A1, COL1A3, TGFB1, VEGFA, CCL5 and CCR5 genes. The hypothesis that NP/FO treatment is superior to FO alone and that it is comparable to NP/MUP was tested. NP/FO treatment increased HP in comparison with both FO alone and NP/MUP (day 14) but decreased (p < 0.05) angiogenesis in comparison with FO alone (day 3). NP/FO increased (p < 0.05) the expression of the CCR5 gene (day 3) and tended (p > 0.05) to increase the expressions of the EGF (day 7, day 14), TGFB1 (day 21) and CCL5 (day 7, day 21) genes as compared with NP/MUP. NP/FO can be suggested as a suitable alternative to NP/MUP in cutaneous wound treatment., info:eu-repo/semantics/openAccess

The utilization of poly(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) with entrapped fish oil (FO) loaded in collagen-based scaffolds for cutaneous wound healing using a porcine model is unique for the present study. Full-depth cutaneous excisions (5 x 5 cm) on the pig dorsa were treated with pure collagen scaffold (control, C), empty PLGA NPs (NP), FO, mupirocin (MUP), PLGA NPs with entrapped FO (NP/FO) and PLGA NPs with entrapped MUP (NP/MUP). The following markers were evaluated on days 0, 3, 7, 14 and 21 post-excision: collagen, hydroxyproline (HP), angiogenesis and expressions of the COX2, EGF, COL1A1, COL1A3, TGFB1, VEGFA, CCL5 and CCR5 genes. The hypothesis that NP/FO treatment is superior to FO alone and that it is comparable to NP/MUP was tested. NP/FO treatment increased HP in comparison with both FO alone and NP/MUP (day 14) but decreased (p < 0.05) angiogenesis in comparison with FO alone (day 3). NP/FO increased (p < 0.05) the expression of the CCR5 gene (day 3) and tended (p > 0.05) to increase the expressions of the EGF (day 7, day 14), TGFB1 (day 21) and CCL5 (day 7, day 21) genes as compared with NP/MUP. NP/FO can be suggested as a suitable alternative to NP/MUP in cutaneous wound treatment.

Acute kidney injury is associated with increased risk of heart failure and mortality. This study demonstrates that acute kidney injury induces remote cardiac dysfunction, damage, injury, and fibrosis via a galectin-3 (Gal-3) dependent pathway. Gal-3 originates from bone marrow-derived immune cells. Cardiac damage could be prevented by blocking this pathway.

Acute kidney injury is associated with increased risk of heart failure and mortality. This study demonstrates that acute kidney injury induces remote cardiac dysfunction, damage, injury, and fibrosis via a galectin-3 (Gal-3) dependent pathway. Gal-3 originates from bone marrow-derived immune cells. Cardiac damage could be prevented by blocking this pathway.

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disease of unknown etiology and dismal prognosis. IPF patients are known to have an increased risk of lung cancer and careful decision-making is required for the treatment of lung cancer associated with IPF. Transforming growth factor (TGF)- signaling plays a central role in tissue fibrosis and tumorigenesis. TGF--mediated pathological changes that occur in IPF lung tissue may promote the process of field cancerization and provide the microenvironment favorable to cancer initiation and progression. This review summarizes the current knowledge related to IPF pathogenesis and explores the molecular mechanisms that underlie the occurrence of lung cancer in the background of IPF, with an emphasis on the multifaceted effects of TGF- signaling.

Metastasis of tumor cells from primary sites of malignancy to neighboring stromal tissue or distant localities entails in several instances, but not in every case, the epithelial-mesenchymal transition (EMT). EMT weakens the strong adhesion forces between differentiated epithelial cells so that carcinoma cells can achieve solitary or collective motility, which makes the EMT an intuitive mechanism for the initiation of tumor metastasis. EMT initiates after primary oncogenic events lead to secondary secretion of cytokines. The interaction between tumor-secreted cytokines and oncogenic stimuli facilitates EMT progression. A classic case of this mechanism is the cooperation between oncogenic Ras and the transforming growth factor (TGF). The power of TGF to mediate EMT during metastasis depends on versatile signaling crosstalk and on the regulation of successive waves of expression of many other cytokines and the progressive remodeling of the extracellular matrix that facilitates motility through basement membranes. Since metastasis involves many organs in the body, whereas EMT affects carcinoma cell differentiation locally, it has frequently been debated whether EMT truly contributes to metastasis. Despite controversies, studies of circulating tumor cells, studies of acquired chemoresistance by metastatic cells, and several (but not all) metastatic animal models, support a link between EMT and metastasis, with TGF, often being a common denominator in this link. This article aims at discussing mechanistic cases where TGF signaling and EMT facilitate tumor cell dissemination.

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disease of unknown etiology and dismal prognosis. IPF patients are known to have an increased risk of lung cancer and careful decision-making is required for the treatment of lung cancer associated with IPF. Transforming growth factor (TGF)- signaling plays a central role in tissue fibrosis and tumorigenesis. TGF--mediated pathological changes that occur in IPF lung tissue may promote the process of field cancerization and provide the microenvironment favorable to cancer initiation and progression. This review summarizes the current knowledge related to IPF pathogenesis and explores the molecular mechanisms that underlie the occurrence of lung cancer in the background of IPF, with an emphasis on the multifaceted effects of TGF- signaling.

Metastasis of tumor cells from primary sites of malignancy to neighboring stromal tissue or distant localities entails in several instances, but not in every case, the epithelial-mesenchymal transition (EMT). EMT weakens the strong adhesion forces between differentiated epithelial cells so that carcinoma cells can achieve solitary or collective motility, which makes the EMT an intuitive mechanism for the initiation of tumor metastasis. EMT initiates after primary oncogenic events lead to secondary secretion of cytokines. The interaction between tumor-secreted cytokines and oncogenic stimuli facilitates EMT progression. A classic case of this mechanism is the cooperation between oncogenic Ras and the transforming growth factor (TGF). The power of TGF to mediate EMT during metastasis depends on versatile signaling crosstalk and on the regulation of successive waves of expression of many other cytokines and the progressive remodeling of the extracellular matrix that facilitates motility through basement membranes. Since metastasis involves many organs in the body, whereas EMT affects carcinoma cell differentiation locally, it has frequently been debated whether EMT truly contributes to metastasis. Despite controversies, studies of circulating tumor cells, studies of acquired chemoresistance by metastatic cells, and several (but not all) metastatic animal models, support a link between EMT and metastasis, with TGF, often being a common denominator in this link. This article aims at discussing mechanistic cases where TGF signaling and EMT facilitate tumor cell dissemination.

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disease of unknown etiology and dismal prognosis. IPF patients are known to have an increased risk of lung cancer and careful decision-making is required for the treatment of lung cancer associated with IPF. Transforming growth factor (TGF)- signaling plays a central role in tissue fibrosis and tumorigenesis. TGF--mediated pathological changes that occur in IPF lung tissue may promote the process of field cancerization and provide the microenvironment favorable to cancer initiation and progression. This review summarizes the current knowledge related to IPF pathogenesis and explores the molecular mechanisms that underlie the occurrence of lung cancer in the background of IPF, with an emphasis on the multifaceted effects of TGF- signaling.

Metastasis of tumor cells from primary sites of malignancy to neighboring stromal tissue or distant localities entails in several instances, but not in every case, the epithelial-mesenchymal transition (EMT). EMT weakens the strong adhesion forces between differentiated epithelial cells so that carcinoma cells can achieve solitary or collective motility, which makes the EMT an intuitive mechanism for the initiation of tumor metastasis. EMT initiates after primary oncogenic events lead to secondary secretion of cytokines. The interaction between tumor-secreted cytokines and oncogenic stimuli facilitates EMT progression. A classic case of this mechanism is the cooperation between oncogenic Ras and the transforming growth factor (TGF). The power of TGF to mediate EMT during metastasis depends on versatile signaling crosstalk and on the regulation of successive waves of expression of many other cytokines and the progressive remodeling of the extracellular matrix that facilitates motility through basement membranes. Since metastasis involves many organs in the body, whereas EMT affects carcinoma cell differentiation locally, it has frequently been debated whether EMT truly contributes to metastasis. Despite controversies, studies of circulating tumor cells, studies of acquired chemoresistance by metastatic cells, and several (but not all) metastatic animal models, support a link between EMT and metastasis, with TGF, often being a common denominator in this link. This article aims at discussing mechanistic cases where TGF signaling and EMT facilitate tumor cell dissemination.

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disease of unknown etiology and dismal prognosis. IPF patients are known to have an increased risk of lung cancer and careful decision-making is required for the treatment of lung cancer associated with IPF. Transforming growth factor (TGF)- signaling plays a central role in tissue fibrosis and tumorigenesis. TGF--mediated pathological changes that occur in IPF lung tissue may promote the process of field cancerization and provide the microenvironment favorable to cancer initiation and progression. This review summarizes the current knowledge related to IPF pathogenesis and explores the molecular mechanisms that underlie the occurrence of lung cancer in the background of IPF, with an emphasis on the multifaceted effects of TGF- signaling.

Metastasis of tumor cells from primary sites of malignancy to neighboring stromal tissue or distant localities entails in several instances, but not in every case, the epithelial-mesenchymal transition (EMT). EMT weakens the strong adhesion forces between differentiated epithelial cells so that carcinoma cells can achieve solitary or collective motility, which makes the EMT an intuitive mechanism for the initiation of tumor metastasis. EMT initiates after primary oncogenic events lead to secondary secretion of cytokines. The interaction between tumor-secreted cytokines and oncogenic stimuli facilitates EMT progression. A classic case of this mechanism is the cooperation between oncogenic Ras and the transforming growth factor (TGF). The power of TGF to mediate EMT during metastasis depends on versatile signaling crosstalk and on the regulation of successive waves of expression of many other cytokines and the progressive remodeling of the extracellular matrix that facilitates motility through basement membranes. Since metastasis involves many organs in the body, whereas EMT affects carcinoma cell differentiation locally, it has frequently been debated whether EMT truly contributes to metastasis. Despite controversies, studies of circulating tumor cells, studies of acquired chemoresistance by metastatic cells, and several (but not all) metastatic animal models, support a link between EMT and metastasis, with TGF, often being a common denominator in this link. This article aims at discussing mechanistic cases where TGF signaling and EMT facilitate tumor cell dissemination.

Metastasis of tumor cells from primary sites of malignancy to neighboring stromal tissue or distant localities entails in several instances, but not in every case, the epithelial-mesenchymal transition (EMT). EMT weakens the strong adhesion forces between differentiated epithelial cells so that carcinoma cells can achieve solitary or collective motility, which makes the EMT an intuitive mechanism for the initiation of tumor metastasis. EMT initiates after primary oncogenic events lead to secondary secretion of cytokines. The interaction between tumor-secreted cytokines and oncogenic stimuli facilitates EMT progression. A classic case of this mechanism is the cooperation between oncogenic Ras and the transforming growth factor (TGF). The power of TGF to mediate EMT during metastasis depends on versatile signaling crosstalk and on the regulation of successive waves of expression of many other cytokines and the progressive remodeling of the extracellular matrix that facilitates motility through basement membranes. Since metastasis involves many organs in the body, whereas EMT affects carcinoma cell differentiation locally, it has frequently been debated whether EMT truly contributes to metastasis. Despite controversies, studies of circulating tumor cells, studies of acquired chemoresistance by metastatic cells, and several (but not all) metastatic animal models, support a link between EMT and metastasis, with TGF, often being a common denominator in this link. This article aims at discussing mechanistic cases where TGF signaling and EMT facilitate tumor cell dissemination.

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disease of unknown etiology and dismal prognosis. IPF patients are known to have an increased risk of lung cancer and careful decision-making is required for the treatment of lung cancer associated with IPF. Transforming growth factor (TGF)- signaling plays a central role in tissue fibrosis and tumorigenesis. TGF--mediated pathological changes that occur in IPF lung tissue may promote the process of field cancerization and provide the microenvironment favorable to cancer initiation and progression. This review summarizes the current knowledge related to IPF pathogenesis and explores the molecular mechanisms that underlie the occurrence of lung cancer in the background of IPF, with an emphasis on the multifaceted effects of TGF- signaling.

Transformišući faktor rasta beta 1 (TGFB1) je važan inflamatorni modulator i profibrotički medijator u astmi. Promene u regulaciji ekspresije TGFB1 bi mogle biti povezane sa patogenezom astme i sa adekvatnim odgovorom na terapiju. Cilj ovog rada je bio ispitivanje promotorskih varijanti gena TGFB1 kao faktora rizika za nastanak astme i modulatora odgovora na terapiju. Studija je uključila pacijente sa astmom i zdrave ispitanike kod kojih je analiziran 5' regulatorni region gena TGFB1. Detektovane su promotorske varijante -509C>T (rs1800469) i -800G>A (rs1800468) za koje asocijativnom studijom nije pokazana povezanost sa patogenezom astme, ali je uočena potencijalna povezanost između promotorske varijante -509C>T i egzacerbacija bolesti. Uticaj ove promotorske varijante na ekspresiju gena TGFB1 pod dejstvom montelukasta, leka za astmu, je ispitivan ex vivo u ćelijama indukovanog sputuma i in vitro u ćelijama BEAS 2B. Analizom ex vivo je pokazana smanjena ekspresija gena TGFB1, pri čemu je efekat bio izraženiji kod astmatičara sa genotipom -509TT (58,9%) u odnosu na astmatičare sa genotipovima -509CC (49,6%) i -509CT (31,8%). U ćelijama BEAS 2B je ekspresija gena bila smanjena za 27%. Nakon transfekcije ćelija BEAS 2B konstruktima sa promotorskim varijantama -509C ili -509T i tretmana montelukastom, zapažena je povećana aktivnost promotora gena TGFB1. Odgovor varijante -509T je bio intenzivniji i za 139,9% veći u odnosu na odgovor varijante -509C. Ova studija je pokazala da promotorska varijanta -509C>T menja ekspresiju gena TGFB1 u odgovoru na montelukast, ali da bi se njena modulatorna uloga razjasnila, u daljim istraživanjima bi trebalo analizirati i druge genetičke i negenetičke faktore., Transforming growth factor beta 1 (TGFB1) is an important inflammatory modulator and profibrotic mediator in asthma. The changes in regulation of TGFB1 expression could be associated with asthma pathogenesis and adequate response to therapy. The aim of this study was to investigate TGFB1 gene promoter variants as asthma risk factors and modulators of response to asthma therapy. The 5' regulatory region of the TGFB1 gene was analyzed in asthmatic patients and healthy individuals included in this study. The promoter variants -509C>T (rs1800469) and -800G>A (rs1800468) were detected and the association between these variants and asthma pathogenesis was not found. The association was observed between the presence of variant -509C>T and occurence of exacerbations in patients. The influence of -509C>T variant on TGFB1 gene expression in response to montelukast, used for asthma treatment, was further investigated ex vivo in induced sputum cells and in vitro in BEAS 2B cells. Analysis ex vivo has shown the decrease in the TGFB1 gene expression, with more pronaunced effect in asthmatics with genotype -509TT (58.9%) in comparison with -509CC (49.6%) and -509CT (31.8%) genotypes. In BEAS 2B cells the gene expression was reduced for 27%. After transfection of BEAS 2B cells with constructs carrying promoter variants -509C or -509T and treatment with montelukast the increased activity was observed for both promoter variants. The response of variant -509T was more intensive and 139,3% higher than response of variant -509C. This study showed that promoter variant -509T alters TGFB1 gene expression in response to montelukast, but to clarify its modulatory role other genetic, and nongenetic factors as well, should be taken into consideration in further studies.

Background: Tendinosis is a painful chronic, degenerative condition characterized by objective changes in the tissue structure of a tendon. Hallmark features in tendinosis tendons include increased number of cells (hypercellularity), extracellular matrix (ECM) degradation and disorganized collagen. The progression of these pathological changes seen in tendinosis is neither well characterized nor fully understood. Studies have suggested that there are biochemical and mechanical elements involved in tendinosis. From a biochemical perspective, studies have shown that the tendon cells, tenocytes, produce a number of neuronal signal substances/neuromodulators, such as substance P (SP) and acetylcholine (ACh), traditionally thought to be confined to the nervous system. Furthermore, it has been shown that the expression of these neuromodulators is elevated in tendinosis tendons as compared to normal healthy tendons. Interestingly, studies on other tissue types have revealed that both SP and ACh can induce tissue changes seen in tendinosis, such as hypercellularity and collagen disorganization. From a mechanical angle, it has been suggested that overload of tendons, including extensive strain on the primary tendon cells (tenocytes), causes the degenerative processes associated with tendinosis. In vivo studies have shown that in overloaded tendons, the presence of neuromodulators is elevated, not least SP, which also precedes the development of the tissue changes seen in tendinosis. This further supports the importance of combining biochemical factors and mechanical factors in the pathogenesis of tendinosis. Hypotheses: In this thesis project, we hypothesize: 1) that neuromodulators, such as SP and ACh when stimulating their preferred receptors, the neurokinin 1 (NK-1 R) and muscarinic receptors (mAChRs), respectively, can cause increased tenocyte proliferation; 2) that the effects of SP and ACh on tenocyte proliferation converge mechanistically via a shared signalling pathw

Background: Tendinosis is a painful chronic, degenerative condition characterized by objective changes in the tissue structure of a tendon. Hallmark features in tendinosis tendons include increased number of cells (hypercellularity), extracellular matrix (ECM) degradation and disorganized collagen. The progression of these pathological changes seen in tendinosis is neither well characterized nor fully understood. Studies have suggested that there are biochemical and mechanical elements involved in tendinosis. From a biochemical perspective, studies have shown that the tendon cells, tenocytes, produce a number of neuronal signal substances/neuromodulators, such as substance P (SP) and acetylcholine (ACh), traditionally thought to be confined to the nervous system. Furthermore, it has been shown that the expression of these neuromodulators is elevated in tendinosis tendons as compared to normal healthy tendons. Interestingly, studies on other tissue types have revealed that both SP and ACh can induce tissue changes seen in tendinosis, such as hypercellularity and collagen disorganization. From a mechanical angle, it has been suggested that overload of tendons, including extensive strain on the primary tendon cells (tenocytes), causes the degenerative processes associated with tendinosis. In vivo studies have shown that in overloaded tendons, the presence of neuromodulators is elevated, not least SP, which also precedes the development of the tissue changes seen in tendinosis. This further supports the importance of combining biochemical factors and mechanical factors in the pathogenesis of tendinosis. Hypotheses: In this thesis project, we hypothesize: 1) that neuromodulators, such as SP and ACh when stimulating their preferred receptors, the neurokinin 1 (NK-1 R) and muscarinic receptors (mAChRs), respectively, can cause increased tenocyte proliferation; 2) that the effects of SP and ACh on tenocyte proliferation converge mechanistically via a shared signalling pathw

Transformišući faktor rasta beta 1 (TGFB1) je važan inflamatorni modulator i profibrotički medijator u astmi. Promene u regulaciji ekspresije TGFB1 bi mogle biti povezane sa patogenezom astme i sa adekvatnim odgovorom na terapiju. Cilj ovog rada je bio ispitivanje promotorskih varijanti gena TGFB1 kao faktora rizika za nastanak astme i modulatora odgovora na terapiju. Studija je uključila pacijente sa astmom i zdrave ispitanike kod kojih je analiziran 5' regulatorni region gena TGFB1. Detektovane su promotorske varijante -509C>T (rs1800469) i -800G>A (rs1800468) za koje asocijativnom studijom nije pokazana povezanost sa patogenezom astme, ali je uočena potencijalna povezanost između promotorske varijante -509C>T i egzacerbacija bolesti. Uticaj ove promotorske varijante na ekspresiju gena TGFB1 pod dejstvom montelukasta, leka za astmu, je ispitivan ex vivo u ćelijama indukovanog sputuma i in vitro u ćelijama BEAS 2B. Analizom ex vivo je pokazana smanjena ekspresija gena TGFB1, pri čemu je efekat bio izraženiji kod astmatičara sa genotipom -509TT (58,9%) u odnosu na astmatičare sa genotipovima -509CC (49,6%) i -509CT (31,8%). U ćelijama BEAS 2B je ekspresija gena bila smanjena za 27%. Nakon transfekcije ćelija BEAS 2B konstruktima sa promotorskim varijantama -509C ili -509T i tretmana montelukastom, zapažena je povećana aktivnost promotora gena TGFB1. Odgovor varijante -509T je bio intenzivniji i za 139,9% veći u odnosu na odgovor varijante -509C. Ova studija je pokazala da promotorska varijanta -509C>T menja ekspresiju gena TGFB1 u odgovoru na montelukast, ali da bi se njena modulatorna uloga razjasnila, u daljim istraživanjima bi trebalo analizirati i druge genetičke i negenetičke faktore., Transforming growth factor beta 1 (TGFB1) is an important inflammatory modulator and profibrotic mediator in asthma. The changes in regulation of TGFB1 expression could be associated with asthma pathogenesis and adequate response to therapy. The aim of this study was to investigate TGFB1 gene promoter variants as asthma risk factors and modulators of response to asthma therapy. The 5' regulatory region of the TGFB1 gene was analyzed in asthmatic patients and healthy individuals included in this study. The promoter variants -509C>T (rs1800469) and -800G>A (rs1800468) were detected and the association between these variants and asthma pathogenesis was not found. The association was observed between the presence of variant -509C>T and occurence of exacerbations in patients. The influence of -509C>T variant on TGFB1 gene expression in response to montelukast, used for asthma treatment, was further investigated ex vivo in induced sputum cells and in vitro in BEAS 2B cells. Analysis ex vivo has shown the decrease in the TGFB1 gene expression, with more pronaunced effect in asthmatics with genotype -509TT (58.9%) in comparison with -509CC (49.6%) and -509CT (31.8%) genotypes. In BEAS 2B cells the gene expression was reduced for 27%. After transfection of BEAS 2B cells with constructs carrying promoter variants -509C or -509T and treatment with montelukast the increased activity was observed for both promoter variants. The response of variant -509T was more intensive and 139,3% higher than response of variant -509C. This study showed that promoter variant -509T alters TGFB1 gene expression in response to montelukast, but to clarify its modulatory role other genetic, and nongenetic factors as well, should be taken into consideration in further studies.

Transformišući faktor rasta beta 1 (TGFB1) je važan inflamatorni modulator i profibrotički medijator u astmi. Promene u regulaciji ekspresije TGFB1 bi mogle biti povezane sa patogenezom astme i sa adekvatnim odgovorom na terapiju. Cilj ovog rada je bio ispitivanje promotorskih varijanti gena TGFB1 kao faktora rizika za nastanak astme i modulatora odgovora na terapiju. Studija je uključila pacijente sa astmom i zdrave ispitanike kod kojih je analiziran 5' regulatorni region gena TGFB1. Detektovane su promotorske varijante -509C>T (rs1800469) i -800G>A (rs1800468) za koje asocijativnom studijom nije pokazana povezanost sa patogenezom astme, ali je uočena potencijalna povezanost između promotorske varijante -509C>T i egzacerbacija bolesti. Uticaj ove promotorske varijante na ekspresiju gena TGFB1 pod dejstvom montelukasta, leka za astmu, je ispitivan ex vivo u ćelijama indukovanog sputuma i in vitro u ćelijama BEAS 2B. Analizom ex vivo je pokazana smanjena ekspresija gena TGFB1, pri čemu je efekat bio izraženiji kod astmatičara sa genotipom -509TT (58,9%) u odnosu na astmatičare sa genotipovima -509CC (49,6%) i -509CT (31,8%). U ćelijama BEAS 2B je ekspresija gena bila smanjena za 27%. Nakon transfekcije ćelija BEAS 2B konstruktima sa promotorskim varijantama -509C ili -509T i tretmana montelukastom, zapažena je povećana aktivnost promotora gena TGFB1. Odgovor varijante -509T je bio intenzivniji i za 139,9% veći u odnosu na odgovor varijante -509C. Ova studija je pokazala da promotorska varijanta -509C>T menja ekspresiju gena TGFB1 u odgovoru na montelukast, ali da bi se njena modulatorna uloga razjasnila, u daljim istraživanjima bi trebalo analizirati i druge genetičke i negenetičke faktore., Transforming growth factor beta 1 (TGFB1) is an important inflammatory modulator and profibrotic mediator in asthma. The changes in regulation of TGFB1 expression could be associated with asthma pathogenesis and adequate response to therapy. The aim of this study was to investigate TGFB1 gene promoter variants as asthma risk factors and modulators of response to asthma therapy. The 5' regulatory region of the TGFB1 gene was analyzed in asthmatic patients and healthy individuals included in this study. The promoter variants -509C>T (rs1800469) and -800G>A (rs1800468) were detected and the association between these variants and asthma pathogenesis was not found. The association was observed between the presence of variant -509C>T and occurence of exacerbations in patients. The influence of -509C>T variant on TGFB1 gene expression in response to montelukast, used for asthma treatment, was further investigated ex vivo in induced sputum cells and in vitro in BEAS 2B cells. Analysis ex vivo has shown the decrease in the TGFB1 gene expression, with more pronaunced effect in asthmatics with genotype -509TT (58.9%) in comparison with -509CC (49.6%) and -509CT (31.8%) genotypes. In BEAS 2B cells the gene expression was reduced for 27%. After transfection of BEAS 2B cells with constructs carrying promoter variants -509C or -509T and treatment with montelukast the increased activity was observed for both promoter variants. The response of variant -509T was more intensive and 139,3% higher than response of variant -509C. This study showed that promoter variant -509T alters TGFB1 gene expression in response to montelukast, but to clarify its modulatory role other genetic, and nongenetic factors as well, should be taken into consideration in further studies.

Background: Tendinosis is a painful chronic, degenerative condition characterized by objective changes in the tissue structure of a tendon. Hallmark features in tendinosis tendons include increased number of cells (hypercellularity), extracellular matrix (ECM) degradation and disorganized collagen. The progression of these pathological changes seen in tendinosis is neither well characterized nor fully understood. Studies have suggested that there are biochemical and mechanical elements involved in tendinosis. From a biochemical perspective, studies have shown that the tendon cells, tenocytes, produce a number of neuronal signal substances/neuromodulators, such as substance P (SP) and acetylcholine (ACh), traditionally thought to be confined to the nervous system. Furthermore, it has been shown that the expression of these neuromodulators is elevated in tendinosis tendons as compared to normal healthy tendons. Interestingly, studies on other tissue types have revealed that both SP and ACh can induce tissue changes seen in tendinosis, such as hypercellularity and collagen disorganization. From a mechanical angle, it has been suggested that overload of tendons, including extensive strain on the primary tendon cells (tenocytes), causes the degenerative processes associated with tendinosis. In vivo studies have shown that in overloaded tendons, the presence of neuromodulators is elevated, not least SP, which also precedes the development of the tissue changes seen in tendinosis. This further supports the importance of combining biochemical factors and mechanical factors in the pathogenesis of tendinosis. Hypotheses: In this thesis project, we hypothesize: 1) that neuromodulators, such as SP and ACh when stimulating their preferred receptors, the neurokinin 1 (NK-1 R) and muscarinic receptors (mAChRs), respectively, can cause increased tenocyte proliferation; 2) that the effects of SP and ACh on tenocyte proliferation converge mechanistically via a shared signalling pathw

Background: Tendinosis is a painful chronic, degenerative condition characterized by objective changes in the tissue structure of a tendon. Hallmark features in tendinosis tendons include increased number of cells (hypercellularity), extracellular matrix (ECM) degradation and disorganized collagen. The progression of these pathological changes seen in tendinosis is neither well characterized nor fully understood. Studies have suggested that there are biochemical and mechanical elements involved in tendinosis. From a biochemical perspective, studies have shown that the tendon cells, tenocytes, produce a number of neuronal signal substances/neuromodulators, such as substance P (SP) and acetylcholine (ACh), traditionally thought to be confined to the nervous system. Furthermore, it has been shown that the expression of these neuromodulators is elevated in tendinosis tendons as compared to normal healthy tendons. Interestingly, studies on other tissue types have revealed that both SP and ACh can induce tissue changes seen in tendinosis, such as hypercellularity and collagen disorganization. From a mechanical angle, it has been suggested that overload of tendons, including extensive strain on the primary tendon cells (tenocytes), causes the degenerative processes associated with tendinosis. In vivo studies have shown that in overloaded tendons, the presence of neuromodulators is elevated, not least SP, which also precedes the development of the tissue changes seen in tendinosis. This further supports the importance of combining biochemical factors and mechanical factors in the pathogenesis of tendinosis. Hypotheses: In this thesis project, we hypothesize: 1) that neuromodulators, such as SP and ACh when stimulating their preferred receptors, the neurokinin 1 (NK-1 R) and muscarinic receptors (mAChRs), respectively, can cause increased tenocyte proliferation; 2) that the effects of SP and ACh on tenocyte proliferation converge mechanistically via a shared signalling pathw

Background: Tendinosis is a painful chronic, degenerative condition characterized by objective changes in the tissue structure of a tendon. Hallmark features in tendinosis tendons include increased number of cells (hypercellularity), extracellular matrix (ECM) degradation and disorganized collagen. The progression of these pathological changes seen in tendinosis is neither well characterized nor fully understood. Studies have suggested that there are biochemical and mechanical elements involved in tendinosis. From a biochemical perspective, studies have shown that the tendon cells, tenocytes, produce a number of neuronal signal substances/neuromodulators, such as substance P (SP) and acetylcholine (ACh), traditionally thought to be confined to the nervous system. Furthermore, it has been shown that the expression of these neuromodulators is elevated in tendinosis tendons as compared to normal healthy tendons. Interestingly, studies on other tissue types have revealed that both SP and ACh can induce tissue changes seen in tendinosis, such as hypercellularity and collagen disorganization. From a mechanical angle, it has been suggested that overload of tendons, including extensive strain on the primary tendon cells (tenocytes), causes the degenerative processes associated with tendinosis. In vivo studies have shown that in overloaded tendons, the presence of neuromodulators is elevated, not least SP, which also precedes the development of the tissue changes seen in tendinosis. This further supports the importance of combining biochemical factors and mechanical factors in the pathogenesis of tendinosis. Hypotheses: In this thesis project, we hypothesize: 1) that neuromodulators, such as SP and ACh when stimulating their preferred receptors, the neurokinin 1 (NK-1 R) and muscarinic receptors (mAChRs), respectively, can cause increased tenocyte proliferation; 2) that the effects of SP and ACh on tenocyte proliferation converge mechanistically via a shared signalling pathw

Epithelial-mesenchymal transition (EMT) refers to plastic changes in epithelial tissue architecture. Breast cancer stromal cells provide secreted molecules, such as transforming growth factor (TGF), that promote EMT on tumor cells to facilitate breast cancer cell invasion, stemness and metastasis. TGF signaling is considered to be abnormal in the context of cancer development; however, TGF acting on breast cancer EMT resembles physiological signaling during embryonic development, when EMT generates or patterns new tissues. Interestingly, while EMT promotes metastatic fate, successful metastatic colonization seems to require the inverse process of mesenchymal-epithelial transition (MET). EMT and MET are interconnected in a time-dependent and tissue context-dependent manner and are coordinated by TGF, other extracellular proteins, intracellular signaling cascades, non-coding RNAs and chromatin-based molecular alterations. Research on breast cancer EMT/MET aims at delivering biomolecules that can be used diagnostically in cancer pathology and possibly provide ideas for how to improve breast cancer therapy.

Aim: To determine the role of TAK1 in acute glomerular inflammation. Background(s): Activation of p38/JNK signalling promotes glomerular inflammation. TAK1 is an upstream kinase in the p38/JNK pathway. TAK1 regulates the response to a range of stimuli which promote glomerular inflammation (TNF-alpha, IL-1 and TLR ligands). Therefore, we investigated whether TAK1 activates p38/JNK signalling in kidney disease. Method(s): Global Tak1 gene deletion was induced in Tak1f/fERCre mice via tamoxifen, while Tak1f/fPodCre mice have conditional Tak1 deletion in podocytes. Accelerated anti-glomerular basement membrane (GBM) disease was induced in Tak1f/fERCre and Tak1f/f littermate controls (n = 8-11) and killed after 3 hr or 24 hr. Disease was also induced in Tak1f/fPodCre and Tak1f/f littermate controls (n = 5) and killed at 24 hr. Result(s): Compared to controls, global deletion of TAK1 had no effect upon the initial inflammatory response at 3 hr in terms of neutrophil and macrophage infiltration and platelet deposition. However, at 24 hr TAK1 deletion caused exacerbation of glomerular injury compared to littermate controls in terms of: proteinuria (11.9 +/- 5 vs 5.4 +/- 0.5 mg/mmol; P = 0.03); thrombosis (59 +/- 11% vs 8 +/- 3% glomeruli; P = 0.001); renal function (1.4-fold increase in serum cystatin c; P < 0.01); p38 and JNK activation (2-fold increase in glomerular p-p38+ and p-c-Jun+ cells; both P < 0.01). The neutrophil infiltrate had largely subsided in control mice at 24 hr; however, prominent neutrophil infiltration and platelet deposition was evident in mice with global TAK1 deletion at 24 hr. This exacerbation of glomerular damage was not due to a direct effect upon podocytes since Tak1f/fPodCre mice showed no difference compared to controls in proteinuria, p38/JNK activation or thrombosis at 24 hr. Conclusion(s): TAK1 is a negative regulator of glomerular p38/JNK signalling and acute renal inflammation, which may operate via limiting neutrophil-mediated glomerula

Цель: определение уровней галектина-3 и TGF-β1, как маркеров фиброзообразования, у детей раннего возраста, больных пиелонефритом на фоне пузырно-мочеточникового рефлюкса (ПМР).Пациенты и методы. Обследовано 55 детей раннего возраста, больных пиелонефритом. Первую подгруппу составили 28 детей с пиелонефритом без признаков ПМР (первичный пиелонефрит — ППН), вторую подгруппу — 27 детей с пиелонефритом на фоне ПМР (вторичный пиелонефрит — ВПН). Контрольную группу составили 24 практически здоровых ребенка. Содержание TGF-β1 и галектина-3 определяли иммуноферментным методом (ELISA).Результаты. Плазменная концентрация TGF-β1 и галектина-3 достоверно выше у детей с пиелонефритом на фоне ПМР, чем у детей с ППН, что указывает на возможное фиброзообразования в почках. С увеличением продолжительности пиелонефрита повышались уровни профибротических показателей. При ВПН у большинства детей (81,4%) установлено уменьшение толщины паренхимы почек по данным показателей ренально-кортикального индекса, что свидетельствует о возможности снижения функциональной способности почек. Высокая чувствительность (TGF-β1 — 85%, галектин-3 — 92%) и специфичность (TGF-β1 — 67%, галектин-3 — 82%) обоих показателей при пиелонефрите на фоне ПМР указывает на необходимость их определения в качестве маркеров раннего фиброзообразования.Выводы. Перспективным остается дальнейшее изучение патогенетических аспектов формирования необратимых изменений в почках при пиелонефрите на фоне ПМР у детей раннего возраста.Ключевые слова: пиелонефрит, дети раннего возраста, трансформирующий фактор роста В1, галектин-3., Мета: визначення рівнів галектину-3 та TGF-β1, як маркерів фіброзоутворення, у дітей раннього віку, хворих на пієлонефрит на тлі міхурово-сечовідного рефлюксу (МСР).Пацієнти і методи. Обстежено 55 дітей раннього віку, хворих на пієлонефрит. Першу підгрупу склали 28 дітей з пієлонефритом без ознак МСР (первинний пієлонефрит — ППН), другу підгрупу — 27 дітей з пієлонефритом на тлі МСР (вторинний пієлонефрит — ВПН). Контрольну групу склали 24 практично здорові дитини. Вміст TGF- β1 та галектину-3 визначали імуноферментним методом (ELISA).Результати. Плазмова концентрація TGF- β1 та галектину-3 достовірно вища у дітей із пієлонефритом на тлі МСР, ніж у малюків із ППН, що вказує на можливе фіброзоутворення у нирках. Зі збільшенням тривалості пієлонефриту підвищувались рівні профібротичних показників. При ВПН у більшості дітей (81,4%) встановлено зменшення товщини паренхіми нирок за даними показників ренально-кортикального індексу, що свідчить про можливість зниження функціональної здатності нирок. Висока чутливість (TGF-β1 — 85%, галектин-3 — 92%) та специфічність (TGF-β1 — 67%, галектин-3 — 82%) обох показників при пієлонефриті на тлі МСР вказує на необхідність їх визначення у якості маркерів раннього фіброзоутворення.Висновки. Перспективним залишається подальше вивчення патогенетичних аспектів формування незворотних змін у нирках при пієлонефриті на тлі МСР у дітей раннього віку.Ключові слова: пієлонефрит, діти раннього віку, трансформуючий фактор росту β1, галектин-3., The objective. To determine the levels of TGF-B1 and galectin 3 pyelonephritis against the background of vesicle-urethral reflux.Patients and methods. The observation of 55 infants of patients with pyelonephritis who were on the treatment course in Khmelnytsky regional children's hospital. The first subgroup consisted of 28 children with pyelonephritis without evidence of vesiculo-urethrally reflux (primary pyelonephritis) and the second subgroup consisted of 27 young children of patients with pyelonephritis on the background of the vesiculo-urethrally reflux (secondary pyelonephritis). The control group consisted of 24 healthy children. The content of TGFB1 and galectin_3 was determined by enzyme immunoassay (ELISA).Results. Plasma concentration of transforming growth factor B1 and galectin-3 were significantly higher in children with pyelonephritis on the background of vesicoureteral reflux than in patients with primary pyelonephritis, indicating a possible fibrosis formation in the kidneys. It is established that with increasing length of pyelonephritis increased level profibrotic indicators. When SPN most children (81.4%) found the reduction of the thickness of the renal parenchyma according to the indicators of renal cortical index, which indicates the possibility of reducing the functional capacity of the kidneys. High sensitivity (TGF-B1 — 85%, the galectin-3 — 92%) and specificity (TGF-B1 — 67%, galectin-3 — 82%) for both indicators in pyelonephritis on the background of vesicoureteral refluxa indicates the need for a definition of these indicators in children of early age, as early markers of fibrosis formation.Conclusion. Perspective is further investigations of the pathogenetic aspects of the formation of irreversible changes in the kidney in pyelonephritis on the background of vesicoureteral reflux in children of early age.Key words: pyelonephritis, infants, transforming growth factor, galectin 3.

Aim: To determine the role of TAK1 in acute glomerular inflammation. Background(s): Activation of p38/JNK signalling promotes glomerular inflammation. TAK1 is an upstream kinase in the p38/JNK pathway. TAK1 regulates the response to a range of stimuli which promote glomerular inflammation (TNF-alpha, IL-1 and TLR ligands). Therefore, we investigated whether TAK1 activates p38/JNK signalling in kidney disease. Method(s): Global Tak1 gene deletion was induced in Tak1f/fERCre mice via tamoxifen, while Tak1f/fPodCre mice have conditional Tak1 deletion in podocytes. Accelerated anti-glomerular basement membrane (GBM) disease was induced in Tak1f/fERCre and Tak1f/f littermate controls (n = 8-11) and killed after 3 hr or 24 hr. Disease was also induced in Tak1f/fPodCre and Tak1f/f littermate controls (n = 5) and killed at 24 hr. Result(s): Compared to controls, global deletion of TAK1 had no effect upon the initial inflammatory response at 3 hr in terms of neutrophil and macrophage infiltration and platelet deposition. However, at 24 hr TAK1 deletion caused exacerbation of glomerular injury compared to littermate controls in terms of: proteinuria (11.9 +/- 5 vs 5.4 +/- 0.5 mg/mmol; P = 0.03); thrombosis (59 +/- 11% vs 8 +/- 3% glomeruli; P = 0.001); renal function (1.4-fold increase in serum cystatin c; P < 0.01); p38 and JNK activation (2-fold increase in glomerular p-p38+ and p-c-Jun+ cells; both P < 0.01). The neutrophil infiltrate had largely subsided in control mice at 24 hr; however, prominent neutrophil infiltration and platelet deposition was evident in mice with global TAK1 deletion at 24 hr. This exacerbation of glomerular damage was not due to a direct effect upon podocytes since Tak1f/fPodCre mice showed no difference compared to controls in proteinuria, p38/JNK activation or thrombosis at 24 hr. Conclusion(s): TAK1 is a negative regulator of glomerular p38/JNK signalling and acute renal inflammation, which may operate via limiting neutrophil-mediated glomerula

Hypothesis: This proposal will address the hypothesis that TGF signals in fibroblasts allow normal mammary gland development; and prevents breast cancer growth and progression. Specific Aims: We will (1) determine the effect of loss of TGF signaling within stromal cells on mammary gland development and (2) determine the effect of loss of TGF signaling within stromal cells on mammary carcinoma development. Study Design: This will be accomplished using transgenic mice with an inducible deletion of the type II TGF receptor (TGFBRII), which is required for TGF signaling. Cre recombinase driven by Fibroblast-Specific Protein (FSP1) and Pro-Collagen I 2 will delete TGFBRII within fibroblasts. To understand the normal biology of fibroblast TGF signaling, we will first examine mammary gland development in floxed TGFBRII, inducible fibroblast specific-Cre mice. Then we will examine carcinogenesis and tumor progression in floxed TGFBRII, inducible fibroblast-specific Cre, MMTV-PyVmT mice. This is the first proposed model to examine the biology of TGF signaling in fibroblasts in the intact mammary gland. Since TGF can promote or suppress cancer, understanding the roles of fibroblast TGF signaling in breast cancer will allow us to identify patients for which anti-TGF therapy is appropriate., The original document contains color images.

Treatment of pseudoglandular stage fetal lungs in vitro with the pan-retinoic acid receptor (pan-RAR) antagonist, BMS493, reduces retinoic acid receptor beta (Rarb) gene expression within the proximal bronchial tubules and increases explant bud formation. Treatment with retinoic acid (RA) increases Rarb expression and reduces explant bud formation through a signaling mechanism involving RARbeta. Together these data suggest that RA through RARbeta provides morphogenetic stabilizing activity to the proximal tubules during lung branching morphogenesis. Here we further investigate RA-mediated morphogenetic stabilization of the proximal respiratory tubules during fetal lung development. We demonstrate that Rarb isoform transcripts are the only known Rar transcripts to specifically localize to the proximal tubules and that RAREhsp68lacZ reporter transgene activity reveals endogenous RA signaling activity within these same proximal tubules. Furthermore, the expression patterns of the RA-producing enzyme retinaldehyde dehydrogenase 1 (Raldh1), as well as of transforming growth factor-3beta (Tgfb3), Foxa2, and the cystic fibrosis transmembrane conductance regulator (Cftr) within the proximal tubules are all altered by the application of either RA or BMS493 in vitro. We therefore discuss an interbud/proximal tubule signaling niche involving feedback between Rarb expression and Raldh1-mediated synthesis of RA. We suggest that this feedback favors interbud morphogenetic stability by increasing expression of morphoregulatory molecules such as TGFbeta3 and Foxa2, thus promoting bronchial tubule formation rather than continual budding and branching. The relationship between this RAR signaling center and the previously described distal bud signaling center is also addressed. © 2003 Elsevier Science Ireland Ltd. All rights reserved.

The capacity of macrophages to influence directly and indirectly fibrinolytic processes in atherosclerosis was studied using macrophages isolated from atherosclerotic plaques of patients undergoing surgical repair of distal aortic and femoral arteries. These cells were characterized by their morphology, adherence, esterase positivity, and expression of CD14 antigen. Production of plasminogen activator inhibitor type-1 (PAI-1) by plaque macrophages (6.7 +/- 2.7 ng/105 cells/24 hours [mean +/- SEM]) was significantly greater than PAI-1 production by blood monocytes isolated simultaneously from the same patients (1.8 +/- 1.5 ng/105 cells/24 hours). Production of tissue type plasminogen activator and urokinase type plasminogen activator by plaque macrophages was not augmented compared to blood monocytes. Conditioned medium from cultured plaque macrophages significantly increased production of PAI-1 by endothelial cells (85 +/- 11% above basal) and vascular smooth muscle cells (25 +/- 10%) in vitro. This response was significantly greater than the response to monocyte-conditioned medium (endothelial cells 38 +/- 11%, vascular smooth muscle cells 2.5 +/- 2.0%). Stimulation of endothelial cell PAI-1 production by macrophage- conditioned medium was partially inhibitable by a monoclonal antibody to transforming growth factor-beta. Tissue type plasminogen activator production by endothelial cells and vascular smooth muscle cells was not affected by plaque macrophage- or monocyte-conditioned medium. Urokinase type plasminogen activator production by endothelial cells and vascular smooth muscle cells was undetectable in control medium and was augmented to similar levels in response to plaque macrophage- and monocyte-conditioned media. These results demonstrate up-regulation of PAI-1 production by macrophages in atheromatous plaques and the capacity of soluble products from plaque macrophages to up- regulate PAI-1 production by endothelial cells and vascular smooth muscle cells

The recent purification, sequencing and molecular cloning of inhibin have led to its redefinition incorporating reference to its structure in order to avoid confusion with other proteins with inhibin-like activity. Thus, inhibin is now defined as a glycoprotein hormone consisting of two dissimilar disulphide linked subunits termed alpha and beta, which acts to inhibit the production and/or secretion of the gonadotrophins, preferentially that of FSH. The trophic action of FSH upon gonadal inhibin production has been confirmed in man and animals. On the other hand, the relative role of inhibin, the sex steroids and perhaps activin, in the physiological regulation of FSH remains unclear. The availability of inhibin radioimmunoassays and very recently, inhibin and activin produced by recombinant DNA technology (Mason et al., 1987), will allow such assessments and will clarify the physiological, pathological and potential therapeutic roles of inhibin. The unique ability of the inhibin subunits to form homo- or heterodimers with divergent biological activities and their homologies with other growth factors indicate that inhibin is a remarkable member of a family of proteins derived from a common ancestral gene. As such, the distinct possibility arises that proteins composed of inhibin alpha and beta subunits will also show growth stimulatory or inhibitory activities. As presented above, tentative evidence exists for such function in respect of human gestation, adrenal cortical function, haemopoietic cellular differentiation and the paracrine regulation of ovarian function. The study of inhibin and related proteins seems certain to have ramifications far beyond the field of reproductive endocrinology.

Treatment of pseudoglandular stage fetal lungs in vitro with the pan-retinoic acid receptor (pan-RAR) antagonist, BMS493, reduces retinoic acid receptor beta (Rarb) gene expression within the proximal bronchial tubules and increases explant bud formation. Treatment with retinoic acid (RA) increases Rarb expression and reduces explant bud formation through a signaling mechanism involving RARbeta. Together these data suggest that RA through RARbeta provides morphogenetic stabilizing activity to the proximal tubules during lung branching morphogenesis. Here we further investigate RA-mediated morphogenetic stabilization of the proximal respiratory tubules during fetal lung development. We demonstrate that Rarb isoform transcripts are the only known Rar transcripts to specifically localize to the proximal tubules and that RAREhsp68lacZ reporter transgene activity reveals endogenous RA signaling activity within these same proximal tubules. Furthermore, the expression patterns of the RA-producing enzyme retinaldehyde dehydrogenase 1 (Raldh1), as well as of transforming growth factor-3beta (Tgfb3), Foxa2, and the cystic fibrosis transmembrane conductance regulator (Cftr) within the proximal tubules are all altered by the application of either RA or BMS493 in vitro. We therefore discuss an interbud/proximal tubule signaling niche involving feedback between Rarb expression and Raldh1-mediated synthesis of RA. We suggest that this feedback favors interbud morphogenetic stability by increasing expression of morphoregulatory molecules such as TGFbeta3 and Foxa2, thus promoting bronchial tubule formation rather than continual budding and branching. The relationship between this RAR signaling center and the previously described distal bud signaling center is also addressed. © 2003 Elsevier Science Ireland Ltd. All rights reserved.

The capacity of macrophages to influence directly and indirectly fibrinolytic processes in atherosclerosis was studied using macrophages isolated from atherosclerotic plaques of patients undergoing surgical repair of distal aortic and femoral arteries. These cells were characterized by their morphology, adherence, esterase positivity, and expression of CD14 antigen. Production of plasminogen activator inhibitor type-1 (PAI-1) by plaque macrophages (6.7 +/- 2.7 ng/105 cells/24 hours [mean +/- SEM]) was significantly greater than PAI-1 production by blood monocytes isolated simultaneously from the same patients (1.8 +/- 1.5 ng/105 cells/24 hours). Production of tissue type plasminogen activator and urokinase type plasminogen activator by plaque macrophages was not augmented compared to blood monocytes. Conditioned medium from cultured plaque macrophages significantly increased production of PAI-1 by endothelial cells (85 +/- 11% above basal) and vascular smooth muscle cells (25 +/- 10%) in vitro. This response was significantly greater than the response to monocyte-conditioned medium (endothelial cells 38 +/- 11%, vascular smooth muscle cells 2.5 +/- 2.0%). Stimulation of endothelial cell PAI-1 production by macrophage- conditioned medium was partially inhibitable by a monoclonal antibody to transforming growth factor-beta. Tissue type plasminogen activator production by endothelial cells and vascular smooth muscle cells was not affected by plaque macrophage- or monocyte-conditioned medium. Urokinase type plasminogen activator production by endothelial cells and vascular smooth muscle cells was undetectable in control medium and was augmented to similar levels in response to plaque macrophage- and monocyte-conditioned media. These results demonstrate up-regulation of PAI-1 production by macrophages in atheromatous plaques and the capacity of soluble products from plaque macrophages to up- regulate PAI-1 production by endothelial cells and vascular smooth muscle cells

The recent purification, sequencing and molecular cloning of inhibin have led to its redefinition incorporating reference to its structure in order to avoid confusion with other proteins with inhibin-like activity. Thus, inhibin is now defined as a glycoprotein hormone consisting of two dissimilar disulphide linked subunits termed alpha and beta, which acts to inhibit the production and/or secretion of the gonadotrophins, preferentially that of FSH. The trophic action of FSH upon gonadal inhibin production has been confirmed in man and animals. On the other hand, the relative role of inhibin, the sex steroids and perhaps activin, in the physiological regulation of FSH remains unclear. The availability of inhibin radioimmunoassays and very recently, inhibin and activin produced by recombinant DNA technology (Mason et al., 1987), will allow such assessments and will clarify the physiological, pathological and potential therapeutic roles of inhibin. The unique ability of the inhibin subunits to form homo- or heterodimers with divergent biological activities and their homologies with other growth factors indicate that inhibin is a remarkable member of a family of proteins derived from a common ancestral gene. As such, the distinct possibility arises that proteins composed of inhibin alpha and beta subunits will also show growth stimulatory or inhibitory activities. As presented above, tentative evidence exists for such function in respect of human gestation, adrenal cortical function, haemopoietic cellular differentiation and the paracrine regulation of ovarian function. The study of inhibin and related proteins seems certain to have ramifications far beyond the field of reproductive endocrinology.

Aim: To define the role of Mast Cells (MCs) in renal fibrosis induced by unilateral ureteric obstruction (UUO). Background(s): MCs are pluripotent innate immune cells, which after degranulation release cytokines, chemokines and pro-fibrotic factors including transforming growth factor beta (TGFbeta). Method(s): To define a role for MCs in renal fibrosis, we performed UUO on C57BL/6 wild type (WT) and mast cell deficient, KitWsh/Wsh mice. Experiments ended 7 days later. We measured collagen deposition, intrarenal mRNA expression of pro-fibrotic factors, TGFbeta and smooth muscle actin (alphaSMA) and interstitial leukocyte recruitment. To confirm these results were exclusively due to MC deficiency we measured renal fibrosis in KitWsh/Wsh mice reconstituted with MCs from WT mice. Finally we assessed renal fibrosis after administration of sodium chromoglycate, a mast cell stabilizer. NEPHROLOGY 2011; 16 (Suppl. 1), 44-69 doi:10.1111/j.1440-1797.2011.01491.x Results: Compared to WT mice, renal fibrosis was decreased in KitWsh/Wsh mice 7 days after UUO. Histological assessment demonstrated decreased collagen deposition (12.7 +/- 1.2 vs. 9.9 +/- 0.9%, P < 0.05). Intrarenal TGFbeta (1.0 +/- 0.1 vs. 0.7 +/- 0.1, P < 0.05) and alphaSMA (1.0 +/- 0.1 vs. 0.7 +/- 0.1, P < 0.05) expression was also decreased in KitWsh/Wsh mice. Fewer interstitial macrophages (score 2.4 +/- 0.1 vs. 1.6 +/- 0.1 P < 0.001) and CD4+T cells (7.1 +/- 0.4 vs. 4.1 +/- 0.6 cells/high power field, P < 0.001) were seen in KitWsh/Wsh mice. Collagen deposition (17.0 +/- 2.5 vs. 9.1 +/- 1.2%, P < 0.05) and intrarenal mRNA expression of TGFbeta and alphaSMA was decreased in (unreconstituted) KitWsh/Wsh mice compared to KitWsh/Wsh mice reconstituted with MCs from WT mice. Pre-emptive administration of sodium chromoglycate to WT mice resulted in decreased collagen deposition (16.3 +/- 1.3 vs. 10.5 +/- 0.7%, P < 0.01) and decreased intrarenal mRNA expression of TGFbeta (1.0 +/- 0.1 vs. 0.7 +/- 0.1, P < 0.01) and a

Aim: To define the role of Mast Cells (MCs) in renal fibrosis induced by unilateral ureteric obstruction (UUO). Background(s): MCs are pluripotent innate immune cells, which after degranulation release cytokines, chemokines and pro-fibrotic factors including transforming growth factor beta (TGFbeta). Method(s): To define a role for MCs in renal fibrosis, we performed UUO on C57BL/6 wild type (WT) and mast cell deficient, KitWsh/Wsh mice. Experiments ended 7 days later. We measured collagen deposition, intrarenal mRNA expression of pro-fibrotic factors, TGFbeta and smooth muscle actin (alphaSMA) and interstitial leukocyte recruitment. To confirm these results were exclusively due to MC deficiency we measured renal fibrosis in KitWsh/Wsh mice reconstituted with MCs from WT mice. Finally we assessed renal fibrosis after administration of sodium chromoglycate, a mast cell stabilizer. NEPHROLOGY 2011; 16 (Suppl. 1), 44-69 doi:10.1111/j.1440-1797.2011.01491.x Results: Compared to WT mice, renal fibrosis was decreased in KitWsh/Wsh mice 7 days after UUO. Histological assessment demonstrated decreased collagen deposition (12.7 +/- 1.2 vs. 9.9 +/- 0.9%, P < 0.05). Intrarenal TGFbeta (1.0 +/- 0.1 vs. 0.7 +/- 0.1, P < 0.05) and alphaSMA (1.0 +/- 0.1 vs. 0.7 +/- 0.1, P < 0.05) expression was also decreased in KitWsh/Wsh mice. Fewer interstitial macrophages (score 2.4 +/- 0.1 vs. 1.6 +/- 0.1 P < 0.001) and CD4+T cells (7.1 +/- 0.4 vs. 4.1 +/- 0.6 cells/high power field, P < 0.001) were seen in KitWsh/Wsh mice. Collagen deposition (17.0 +/- 2.5 vs. 9.1 +/- 1.2%, P < 0.05) and intrarenal mRNA expression of TGFbeta and alphaSMA was decreased in (unreconstituted) KitWsh/Wsh mice compared to KitWsh/Wsh mice reconstituted with MCs from WT mice. Pre-emptive administration of sodium chromoglycate to WT mice resulted in decreased collagen deposition (16.3 +/- 1.3 vs. 10.5 +/- 0.7%, P < 0.01) and decreased intrarenal mRNA expression of TGFbeta (1.0 +/- 0.1 vs. 0.7 +/- 0.1, P < 0.01) and a

While the central nervous system (CNS) was once thought to be excluded from surveillance by immune cells, a concept known as “immune privilege,” it is now clear that immune responses do occur in the CNS—giving rise to the field of neuroimmunology. These CNS immune responses can be driven by endogenous (glial) and/or exogenous (peripheral leukocyte) sources and can serve either productive or pathological roles. Recent evidence from mouse models supports the notion that infiltration of peripheral monocytes/macrophages limits progression of Alzheimer's disease pathology and militates against West Nile virus encephalitis. In addition, infiltrating T lymphocytes may help spare neuronal loss in models of amyotrophic lateral sclerosis. On the other hand, CNS leukocyte penetration drives experimental autoimmune encephalomyelitis (a mouse model for the human demyelinating disease multiple sclerosis) and may also be pathological in both Parkinson's disease and human immunodeficiency virus encephalitis. A critical understanding of the cellular and molecular mechanisms responsible for trafficking of immune cells from the periphery into the diseased CNS will be key to target these cells for therapeutic intervention in neurodegenerative diseases, thereby allowing neuroregenerative processes to ensue.

While the central nervous system (CNS) was once thought to be excluded from surveillance by immune cells, a concept known as “immune privilege,” it is now clear that immune responses do occur in the CNS—giving rise to the field of neuroimmunology. These CNS immune responses can be driven by endogenous (glial) and/or exogenous (peripheral leukocyte) sources and can serve either productive or pathological roles. Recent evidence from mouse models supports the notion that infiltration of peripheral monocytes/macrophages limits progression of Alzheimer's disease pathology and militates against West Nile virus encephalitis. In addition, infiltrating T lymphocytes may help spare neuronal loss in models of amyotrophic lateral sclerosis. On the other hand, CNS leukocyte penetration drives experimental autoimmune encephalomyelitis (a mouse model for the human demyelinating disease multiple sclerosis) and may also be pathological in both Parkinson's disease and human immunodeficiency virus encephalitis. A critical understanding of the cellular and molecular mechanisms responsible for trafficking of immune cells from the periphery into the diseased CNS will be key to target these cells for therapeutic intervention in neurodegenerative diseases, thereby allowing neuroregenerative processes to ensue.

Through differential screening between oyster families selected for high and low summer survival, we have characterized a new transforming growth factor-beta (TGF-beta) superfamily member. This novel factor, named oyster-gonadal-TGF beta-like (og-TGF beta-like), is synthesized as a 307 amino acid precursor and displays 6 of the 7 characteristic cysteine residues of the C-terminal, mature peptide. Sequence comparison revealed that og-TGF beta-like has a low percentage of identity with other known TGF-beta superfamily members, suggesting that og-TGF beta-like is a derived member of this large superfamily. Real-time PCR (RT-PCR) analysis in different oyster tissues showed that og-TGF beta-like is specifically expressed in both male and female gonads, at distinct levels according to the reproductive stage. Og-TGF beta-like relative expression was the lowest at the initiation of the reproductive cycle and increased as maturation proceeded to achieve a maximal level in fully mature female and male oysters. In situ hybridisation demonstrated that expression was exclusively detected in the somatic cells surrounding oocytes and spermatocytes. The role of this newly-characterized TGF beta member in the reproduction of cupped oyster is discussed in regard to the specificity and the localization of its expression, which singularly contrasts with the pleiotropic roles in a variety of physiological processes commonly ascribed to most TGF-beta family members identified so far. (C) 2007 Elsevier B.V. All rights reserved.

Through differential screening between oyster families selected for high and low summer survival, we have characterized a new transforming growth factor-beta (TGF-beta) superfamily member. This novel factor, named oyster-gonadal-TGF beta-like (og-TGF beta-like), is synthesized as a 307 amino acid precursor and displays 6 of the 7 characteristic cysteine residues of the C-terminal, mature peptide. Sequence comparison revealed that og-TGF beta-like has a low percentage of identity with other known TGF-beta superfamily members, suggesting that og-TGF beta-like is a derived member of this large superfamily. Real-time PCR (RT-PCR) analysis in different oyster tissues showed that og-TGF beta-like is specifically expressed in both male and female gonads, at distinct levels according to the reproductive stage. Og-TGF beta-like relative expression was the lowest at the initiation of the reproductive cycle and increased as maturation proceeded to achieve a maximal level in fully mature female and male oysters. In situ hybridisation demonstrated that expression was exclusively detected in the somatic cells surrounding oocytes and spermatocytes. The role of this newly-characterized TGF beta member in the reproduction of cupped oyster is discussed in regard to the specificity and the localization of its expression, which singularly contrasts with the pleiotropic roles in a variety of physiological processes commonly ascribed to most TGF-beta family members identified so far. (C) 2007 Elsevier B.V. All rights reserved.

Through differential screening between oyster families selected for high and low summer survival, we have characterized a new transforming growth factor-beta (TGF-beta) superfamily member. This novel factor, named oyster-gonadal-TGF beta-like (og-TGF beta-like), is synthesized as a 307 amino acid precursor and displays 6 of the 7 characteristic cysteine residues of the C-terminal, mature peptide. Sequence comparison revealed that og-TGF beta-like has a low percentage of identity with other known TGF-beta superfamily members, suggesting that og-TGF beta-like is a derived member of this large superfamily. Real-time PCR (RT-PCR) analysis in different oyster tissues showed that og-TGF beta-like is specifically expressed in both male and female gonads, at distinct levels according to the reproductive stage. Og-TGF beta-like relative expression was the lowest at the initiation of the reproductive cycle and increased as maturation proceeded to achieve a maximal level in fully mature female and male oysters. In situ hybridisation demonstrated that expression was exclusively detected in the somatic cells surrounding oocytes and spermatocytes. The role of this newly-characterized TGF beta member in the reproduction of cupped oyster is discussed in regard to the specificity and the localization of its expression, which singularly contrasts with the pleiotropic roles in a variety of physiological processes commonly ascribed to most TGF-beta family members identified so far. (C) 2007 Elsevier B.V. All rights reserved.

The isoflavonoid genistein inhibits mitosis and increases apoptosis in a variety of tumour cell lines in vitro, and may exert anticarcinogenic effects in vivo. To assess its effects on the colon, rats were fed a semi-synthetic control diet, or similar diets enriched with genistein (0.25 g/kg), either as the pure isoflavone or as part of a soya protein isolate, for 7 days before receiving subcutaneous injections of saline or 1,2-dimethylhydrazine (DMH). After 48 h, rats given saline were killed and samples of their small and large intestinal mucosa were obtained for assessment of crypt cell mitosis and apoptosis by visual analysis of isolated intact crypts. Rats given DMH were fed control diet and killed after 48 h for assessment of crypt cytokinetics or maintained for 42 days then killed and their colonic mucosa analysed for aberrant crypt foci (ACF). Two further groups were given control diet before DMH, followed by the genistein or soya-based diet for 42 days before assessment of ACF. Neither genistein nor soya protein isolate had a significant effect on crypt cell mitosis or apoptosis in untreated rats, or on the proliferative response to treatment with DMH. However, consumption of pure genistein or the soya protein isolate before treatment with DMH was associated with a 3-fold (P < 0.001) or 2-fold (P < 0.05) increase, respectively, in ACF in the distal colon. There was no significant effect of genistein or soya protein isolate given after DMH treatment. We conclude that genistein has no detectable effect on colonic crypt mitosis or apoptosis in the rat in vivo, but that it promotes induction of ACF by an as yet undefined mechanism when fed immediately before treatment with DMH.

Transforming growth factor-beta (TGF-beta) is a cytokine with antiatherogenic propretis. The active form of TGF-beta inhibits the activation of endothelial cells, and acts on vascular smooth muscle cells by inhibiting their migration, dediferentiation and proliferation. Lipoprotein (a) i PAI-1 interfere with plasmin-mediated TGF-beta activation by inhibiting the conversion of plasminogen to plasmin. Genetic polymorphism in the TGF beta(1) gene promoter were shown to be correlated with total TGF-beta concentrations, as measured by ELISA after acetic acid/urea activation. Althought many patients with coronary artery disease (CAD) have total TGF-beta concentration below the detection limit, the ability of total TGF-beta assay in discriminating between patients with CAD and individuals with normal coronary arteries is rather limited. Very significant inverse relationship between the concentration of active TGF-beta and CAD was found in several retrospective studies. Minor overlapping between the values in CAD patients and controls was obtained, demostrating the potential of the active TGF-beta as a new marker of CAD, superior to any of the conventional risk factors.