High iNOS mRNA and protein levels during early third trimester suggest a role for NO in prelabor cervical ripening in the bovine

Aalberts, Marian; Dissel‐Emiliani, F. M. F. Van; Tol, Helena T A van; Taverne, M.A.M.; Breeveld-Dwarkasing, V.N.A.

doi:10.1002/mrd.20546

Cited by 19 publications

(12 citation statements)

References 42 publications

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“…According to this principle, we could type the NOS easily. Here, we used the NOS examining kit (The first substation of Jiang Cheng bioengineering institute, Nanking) to detect the serum iNOS activity in differential samples, the experimental process was performed according to the instruction manual and previously reported [23]. Briefly, under the 530 nm wavelength, we first detected the absorbance of differential samples with the chromatometry method, then according the following formula to calculate the iNOS activity.…”

Section: Methodsmentioning

confidence: 99%

Analysis of Inducible Nitric Oxide Synthase Gene Polymorphisms in Vitiligo in Han Chinese People

Zhang

et al. 2011

PLoS ONE

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BackgroundVitiligo is a chronic depigmented skin disorder with regional melanocytes depletion. The pathogenesis was not completely clarified. Recently, more and more evidence suggested that polymorphisms of some genes are associated with vitiligo risk. Here, we want to examine the association between the inducible nitric oxide synthase (iNOS) gene polymorphisms and the risk of vitiligo in Chinese populations.Methods and Principal FindingsIn a hospital-based case-control study of 749 patients with vitiligo and 763 age- and sex-matched healthy controls, three polymorphisms of iNOS gene were genotyped by using the PCR-restriction fragment length polymorphism (PCR-RFLP) and mutagenically separated PCR (MS-PCR) methods, respectively. We found the iNOS-954 polymorphism was associated with a significantly higher risk of vitiligo (adjusted OR = 1.36, 95% CI = 1.02–1.81). Furthermore, this association is more pronounced in vulgaris vitiligo, active vitiligo and vitiligo without other autoimmune diseases in the stratification study. Analysis of haplotypes showed increased risk for the C -1173 C -954 CEx 16+14 (OR = 1.44, 95% CI = 1.01–1.74). In addition, the serum iNOS activity is significantly associated with iNOS-954 combined genotype (GC+CC) and is much higher in vitiligo patients than in the controls (P<0.01). Logistic regression analysis of iNOS activity showed increased risk between higher activity and iNOS-954 G→C variant genotype carriers (Ptrend<0.001).Conclusions and Significance INOS gene polymorphisms may play an important role in the genetic susceptibility to the development of vitiligo.

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Section: Methodsmentioning

confidence: 99%

Analysis of Inducible Nitric Oxide Synthase Gene Polymorphisms in Vitiligo in Han Chinese People

Zhang

et al. 2011

PLoS ONE

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“…2002). An earlier study of the cervix of pregnant and parturient cows showed that the expression of GAPDH was highly correlated ( r = 0.9) with the expression of actin‐ β (Aalberts et al. 2007), therefore GAPDH was regarded as a household gene and used as reference.…”

Section: Methodsmentioning

confidence: 99%

“…Quantification was performed using a mathematical model for the relative expression ratio in real-time PCR, the 2 -DDCt -method (Livak and Schmittgen 2001;De Preter et al 2002). An earlier study of the cervix of pregnant and parturient cows showed that the expression of GAP-DH was highly correlated (r = 0.9) with the expres-sion of actin-b (Aalberts et al 2007), therefore GAPDH was regarded as a household gene and used as reference.…”

Section: Rt-pcr For Sma Connexin-43 Vimentin and Desminmentioning

confidence: 99%

“…The distribution of smooth muscle cells over an outer muscular layer and within the deeper part of the stromal layer has been demonstrated using immunohistochemical staining for smooth muscle actin α (SMA) (Breeveld‐Dwarkasing et al. 2000; Aalberts et al. 2007).…”

Section: Introductionmentioning

confidence: 99%

“…The bovine cervix has a high muscular content (Hafez 1973). The distribution of smooth muscle cells over an outer muscular layer and within the deeper part of the stromal layer has been demonstrated using immunohistochemical staining for smooth muscle actin a (SMA) (Breeveld-Dwarkasing et al 2000;Aalberts et al 2007). The smooth muscle cells within the stromal layer are grouped in small bundles with a random orientation, embedded in a connective tissue meshwork (Breeveld-Dwarkasing et al 2000) and it has been observed that these smooth muscle bundles contract in response to oxytocin in vitro (van Engelen et al 2007).…”

Section: Introductionmentioning

confidence: 99%

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Smooth Muscle Cells of the Bovine Cervical Stroma may have a Secretory, rather than a Contractile Function during Parturition

Engelen¹,

Breeveld-Dwarkasing²,

Everts³

et al. 2009

Reprod Domestic Animals

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The bovine cervix contains a large amount of smooth muscle cells distributed over an outer muscular layer and within a stromal layer. The stromal layer exhibits no electromyographic (EMG) activity at parturition. This leads to the question whether the stromal smooth muscle cells of the bovine cervix are prepared to contract with parturition, or whether they have another function. To this end, cervical biopsies were repeatedly taken from 10 pregnant cows at day-185 and -275 of gestation, at spontaneous, uncomplicated calving and at 30 days after calving. The smooth muscle bundles of the stroma were immunohistochemically analysed (n = 5) with regard to their integrity and cellular density, and the degree of staining for connexin-43, smooth muscle actin alpha (SMA), desmin and vimentin. Additionally, the mRNA expression for connexin-43, SMA, desmin and vimentin was determined with RT-PCR (n = 5). The smooth muscle tissue was arranged in bundles, also at parturition. However, the cellular density of these bundles and the SMA mRNA expression were decreased at parturition. Additionally, the SMA staining and connexin-43 expression and staining remained constant during pregnancy and at parturition. This might indicate that stromal smooth muscle cells are not prepared to contract with parturition, in contrast to the myometrial smooth muscle cells. The smooth muscle cells, stained for SMA, also expressed vimentin, and the proportion of co-expression was increased at day-275 of pregnancy. This suggests that the stromal smooth muscle cells predominantly have a secretory function in cows.

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MMP‐2 expression precedes the final ripening process of the bovine cervix

Engelen

Breeveld-Dwarkasing

Taverne

et al. 2008

Molecular Reproduction Devel

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Collagen is denatured in the gradual cervical ripening process during late pregnancy, already before the onset of final cervical ripening at parturition. Matrix Metallo Proteinases (MMPs) might be responsible for this process. To investigate the presence and potential function of MMPs at the different stages of the ripening process, serial cervical biopsies were obtained from 10 cows at Days 185 and 275 of pregnancy (approximately 5 days before calving), at parturition and at 30 days after parturition. The mRNA and protein expression of MMP-1, MMP-2, and MMP-9 and of the tissue inhibitors of MMPs (TIMP)-1 and TIMP-2 were semi-quantitatively determined using RT-PCR, respectively, zymography, Westernblot, and ELISA techniques and the localization of MMP-2 protein and presence of granulocytes by immunohistochemistry and Luna staining. At parturition compared to 185 days pregnancy the MMP-1 protein expression and the numbers of granulocytes were significantly increased by 3 and 26-fold respectively. MMP-2 mRNA and protein expression had already increased 2.5 (P < 0.05) and twofold (P < 0.05) at 5 days before parturition, prior to final ripening. At that time, MMP-2 was present in smooth muscle cells and extra cellular matrix. TIMP-1 mRNA expression was significantly increased at parturition and TIMP-2 mRNA expression peaked at 5 days before parturition. The increased expression of MMP-2 at 5 days before parturition, suggests that in the cow MMP-2 is responsible for collagen denaturation in the last part of gradual cervical ripening, while MMP-1 and MMP-9 are only active during the final cervical ripening process at parturition.

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High iNOS mRNA and protein levels during early third trimester suggest a role for NO in prelabor cervical ripening in the bovine

Cited by 19 publications

References 42 publications

Analysis of Inducible Nitric Oxide Synthase Gene Polymorphisms in Vitiligo in Han Chinese People

Analysis of Inducible Nitric Oxide Synthase Gene Polymorphisms in Vitiligo in Han Chinese People

Smooth Muscle Cells of the Bovine Cervical Stroma may have a Secretory, rather than a Contractile Function during Parturition

MMP‐2 expression precedes the final ripening process of the bovine cervix

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