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Various local and systemic factors compromise oral wound healing and may lead to wound dehiscence, inflammation, or ulcers. Currently, there is a lack of topical therapeutical options. Thus, this study aimed to investigate the effect of

The binding of nitric oxide (NO) to heme in the β1 subunit of soluble guanylyl cyclase (sGC) activates both the heterodimeric α1β1 and α2β1 isoforms of the enzyme, leading to the increased production of cGMP from GTP. In cultured human mast cells, exogenous NO is able to inhibit mast cell degranulation via NO-cGMP signaling. However, under inflammatory oxidative or nitrosative stress, sGC becomes insensitive to NO. The occurrence of mast cells in healthy and inflamed human tissues and the in vivo expression of the α1 and β1 subunits of sGC in human mast cells during inflammation remain largely unresolved and were investigated here. Using peroxidase and double immunohistochemical incubations, no mast cells were found in healthy dental pulp, whereas the inflammation of dental pulp initiated the occurrence of several mast cells expressing the α1 and β1 subunits of sGC. Since inflammation-induced oxidative and nitrosative stress oxidizes Fe2+ to Fe3+ in the β1 subunit of sGC, leading to the desensitization of sGC to NO, we hypothesize that the NO- and heme-independent pharmacological activation of sGC in mast cells may be considered as a regulatory strategy for mast cell functions in inflamed human dental pulp.

VEGF signaling regulated by the vascular endothelial growth factor receptor 2 (VEGFR2) plays a decisive role in tumor angiogenesis, initiation and progression in several tumors including HNSCC. However, the impact of HPV-status on the expression of VEGFR2 in OPSCC has not yet been investigated, although HPV oncoproteins E6 and E7 induce VEGF-expression. In a series of 56 OPSCC with known HPV-status, VEGFR2 expression patterns were analyzed both in blood vessels from tumor-free and tumor-containing regions and within tumor cells by immunohistochemistry using densitometry. Differences in subcellular colocalization of VEGFR2 with endothelial, tumor and stem cell markers were determined by double-immunofluorescence imaging. Immunohistochemical results were correlated with clinicopathological data. HPV-infection induces significant downregulation of VEGFR2 in cancer cells compared to HPV-negative tumor cells (p = 0.012). However, with respect to blood vessel supply, the intensity of VEGFR2 staining differed only in HPV-positive OPSCC and was upregulated in the blood vessels of tumor-containing regions (p &lt, 0.0001). These results may suggest different routes of VEGFR2 signaling depending on the HPV-status of the OPSCC. While in HPV-positive OPSCC, VEGFR2 might be associated with increased angiogenesis, in HPV-negative tumors, an autocrine loop might regulate tumor cell survival and invasion.

Background The primary dentin, secondary dentin, and reactive tertiary dentin are formed by terminal differentiated odontoblasts, whereas atubular reparative tertiary dentin is formed by odontoblast-like cells. Odontoblast-like cells differentiate from pulpal stem cells, which express the neural stem cell markers nestin, S100β, Sox10, and P0. The denticle (pulp stone) is an unique mineralized extracellular matrix that frequently occurs in association with the neurovascular structures in the dental pulp. However, to date, the cellular origin of denticles in human dental pulp is unclear. In addition, the non-collagenous extracellular dentin matrix proteins dentin matrix protein 1 (DMP1), dentin sialoprotein (DSP), and dentin phosphoprotein (DPP) have been well characterized in the dentin matrix, whereas their role in the formation and mineralization of the denticle matrix remains to be clarified. Methods To characterize the formation of denticle, healthy human third molars (n = 59) were completely sectioned and evaluated by HE staining in different layers at 720 µm intervals. From these samples, molars with (n = 5) and without denticles (n = 8) were selected. Using consecutive cryo-sections from a layer containing denticles of different sizes, we examined DMP1, DSP, and DPP in denticle lining cells and tested their co-localizations with the glial stem cell markers nestin, S100β, Sox10, and P0 by quantitative and double staining methods. Results DMP1, DSP and DPP were found in odontoblasts, whereas denticle lining cells were positive only for DMP1 and DSP but not for DPP. Nestin was detected in both odontoblasts and denticle lining cells. S100β, Sox10, and P0 were co-localized with DMP1 and DSP in different subpopulations of denticle lining cells. Conclusions The co-localization of S100β, Sox10, and P0 with DMP1 and DSP in denticle lining cells suggest that denticle lining cells are originated from glial and/or endoneurial mesenchymal stem cells which are involved in biomineralization of denticle matrix by secretion of DMP1 and DSP. Since denticles are atubular compared to primary, secondary, reactionary tertiary dentin and denticle formed by odontoblasts, our results suggest that DPP could be one of the proteins involved in the complex regulation of dentinal tubule formation.

The macroscopic and microscopic anatomy of the oral cavity is complex and unique in the human body. Soft-tissue structures are in close interaction with mineralized bone, but also dentine, cementum and enamel of our teeth. These are exposed to intense mechanical and chemical stress as well as to dense microbiologic colonization. Teeth are susceptible to damage, most commonly to caries, where microorganisms from the oral cavity degrade the mineralized tissues of enamel and dentine and invade the soft connective tissue at the core, the dental pulp. However, the pulp is well-equipped to sense and fend off bacteria and their products and mounts various and intricate defense mechanisms. The front rank is formed by a layer of odontoblasts, which line the pulp chamber towards the dentine. These highly specialized cells not only form mineralized tissue but exert important functions as barrier cells. They recognize pathogens early in the process, secrete antibacterial compounds and neutralize bacterial toxins, initiate the immune response and alert other key players of the host defense. As bacteria get closer to the pulp, additional cell types of the pulp, including fibroblasts, stem and immune cells, but also vascular and neuronal networks, contribute with a variety of distinct defense mechanisms, and inflammatory response mechanisms are critical for tissue homeostasis. Still, without therapeutic intervention, a deep carious lesion may lead to tissue necrosis, which allows bacteria to populate the root canal system and invade the periradicular bone via the apical foramen at the root tip. The periodontal tissues and alveolar bone react to the insult with an inflammatory response, most commonly by the formation of an apical granuloma. Healing can occur after pathogen removal, which is achieved by disinfection and obturation of the pulp space by root canal treatment. This review highlights the various mechanisms of pathogen recognition and defense of dental pulp cells and periradicular tissues, explains the different cell types involved in the immune response and discusses the mechanisms of healing and repair, pointing out the close links between inflammation and regeneration as well as between inflammation and potential malignant transformation.

Nitric oxide (NO) binds to soluble guanylyl cyclase (sGC), activates it in a reduced oxidized heme iron state, and generates cyclic Guanosine Monophosphate (cGMP), which results in vasodilatation and inhibition of osteoclast activity. In inflammation, sGC is oxidized and becomes insensitive to NO. NO- and heme-independent activation of sGC requires protein expression of the &alpha, 1- and &beta, 1-subunits. Inflammation of the periodontium induces the resorption of cementum by cementoclasts and the resorption of the alveolar bone by osteoclasts, which can lead to tooth loss. As the presence of sGC in cementoclasts is unknown, we investigated the &alpha, 1-subunits of sGC in cementoclasts of healthy and inflamed human periodontium using double immunostaining for CD68 and cathepsin K and compared the findings with those of osteoclasts from the same sections. In comparison to cementoclasts in the healthy periodontium, cementoclasts under inflammatory conditions showed a decreased staining intensity for both &alpha, 1-subunits of sGC, indicating reduced protein expression of these subunits. Therefore, pharmacological activation of sGC in inflamed periodontal tissues in an NO- and heme-independent manner could be considered as a new treatment strategy to inhibit cementum resorption.

The Merkel cell-neurite complex initiates the perception of touch and mediates Aβ slowly adapting type I responses. Lichen planus is a chronic inflammatory autoimmune disease with T-cell-mediated inflammation, whereas hyperkeratosis is characterized with or without epithelial dysplasia in the oral mucosa. To determine the effects of lichen planus and hyperkeratosis on the Merkel cell-neurite complex, healthy oral mucosal epithelium and lesional oral mucosal epithelium of lichen planus and hyperkeratosis patients were stained by immunohistochemistry (the avidin-biotin-peroxidase complex and double immunofluorescence methods) using pan cytokeratin, cytokeratin 20 (K20, a Merkel cell marker), and neurofilament 200 (NF200, a myelinated Aβ- and Aδ-nerve fibre marker) antibodies. NF200-immunoreactive (ir) nerve fibres in healthy tissues and in the lesional oral mucosa epithelium of lichen planus and hyperkeratosis were counted and statistically analysed. In the healthy oral mucosa, K20-positive Merkel cells with and without close association to the intraepithelial NF200-ir nerve fibres were detected. In the lesional oral mucosa of lichen planus and hyperkeratosis patients, extremely rare NF200-ir nerve fibres were detected only in the lamina propria. Compared with healthy tissues, lichen planus and hyperkeratosis tissues had significantly decreased numbers of NF200-ir nerve fibres in the oral mucosal epithelium. Lichen planus and hyperkeratosis were associated with the absence of Aβ-nerve endings in the oral mucosal epithelium. Thus, we conclude that mechanosensation mediated by the Merkel cell-neurite complex in the oral mucosal epithelium is impaired in lichen planus and hyperkeratosis.

Introduction Odontoblasts are terminally differentiated cells of ectomesenchymal origin that produce the dentin. Differentiated odontoblasts cannot be identified yet by a single phenotypic marker protein; therefore, a combination of markers is currently used. Up-regulation of the cyclin-dependent kinase inhibitor p27 Kip1 has been associated with exit from the cell cycle and terminal differentiation of mammalian cells. Immunoreactivity for p27 Kip1 protein was shown in many adult mouse tissues, but no information is available on the expression of p27 Kip1 in mammalian dental pulp. Methods Healthy and carious adult human molars with reparative dentin formation were decalcified, cryoprotected, frozen embedded, and frozen sectioned. The expression of p27 Kip1 and nestin in cells of adult human dental pulp was analyzed by immunohistochemistry using free floating sections. Results p27 Kip1 showed strong nuclear expression in many differentiated human molar odontoblasts at the odontoblastic layer. Most cells of the cell-rich zone displayed low levels of p27 Kip1 despite the fact that preodontoblasts localized in the cell-rich zone of the subodontoblastic layer have been identified as quiescent cells. The nuclear expression of p27 Kip1 in stromal cells of the dental pulp was variable, indicating that subpopulations of these cells were in distinct states of differentiation. Odontoblasts generating reparative dentin showed comparable nuclear expression of p27 Kip1 in comparison with odontoblasts synthesizing primary/secondary dentin. This result indicates that odontoblasts synthesizing primary/secondary or reparative dentin exhibit a similar differentiation status. Conclusions Our findings show that increased expression of nuclear p27 Kip1 occurred during differentiation from preodontoblasts to odontoblasts in adult healthy and carious molars. p27 Kip1 can be used as a novel nuclear marker protein for differentiated human odontoblasts in vivo .

Aims/hypothesis Pathological cardiac hypertrophy is an early phenotype in both types 1 and 2 diabetes. The primary stimulus for hypertrophic growth in diabetes is yet unknown and may involve neurohumoral stimulation of Gq-coupled receptors as well as direct glucose-dependent mechanisms. To discriminate between these hypertrophic stimuli we analyzed hypertrophic signalling pathways in wildtype and Gα 11 -knockout mice. Methods Experimental diabetes was induced in wildtype and knockout mice by intraperitoneal injection of streptozotocin. 8weeks after induction of diabetes myocardial function and structure was assessed by echocardiography before sacrifice. To identify prohypertrophic signalling pathways expression and translocation of protein kinase C isoforms α, β II , δ, e and ζ were analyzed by immunohistochemical staining and immunoblot analysis after tissue fractionation. Changes in calcineurin signalling were identified by immunoblot analysis and functional assays. Expression levels of transcription factors GATA4 and NF-κB were quantified by real-time RT-PCR. Results Diabetic wildtype mice developed myocardial hypertrophy with preserved cardiac function. Calcineurin signalling was not different between the two groups. However, diabetic wildtype mice showed increased protein levels of PKC-α and PKC-ζ, translocation of PKC-α, -δ and -e to cellular membranes and higher levels of NF-κB expression. In contrast, diabetic Gα 11 -knockout mice showed no altered phenotype and no changes in NF-κB or PKC expression, although translocation of PKC-e occurred as in wildtypes. Conclusions Gα 11 is essential for the development of cardiac hypertrophy in type 1-diabetes. Stimulation of hypertrophic signalling through PKC-α, PKC-δ, PKC-ζ, and NF-κB appears to be receptor-dependent, whereas PKC-e is activated by hyperglycemia, independent of Gα 11 .

Histone deacetylases (HDACs) are components of nuclear multiprotein complexes that deacetylate histones and perform important roles in repression of transcription. Using specific rabbit mAbs, we analyzed by immunohistochemistry and confocal immunofluorescence analysis the expression and subcellular localization of HDAC1–4 and HDAC9 in sections of adult human third molars. HDAC2 and HDAC9 were expressed in some pulpal cells and strongly expressed in the majority of mature odontoblasts. In contrast, only weak expression of HDAC1, HDAC3 and HDAC4 was observed. Confocal immunofluorescence analysis together with the DNA stain DRAQ5 revealed that HDAC2 and HDAC9 were coexpressed within the odontoblast nucleus, but localized to distinct subnuclear structures. In contrast to the current point of view, HDAC2 is strongly expressed in a terminally differentiated cell type. Our results imply that class I and II HDACs are involved in the transcriptional regulation of human odontoblasts in vivo.

The nitric oxide (NO) receptor enzyme soluble guanylate cyclase (sGC) contains one prosthetic heme group as an αβ heterodimer, and two heterodimer isoforms (α1β1, α2β1) were characterized to have enzyme activity. To test the irreversible inflammation-dependent regulation of sGC in odontoblasts, we incubated decalcified frozen sections of healthy and inflamed human third molars with antibodies against β-actin, nitrotyrosine, inducible nitric oxide synthase (iNOS), α1-, β1-, and α2-subunits of sGC and analyzed them at protein levels by quantitative immunohistochemistry. The irreversible inflammation induced an increase in the signal intensities for nitrotyrosine and iNOS and a decrease for the α1-, β1-, and α2-subunits of sGC in odontoblasts. Inflammatory mediators, reactive oxygen, and nitrogen species may impair the expression of the α1-, β1-, and α2-subunits in odontoblasts. The decrease of sGC at the protein level in inflamed odontoblasts is compatible with a critical role for sGC to mediate biological effects of NO in health.

The extracellular signal-regulated kinases 1 and 2 (ERK1/2) have been implicated in the inflammation-dependent sensitization of nociceptors. Because the periodontal ligament (PDL) contains numerous nociceptors and mechanoceptors, phosphorylation of ERK1/2 was investigated in nerve fibers of the PDL to elucidate the role of constitutive local activation of ERK1/2 in peripheral sensitization.Decalcified free-floating sections of rat molars with PDL were incubated using total (t)-ERK1/2 and phosphorylated (p)-ERK1/2 antibodies. For identification of nerve fibers in the PDL, double staining was performed using protein gene product 9.5 (PGP 9.5) with p-ERK1/2. To test whether p-ERK1/2 activated in sensory and mechanoreceptive terminals, double incubations were performed using p-ERK1/2 with calcitonin gene-related peptide (CGRP) and with calretinin. Labeled nerve fibers were quantified by the point-counting method.In cervical, midroot, and apical zones of the PDL, t-ERK1/2- and p-ERK1/2-labeled nerve fibers were found in close association with blood vessels. The p-ERK1/2-labeled free nerve fibers were often detected in cervical and apical areas of the PDL. In nerve fibers of the PDL, p-ERK1/2 was colocalized with PGP 9.5, CGRP, and calretinin.The perivascular distribution of t-ERK1/2 and p-ERK1/2 in nerve fibers in the PDL is compatible with a role for the constitutive activation of ERK1/2 in the neural regulation of blood vessels in the PDL. The colocalizations of p-ERK1/2 with CGRP and calretinin indicate that ERK1/2 is constitutively activated in a subpopulation of sensory and mechanoreceptive nerve terminals in the PDL.

The objective of this study was to evaluate the role of collagen membrane and Bio-Oss® coverage in healing of an onlay graft to the mandible. Twelve adult sheep each received an onlay bone graft (experiment 1), bone graft + Bio-Gide® (experiment 2), and bone graft + Bio-Oss®/Bio-Gide® (experiment 3) on the lateral surface of the mandible. The animals were euthanized at 4, 8, 12 or 16 weeks after surgery, and findings were analysed by routine microscopy and immunohistochemistry for proliferation (Ki67) and apoptotic (Caspase-3) markers. Grafts were fully incorporated in all specimens. Pronounced resorption was observed in experiment 1. Minimal loss of graft volume was seen in experiment 2 specimens without membrane displacement. A remarkable increase in the augmented region of the mandible was observed in experiment 3. A high number of osteoclasts were expressed within the grafts during the early healing period, and thereafter declined markedly. Osteoblasts within the grafts expressed a moderate level of Ki67 at 8 weeks, which thereafter declined markedly. The strongest expression of Caspase-3 on the bone surface was observed after 16 weeks. In conclusion, the effect of collagen membrane coverage on bone graft volume maintenance was dependent on membrane stability during healing. An autogenous bone graft covered with Bio-Oss® particles resulted in a remarkable increase in augmented lateral surface of the mandible. The late stage of bone graft healing was associated with a high apoptotic induction pathway of osteoblasts lining the surfaces of the new bone, demonstrated by strong positive Caspase-3 immunoreactivity.

The aim of this study was to evaluate the role of host periosteum and recipient bed perforation on the healing of an onlay graft to the mandible based on histologic and immunohistochemical analysis. Each of the 12 adult sheep used in the study received four iliac corticocancellous onlay bone grafts on the lateral surface of the mandible. In experiment 1, the block graft was placed in direct contact with the recipient bed and fixed with micro-screws, and in experiment 2, the recipient cortical bed was perforated before graft placement. The host periosteum around the graft was excised before flap replacement in experiment 3, and in experiment 4, a sheet of silicone membrane was placed between the graft and the recipient bed. The animals were euthanised at 4, 8, 12 and 16 weeks after surgery and the findings were analysed by routine microscopy (haematoxylin and eosin and tartrate-resistant acid phosphatase [TRAP] stains) and immunohistochemistry for proliferation and apoptotic markers (Ki67, caspase-3 and TUNEL stains). After 8 weeks, full graft–host integration was seen in all the experimental groups except experiment 4. After 16 weeks, pronounced graft resorption and volume reduction was observed in experiments 1 and 2, whilst experiment 3 was characterised with extensive connective tissue infiltration and severe resorption of the graft. The number of osteoclasts expressed peaked at 4 weeks in experiments 1 and 2 and at 16 weeks in experiment 3. Immunoreactivity for Ki67 by osteoblasts lining the trabecular bone of the graft’s spongiosa expressed moderate level of Ki67 at 8 weeks, and thereafter declined markedly. The strongest expression of caspase-3 on the bone surface was observed after 16 weeks. Recipient cortical bed perforation offered no advantage over non-perforated bed regarding healing and integration of a bone graft. Excision of the host overlying the periosteum was accompanied with rapid absorption of the grafts and partial or complete replacement with fibrous connective tissue. This study also demonstrated that cell death by apoptosis is a fundamental component of osteoblastic phenotypic differentiation during healing of corticocancellous bone graft.

Havacılık alanında İnsansız Hava Araçlarına (İHA) yönelik talep ve çalışmalar tüm dünyada ve ülkemizde gün geçtikçe artmaktadır. İHA’lar, kumanda ya da uydu sistemleri aracılığıyla kontrol edilen, dikey olarak kalkış ve iniş yapabilen, uzun süreli uçuşları gerçekleştirebilen araçlardır. İHA’ların kullanım alanları arasında gözetleme, takip, nesne tanıma ve uzaktan müdahale sayılabilir. Günümüzde, çok rotorlu insansız hava araçları konusunda yapılan çalışmalar daha çok önem kazanmıştır. Dikey kalkış-iniş yapabilmesi nedeniyle piste ihtiyaç duyulmaması, iç ve dış ortamlarda geniş hareket kabiliyetine sahip olması, havada belli bir yükseklikte asılı kalarak uçabilmesi ve uçuş için tek rotorlu helikopterlerdeki kompleks mekanizmalara ihtiyaç duymaması bu araçların yaygınlaşmasında ana neden olmuştur. Bu çalışmada, son 20 yılda gerçekleştirilen İHA araştırmaları gözden geçirilmiştir. Genel olarak İHA’lar askeri ve sivil misyonlar için ikiye ayrılmış olup, bu çalışmada özellikle sivil amaçlı İHA’ların uygulama alanları incelenmiştir. İHA’yı oluşturan temel elemanlar irdelendikten sonra, uygulanan mekanik testler, kaldırma kuvveti ve aerodinamik testler özetlenmiştir. Son olarak da, İHA’ların avantaj ve dezavantajları vurgulanarak, gelecekte yapılabilecek çalışmalara ışık tutulmuştur, CaCO3 as a filler has been used in the PVC. In this study, industrial waste was used instead of CaCO3 and were investigated changes in the thermal properties. Used in cutting waste was emerged during the produced in GRP pipe production from wet cutting. PVC, GRP waste powder and process aids were mixtured then produced in extrusion system for polymer matrix composite production. Thirteen different weight (%5-%70) percentages of cutting waste as a filler were reinforced to PVC. CaCO3 reinforced and pure PVC were produced in extrusion system to make comparison. All PVC samples were evaluated through DTA-TG analysis.

The destabilizing effect of endodontic treatment upon teeth is still controversial. The purpose of this study was to investigate the effects of different steps of endodontic treatments upon the rigidity of teeth. Extracted untreated central maxillary anterior teeth were loaded (3.75 N), and deformations of the root were assessed by Speckle pattern interferometry. The following treatments (with subsequent determination of deformability) were conducted sequentially: access preparation, manual instrumentation (Kerr files ISO-40, ISO-60, ISO-80, ISO-110), and tapered and parallel-sided post preparation. It was found that the teeth were increasingly destabilized by any treatment. While the increased deformability was not significant with the manual enlargement (p > 0.05), we found a significant destabilization after access preparation and post preparation (p < 0.05). A corresponding difference was found after conversion of the post preparation from tapered to parallel-sided (p < 0.05). Both substance loss and modifications of the natural root canal geometry play an important role in tooth rigidity.

The epithelial rests of Malassez (ERM) are derived from the Hertwig's epithelial root sheath during tooth development. The ERM contain endothelial nitric oxide synthase (eNOS), but the existence of phosphorylation site/s of eNOS in the ERM is unclear.Rat molars with periodontium were perfusion- and post-fixed, decalcified, and frozen-sectioned. Free-floating sections were incubated using antisera against total eNOS, eNOS phosphorylated at serine (Ser)1177, Ser116, and threonine (Thr)495. The signal intensities of t-eNOS, p-eNOS at Ser1177 and Ser116 in the ERM were measured by densitometry and statistically analyzed.In the ERM, localization of total eNOS and the phosphorylation sites of eNOS at Ser1177 and Ser116 were detected, while a basal localization of eNOS phosphorylated at Thr495 in the ERM was undetectable. For p-eNOS at Ser116 regional differences in phosphorylation were detected.The basal production of NO by eNOS in the ERM is modulated by phosphorylation of eNOS at Ser1177 and Ser116 residues, while the basal activity of the eNOS is not influenced by phosphorylation of eNOS at Thr495 residue. This provides evidence that phosphorylation plays a key role for regulation of the catalytic activity of eNOS.

Nitric oxide (NO) mediates different cellular functions by activating soluble guanylate cyclase (sGC) that converts guanosine-5'-triphosphate (GTP) to cyclic guanosine-3',5'-monophosphate (cGMP). Membrane-bound GCs produce cGMP in response to natriuretic peptides in osteoblasts, but neither the NO-target enzyme sGC, nor the phosphorylation sites of NOS III, nor their regulation by extracellular signal-regulated kinases 1 and 2 (ERK1/2) and Akt/protein kinase B (Akt/PKB) in osteoclasts have been established.Rat molars with periodontium were perfusion- and post-fixed, decalcified, and frozen-sectioned. Free-floating sections were stained using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) and tartrate-resistant acid phosphatase (TRAP) histochemical techniques and immunoreacted with antisera against NO-synthase (NOS) I-III, NOS III phoshorylated at Thr495, NOS III phoshorylated at Serine1177 (Ser1177), ERK1/2, phosphorylated ERK1/2, Akt/PKB, phosphorylated Akt/PKB, sGC (alpha2/beta1), and cGMP.NADPH-d staining and immunostaining of NOS I-III, NOS III phosphorylated at Ser1177, ERK1/2, Akt/PKB, phosphorylated Akt/PKB, sGC (alpha2 and beta1-subunits), and cGMP were detected in osteoclasts. Immunohistochemical reaction products for NOS III phosphorylated at threonine495 (Thr495) and phosphorylated ERK1/2 could not be identified in osteoclasts. Comparison of TRAP activity and immunostaining for sGC beta1-subunit revealed that sGC beta1-subunit is only expressed in a sub-population of osteoclasts.NO is likely to be generated by NOS I and NOS III in osteoclasts. The inconstant expression of NOS II in some osteoclasts may be explained with inducible expression of NOS II upon physiological cell activation. Localization of the sGC alpha2- and beta1-subunits and cGMP in osteoclasts is compatible with an involvement of NO-sGC signaling in the homeostasis of osteoclasts. The phosphorylation site of NOS III at Ser1177 and phosphorylated Akt/PKB are involved in regulation of NO production by NOS III in osteoclasts under basal conditions.

It is known that total endothelial nitric oxide synthase (eNOS) is localized in peripheral nerve fibers, but the existence of the phosphorylation site/s of eNOS in peripheral nerve fibers is unknown. In the perfusion-fixed and decalcified sections of rat palates, eNOS and eNOS phosphorylated at Ser(116) were identified in the nerve fibers of the glandulae palatinae. The localization of eNOS phosphorylated at Ser(1177) and Thr(495) in nerve fibers was undetectable. It is concluded that eNOS is phosphorylated at Ser(116) in nerve fibers of the glandulae palatinae under basal conditions. The basal Ser(1177) and Thr(495) phosphorylation of eNOS are missing in nerve fibers of the glandulae palatinae.

Knowledge of the histochemical substrates of cellular and neurovascular connections in the gingiva is essential in order to understand the initial mechanisms of inflammation in the periodontium. Since the localization of the neuroendocrine peptide galanin in the gingiva is still unclear, we used immunohistochemical, in situ hybridization and immunoblot techniques to assess the localization of galanin in the gingiva of rat molars. Galanin-immunoreactive nerve fibers were located around blood vessels in the lamina propria, beneath the epithelium, in the epithelial-proprial junction and in the basal layer of the epithelium. Galanin was highly expressed in the suprabasal keratinocytes of the gingival epithelium. The localization of galanin in gingival nerve fibers and the expression of galanin in keratinocytes of the gingival epithelium indicate that galanin may be a possible regulator of different cellular functions in the gingiva.

Özet Türbin kanatlarının soğutulması farklı bölgelerinde farklı soğutma teknikleri kullanarak yapılmaktadır. Kompresörde üretilen havanın yaklaşık %20’si soğutma sistemlerinde kullanılmaktadır. Kanat içine gönderilen bu hava önce iç kanallardan daha sonrada kanat dış profili üzerinden ve kanat uç kısmından dışarıya atılarak soğutma gerçekleştirilir. Bu soğutma tekniklerinin iç soğutma ve yüzeyde hava filmi meydana getirme ve çarpmalı jetle soğutma teknikleri uygulanmaktadır. Kanatta iç soğutma yapılması için bazen U şeklinde soğutma kanalları kullanılmaktadır. Bu kanallarda çeşitli açılarda kanatçıklar (ripler) kullanılarak soğutma performansları arttırılmaya çalışılmaktadır. Bu çalışmada U şeklindeki 180° dönüşlü düz bir kanalın ve alt ve üst yüzeylerine 90° ve 45° yerleştirilen kanatçıkların yerleştirildiği kanallardaki soğutma performansına etkisi 3 farklı Reynolds sayısı (22000, 27500 ve 33000) için deneysel olarak sıvı kristal termografisi metoduyla kanal yüzeyindeki ısı geçiş katsayısı ve Nu sayısının değişimi araştırılmıştır. Elde edilen sonuçlar jet geometrisinin etkisi, Reynolds sayısının etkisi, kanal içindeki hız dağılımı ve ısı geçiş katsayısı dağılımları gösterilmiştir.

A wide variety of stimuli can trigger activation of the transcription factor CREB (cAMP-responsive element binding protein), pointing toward a central role for CREB in the integration of various signaling inputs. No data are available on the expression and phosphorylation of CREB in mammalian teeth. Using immunohistochemical analysis of free-floating sections, we show here that CREB was strongly expressed and phosphorylated at Ser-133 within the nucleus of a subpopulation of adult human molar odontoblasts. Many dental pulp stromal cells and periodontal ligament fibroblasts expressed CREB and showed phosphorylation of CREB at Ser-133. In addition, cementoblasts displayed nuclear expression and phosphorylation of CREB at Ser-133. The epithelial rests of Malassez revealed strong nuclear expression of CREB, but phosphorylation at Ser-133 was variable. Our results provide the first evidence that the constitutively phosphorylated transcription factor CREB is involved in the biomineralization process of adult human molar odontoblasts and cementoblasts.

Bu çalışma, Erzurum ilinde toplu yemek üretimi yapan üç farklı işletmenin üretim hattında yer alan bazı kritik noktalarında S.aureus ve enterotoksinin varlığı ile bazı indikatörler kullanılarak hijyenik standartlara uygunluğunun belirlenmesi amacıyla yapılmıştır.Analiz edilen gıda örneklerinin bütününde S.aureus sayısı

BackgroundWhile ryanodine receptor 1 (RyR1) critically contributes to skeletal muscle contraction abilities by mediating Ca²⁺ion oscillation between sarcoplasmatic and myofibrillar compartments, AMP-activated protein kinase (AMPK) senses contraction-induced energetic stress by phosphorylation at Thr¹⁷². Phosphorylation of RyR1 at serine²⁸⁴³ (pRyR1Ser²⁸⁴³) results in leaky RyR1 channels and impaired Ca²⁺homeostasis. Because acute resistance exercise exerts decreased contraction performance in skeletal muscle, preceded by high rates of Ca²⁺-oscillation and energetic stress, intense myofiber contractions may induce increased RyR1 and AMPK phosphorylation. However, no data are available regarding the time-course and magnitude of early RyR1 and AMPK phosphorylation in human myofibers in response to acute resistance exercise.PurposeDetermine the effects and early time-course of resistance exercise on pRyR1Ser²⁸⁴³ and pAMPKThr¹⁷² in type I and II myofibers.Methods7 male subjects (age 23±2 years, height: 185±7 cm, weight: 82±5 kg) performed 3 sets of 8 repetitions of maximum eccentric knee extensions. Muscle biopsies were taken at rest, 15, 30 and 60 min post exercise. pRyR1Ser²⁸⁴³ and pAMPKThr¹⁷² levels were determined by western blot and semi-quantitative immunohistochemistry techniques.ResultsWhile total RyR1 and total AMPK levels remained unchanged, RyR1 was significantly more abundant in type II than type I myofibers. pRyR1Ser²⁸⁴³ increased 15 min and peaked 30 min (pConclusionResistance exercise induces acutely increased pRyR1Ser²⁸⁴³ and concomitantly pAMPKThr¹⁷² levels for up to 30 min in resistance exercised myofibers. This provides a time-course by which pRyR1Ser²⁸⁴³ can mechanistically impact Ca²⁺handling properties and consequently induce reduced myofiber contractility beyond immediate fatiguing mechanisms.

During tooth development, the inner and outer enamel epithelia fuse by mitotic activity to produce a bilayered epithelial sheath termed Hertwig's epithelial root sheath (HERS). The epithelial rests of Malassez (ERM) are the developmental residues of HERS and remain in the adult periodontal ligament (PDL). Although the cellular regulation of the Ca(2+)-binding proteins parvalbumin, calbindin-D28k, and calretinin has been reported in the inner and outer enamel epithelia during tooth development, an involvement of Ca(2+)-binding proteins in the ERM has not so far been characterized. Among the three Ca(2+)-binding proteins tested (calbindin D28k, parvalbumin, calretinin), we have only been able to detect calretinin in a subpopulation of adult rat molar ERM, by using quantitative immunohistochemical and confocal immunofluorescence techniques. TrkA (a marker for ERM) is present in numerous epithelial cell clusters, whereas calretinin has been localized in the cytosol and perinuclear region of a subpopulation of TrkA-positive cells. We conclude that, in inner and outer enamel epithelial cells, Ca(2+) is regulated by calbindin, parvalbumin, and calretinin during tooth development, whereas in the ERM of adult PDL, Ca(2+) is regulated only by calretinin. The expression of Ca(2+)-binding proteins is restricted in a developmental manner in the ERM.

The cell specific detection of enzyme activation in response to the physiological contractile load within muscle-tendon-bone unit is essential for understanding of the mechanical forces transmission from muscle cells via tendon to the bone. The hypothesis that the physiological mechanical loading regulates activation of Akt1/PKBαat Thr308 and at Ser473 in muscle fibers within muscle-tendon-bone unit was tested using quantitative immunohistochemistry, confocal double fluorescence analysis, and immunoblot analysis. In comparison to the staining intensities in peripheral regions of the muscle fibers, Akt1/PKBαwas detected with a higher staining intensity in muscle fibers at the myotendinous junction (MTJ) areas. In muscle fibers at the MTJ areas, Akt1/PKBαis dually phosphorylated at Thr308 and Ser473. The immunohistochemical results were confirmed by immunoblot analysis. We conclude that contractile load generated by masticatory muscles induces local domain-dependent expression of Akt1/PKBαas well as activation by dually phosphorylation at Thr308 and Ser473 in muscle fibers at the MTJ areas within muscle-tendon-bone unit.

The extracellular signal-regulated kinases 1 and 2 (ERK1/2) have been implicated in the inflammation-dependent sensitization of nociceptors, and the inflammatory mediator bradykinin (BK) led to a reduced threshold in the nociceptor terminals, activating intracellular signaling by phosphorylating receptors and ion channels. The effects of BK on the non-transcriptional modulation of the ERK1/2 in the peripheral nociceptor terminals, including in nerve endings of the dentin-pulp complex, are unknown. The time-dependent effects of BK (10−7 M) on the ERK1/2 phosphorylation in nerve terminals of the dentin-pulp complex were investigated by quantitative and double immunolabeling with organ bath experiments. In nerve terminals, total and p-ERK1/2 were detected. In comparison with the controls, the numbers of p-ERK1/2-positive nerve endings increased after 1 and 3 min and decreased after 10 min of BK treatment. Analysis of the data indicates that BK induces phosphorylation-mediated local activation of ERK1/2 in nerve terminals modulating nociception in the dentin-pulp complex.

Although prion protein’s (PrP) involvement in transmission of degenerative neurological diseases has been subjected to considerable scrutiny, its physiological role is still obscure. The distribution of PrP in dental tissues was investigated using three different methods: immunohistochemistry, cell culture, and scanning electron microscopy. PrP knockout mice were found to have marked anomalies in dentin structure. In human teeth, cementoblasts and odontoblasts showed prominent staining for PrP at levels comparable to those of nerve fibers. Epithelial rests of Malassez, which are remnants of a cell type formerly forming enamel, were also positive. Thus, all PrP-positive cells in human dentition are in some way involved in calcified tissue formation. This suggests a previously undetected function of prion protein in healthy vertebrates as evidenced by an obvious phenotype in PrP knockout mice. Periodontal and pulpal tissue exposed by disease or trauma might represent a clinically relevant entry point for prions incorporated orally and thus a possible mode of infection.

While the activation of eNOS by Akt/PKB-dependent phosphorylation, leading to NO release, and the inhibition of enzyme activity by bradykinin (BK)-mediated phosphorylation of eNOS in endothelial cells are established, the phosphorylation of eNOS in odontoblasts is unknown. To clarify the regulation of eNOS in odontoblasts by BK, we examined the phosphorylation of eNOS, Akt/PKB, and ERK1/2 in odontoblasts of rat molars. BK (10−7 M) transiently induced the phosphorylation of eNOS at Ser1177, Akt/PKB in odontoblasts, while it induced the phosphorylation of eNOS at Thr495 throughout the entire period of BK treatment. BK receptor 2 antagonist HOE 140 (10−6 M) significantly reduced signal intensities of phosphorylated-eNOS at Ser1177, Thr495, and phosphorylated-Akt/PKB. These results suggest that BK has dual effects on the activation of eNOS in odontoblasts, the Akt/PKB-dependent up-regulation of eNOS by the transient phosphorylation at Ser1177, and the ERK1/2-independent down-regulation of eNOS by the phosphorylation at Thr495.

Knowledge of the histochemical substrates of cellular and neurovascular connections in the gingiva is essential in order to understand the initial mechanisms of inflammation in the periodontium. Since the localization of the neuroendocrine peptide galanin in the gingiva is still unclear, we used immunohistochemical, in situ hybridization and immunoblot techniques to assess the localization of galanin in the gingiva of rat molars. Galanin-immunoreactive nerve fibers were located around blood vessels in the lamina propria, beneath the epithelium, in the epithelial-proprial junction and in the basal layer of the epithelium. Galanin was highly expressed in the suprabasal keratinocytes of the gingival epithelium. The localization of galanin in gingival nerve fibers and the expression of galanin in keratinocytes of the gingival epithelium indicate that galanin may be a possible regulator of different cellular functions in the gingiva.

Large clinical studies provided evidence of a lower incidence of a reduced cardiac morbidity in diabetic patients treated with angiotensin type 1 (AT1) receptor blockers. The AT1 receptor is coupled to the Gq class of G-proteins, which stimulate proteinkinase C (PKC) via activation of phospholipase Cβ. To study the role of the Gq protein Gα11 and its signaling through PKC in diabetic heart disease, we induced diabetes in wildtype and Gα11 knockout mice using streptozotocin.After eight weeks of stable hyperglycemia, cardiac morphology and function were assessed by echocardiography, myocardial expression and translocation of PKC isoforms by immunohistochemistry and immunoblot after tissue fractionation.Wildtype mice (n=8) but not Gα11 knockout animals (n=8) developed ventricular hypertrophy upon induction of hyperglycemia. PKC isoforms α, βII, δ, e, ζ and θ were all detected in myocardium of wildtype animals. Compared to normoglycemic control animals, PKC isoforms α and ζ showed increased expression levels in diabetic wildtype mice. In addition, PKC ζ was phosphorylated at Thr410/403 and showed strong translocation to nuclear membranes in cardiomyocytes of diabetic but not of control animals. Hearts from normoglycemic Gα11-knockout mice showed lower expression levels of PKC α and δ compared to wildtype animals. Upon induction of hyperglycemia in knockouts, expression of PKC δ was only slightly increased to the level found in non-diabetic wildtypes. Expression, phosphorylation and translocation of PKC isoforms, however, remained unaffected by the induction of diabetes in Gα11-deficient mice.We conclude that Gα11 is central to the induction of myocardial hypertrophy in type 1 diabetes. Activation of PKC α and ζ appear to be important pathways in hypertrophic signaling via Gα11. The inhibition of this pathway may in part explain the strong therapeutic benefit of AT1 receptor blockade in diabetic patients.

By the formation of cyclic guanosine 3′,5′-monophosphate (cGMP), nitric oxide (NO)-sensitive enzyme-soluble guanylate cyclase (sGC) plays a receptor role for NO within the NO-cGMP signaling cascade, which is involved in vasodilatation and neurotransmission. The hypothesis that NO-cGMP signaling molecules modulate cells of the dentin-pulp complex was investigated in rat molars by histochemical, immunohistochemical, immuno-ultrastructural, and organ bath techniques. NO synthase (NOS) I-III, the sGC α2-subunit/β1-subunit, and cGMP were detected in odontoblasts and blood vessels. NOS I, sGC α2, and cGMP were identified in nerve fibers. Treatment of rat molars with the NO donor NONOate (10−5 M) increased cGMP staining intensities in blood vessels and odontoblasts, while NO synthase inhibitor L-NAME (10−4 M) attenuated intensity of the reaction products for cGMP, suggesting an effect of endogenous NO on sGC. These correlations of patterns and alterations of cGMP staining intensities after treatment with the NO donor or NO inhibitor might represent an NO-sGC-cGMP signaling-dependent modulation of odontoblasts, blood vessels, and nerve fibers in the dentin-pulp complex.