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TGF-β3 mediates epithelial-mesenchymal transformation during normal fusion of lip and palate, but how TGF-β3 functions during cleft lip repair remains unexplored. We hypothesize that TGF-β3 promotes fetal cleft lip repair and fusion by increasing the availability of mesenchymal cells. In this investigation, we demonstrated that cleft lips in mouse fetuses were repaired by fetal surgery, producing scarless fusion. At the site of the operation, we first observed an infusion of platelets expressing TGF-β3, followed by increased expression of cyclin D1 and tenascin-C, and coupled with increased mesenchymal cell proliferation. In an ex vivo serumless culture system, cleft lip explants fused in the presence of exogenous TGF-β3. Cultured lips also showed upregulation in cyclin D1 and tenascin-C expression. These findings suggest that microsurgical repair of cleft lip in the fetus that produced scarless fusion is mediated by TGF-β3 regulation of mesenchymal cell proliferation and migration at the site of repair. [ABSTRACT FROM AUTHOR]

Wounds that occur in adults form scars due to fibrosis, whereas those in embryos regenerate. If wound healing in embryos is mimicked in adults, scarring can be reduced. We found that mouse fetuses could regenerate tissues up to embryonic day (E) 13, but visible scars remained thereafter. This regeneration pattern requires actin cable formation at the epithelial wound margin via activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK). Here, we investigated whether the AMPK-activating effect of salicylate, an anti-inflammatory drug, promotes regenerative wound healing. Salicylate administration resulted in actin cable formation and complete wound regeneration in E14 fetuses, in which scarring should have normally occurred, and promoted contraction of the panniculus carnosus muscle, resulting in complete wound regeneration. In vitro, salicylate further induced actin remodeling in mouse epidermal keratinocytes in a manner dependent on cell and substrate target-specific AMPK activation and subsequent regulation of Rac1 signaling. Furthermore, salicylate promoted epithelialization, enhanced panniculus carnosus muscle contraction, and inhibited scar formation in adult mice. Administration of salicylates to wounds immediately after injury may be a novel method for preventing scarring by promoting a wound healing pattern similar to that of embryonic wounds. [ABSTRACT FROM AUTHOR]

P1 Altered Expression of Gene Products Involved as a Complex in the Endogenous Hepatocarcinogenesis of Rats Fed a Choline-Deficient, L-Amino Acid-Defined Diet Dai Nakae, 1 Fumiyuki Uematsu, 1 Yutaka Hatanaka, 2 Yoichi Konishi, 3 Akihiko Maekawa, 3 Masakazu Takahashi, 3 and Midori Yoshida, 3 1 Sasaki Institute, Sasaki Foundation, Chiyoda, Tokyo, Japan, 2 DakoCytometaion Company Limited, City of Kyoto, Japan, and 3 Nara Medical University, Sakai, Osaka, Japan Chronic feeding, for up to 2 years, of a choline-deficient, L-amino acid-defined (CDAA) diet results in a high incidence of hepatocellular carcinomas (HCCs) in male rats through endogenous mechanisms. Its morphological course consists of sequentially developed (pre)neoplastic and nonneoplastic liver lesions, and its underlying mechanisms are associated with oxidative stress, multiple signaling alterations, and genetic and epigenetic changes of specific genes. These characteristics are closely similar to those seen in human hepatocarcinogenic cases including those due to an infection of hepatitis viruses. Furthermore, this has recently been recognized as a good animal model for human nonalcoholic steatohepatitis (famous as NASH). The present study was conducted as a part of our work to explore details of molecular events occurring in this model with our aim to obtain information contributable to the control of human cancers, especially HCC, a deadly human neoplasm. Male Fischer 344 rats (6 weeks old) were fed the CDAA diet or a control diet for up to 70 weeks and serially sacrificed to obtain livers. Using the livers obtained up to the end of week 12, mRNA and protein expressions of 18 cytokines were assessed by a ribonuclease protection assay and an immunohistochemical technique, respectively. Using the livers obtained at the end of week 70, comprehensive gene expression profiles were assessed for HCCs, their surrounding tissues and the normal tissues by an oligonucleotide microarray technique for 3757 genes. All assessments were conducted using at least 5 samples per group from individual animals. mRNAs of the cytokines including IL-1-alfa, IL-1-beta, TNF-alfa, TGF-beta-1, TGF-beta-2, and TGF-beta-3 were overexpressed in differentially time-dependent manners in the early phase within the first 12 weeks. At the end of week 12, proteins of such cytokines were also overexpressed. IL-1-alfa and TNF-alfa proteins were overexpressed more remarkably in preneoplastic liver lesions than in their surroundings. IL-1-beta and TGF-beta-1 proteins behaved similarly, but the differences between inside and outside of preneoplastic lesions were less evident. TGF-beta-2 protein was overexpressed less remarkably in preneoplastic liver lesions than in their surroundings. TGF-beta-3 protein behaved similarly, but the difference between inside and outside of preneoplastic lesions was less evident. Comparing profiles among HCCs, their surroundings and normal liver tissues, 147 genes were differentially expressed, which were classified according to the expression patterns into 4 identical clusters by different methods. These contained conspicuous changes for notable genes interacting each other within a bio-signaling complex. Such genes included transcription factors (e.g., hepatic nuclear factor-1 [Tcf1], hepatic nuclear factor-4 and Rheb), signaling factors (e.g., IL-1 receptor, IL-2 receptor, PDGF receptor, beta-, NGF, FGF-5, FGF-9, and COX-1), apoptosis and cell proliferation regulatory factors (e.g., caspase 3, inhibin, lifeguard, c-fos, src-related tyrosine kinase, and Tsc2), pre-mRNA processing factors (e.g., cyclin L and zinc finger protein 265), metabolic enzymes (e.g., CYP1B1) and anti-oxidant defense system members (e.g., phospholipase C and superoxide dismutase). The present results indicate that altered expression of numerous gene products is involved as a complex in the endogenous hepatocarcinogenesis of rats fed the CDAA diet. [ABSTRACT FROM AUTHOR]

Purpose: To review the atypical development of Salzmann's nodular degeneration (SND) after two cases of laser in situ keratomileusis (LASIK) and one case of photorefractive keratomileusis (PRK), and to highlight the pathophysiology of SND and its treatment. Methods: Three cases of SND (two following LASIK performed with microkeratomes and one following PRK) were reviewed and Pubmed.gov and internet searches were performed. Results: SND is myofibroblast-generated fibrosis in the subepithelial space between the epithelium and Bowman's layer that develops years or decades after traumatic, surgical, infectious, or inflammatory injuries to the cornea in which the epithelial basement membrane is damaged in one or more locations and does not fully regenerate. It is hypothesized based on these cases, and the previous immunohistochemistry of other investigators, that myofibroblast precursors, such as fibrocytes or corneal fibroblasts, that enter the subepithelial space are driven to develop into myofibroblasts, which slowly proliferate and extend the fibrosis, by transforming growth factor-beta from epithelium and tears that passes through the defective epithelial basement membrane. These myofibroblasts and the disordered collagens, and other extracellular matrix components they produce, make up the subepithelial opacity characteristic of SND. Nodules are larger accumulations of myofibroblasts and disordered extracellular matrix. If the injury is associated with damage to the underlying Bowman's layer and stroma, as in LASIK flap generation, then the myofibroblasts and fibrosis can extend into Bowman's layer and the underlying anterior stroma. Conclusions: SND fibrosis often extends into Bowman's layer and the anterior stroma if there are associated Bowman's defects, such as incisions or lacerations. In the latter cases, SND frequently cannot be removed by simple scrape and peel, as typically performed for most common SND cases, but can be trimmed to remove the offending tissue. This condition is more accurately termed Salzmann's subepithelial fibrosis. [J Refract Surg. 2024;40(5):e279–e290.] [ABSTRACT FROM AUTHOR]

Sepsis is a crucial public health problem with a high mortality rate caused by a dysregulated host immune response to infection. Vascular endothelial cell injury is an important hallmark of sepsis, which leads to multiple organ failure and death. Early biomarkers to diagnose sepsis may provide early intervention and reduce risk of death. Damage-associated molecular patterns (DAMPs) are host nuclear or cytoplasmic molecules released from cells following tissue damage. We postulated that DAMPs could potentially be a novel sepsis biomarker. We used an in vitro model to determine suitable protein–DAMPs biomarkers for early sepsis diagnosis. Low and high lipopolysaccharide (LPS) doses were used to stimulate the human umbilical vein endothelial cell line EA.hy926 for 24, 48, and 72 h. Results showed that cell viability was reduced in both dose-dependent and time-dependent manners. Cell injury was corroborated by a significant increase in lactate dehydrogenase (LDH) activity within 24 h in cell-conditioned medium. Secreted protein–DAMPs in the supernatant, collected at different time points within 24 h, were characterized using shotgun proteomics LC-MS/MS analysis. Results showed that there were 2233 proteins. Among these, 181 proteins from the LPS-stimulated EA.hy926 at 1, 12, and 24 h were significantly different from those of the control. Twelve proteins were up-regulated at all three time points. Furthermore, a potential interaction analysis of predominant DAMPs-related proteins using STITCH 5.0 revealed the following associations with pathways: response to stress; bacterium; and LPS (GO:0080134; 0009617; 0032496). Markedly, alpha-2-HS-glycoprotein (AHSG or fetuin-A) and lactotransferrin (LTF) potentially presented since the first hour of LPS stimulation, and were highly up-regulated at 24 h. Taken together, we reported proteomic profiling of vascular endothelial cell-specific DAMPs in response to early an in vitro LPS stimulation, suggesting that these early damage-response protein candidates could be novel early biomarkers associated with sepsis. [ABSTRACT FROM AUTHOR]

Oral submucous fibrosis (OSF) is a potentially malignant disorder characterised by inflammation and progressive fibrosis. Transforming growth factor-β (TGF-β) has been established as a master regulator of fibrosis in various organs; however, lack of systematic review on expression of TGF-β and its isoforms in OSF restrict the understanding of their behaviour in its pathogenesis. Online electronic databases, such as PubMed Medline, Cochrane Library, Embase, and Scopus, were searched from their respective dates of inception till 31st March 2022. Human studies related to TGF-β expression in histopathologically diagnosed OSF cases, with or without malignant transformation, were included and assessed using a Cochrane risk of bias assessment tool: For non randomised studies of interventions (ACROBAT NRSI). The electronic literature search yielded 394 articles. Of those, ten articles met the inclusion criteria and involved total of 579 OSF patients. The risk of bias (RoB) was low to moderate. These studies demonstrated a significant positive expression of TGF-β and its isoforms in OSF compared to that in normal tissue samples. An increased pan TGF-β expression was observed in the early stages of OSF, and an increased expression of TGF-β1 and TGF-β2 were seen in advanced stages of OSF. Stage wise expression of TGF-β3 has not been discussed in the included studies. No significant relationship was observed between epithelial dysplasia and TGF-β expression in OSF. The distinct pattern in the expression of pan TGF-β, TGF-β1 and TGF-β2 in various stages of OSF indicates their different roles in OSF progression. We believe isoform targeted studies exploring stage wise expression of the marker will open new treatment avenues for OSF. [ABSTRACT FROM AUTHOR]

Due to their immunomodulatory and regenerative capacity, human bone marrow-derived mesenchymal stem cells (hBMSCs) are promising in the treatment of patients suffering from polytrauma. However, few studies look at the effects of sera from polytraumatized patients on hBMSCs. The aim of this study was to explore changes in hBMSC properties in response to serum from polytrauma patients taken at different time points after the trauma incident. For this, sera from 84 patients with polytrauma (collected between 2010 and 2020 in our department) were used. In order to test the differential influence on hBMSC, sera from the 1st (D1), 5th (D5), and 10th day (D10) after polytrauma were pooled, respectively. As a control, sera from three healthy donors (HS), matched with respect to age and gender to the polytrauma group, were collected. Furthermore, hBMSCs from four healthy donors were used in the experiments. The pooled sera of HS, D1, D5, and D10 were analyzed by multicytokine array for pro-/anti-inflammatory cytokines. Furthermore, the influence of the different sera on hBMSCs with respect to cell proliferation, colony forming unit-fibroblast (CFU-F) assay, cell viability, cytotoxicity, cell migration, and osteogenic and chondrogenic differentiation was analyzed. The results showed that D5 serum significantly reduced hBMSC cell proliferation capacity compared with HS and increased the proportion of dead cells compared with D1. However, the frequency of CFU-F was not reduced in polytrauma groups compared with HS, as well as the other parameters. The serological effect of polytrauma on hBMSCs was related to the time after trauma. It is disadvantageous to use BMSCs in polytraumatized patients at least until the fifth day after polytrauma as obvious cytological changes could be found at that time point. However, it is promising to use hBMSCs to treat polytrauma after five days, combined with the concept of "Damage Control Orthopedics" (DCO). [ABSTRACT FROM AUTHOR]

Background: The injured flexor tendon has poor healing ability, which is easy to cause tendon adhesion. It can affect the recovery of tendon function, which is still a long-term and difficult task for surgeons. Transforming growth factor β (TGF-β) has been widely considered to play an important role in flexor tendon repair in recent years.Aim: This work was to investigate the anti-adhesion and anti-inflammatory effects of TGF-β3 on flexor digitorum longus (FDL) tendon repair rats.Method: Anastomosis models of tendon laceration in the flexion toes of rats were delivered with no treatment, vehicle, or TGF-β3 -overexpressed adenovirus vector (ad-TGF-β3) locally to the injured tendon area from day 3 to 8. Subsequently, the expression of TGF-β3, TGF-β1/2, Smad3, Smad7, JNK, phosphorylation (p)-JNK, c-Jun, and phosphorylation (p)-c-Jun were detected by western blot, the expression of Mmp9 and Mmp2 by RT-qPCR, the Range of motion (ROM) and gliding resistance by adhesion formation testing, the mechanical strength of tendon healing by biomechanical testing, the pathologic changes of flexor tendon tissues by HE staining, the expression of collagen type III by immunohistochemical staining, and the levels of IL-6, TNF-α, COX2 and IL-1β in serum by ELISA, respectively.Results: Rat models treated with no treatment showed a lower elevation of TGF-β3 and Smad7 expression, and a higher elevation of TGF-β1/2 and Smad3 expression, during day 14 to day 28. Besides, under the treatment of ad-TGF-β3, a significantly increase was reflected in the expression of TGF-β3 and Smad7, ROM, as well as mechanical strength of flexor tendon, whereas significantly reduction was shown in gliding resistance, the content of inflammatory cytokines, the ratio of p-JNK/JNK, p-c-Jun/c-Jun, as well as the expression of TGF-β1/2, Smad3, Mmp9, and Mmp2 genes, as compared to those from vehicle treatment. Meanwhile, TGF-β3 demonstrated a better pathologic recovery process with no obvious necrosis or fracture of collagen fibers. Besides, TGF-β3 revealed a significant reduction of collagen type-III expression in the flexor tendon healing tissues.Conclusion: These findings suggested that TGF-β3 effectively protected against flexor tendon injury via regulating adhesion formation. [ABSTRACT FROM AUTHOR]

Chondrocytes, comparable to many cells from the connective tissue, dedifferentiate and end up in a similar fibroblastoid cell type, marked by the loss of the specific expression pattern. Here, chondrocytes isolated from osteoarthritic (OA) patients were investigated. The OA chondrocytes used in this work were not affected by the loss of specific gene expression upon cell culture. The mRNA levels of known cartilage markers, such as SOX5, SOX6, SOX9, aggrecan and proteoglycan 4, remained unchanged. Since chondrocytes from OA and healthy tissue show different DNA methylation patterns, the underlying mechanisms of cartilage marker maintenance were investigated with a focus on the epigenetic modification by DNA methylation. The treatment of dedifferentiated chondrocytes with the DNA methyltransferase inhibitor 5-aza-2´-deoxycytidine (5-aza-dC) displayed no considerable impact on the maintenance of marker gene expression observed in the dedifferentiated state, while the chondrogenic differentiation capacity was compromised. On the other hand, the pre-cultivation with 5-aza-dC improved the osteogenesis and adipogenesis of OA chondrocytes. Contradictory to these effects, the DNA methylation levels were not reduced after treatment for four weeks with 1 μM 5-aza-dC. In conclusion, 5-aza-dC affects the differentiation capacity of OA chondrocytes, while the global DNA methylation level remains stable. Furthermore, dedifferentiated chondrocytes isolated from late-stage OA patients represent a reliable cell source for in vitro studies and disease models without the need for additional alterations. [ABSTRACT FROM AUTHOR]

Transforming growth factor-β signaling through intracellular Smad3 has been implicated in Parkinson's disease ( PD) and it fulfills an important role in the neurogenesis and synaptic plasticity that occurs in the adult dentate gyrus ( DG). The long-term potentiation ( LTP) induced in the DG by high-frequency stimulation of the medial perforant pathway is abolished in the DG of Smad3-deficient mice, but not in the CA1 hippocampal region. Here, we show that NMDA- and AMPA-type glutamate receptors do not participate in the inhibition of LTP associated with Smad3 deficiency. Moreover, there is no difference in the hippocampal GAD65 and GAD67 content, suggesting that GABA biosynthesis remains unaffected. Increased conductance and higher action potential firing thresholds were evident in intracellular recordings of granule cells from Smad3 deficient mice. Interestingly, phasic and tonic GABAA receptor ( GABAAR) -mediated neurotransmission is enhanced in the DG of Smad3-deficient mice, and LTP induction can be rescued by inhibiting GABAAR with picrotoxin. Hence, Smad3 signaling in the DG appears to be necessary to induce LTP by regulating GABAA neurotransmission, suggesting a central role of this intracellular signaling pathway in the hippocampal brain plasticity related to learning and memory. [ABSTRACT FROM AUTHOR]

The single nucleotide polymorphism (SNP) rs2275294 of theZNF512Bgene has been reported to be associated with a risk of ALS in the Japanese population. Here we conducted a case-control study examining the possible association of rs2275294 with risk of sporadic ALS and PD in Han Chinese. Our study included 301 patients with ALS and 457 age- and gender-matched controls, as well as 555 patients with PD and 473 age- and gender-matched controls. Subjects were genotyped at rs2275293 using the ligase detection reaction. The genotype distribution of rs2275294 shows significant difference between patients with ALS and the control group according to the dominant model (OR 1.518, 95% CI 1.074–2.145,p= 0.018) and based on alleles (OR 1.249, 95% 1.016–1.534,p= 0.035). Stratification analysis showed a significant difference between females with ALS and female controls based on the dominant model (OR 3.285, 95% CI 1.856–5.815,p< 0.001) or alleles (OR 1.697, 95% CI 1.208–2.383,p= 0.002). In contrast, no significant differences were identified between rs2275294 and patients with PD. In conclusion, our case-control study suggests that the CC genotype and C allele at rs2275294 are associated with increased risk of ALS in Han Chinese, particularly females. [ABSTRACT FROM AUTHOR]

Objective: In this study, we describe placental gene expression patterns of endoglin in pregnancies with intrauterine growth restriction (IUGR) compared to normal pregnancies. Methods: Placental samples were obtained from 101 pregnancies with IUGR using 140 normal pregnancy cases as control. Gene expression patterns and protein levels of the endoglin were compared between the two groups. For the gene expression analysis real-time PCR was applied, while for the estimation of placental protein level we performed Western analysis. Results: The placental endoglin gene was significantly overexpressed in the IUGR group versus the control group (Ln2α: 1.69). The placental endoglin protein level proved to be significantly higher in case of IUGR (endoglin/β-actin ratio: 13.8 ± 2.3) versus the control cases (5.3 ± 1.1). The placental gene expression as well as the protein levels of endoglin showed no significant difference between female and male newborns. Concerning the placental gene expression and protein level, no significant difference was justified between the more (0-5 percentile) and less (5-10 percentile) severe cases of IUGR. Conclusion: Increased placental gene expression of endoglin may result in vascular dysfunction leading to chronic fetal hypoxia, which may induce VEGF-A to stimulate angiogenesis. This can be explained as feed back response to restore fetal placental circulation. [ABSTRACT FROM AUTHOR]

The article presents abstracts on translational science topic which include assessment of the knowledge, attitude and behavior of Georgia clinicians regarding best practices for disclosing HIV positive and providing HIV/AIDS care to sexual and needle-sharing partners, investigation of the validity of the Autism Mental Status Exam (AMSE) in a community-based population of children at risk of autism spectrum disorders (ASD), and assessment of hepatic de novo lipogenesis (DNL) in youth.

Copyright of Magyar Traumatológia, Ortopédia, Kézsebészet, Plasztikai Sebészet is the property of Matrokplaszt Folyoirat Alapitvany and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Advanced hepatocellular carcinoma (HCC) is an important cause of cancer mortality. Epithelial-mesenchymal transition (EMT) has been shown to be an important biological process in cancer progression and metastasis. We have focused on elucidating factors that induce EMT to promote carcinogenesis and subsequent metastasis in HCC using the BNL CL.2 (BNL) and BNL 1ME A. 7R.1 (1MEA) cell lines. BNL cells are normal hepatocytes whereas the 1MEA cells are HCC cells derived from chemical transformation of the BNL cells. Their morphological characteristics were examined. Expression levels of hepatocyte growth factor (HGF), markers of EMT and mediators of HGF signaling were determined and functional characteristics were compared. BNL cells were treated with HGF and effects on EMT-marker and mediators of HGF signaling were analyzed. BNL cells display characteristic epithelial morphology whereas 1MEA cells display mesenchymal characteristics. 1MEA cells express and secrete more HGF than BNL cells. There was significantly decreased expression of E-cadherin, albumin, AAT and increased expression of fibronectin, collagen-1, vimentin, snail and slug in 1MEA cells. There was also increased expression of cyclooxygenase-2 (COX-2), Akt and phosphorylated Akt (pAkt) in 1MEA cells. Moreover, 1MEA cells had increased migratory capacity inhibited by inhibition of COX-2 and Akt but not extracellular signal regulated kinase (ERK). Molecular mesenchymal characteristics of 1MEA cells were reversed by inhibition of COX-2, Akt and ERK. Treatment of BNL cells with HGF led to decreased expression of E-cadherin and increased expression of fibronectin, vimentin, snail, slug, COX-2, Akt, pAkt and increased migration, invasiveness and clonogenicity. We conclude that development of HCC is associated with upregulation of HGF which promotes EMT and carcinogenesis via upregulation of COX-2 and Akt. Consequently, HGF signaling may be targeted for therapy in advanced and metastatic HCC. [ABSTRACT FROM AUTHOR]

Bronson R. Biology of the male reproductive tract: Its cellular and morphological considerations. Am J Reprod Immunol 2011; 65: 212-219 For many years, the focus of attention in the study of semen has been on spermatozoa, its major cellular component, given their importance in the process of reproduction, and the role of the seminal fluid as their transport medium. More recently, evidence has accumulated of the complexity of seminal fluid, its components that perturb the female reproductive tract in ways promoting both survival of spermatozoa there-in and facilitating the implantation of embryos within the endometrium, hence initiating pregnancy. These same factors, however, may also make the female reproductive tract susceptible to invasion not only by spermatozoa but viruses, playing a significant role in the male-to-female transmission of HIV. Knowledge of the histology, anatomy, and immunology of the male reproductive tract is essential in understanding its role in HIV pathogenesis. [ABSTRACT FROM AUTHOR]

The article cites a research study that investigates the mesostriatal system in adult mice deficient in Smad3, a molecule involved in the intracellular transforming growth factor-beta1 (TGF-b1) signalling cascade. It is said that TGF-b1 levels increase in patients with Parkinson's disease (PD), characterized by dopaminergic neurodegeneration in the substantia nigra (SN), although the effects of this increment remain to be further studied.

An abstract of the article "Actin Modifying Protein CLP36 as a New Marker for Villous Trophoblasts and Its Role in Pregnancies Complicated by Preeclampsia," by Ulrich Miehe, Berthold Huppertz, Peruka Neumaier-Wagner, Mamed Kadyrov, and Werner Rath is presented.

An abstract of the article "Transforming Growth Factor Beta 3 In The Hellp Syndrome: A Placental Analysis," by A. Corradetti, M. Emanuelli, B. Landi, D. Sartini, F. Pierella, S. R. Giannubilo, and A. L. Tranquilli is presented.

Background/aims: Hepatic fibrosis is one kind of common wound-healing response to chronic liver injury. Transforming growth factor (TGF)-β(3) performs an anti-fibrosis function under certain conditions such as pancreatic fibrosis and wound healing. This study aimed at investigating the effect of TGF-β(3) on the histology in the liver of rat with liver fibrosis.Methods: Recombinantadeno-associated virus (rAAV) 2-TGF-β(3) and rAAV2-EGFP were constructed. Rats were randomly divided into normal control group, model group, negative control group and TGF-β(3) treated group. The hepatic fibrosis model was induced by CCl(4) administration. We injected a single dose of either rAAV2-TGF-β(3) or rAAV2-EGFP into the TGF-β(3) group and the negative control group. The histopathologic changes of liver were determined by hematoxylin and eosin (HE) staining and Masson staining. The expressions of type I collagen, MMP-9, MMP-2, and TIMP-1 in liver were detected by Immunohistochemical staining.Results: With the treatment of TGF-β(3), the degree of fibrosis and the deposition of collagen fiber in liver were markedly reduced, and the expression of MMP-9 was obviously increased (P < 0.001), while type I collagen and TIMP-1 were decreased (P = 0.004, P = 0.001) compared with the model group, but the expressed difference of MMP-2 had no statistical significance (P = 0.180).Conclusion: rAAV2-TGF-β(3) reduces the histopathologic damage of liver fibrosis on rats, and it may suppress the synthesis and deposition of type I collagen by regulating the expressions of matrix metalloproteinases and their inhibitors. Potentially, our findings might help with the design of a new TGF-β(3)-based therapy for hepatic fibrosis. [ABSTRACT FROM AUTHOR]

Copyright of Anatolian Journal of Cardiology / Anadolu Kardiyoloji Dergisi is the property of KARE Publishing and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) has been shown to induce cleft palate, in which the molecular etiology of the defect is poorly characterized. Recently, transforming growth factor-β3 (TGF-β3) has been indicated to play an essential role in the development of palatal shelves. In this developmental toxicity study, we investigated the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the expression of TGF-β3 in fetal mice. Pregnant C57BL/6 mice were exposed to corn oil or TCDD (32 μg/kg/day 64 μg/kg/day, per os) at embryonic day 10 (ED10), a drastic inhibition of palatal shelves was induced. By using RT-PCR (reverse transcription-polymerase chain reaction) and Western blot, the expressions of TGF-β3 was investigated. We found that the expression of TGF-β3 was gradually up-regulated in TCDD-treated group. These results suggest that cleft palate can be induced by TCDD exposure, the modification of TGF-β3 is related to its pathogenesis. [ABSTRACT FROM AUTHOR]

This article reviews our understanding of the mechanisms involved in wound healing following injury. Models of preferential healing are described in the foetal skin, aged skin and the oral mucosa. The mechanisms underpinning the improved scarring observed in the oral mucosa are described and the role of resident fibroblasts, in mediating reduced scarring, is discussed. [ABSTRACT FROM AUTHOR]

Background and purpose:Several selective oestrogen receptor modulators (SERMs) with oestrogen agonist effects in bone cells and without increased risk of breast and endometrial cancer have been developed. Here, we have investigated the effects of different types of SERMs on osteoclast differentiation, bone resorption and apoptosis in vitro.Experimental approach:Human peripheral blood-derived CD14+ monocytes were cultured on bovine bone slices in the presence of RANKL, M-CSF, TNF-α and dexamethasone for seven days. Also, CD14+ monocytes were co-cultured either with human SaOS-2 or MG-63 osteosarcoma cells, in the presence of parathyroid hormone. Osteoclast cultures were treated with different SERMs. TRACP+ multinucleated cells and C-terminal telopeptide of type I collagen were used as markers for osteoclast formation and bone resorption, respectively.Key Results:In CD14+ monocyte cultures, tamoxifen directly inhibited human osteoclast formation and bone resorption, while raloxifene and ospemifene had no inhibitory effect. In the co-cultures either with SaOS-2 or MG-63 cells, ospemifene and raloxifene as well as tamoxifen inhibited osteoclast formation in a concentration-dependent manner. The inhibitory effect was associated with an increased production of osteoprotegerin. The anti-oestrogen ICI 182 780 completely reversed the effects of these SERMs.Conclusion and Implications:Tamoxifen had an oestrogen receptor dependent, direct, inhibitory effect on human osteoclast differentiation and bone resorption, whereas ospemifene and raloxifene required osteoblastic cells to achieve a similar inhibition. The effects of ospemifene and raloxifene were mediated by oestrogen receptors by a mechanism involving paracrine induction of osteoprotegerin in cultures with osteoblast derived osteosarcoma cells.British Journal of Pharmacology (2007) 151, 384–395; doi:10.1038/sj.bjp.0707232; published online 10 April 2007 [ABSTRACT FROM AUTHOR]

Epithelial-mesenchymal transition (EMT) is a highly conserved and fundamental process that governs morphogenesis in development and may also contribute to cancer metastasis. Transforming growth factor (TGF-β) is a potent inducer of EMT in various developmental and tumor systems. The analysis of TGF-β signal transduction pathways is now considered a critically important area of biology, since many defects occur in these pathways in embryonic development. The complexity of TGF-β signal transduction networks is overwhelming due to the large numbers of interacting constituents, complicated feedforward, feedback and crosstalk circuitry mechanisms that they involve in addition to the cellular kinetics and enzymatics that contribute to cell signaling. As a result of this complexity, apparently simple but highly important questions remain unanswered, that is, how do epithelial cells respond to such TGF-β signals? System biology and cellular kinetics play a crucial role in cellular function; omissions of such a critical contributor may lead to inaccurate understanding of embryonic EMT. In this review, we identify and explain why certain conditions need to be considered for a true representation of TGF-β signaling in vivo to better understand the controlled, yet delicate mechanism of embryonic EMT. Copyright © 2007 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Nonsyndromic or isolated cleft lip with or without cleft palate (CL/P) occurs in wide geographic distribution with an average birth prevalence of 1/700. We used direct sequencing as an approach to study candidate genes for CL/P. We report here the results of sequencing on 20 candidate genes for clefts in 184 cases with CL/P selected with an emphasis on severity and positive family history. Genes were selected based on expression patterns, animal models, and/or role in known human clefting syndromes. For seven genes with identified coding mutations that are potentially etiologic, we performed linkage disequilibrium studies as well in 501 family triads (affected child/mother/father). The recently reported MSX1 P147Q mutation was also studied in an additional 1,098 cleft cases. Selected missense mutations were screened in 1,064 controls from unrelated individuals on the Centre d'Étude du Polymorphisme Humain (CEPH) diversity cell line panel. Our aggregate data suggest that point mutations in these candidate genes are likely to contribute to 6% of isolated clefts, particularly those with more severe phenotypes (bilateral cleft of the lip with cleft palate). Additional cases, possibly due to microdeletions or isodisomy, were also detected and may contribute to clefts as well. Sequence analysis alone suggests that point mutations in FOXE1, GLI2, JAG2, LHX8, MSX1, MSX2, SATB2, SKI, SPRY2, and TBX10 may be rare causes of isolated cleft lip with or without cleft palate, and the linkage disequilibrium data support a larger, as yet unspecified, role for variants in or near MSX2, JAG2, and SKI. This study also illustrates the need to test large numbers of controls to distinguish rare polymorphic variants and prioritize functional studies for rare point mutations. [ABSTRACT FROM AUTHOR]

The transforming growth factor (TGF) superfamily encompasses about 30 members in mammals. The effect of TGF- subfamily members is exerted and regulated via selective pathways of synthesis and signaling that involve activation of latent TGF-, specific and high-affinity binding to cell membrane serine/threonine kinase receptors, activation of intracellular cascades that include Smad molecules and mitogen-activated protein kinases, and regulated termination of the effect by diverse mechanisms including protein degradation and transcriptional activation. Several comprehensive reviews on TGF- biology in general and on the role of this cytokine in other diseases have been published recently. In recent years an unexpected role of TGF- on lung homeostasis has been revealed. Here, we discuss the contribution of TGF- to the pathogenesis of asthma and chronic obstructive pulmonary disease, two common illnesses of the lung, as well as of lymphangioleiomyomatosis, a rare disease in women. The information we collate and integrate places TGF- at a pivotal point within complex networks that control lung physiology as well as the physiopathology of these lung diseases. [ABSTRACT FROM AUTHOR]

Several members of the transforming growth factor (TGF)-β superfamily are expressed in developing teeth from the initiation stage through adulthood. Of those, TGF-β1 regulates odontoblast differentiation and dentin extracellular matrix synthesis. However, the molecular mechanism of TGF-β3 in dental pulp cells is not clearly understood. In the present study, beads soaked with human recombinant TGF-β3 induced ectopic mineralization in dental pulp from fetal mouse tooth germ samples, which increased in a dose-dependent manner. Further, TGF-β3 promoted mRNA expression, and increased protein levels of osteocalcin (OCN) and type I collagen (COL I) in dental pulp cells. We also observed that the expression of dentin sialophosphoprotein and dentin matrix protein 1 was induced by TGF-β3 in primary cultured dental pulp cells, however, not in calvaria osteoblasts, whereas OCN, osteopontin and osteonectin expression was increased after treatment with TGF-β3 in both dental pulp cells and calvaria osteoblasts. Dentin sialoprotein was also partially detected in the vicinity of TGF-β3 soaked beadsin vivo. These results indicate for the first time that TGF-β3 induces ectopic mineralization through upregulation of OCN and COL I expression in dental pulp cells, and may regulate the differentiation of dental pulp stem cells to odontoblasts. [ABSTRACT FROM AUTHOR]

Objective --Bone morphogenetic protein-2 offers potential benefits for cartilage regeneration. We investigated the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on the regeneration of laryngeal cartilage and respiratory epithelium in a rabbit model. Material and Methods --We used a cricoid defect rabbit model. Twenty-four rabbits were randomly divided into four equal groups. Two groups were treated with 5 μg of rhBMP-2 delivered on an absorbable collagen sponge and the other two groups were used as controls. One group of treated rabbits and one group of control rabbits were euthanized 1 week after surgery, while the others were euthanized 4 weeks after surgery. The healing pattern of the laryngeal wound was evaluated by means of histomorphometry. Results --Regeneration of both the epithelial layer and cartilage was significantly better in rabbits treated with rhBMP-2. Four weeks after surgery, the cricoid cartilage defect was completely repaired by new cartilage and new bone in rabbits treated with rhBMP-2. Furthermore, the lining respiratory epithelium healed more rapidly in treated rabbits. Conclusion --rhBMP-2, delivered via an absorbable collagen sponge, induces complete regeneration and repair of rabbit cricoid cartilage defects. It also induces faster relining and regeneration of airway epithelium than in control rabbits. [ABSTRACT FROM AUTHOR]

Objectives: Determine the model of inheritance of non-syndromic cleft palate in humans.Design: Complex segregation analysis performed in families of consecutive newborns affected with non-syndromic cleft palate.Setting and Sample Population: The Latin American Collaborative Study of Congenital Malformations (ECLAMC). Four hundred and seven consecutive newborns affected with non-syndromic cleft palate registered during the period 1967-97.Outcome Measure: Likelihood ratio test and Akaike information criterion (AIC) values.Results: The single major locus recessive model provided a significantly better explanation of the data. It was the most parsimonious and had the smallest AIC value of the six models tested with approximately the same likelihood as the general model (chi2 = 2.44, p = 0.5).Conclusions: To have defined a genetic model for non-syndromic cleft palate and provided evidence for a single major locus inheritance suggests that genetic linkage studies could be implemented. [ABSTRACT FROM AUTHOR]

The aim of the present work was to investigate the possibility of achieving buccal delivery of a problematic drug, acyclovir, from films based on chitosan hydrochloride (HCS) and polyacrylic acid sodium salt (PAA). At first, the ionic interaction between HCS and PAA in distilled water was investigated by means of rheological and turbidimetric analysis. Films containing 1 mg/cm2 of acyclovir and based on pure HCS and on HCS and PAA mixed in different ratios were prepared by casting technique. The films were subjected to hydration, rheological, mucoadhesion, drug release, “wash away,” and permeation/penetration measurements. A commercial cream containing acyclovir and an aqueous acyclovir suspension were used as references. The addition of PAA to HCS produced a decrease in film hydration. Films based on HCS/PAA weight ratio close to interaction product stoichiometry were characterized by higher rigidity and better “wash away” properties with respect to the other films and the reference formulation. The worst mucoadhesive properties were shown by films based on mixing ratios close to interaction product stoichiometry. The addition of PAA to HCS produced a lowering in drug release profile. All the films examined promoted the permeation of acyclovir across porcine cheek epithelium when compared with acyclovir suspension and the commercial cream. The penetration enhancement properties were affected by the mixing ratio of the two polymers. The film based on 1/1.3 HCS/PAA weight ratio, besides possessing the best resilience properties on the mucosa, was also characterized by the highest permeation profile and, therefore, represents a promising formulation for buccal delivery of acyclovir. [ABSTRACT FROM AUTHOR]

The surrounding extracellular matrix of airway wall tissues changes in response to mechanical stresses and hypoxia. The presence of matrix metalloproteinase-9 (MMP-9) and its inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), is correlated with collagen degradation and tissue repair in lung disorders. The aim of this study was to evaluate the expression of MMP-9 and TIMP-1 in the lung of fetal rats with nitrofen-induced congenital diaphragmatic hernia (CDH). Administering 100 mg of nitrofen dissolved in 1 ml olive oil to pregnant Wistar rats on day 9 of gestation induced left-sided CDH in fetal rats. In control animals, the same dose of olive oil was given without nitrofen. Cesarean section was performed on day 21 of gestation. The fetuses were divided into two groups: normal controls (n = 10) and nitrofen-induced left-sided CDH (n = 10). Immunoreactivity of the staining for MMP-9 and TIMP-1 in the lung tissues was semiquantitatively analyzed using the staining scores. The relative amount of MMP-9 or TIMP-1 divided by the amount of β-actin for each lung sample was measured by using the real-time reverse-transcriptase polymerase chain reaction. The immunoreactivity of MMP-9 was significantly increased in the CDH group (n = 5) compared with the control group (n = 5) (p = 0.031). On the other hand, the immunoreactivity of TIMP-1 in the two groups was not significantly different (n = 0.134). The relative amount of MMP-9 (or TIMP-1) in the CDH group (n = 5) does not differ significantly from that in the control group (n = 5) (p = 0.059, 0.596, respectively), but the relative amount of MMP-9 is higher in the CDH group, although it is not significantly higher. On the other hand, the ratios of MMP-9 to TIMP-1 were significantly higher in the CDH group (p = 0.028). In conclusion, fetal rats with nitrofen-induced CDH, a model of respiratory disorders, manifested the excess of MMP-9 activity due to the absence of TIMP-1 that would suggest a trend toward disruption of the extracellular matrix in the CDH lung tissues. [ABSTRACT FROM AUTHOR]

Calcium has an established role in the normal homeostasis of mammalian skin and serves as a modulator in keratinocyte proliferation and differentiation. Gradients of calcium concentration increasing from 0.5 mM in the basal layer to > 1.4 mM in the stratum granulosum are consistent with migration patterns in response to minor abrasion (normal wear). Dermal fibroblasts require calcium but are approximately 100 times less sensitive than keratinocytes. Normal calcium metabolism in the skin is dependent on cell membrane and cytosolic calcium binding proteins (calmodulin, cadherins, etc.), but their modulation through parathyroid hormone, vitamin D or growth factors in normal or damaged tissue is not well documented. In wound repair, calcium is predominantly involved as Factor IV in the hemostatic phase, but it is expected to be required in epidermal cell migration and regeneration patterns in later stages of healing. Calcium alginate dressings are designed to liberate calcium early in the acute phase to promote hemostasis, but it is presently unclear whether the supplementary calcium influences the intracellular environment at later stages of wound repair, notably during the remodeling phase. Although experimental studies suggest that control of calcium is obligatory in wound management, we know very little as to how calcium in the wound bed is modulated through hormones, vitamin D, or various growth factors. Also, there is limited information as to how calcium released either from dressings, platelets, or from the circulation through the action of parathyroid hormone, growth factors or other modulators influences cell migration and remodeling in skin wounds, although experimental models suggest that management of calcium is essential in wound management. (WOUND REP REG 2002;10:271–285). [ABSTRACT FROM AUTHOR]

Laryngeal wound healing is essential following laryngotracheal surgery. Patients with poor wound healing develop poor epithelial closure and increased granulation tissue which cause a stenosis of the repaired airway. Transforming growth factor-β3 has been shown to enhance wound healing in cutaneous wounds, but has never been used in the airway. This study utilized a rabbit laryngeal wound-healing model that has been shown to be reproducible with limited morbidity. Thirty-four rabbits underwent a cricoid-split operation with collagen sponge insertion. All animals were classified randomly into three groups: local administration of placebo (Group G1, n = 13), 0.18 μg transforming growth factor-β3 (Group G2, n = 11) and of 1.0 μg transforming growth factor-β3 (Group G3, n = 10). All animals survived the postoperative period without respiratory distress. The airway was harvested six days after surgery and assessed by light microscopy. Histologic evidence for healing was subjectively graded by two blinded, independent investigators, and the results were statistically analyzed for significance. A significant improvement in the epithelial closure (p < 0.01) and subepithelial connective tissue closure (p < 0.005) was found in the 1.0 μg transforming growth factor-β3 group (G3) compared with the placebo group (G1). Analysis of the 0.18 μg transforming growth factor-β3 group (G2) did not reveal any significant differences compared with the placebo group (G1). These results suggest an application for transforming growth factor-β3 in accelerating wound healing in the larynx. [ABSTRACT FROM AUTHOR]

Pituitary prolactin (PRL) cell gene expression and proliferation are regulated by various hormones and growth factors. Transforming growth factor beta (TGFβ) and basic fibroblast growth factor (bFGF) have been implicated in the regulation of anterior pituitary function. To study the roles of TGFβ and bFGF in anterior pituitary cell function, we analyzed normal and neoplastic pituitary cells in serum-free media. The various isoforms of TGFβ and TGFβ receptor types I, II, and III were also analyzed by reverse transcriptionpolymerase chain reaction (RT-PCR) in pituitary cells. Transforming growth factor beta 1 (TGFβ1) stimulated PRL expression and PRL cell proliferation in normal pituitary. TGFβ1 stimulated PRL expression, but inhibited proliferation in the growth hormone (GH) and PRL-producing GH cells. Estradiol 17β (E) and bFGF stimulated PRL gene expression in normal pituitary and GH cells, whereas E inhibited and bFGF stimulated TGFβ1 mRNA levels in normal pituitary PRL cells, but not in GH cells. Both normal pituitary and GH cells expressed the mRNAs for TGFβ1, TGFβ2, and TGFβ3 isoforms and for TGFβ receptors I, II, and III. These results indicate that there is a relative loss of regulatory control by growth factors in neoplastic GH cells compared to normal pituitary PRL cells. [ABSTRACT FROM AUTHOR]