Development of Pilomatrixoma at the Vaccination Site: A Rare Complication of COVID-19 Vaccination - A Case Report.

BACKGROUND Pilomatrixoma, pilomatricoma, or calcifying epithelioma of Malherbe, is a common benign tumor that arises from the base of the hair follicle. Pilomatrixoma has previously been reported at vaccination sites. This report is of a 65-year-old man with an 18-month history of an enlarging pilomatrixoma of the left upper arm at the vaccination site, following a first COVID-19 vaccination. CASE REPORT The case involves a 65-year-old man who developed a left shoulder mass 1.5 years ago. The mass appeared at his COVID-19 vaccine site 3 months after receiving the first dose. The mass measures 3 cm in diameter, was mobile, and exhibited no signs of infection in the physical examination. Surgical excision was performed, and pathology confirmed the mass as a pilomatrixoma, characterized by basaloid cells and keratinization. Three months after surgery, no recurrence was observed. CONCLUSIONS This report has presented an association between vaccination injection sites and pilomatrixoma aligning with previous findings. Enhanced awareness about this condition can substantially improve pilomatrixoma diagnosis accuracy and reduce unnecessary examinations and treatments. Furthermore, we recommend that, along with clinical symptoms, ultrasound imaging be considered a valuable diagnostic tool for pilomatrixoma, with histopathological results to confirm the diagnosis.

Effects of Mn-Depleted Zone Formation on Acicular Ferrite Transformation in Weld Metals under High Heat Input Welding.

In this present work, during high heat input welding of the weld metal, different types of Mn-depleted zones were achieved by different cooling rates. The effects of cooling rates on Mn-depleted zone formation and acicular ferrite (AF) transformation were analyzed. The Mn-depleted zone around the inclusions, as well as the interface concentration of Mn atoms, are found to be significantly different with different cooling rates. When the cooling rate is 10 °C/s, the interface concentration of Mn atoms around the inclusions is the lowest, the area of Mn-depleted zone is the smallest, and the proportion of AF in the weld metal is the highest. As the cooling rate decreases further, the interface concentration of Mn begins to rise, the area of the Mn-depleted zone gradually expands, and the proportion of AF decreases. However, when the cooling rate reaches 100 °C/s, only a very small amount of MnS precipitates, no Mn-depleted zone forms around the inclusions, and acicular ferrite cannot be produced effectively in the weld metal.

The Challenges of Enrollment and Retention: A Systematic Review of Psychosocial Behavioral Interventions for Patients With Cancer and Their Family Caregivers.

CONTEXT: Psychosocial behavioral interventions (PBIs) that target patients with cancer and their caregivers face challenges in participant enrollment and retention. OBJECTIVES: 1) Describe characteristics of the patient-caregiver PBI studies; 2) examine participant enrollment and retention rates; 3) identify factors influencing participant enrollment and retention rates; and 4) explore the strategies to promote enrollment and retention rates. METHODS: We identified randomized controlled trials that tested PBIs among adult patients with cancer and caregivers in five electronic databases. We conducted narrative and quantitative analyses to synthesize our findings. RESULTS: Among 55 qualified studies reviewed, most tested the efficacy of PBIs (n = 42) and used two study arms (n = 48). In-person meeting was the most common PBI delivery mode. The primary outcomes included quality of life, physical health, and symptoms. The average of enrollment rates of patient-caregiver dyads was 33% across studies (range 8%-100%; median = 23%). The average retention rate at the end of follow-ups was 69% (range 16%-100%; median = 70%). The number of study arms, recruitment method, type of patient-caregiver relationship, and intervention duration influenced enrollment rates. Study design (efficacy vs. pilot), follow-up duration, mode of delivery, type of relationship, and intervention duration influenced retention rates. Sixteen studies reported retention strategies, including providing money/gift cards upon study completion and/or after follow-up survey, and excluding patients with advanced cancer. CONCLUSION: Researchers need to incorporate effective strategies to optimize enrollment and retention in patient-caregiver PBI trials. Researchers need to report detailed study processes and PBI information to improve research transparency and increase consistency.

Cardiovascular disease, risk factors, and health behaviors among cancer survivors and spouses: A MEPS Study.

PURPOSE: The purpose of this study was to examine the prevalences of CVD, CVD risk factors. and health behaviors among cancer survivor-spouse dyads, assess how these prevalences differ by role (survivor vs spouse) and gender, and report congruences in health behaviors between survivors and their spouses. METHODS: We identified 1026 survivor-spouse dyads from the 2010-2015 Medical Expenditure Panel Survey. We used weighted multivariable logistic and linear regressions to analyze the data related to CVD, CVD risk factors, and health behaviors. RESULTS: Survivors and spouses reported high prevalences of CVD and CVD risk factors but low engagement in healthy behaviors, including non-smoking, physical activity, and maintaining a healthy weight (proxy for healthy diet). Gender and role differences were significantly related to the prevalence of CVD, CVD risk factors, and health behaviors among survivors and spouses. From 39% to 88% of survivors and spouses were congruent in their current smoking status, physical activity engagement/disengagement, and BMI. CONCLUSION: Cancer survivors and spouses have high rates of CVD and CVD risk factors and poor engagement in healthful lifestyle behaviors. A high proportion of survivors and spouses were congruent in their current smoking status, physical activity engagement/disengagement, and BMI. Effective lifestyle interventions are needed for this high-risk population. Couple-focused interventions may be well-suited for these dyads and warrant further study. IMPLICATIONS FOR CANCER SURVIVORS: Both cancer survivors and their spouses need to be non-moking, more physically active, and maintain normal BMI in order to reduce their high risk of CVD and CVD risk factors.

The Axolotl Fibula as a Model for the Induction of Regeneration across Large Segment Defects in Long Bones of the Extremities.

We tested the ability of the axolotl (Ambystoma mexicanum) fibula to regenerate across segment defects of different size in the absence of intervention or after implant of a unique 8-braid pig small intestine submucosa (SIS) scaffold, with or without incorporated growth factor combinations or tissue protein extract. Fractures and defects of 10% and 20% of the total limb length regenerated well without any intervention, but 40% and 50% defects failed to regenerate after either simple removal of bone or implanting SIS scaffold alone. By contrast, scaffold soaked in the growth factor combination BMP-4/HGF or in protein extract of intact limb tissue promoted partial or extensive induction of cartilage and bone across 50% segment defects in 30%-33% of cases. These results show that BMP-4/HGF and intact tissue protein extract can promote the events required to induce cartilage and bone formation across a segment defect larger than critical size and that the long bones of axolotl limbs are an inexpensive model to screen soluble factors and natural and synthetic scaffolds for their efficacy in stimulating this process.

Proteomic analysis of fibroblastema formation in regenerating hind limbs of Xenopus laevis froglets and comparison to axolotl.

BACKGROUND: To gain insight into what differences might restrict the capacity for limb regeneration in Xenopus froglets, we used High Performance Liquid Chromatography (HPLC)/double mass spectrometry to characterize protein expression during fibroblastema formation in the amputated froglet hindlimb, and compared the results to those obtained previously for blastema formation in the axolotl limb. RESULTS: Comparison of the Xenopus fibroblastema and axolotl blastema revealed several similarities and significant differences in proteomic profiles. The most significant similarity was the strong parallel down regulation of muscle proteins and enzymes involved in carbohydrate metabolism. Regenerating Xenopus limbs differed significantly from axolotl regenerating limbs in several ways: deficiency in the inositol phosphate/diacylglycerol signaling pathway, down regulation of Wnt signaling, up regulation of extracellular matrix (ECM) proteins and proteins involved in chondrocyte differentiation, lack of expression of a key cell cycle protein, ecotropic viral integration site 5 (EVI5), that blocks mitosis in the axolotl, and the expression of several patterning proteins not seen in the axolotl that may dorsalize the fibroblastema. CONCLUSIONS: We have characterized global protein expression during fibroblastema formation after amputation of the Xenopus froglet hindlimb and identified several differences that lead to signaling deficiency, failure to retard mitosis, premature chondrocyte differentiation, and failure of dorsoventral axial asymmetry. These differences point to possible interventions to improve blastema formation and pattern formation in the froglet limb.

Effects of resolvin D1 on cell survival and cytokine expression of human gingival fibroblasts.

BACKGROUND: Tissue breakdown in periodontitis is initiated by bacteria, such as Porphyromonas gingivalis, and is caused largely by host responses. Resolvins protect the host against acute inflammation by blocking the migration of polymorphonuclear neutrophils to initiate resolution. The effects of resolvins on human gingival fibroblasts (HGFs) are unknown. This study examines the effects of resolvin D1 on HGF survival and cytokine expression when treated with or without P. gingivalis supernatant. METHODS: Cytotoxicity of resolvin D1 on HGFs with or without a toxic level of P. gingivalis supernatant was measured with lactate dehydrogenase assays. Cytokine arrays were performed on HGF-conditioned media treated with or without resolvin D1 and with or without P. gingivalis supernatant. RESULTS: Resolvin D1 had no cytotoxic effects on HGFs at concentrations between 1 and 1,000 nM (all P > 0.05). Resolvin D1 (1,000 nM) significantly inhibited the toxic effects of 13.5% (v/v) P. gingivalis supernatant on HGFs (P = 0.002). Resolvin D1 significantly reduced the expression of interleukin (IL)-6 (P = 0.010) and monocyte chemoattractant protein (MCP)-1 (P = 0.04) in untreated fibroblasts. P. gingivalis (10%) supernatant significantly increased the expression levels of granulocyte-macrophage colony-stimulating factor (CSF), granulocyte CSF, growth-regulated oncogene (GRO), IL-5, IL-6, IL-7, IL-8, IL-10, MCP-1, MCP-2, MCP-3, and monokine induced by γ-interferon. Resolvin D1 significantly reduced the expression of GRO (P = 0.04), marginally reduced the levels of MCP-1 (P = 0.10), and marginally increased the levels of transforming growth factor (TGF)-ß1 (P = 0.07) from HGFs treated with P. gingivalis supernatant. CONCLUSIONS: Resolvin D1 altered the cytotoxicity of P. gingivalis supernatant on HGFs. Resolvin D1 significantly reduced GRO, marginally reduced MCP-1, and marginally increased TGF-ß1 from P. gingivalis-treated HGFs, which could alter the ability of P. gingivalis to induce inflammation.

Cell cycle regulation and regeneration.

Regeneration of ear punch holes in the MRL mouse and amputated limbs of the axolotl show a number of similarities. A large proportion of the fibroblasts of the uninjured MRL mouse ear are arrested in G2 of the cell cycle, and enter nerve-dependent mitosis after injury to form a ring-shaped blastema that regenerates the ear tissue. Multiple cell types contribute to the establishment of the regeneration blastema of the urodele limb by dedifferentiation, and there is substantial reason to believe that the cells of this early blastema are also arrested in G2, and enter mitosis under the influence of nerve-dependent factors supplied by the apical epidermal cap. Molecular analysis reveals other parallels, such as; (1) the upregulation of Evi5, a centrosomal protein that prevents mitosis by stabilizing Emi1, a protein that inhibits the degradation of cyclins by the anaphase promoting complex and (2) the expression of sodium channels by the epidermis. A central feature in the entry into the cell cycle by MRL ear fibroblasts is a natural downregulation of p21, and knockout of p21 in wild-type mice confers regenerative capacity on non-regenerating ear tissue. Whether the same is true for entry into the cell cycle in regenerating urodele limbs is presently unknown.

Suture compression induced bone resorption with intensified MMP-1 and 13 expressions.

UNLABELLED: Suture compression is a widely used approach to inhibit maxillary growth; however, biological responses in sutures to compressive force are still unclear. The objective of this pilot study was to investigate the matrix metalloproteinase (MMP) expression and osteoclast activities during the midpalatal suture compression. METHODS: 56 six-week old male C57BL/6 mice were randomly assigned to the control and compression groups. The mice in the compression and control groups received helix springs bonded to the maxillary molars delivering initial compressive forces of 0.20 and 0N (no activation), respectively. On Days 1, 4, 7 and 14, animals were sacrificed and scanned using micro-computed tomography to quantify suture width and bone mineral density. Serial histological sections were stained with HE, TRAP, and immunohistochemistry to observe changes in bone resorption, osteoclast activities, and MMP-1, 8, and 13 expressions. Bone volume/total volume (Bv/Tv) ratio, osteoclast count, osteoclast covering area, and MMP expression intensity were measured. The Mann-Whitney and the Kruskal-Wallis tests with Bonferroni post-hoc corrections were performed to compare differences between groups and between time points in the same group at significant level of P<0.05. RESULTS: Compared to the control, suture width in the compression group was significantly reduced on Day 1, but continuously widened with reduced bone mineral density afterwards. With MMP-1 and -13 evidently intensified expressions, osteoclast number and activities significantly increased, leading to reduced Bv/Tv ratio and progressive bone resorption from Days 4 to 14. CONCLUSIONS: Suture compression elevated the MMP-1 and 13 expressions, activated osteoclasts, reduced bone density, and induced bone resorption adjacent to the suture. It suggests that suture compression can be used for bone volume reduction.

Effects of Calendula officinalis on human gingival fibroblasts.

BACKGROUND: Calendula officinalis is commonly called the marigold. It is a staple topical remedy in homeopathic medicine. It is rich in quercetin, carotenoids, lutein, lycopene, rutin, ubiquinone, xanthophylls, and other anti-oxidants. It has anti-inflammatory properties. Quercetin, one of the active components in Calendula, has been shown to inhibit recombinant human matrix metalloproteinase (MMP) activity and decrease the expression of tumor necrosis factor-α, interleukin-1ß (IL), IL-6 and IL-8 in phorbol 12-myristate 13-acetate and calcium ionophore-stimulated human mast cells. OBJECTIVES: To examine the effects of Calendula on human gingival fibroblast (HGF) mediated collagen degradation and MMP activity. MATERIAL AND METHODS: Lactate dehydrogenate assays were performed to determine the non-toxic concentrations of Calendula, doxycycline and quercetin. Cell-mediated collagen degradation assays were performed to examine the inhibitory effect on cell-mediated collagen degradation. Gelatin zymography was performed to examine their effects on MMP-2 activity. The experiments were repeated three times and ANOVA used for statistical analyses. RESULTS: Calendula at 2-3% completely inhibited the MMP-2 activity in the zymograms. Doxycycline inhibited HGF-mediated collagen degradation at 0.005, 0.01, 0.02 and 0.05%, and MMP-2 activity completely at 0.05%. Quercetin inhibited HGF-mediated collagen degradation at 0.005, 0.01 and 0.02%, and MMP-2 activity in a dose-dependent manner. Calendula inhibited HGF-mediated collagen degradation and MMP-2 activity more than the same correlated concentration of pure quercetin. CONCLUSION: Calendula inhibits HGF-mediated collagen degradation and MMP-2 activity more than the corresponding concentration of quercetin. This may be attributed to additional components in Calendula other than quercetin.

Effects of alendronate on human osteoblast-like MG63 cells and matrix metalloproteinases.

OBJECTIVE: The objective of this study was to examine the effects of alendronate on the expression and activity of matrix metalloproteinases (MMPs) and the expression of the tissue inhibitors of MMPs (TIMPs) from human osteoblast-like MG63 cells. MATERIALS AND METHODS: MG63 cells were exposed to various concentrations of alendronate. Cell proliferation and cytotoxicity were evaluated by water-soluble tetrazolium-1 and lactate dehydrogenase, respectively. MG63-mediated collagen degradation was assessed utilising Type I collagen assays. Conditioned media and membrane extracts were collected for Western blot analyses of select MMPs and TIMPs. Gelatin zymography gels were incubated with alendronate to assess its effects on MMP-2 activity. RESULTS: Alendronate affected MG63 proliferation and cytotoxicity at concentrations equal to/or greater than 10(-5) M (all p < 0.05). There were no significant differences in the collagen degrading ability of treated cells at non-toxic levels vs. untreated cells. Alendronate had no effects on the expression of MMP-2 or MT1-MMP (membrane type-1 MMP) in the conditioned media or membrane extracts, and of MMP-1 or TIMP-2 in the conditioned media. TIMP-2 in the membrane extracts was not detectable. MMP-2 activity in the zymograms was inhibited by 10(-3) and 10(-2) M alendronate. CONCLUSION: Alendronate at 10(-5) M or higher was toxic to the cells. Alendronate at 10(-8) to 10(-6) M did not alter the expression of MMP-1, MMP-2, MT1-MMP or TIMP-2, as well as did not alter collagen degradation. Alendronate inhibited MMP-2 activity at 10(-3) and 10(-2) M in the zymograms. In conclusion, non-toxic levels of alendronate (10(-8) to 10(-6) M) did not alter MMP expression in MG63 cells or inhibit MMP-2 activity.

Effects of cigarette smoke condensate on oral squamous cell carcinoma cells.

OBJECTIVE: Epidemiological studies have reported that tobacco use is a major etiological factor for oral cancer. Several matrix metalloproteinases (MMPs) have been shown to play important roles in the invasion and metastasis of oral squamous cell carcinomas, especially MMP-2 and MMP-9. This study examined the effects of cigarette smoke condensate (CSC) on oral cancer cells. DESIGN: Two oral squamous cell carcinoma cell lines, SCC-25 (metastatic) and CAL-27 (non-metastatic), were exposed to different concentrations of CSC and examined for their collagen degrading ability and MMP production using collagen degradation assays, zymograms and Western blots. RESULTS: Exposure to CSC increased the collagen degrading ability of the metastasizing cell line (SCC-25) by a mechanism involving increased MMP-2 and MMP-9 production. CONCLUSION: CSC increased the collagen degrading ability of SCC-25 by increasing the MMP-2 and MMP-9 protein levels. Continued cigarette smoking in oral cancer patients may result in decreased survival rates due to enhanced metastatic potential of the cancer cells.

Network based transcription factor analysis of regenerating axolotl limbs.

BACKGROUND: Studies on amphibian limb regeneration began in the early 1700's but we still do not completely understand the cellular and molecular events of this unique process. Understanding a complex biological process such as limb regeneration is more complicated than the knowledge of the individual genes or proteins involved. Here we followed a systems biology approach in an effort to construct the networks and pathways of protein interactions involved in formation of the accumulation blastema in regenerating axolotl limbs. RESULTS: We used the human orthologs of proteins previously identified by our research team as bait to identify the transcription factor (TF) pathways and networks that regulate blastema formation in amputated axolotl limbs. The five most connected factors, c-Myc, SP1, HNF4A, ESR1 and p53 regulate ~50% of the proteins in our data. Among these, c-Myc and SP1 regulate 36.2% of the proteins. c-Myc was the most highly connected TF (71 targets). Network analysis showed that TGF-ß1 and fibronectin (FN) lead to the activation of these TFs. We found that other TFs known to be involved in epigenetic reprogramming, such as Klf4, Oct4, and Lin28 are also connected to c-Myc and SP1. CONCLUSIONS: Our study provides a systems biology approach to how different molecular entities inter-connect with each other during the formation of an accumulation blastema in regenerating axolotl limbs. This approach provides an in silico methodology to identify proteins that are not detected by experimental methods such as proteomics but are potentially important to blastema formation. We found that the TFs, c-Myc and SP1 and their target genes could potentially play a central role in limb regeneration. Systems biology has the potential to map out numerous other pathways that are crucial to blastema formation in regeneration-competent limbs, to compare these to the pathways that characterize regeneration-deficient limbs and finally, to identify stem cell markers in regeneration.

Effects of cigarette smoke condensate and nicotine on human gingival fibroblast-mediated collagen degradation.

BACKGROUND: Members of the matrix metalloproteinase (MMP) family have been shown to be involved in periodontal disease. Risk factors for periodontal disease include tobacco smoking. Cigarette smoke condensate (CSC) is comprised of thousands of chemicals. Nicotine is one of the active components in tobacco. This study compares the effects of CSC and nicotine at the level in CSC on the collagen-degrading ability of human gingival fibroblasts (HGFs) and the expression of selected MMPs and tissue inhibitors of metalloproteinases (TIMPs). METHODS: HGFs were seeded in six-well collagen-coated plates, exposed to 100 µg/mL (2.4 µg/mL nicotine) of CSC or 2.4 µg/mL nicotine for 3 days, and then collagen degradation was analyzed. After 3 days exposure to CSC or nicotine, the conditioned media from HGFs was collected and the membrane proteins were extracted for gelatin zymography and Western blot analyses. The mRNA levels of MMP-2, MMP-14, and TIMP-2 were measured by reverse transcription-polymerase chain reaction. RESULTS: The CSC increased collagen degradation, and increased the levels of TIMP-2, MMP-14, and the active MMP-2 in the membrane extracts, and their mRNA levels. CSC also increased the level of active MMP-2 in the conditioned media. Nicotine at the level in CSC (2.4 µg/mL) had little influence on collagen degradation, as well as on the protein and mRNA levels of MMP-2, MMP-14, and TIMP-2. CONCLUSIONS: CSC may increase HGF-mediated collagen degradation by affecting membrane-associated MMPs and TIMPs.

Matrix metalloproteinase expression during blastema formation in regeneration-competent versus regeneration-deficient amphibian limbs.

We used an antibody array to compare the protein expression of matrix metalloproteinases (MMPs)-1, -2, -3, -8, -9, -10, and -13, as well as the tissue inhibitors of metalloproteinases (TIMPs)-1, -2, and -4 during blastema formation in amputated hindlimbs of regeneration-competent wild-type axolotls and stage-54 Xenopus, and regeneration-deficient short-toes axolotls and Xenopus froglets. Expression of MMP-9 and -2 was also compared by zymography. Both short-toes and froglet failed to up-regulate MMPs in a pattern comparable to the wild-type axolotl, suggesting that subnormal histolysis is at least in part responsible for the poor blastema formation characteristic of both short-toes and froglet. MMP levels were much lower in amputated stage-54 Xenopus limb buds than in the other animals, suggesting that blastema formation in these limb buds requires much less extracellular matrix degradation than in fully differentiated limbs. TIMP expression patterns followed the same trends as the MMP's in each group of animals.

Ginsenoside Rb1 and paeoniflorin inhibit transient receptor potential vanilloid-1-activated IL-8 and PGE2 production in a human keratinocyte cell line HaCaT.

Ginsenoside Rb1 (GRb1) and paeoniflorin (PF) are active substances of Chinese traditional herbs and have been commonly used to treat skin inflammation diseases, but little is known about the mechanisms involved. In the present study, the effects of GRb1 and PF on the production of inflammatory mediators and the possible mechanisms of transient receptor potential vanilloid-1 (TRPV1) in these mediators induction were explored. It has been shown that GRb1 and PF inhibited the productions of IL-8 and PGE2 induced by capsaicin (CAP) in HaCaT cells and HEK 293T-TRPV1 cells (which were transgenic and overexpressed TRPV1) but had no effect on HEK 293T mock cells (p<0.05). Besides, CAP was able to induce calcium influx and nuclear factor kappa B(NF-κB) transcriptional activity in HaCaT cells and HEK 293T-TRPV1 cells, but had no effect on HEK 293T mock cells. Furthermore, GRb1 inhibited CAP-induced calcium influx and NF-κB transcriptional activity in both HaCaT cells and HEK 293T-TRPV1 cells. However, PF decreased CAP-induced calcium influx and NF-κB transcriptional activity only in HaCaT cells. This would suggest that GRb1 inhibits CAP-induced calcium influx and NF-κB activity through TRPV1 signal, while calcium influx and NF-κB activity might not be involved in the inhibitory effect of PF on TRPV1 signal. Furthermore, the inhibitory rates of GRb1 and PF on IL-8 and PGE2 production were higher than those caused by capsazepine, an antagonist of TRPV1, suggesting that GRb1 and PF have great potential in clinical treatment of skin diseases.

Amphibians as research models for regenerative medicine.

The ability to regenerate bone across a critical size defect would be a marked clinical advance over current methods for dealing with such structural gaps. Here, we briefly review the development of limb bones and the mandible, the regeneration of urodele limbs after amputation, and present evidence that urodele and anuran amphibians represent a valuable research model for the study of segment defect regeneration in both limb bones and mandible.

Proteomic analysis of blastema formation in regenerating axolotl limbs.

BACKGROUND: Following amputation, urodele salamander limbs reprogram somatic cells to form a blastema that self-organizes into the missing limb parts to restore the structure and function of the limb. To help understand the molecular basis of blastema formation, we used quantitative label-free liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS)-based methods to analyze changes in the proteome that occurred 1, 4 and 7 days post amputation (dpa) through the mid-tibia/fibula of axolotl hind limbs. RESULTS: We identified 309 unique proteins with significant fold change relative to controls (0 dpa), representing 10 biological process categories: (1) signaling, (2) Ca2+ binding and translocation, (3) transcription, (4) translation, (5) cytoskeleton, (6) extracellular matrix (ECM), (7) metabolism, (8) cell protection, (9) degradation, and (10) cell cycle. In all, 43 proteins exhibited exceptionally high fold changes. Of these, the ecotropic viral integrative factor 5 (EVI5), a cell cycle-related oncoprotein that prevents cells from entering the mitotic phase of the cell cycle prematurely, was of special interest because its fold change was exceptionally high throughout blastema formation. CONCLUSION: Our data were consistent with previous studies indicating the importance of inositol triphosphate and Ca2+ signaling in initiating the ECM and cytoskeletal remodeling characteristic of histolysis and cell dedifferentiation. In addition, the data suggested that blastema formation requires several mechanisms to avoid apoptosis, including reduced metabolism, differential regulation of proapoptotic and antiapoptotic proteins, and initiation of an unfolded protein response (UPR). Since there is virtually no mitosis during blastema formation, we propose that high levels of EVI5 function to arrest dedifferentiated cells somewhere in the G1/S/G2 phases of the cell cycle until they have accumulated under the wound epidermis and enter mitosis in response to neural and epidermal factors. Our findings indicate the general value of quantitative proteomic analysis in understanding the regeneration of complex structures.

Effects of provisional acrylic resins on gingival fibroblast cytokine/growth factor expression.

STATEMENT OF PROBLEM: Several studies have reported that polymerized resin materials may release agents into surrounding tissues. These agents could alter cytokine/growth factor expression. PURPOSE: The purpose of this study was to determine the effects that provisional acrylic resins have on cell toxicity and the expression of cytokines/growth factors from human gingival fibroblasts (HGFs). MATERIAL AND METHODS: The materials used in this study were chemically activated bis-acryl composite (Chem-Bis), chemically activated polyethyl methacrylate (Chem-PEMA), chemically activated polymethyl methacrylate (Chem-PMMA), and heat-activated polymethyl methacrylate (Heat-PMMA) resins. HGFs were incubated for 72 hours in the presence of eluate from each resin and in the absence of any eluate (negative control). The conditioned media were then collected and stored at -70 degrees C. Cell toxicity was determined using a lactate dehydrogenase method. Cytokine/growth factor expression was examined using cytokine antibody arrays. The experiments were repeated 3 times. The data were analyzed with 1-way ANOVA, Mann-Whitney test, and 1-sample t test (alpha=.05). RESULTS: There was no significant cell toxicity observed from the eluates. The cytokine/growth factor expression induced by Chem-Bis was significantly greater than the control for growth-regulated oncogene (GRO) (P<.001), monocyte chemoattractant protein-1 (MCP-1) (P=.031), and tumor necrosis factor-beta (TNF)-beta (P=.009). For Chem-PEMA, the cytokine/growth factor expression was significantly greater than the control for GRO-alpha (P=.022), interleukin (IL)-13 (P=.031), and TNF-alpha (P=.017). The cytokines/growth factors induced by Chem-PEMA were significantly less than the control (P=.008) and Chem-Bis for IL-8 (P=.042). The expression induced by Chem-PMMA was significantly greater than the control for IL-13 (P=.036), IL-1 alpha (P=.003), IL-2 (P=.020), and IL-5 (P=.045). Finally, Heat-PMMA induced significantly greater levels than the control for GRO (P<.001) and IL-13 (P=.008). CONCLUSIONS: This study demonstrated that the resins evaluated were nontoxic to the HGFs. There were changes in the cytokine/growth factor levels that were statistically significant, but may not be clinically significant.

An evaluation of DNA yield, DNA quality and bite registration from a dental impression wafer.

BACKGROUND: The authors determined the amount and quality of the DNA captured by a bite impression wafer and analyzed any inaccuracies in the impression wafer. METHODS: The authors made bite registrations for subjects aged 7 to 12 years by using a dental impression wafer (Toothprints, Kerr, Orange, Calif.), obtained an oral rinse sample, took cheek cells by using buccal swabs and made an alginate impression to pour a stone model. They extracted and quantified the DNA from the dental impression wafer, mouthwash and buccal swabs by using the Quant-iT PicoGreen (Invitrogen, Carlsbad, Calif.) assay and a real-time polymerase chain reaction (RT-PCR) assay. They compared the stone models and imprints from the wafer. RESULTS: The average amounts of DNA determined by using Quant-iT PicoGreen from the buccal swab, mouthwash and dental impression wafer samples were 113.61, 509.57 and 1.03 micrograms, respectively. The average amounts of DNA determined by using RT-PCR from the buccal swab, mouthwash and dental impression wafer samples were 11.5240, 22.2540 and 0.0279 mug, respectively. The bite registrations and stone models had an average of 14 percent of mismatches. CONCLUSION: The dental impression wafers captured DNA but not in high quantities. They did not produce an accurate representation of the dentition. CLINICAL IMPLICATIONS: The dental impression wafers captured enough DNA to permit amplification. The accuracy of the bite registration was not sufficient for identification purposes. Therefore, dental impression wafers may be useful only as a reservoir for DNA.