bluuebunny/crossref_metadata_2025 huggingface dataset:
Plasma decorin predicts the presence of esophageal squamous cell carcinoma
Authors: Wu I‐Chen , Wu Deng‐Chyang , Huang Chun‐Chi , Lin Hung‐Shun , Chen Yu‐Kuei , Tsai Hui‐Jen , Lu Chieh‐Yu , Chou Shah‐Hwa , Chou Yi‐Ping , Li Ling‐Hui , Tai Shu‐Yu , Wu Ming‐Tsang
Published: February 2010

Using a microarray technique, we found decorin to be underexpressed, but osteopontin (OPN) to be overexpressed, in esophageal squamous cell carcinoma (ESCC). This study aims to examine whether plasma decorin and OPN plus personal substances use (tobacco, alcohol and areca) can serve as suitable clinical markers to predict the presence of ESCC. In total, 570 archived plasma specimens (275 patients and 295 controls) were collected from 2 medical centers in Taiwan between 2000 and 2008. Decorin and OPN protein levels were measured by ELISA. Means and standard deviation of plasma decorin were 5.6 ± 3.6 ng/ml in case patients, which were significantly lower than those in controls (7.8 ± 3.1, p < 0.0001). Plasma OPN levels in case patients were not significantly different from controls (p = 0.33). When compared to subjects with the lowest quartile of plasma decorin, those with the highest quartile one had a significantly lower risk to have ESCC (Adjusted OR = 0.03, p < 0.001). Receiver operator characteristic (ROC) analysis was performed for the combination of plasma decorin and 3 substances use (smoke, alcohol and areca) for the patients compared with the controls. The area under the curve was 88.6% and the optimal cut‐point of ROC curve (any 3 factors) had 73.5% sensitivity and 90.2% specificity with ∼82% of corrected classification. Plasma decorin, but not OPN, is a potential clinical marker for the detection of ESCC. When plasma decorin plus the use of the 3 substances are combined, this factor cluster could be used to detect the presence of ESCC.
Allergies, obesity, other risk factors and survival from pancreatic cancer
Authors: Olson Sara H. , Chou Joanne F. , Ludwig Emmy , O'Reilly Eileen , Allen Peter J. , Jarnagin William R. , Bayuga Sharon , Simon Jennifer , Gonen Mithat , Reisacher William R. , Kurtz Robert C.
Published: February 2010

Survival from pancreatic adenocarcinoma remains extremely poor, approximately 5% at 5 years. Risk factors include smoking, high body mass index (BMI), family history of pancreatic cancer, and long‐standing diabetes; in contrast, allergies are associated with reduced risk. Little is known about associations between these factors and survival. We analyzed overall survival in relation to risk factors for 475 incident cases who took part in a hospital based case–control study. Analyses were conducted separately for those who did (160) and did not (315) undergo tumor resection. Kaplan‐Meier methods were used to describe survival according to smoking, BMI, family history, diabetes, and presence of allergies. Cox proportional hazards models were used to adjust for covariates. There was no association with survival based on smoking, family history, or history of diabetes in either group. Among patients with resection, those with allergies showed nonstatistically significant longer survival, a median of 33.1 months (95% CI: 19.0–52.5) vs. 21.8 months (95% CI: 18.0–33.1), p = 0.25. The adjusted hazard ratio (HR) was 0.72 (95% CI: 0.43–1.23), p = 0.23. Among patients without resection, those with self‐reported allergies survived significantly longer than those without allergies: 13.3 months (95% CI: 10.6–16.9) compared to 10.4 months (95% CI: 8.8–11.0), p = 0.04, with an adjusted HR of 0.68 (95% CI: 0.49–0.95), p = 0.02. Obesity was nonsignificantly associated with poorer survival, particularly in the resected group (HR = 1.62, 95% CI: 0.76–3.44). The mechanisms underlying the association between history of allergies and improved survival are unknown. These novel results need to be confirmed in other studies.
Check your cultures! A list of cross‐contaminated or misidentified cell lines
Authors: Capes‐Davis Amanda , Theodosopoulos George , Atkin Isobel , Drexler Hans G. , Kohara Arihiro , MacLeod Roderick A.F. , Masters John R. , Nakamura Yukio , Reid Yvonne A. , Reddel Roger R. , Freshney R. Ian
Published: February 2010

Continuous cell lines consist of cultured cells derived from a specific donor and tissue of origin that have acquired the ability to proliferate indefinitely. These cell lines are well‐recognized models for the study of health and disease, particularly for cancer. However, there are cautions to be aware of when using continuous cell lines, including the possibility of contamination, in which a foreign cell line or microorganism is introduced without the handler's knowledge. Cross‐contamination, in which the contaminant is another cell line, was first recognized in the 1950s but, disturbingly, remains a serious issue today. Many cell lines become cross‐contaminated early, so that subsequent experimental work has been performed only on the contaminant, masquerading under a different name. What can be done in response—how can a researcher know if their own cell lines are cross‐contaminated? Two practical responses are suggested here. First, it is important to check the literature, looking for previous work on cross‐contamination. Some reports may be difficult to find and to make these more accessible, we have compiled a list of known cross‐contaminated cell lines. The list currently contains 360 cell lines, drawn from 68 references. Most contaminants arise within the same species, with HeLa still the most frequently encountered (29%, 106/360) among human cell lines, but interspecies contaminants account for a small but substantial minority of cases (9%, 33/360). Second, even if there are no previous publications on cross‐contamination for that cell line, it is essential to check the sample itself by performing authentication testing.
The calcium-sensing receptor and 25-hydroxyvitamin D–1α-hydroxylase interact to modulate skeletal growth and bone turnover
Authors: Richard Christian , Huo Rujuan , Samadfam Rana , Bolivar Isabel , Miao Dengshun , Brown Edward M , Hendy Geoffrey N , Goltzman David
Published: February 2010

We examined parathyroid and skeletal function in 3-month-old mice expressing the null mutation for 25-hydroxyvitamin D–1α-hydroxylase [1α(OH)ase−/−] and in mice expressing the null mutation for both the 1α(OH)ase and the calcium-sensing receptor [Casr−/−1α(OH)ase−/−] genes. On a normal diet, all mice were hypocalcemic, with markedly increased parathyroid hormone (PTH), increased trabecular bone volume, increased osteoblast activity, poorly mineralized bone, enlarged and distorted cartilaginous growth plates, and marked growth retardation, especially in the compound mutants. Osteoclast numbers were reduced in the Casr−/−1α(OH)ase−/− mice. On a high-lactose, high-calcium, high-phosphorus “rescue” diet, serum calcium and PTH were normal in the 1α(OH)ase−/− mice but increased in the Casr−/−1α(OH)ase−/− mice with reduced serum phosphorus. Growth plate architecture and mineralization were improved in both mutants, but linear growth of the double mutants remained abnormal. Mineralization of bone improved in all mice, but osteoblast activity and trabecular bone volume remained elevated in the Casr−/−1α(OH)ase−/− mice. These studies support a role for calcium-stimulated maturation of the cartilaginous growth plate and mineralization of the growth plate and bone and calcium-stimulated CaSR-mediated effects on bone resorption. PTH-mediated bone resorption may require calcium-stimulated CaSR-mediated enhancement of osteoclastic activity. © 2010 American Society for Bone and Mineral Research. © 2010 American Society for Bone and Mineral Research
Progressive ankylosis protein (ANK) in osteoblasts and osteoclasts controls bone formation and bone remodeling
Authors: Kim Hyon Jong , Minashima Takeshi , McCarthy Edward F , Winkles Jeffrey A , Kirsch Thorsten
Published: February 2010

The progressive ankylosis gene (ank) encodes a transmembrane protein that transports intracellular inorganic pyrophosphate (PPi) to the extracellular milieu. ank/ank mice, which express a truncated nonfunctional ANK, showed a markedly reduced bone mass, bone-formation rate, and number of tartrate-resistant acid phosphatase–positive (TRAP+) multinucleated osteoclasts. ANK function deficiency suppressed osteoblastic differentiation of ank/ank bone marrow stromal cells, as indicated by the decrease in the expression of bone marker genes, including osterix, reduced alkaline phosphatase activity, and mineralization. Runx2 gene expression levels were not altered. Conversely, overexpression of ANK in the preosteoblastic cell line MC3T3-E1 resulted in increased expression of bone marker genes, including osterix. Whereas runx2 expression was not altered in ANK-overexpressing MC3T3-E1 cells, runx2 transcriptional activity was increased. Extracellular PPi or Pi stimulated osteoblastogenic differentiation of MC3T3-E1 cells or partially rescued delayed osteoblastogenic differentiation of ank/ank bone marrow stromal cells. A loss of PPi transport function ANK mutation also stimulated osteoblastogenic differentiation of MC3T3-E1 cells. Furthermore, ANK function deficiency suppressed the formation of multinucleated osteoclasts from ank/ank bone marrow cells cultured in the presence of macrophage colony-stimulating factor and receptor activator of nuclear factor-κB ligand. In conclusion, ANK is a positive regulator of osteoblastic and osteoclastic differentiation events toward a mature osteoblastic and osteoclastic phenotype. © 2010 American Society for Bone and Mineral Research
Leptin stimulates fibroblast growth factor 23 expression in bone and suppresses renal 1α,25-dihydroxyvitamin D3 synthesis in leptin-deficient ob/ob Mice
Authors: Tsuji Kiyomi , Maeda Toyonobu , Kawane Tetsuya , Matsunuma Ayako , Horiuchi Noboru
Published: February 2010

Leptin is the LEP (ob) gene product secreted by adipocytes. We previously reported that leptin decreases renal expression of the 25-hydroxyvitamin D3 1α-hydroxylase (CYP27B1) gene through the leptin receptor (ObRb) by indirectly acting on the proximal tubules. This study focused on bone-derived fibroblast growth factor 23 (FGF-23) as a mediator of the influence of leptin on renal 1α-hydroxylase mRNA expression in leptin-deficient ob/ob mice. Exposure to leptin (200 ng/mL) for 24 hours stimulated FGF-23 expression by primary cultured rat osteoblasts. Administration of leptin (4 mg/kg i.p. at 12-hour intervals for 2 days) to ob/ob mice markedly increased the serum FGF-23 concentration while significantly reducing the serum levels of calcium, phosphate, and 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3]. Administration of FGF-23 (5 µg i.p. at 12-hour intervals for 2 days) to ob/ob mice suppressed renal 1α-hydroxylase mRNA expression. The main site of FGF-23 mRNA expression was the bone, and leptin markedly increased the FGF-23 mRNA level in ob/ob mice. In addition, leptin significantly reduced 1α-hydroxylase and sodium-phosphate cotransporters (NaPi-IIa and NaPi-IIc) mRNA levels but did not affect Klotho mRNA expression in the kidneys of ob/ob mice. Furthermore, the serum FGF-23 level and renal expression of 1α-hydroxylase mRNA were not influenced by administration of leptin to leptin receptor–deficient (db/db) mice. These results indicate that leptin directly stimulates FGF-23 synthesis by bone cells in ob/ob mice, suggesting that inhibition of renal 1,25(OH)2D3 synthesis in these mice is at least partly due to elevated bone production of FGF-23. © 2010 American Society for Bone and Mineral Research
Near-infrared fluorescent probe traces bisphosphonate delivery and retention in vivo
Authors: Kozloff Kenneth M , Volakis Leo I , Marini Joan C , Caird Michelle S
Published: February 2010

Bisphosphonate use has expanded beyond traditional applications to include treatment of a variety of low-bone-mass conditions. Complications associated with long-term bisphosphonate treatment have been noted, generating a critical need for information describing the local bisphosphonate-cell interactions responsible for these observations. This study demonstrates that a fluorescent bisphosphonate analogue, far-red fluorescent pamidronate (FRFP), is an accurate biomarker of bisphosphonate deposition and retention in vivo and can be used to monitor site-specific local drug concentration. In vitro, FRFP is competitively inhibited from the surface of homogenized rat cortical bone by traditional bisphosphonates. In vivo, FRFP delivery to the skeleton is rapid, with fluorescence linearly correlated with bone surface area. Limb fluorescence increases linearly with injected dose of FRFP; injected FRFP does not interfere with binding of standard bisphosphonates at the doses used in this study. Long-term FRFP retention studies demonstrated that FRFP fluorescence decreases in conditions of normal bone turnover, whereas fluorescence was retained in conditions of reduced bone turnover, demonstrating preservation of local FRFP concentration. In the mandible, FRFP localized to the alveolar bone and bone surrounding the periodontal ligament and molar roots, consistent with findings of osteonecrosis of the jaw. These findings support a role for FRFP as an effective in vivo marker for bisphosphonate site-specific deposition, turnover, and long-term retention in the skeleton. © 2010 American Society for Bone and Mineral Research
Taking vitamin D with the largest meal improves absorption and results in higher serum levels of 25-hydroxyvitamin D
Authors: Mulligan Guy B , Licata Angelo
Published: February 2010

Many patients treated for vitamin D deficiency fail to achieve an adequate serum level of 25-hydroxyvitamin D [25(OH)D] despite high doses of ergo- or cholecalciferol. The objective of this study was to determine whether administration of vitamin D supplement with the largest meal of the day would improve absorption and increase serum levels of 25(OH)D. This was a prospective cohort study in an ambulatory tertiary-care referral center. Patients seen at the Cleveland Clinic Foundation Bone Clinic for the treatment of vitamin D deficiency who were not responding to treatment make up the stugy group. Subjects were instructed to take their usual vitamin D supplement with the largest meal of the day. The main outcome measure was the serum 259(OH)D level after 2 to 3 months. Seventeen patients were analyzed. The mean age (±SD) and sex (F/M) ratio were 64.5 ± 11.0 years and 13 females and 4 males, respectively. The dose of 25(OH)D ranged from 1000 to 50,000 IU daily. The mean baseline serum 25(OH)D level (±SD) was 30.5 ± 4.7 ng/mL (range 21.6 to 38.8 ng/mL). The mean serum 25(OH)D level after diet modification (±SD) was 47.2 ± 10.9 ng/mL (range 34.7 to 74.0 ng/mL, p < .01). Overall, the average serum 25(OH)D level increased by 56.7% ± 36.7%. A subgroup analysis based on the weekly dose of vitamin D was performed, and a similar trend was observed.

Thus it is concluded that taking vitamin D with the largest meal improves absorption and results in about a 50% increase in serum levels of 25(OH)D levels achieved. Similar increases were observed in a wide range of vitamin D doses taken for a variety of medical conditions. © 2010 American Society for Bone and Mineral Research
Splice variant specific increase in Ca2+/calmodulin‐dependent protein kinase 1‐gamma mRNA expression in response to acute pyrethroid exposure
Authors: Harrill Joshua A. , Knapp Geremy W. , Crofton Kevin M.
Published: February 2010

In mammals, pyrethroids are neurotoxicants that interfere with ion channel function in excitable neuronal membranes. Previous work demonstrated increases in the expression of Ca2+/calmodulin‐dependent protein kinase 1‐gamma (Camk1g) mRNA following acute deltamethrin and permethrin exposure. In the rat, this gene is expressed as two distinct splice variants,Camk1g1andCamk1g2. The present study tests the hypothesis that changes inCamk1gmRNA expression in the rat following acute pyrethroid exposure are due to a specific increase in theCamk1g1splice variant and not theCamk1g2splice variant. Long‐Evans rats were acutely exposed to permethrin, deltamethrin, or corn oil vehicle. Frontal cortex was collected at 6 h postdosing. In addition, rats were exposed to permethrin (100 mg/kg) or deltamethrin (3 mg/kg), and frontal cortex was collected at 1, 3, 6, 9, 12, or 24 h along with time‐matched vehicle controls. Expression ofCamk1g1andCamk1g2mRNA was measured by quantitative real‐time RT‐PCR and quantified using the 2‐Δ ΔCTmethod. Dose‐dependent increases inCamk1g1mRNA expression were observed for both pyrethroids at 6 h. In addition, a dose‐dependent increase inCamk1g2was observed at 6 h although it was very small in magnitude. The increases inCamk1g1expression for deltamethrin and permethrin peak between 3 and 6 h postexposure and returns to control levels by 9 h. There was no increase in CAMK1G1 protein as measured with Western blots. The present data demonstrate that pyrethroid‐induced changes inCamk1gare driven mainly by increased expression of theCamk1g1splice variant. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:174–186, 2010; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20324
Young mothers' involvement in a prenatal and postpartum support program
Authors: Wen Xiaoli , Korfmacher Jon , Hans Sydney L. , Henson Linda G.
Published: February 2010

The involvement of 124 young mothers in a doula support program was measured in two dimensions—quantity of program contact and quality of mother–doula helping relationship. The study examined each dimension's differential associations with maternal outcomes, as well as the moderating effects of mother characteristics on these associations. Quantity of program contact was related to the quality of helping relationship, especially when rated by doulas. Both quantity and quality of involvement predicted the maternal outcomes, although not always in the expected direction. For mothers with limited vocabulary skills and better connections with the baby's father, program contact was more likely to promote positive birth experiences. A negative association between program contact and parenting behaviors was observed among mothers who were less mature or had less positive social relationships. The study suggests the multidimensionality of program involvement and the complexity of interactions between program and participant factors in producing program outcomes. © 2010 Wiley Periodicals, Inc.
Measuring perceived procedural justice and coercion among persons with mental illness in police encounters: the Police Contact Experience Scale
Authors: Watson Amy C. , Angell Beth , Vidalon Theresa , Davis Kristin
Published: February 2010

Despite increased recent attention to improving the quality of encounters between police officers and people with serious mental illness, there are no measures available for assessing how consumers perceive their interactions with police officers. Drawing upon conceptual frameworks developed within social psychology, this study reports the development and testing of a new measure, the Police Contact Experience Scale (PCES), which yields indicators of procedural justice and coercion as evaluated by persons with mental illness in relation to specific encounters with the police. The PCES was administered to 154 individuals with mental illness that had police contact within the prior 12 months. Rasch rating scale analysis supported construct validity of a 10‐item procedural justice and a 5‐item coercion scale. Correlational analysis supported convergent validity for both scales. © 2010 Wiley Periodicals, Inc.
Predictors of family participation in a multiple family group intervention for aggressive middle school students
Authors: Quinn William H. , Hall Dan B. , Smith Emilie P. , Rabiner David
Published: February 2010

The authors examine predictors of family participation in the G.R.E.A.T. Families Program of the Multisite Violence Prevention Project (MVPP), a four‐site collaboration examining student, teacher, and family interventions for middle school students. Teachers recruited two cohorts of sixth grade students, recognized as being aggressive and influential with their peers, and their families into a voluntary, 15‐session weekly program. Among the 643 families that consented (66%), the mean number of sessions attended was 8.13, with almost half (48.3%) attending 11 or more sessions. Linear mixed models (LMMs) were developed to predict the number of sessions attended based on a cluster of demographic variables and scales designed to measure aggression, problem behaviors, family factors, and other psychosocial constructs. Three of the nine clusters held significance when associated with attendance: level of aggression as rated by the child (negative association), parent–child bond (negative association), and level of child victimization (positive association). Somewhat surprisingly, the variance component due to interventionist turned out to be small and to constitute a nonsignificant component of the overall variability in attendance. Results suggest that family recruitment for multiple family group programs can be achieved with substantial effort and that resources available for recruitment might be differentially applied across families based on predictors of attendance. © 2010 Wiley Periodicals, Inc.
Implementation of the Communities That Care prevention system by coalitions in the Community Youth Development Study
Authors: Arthur Michael W. , Hawkins J. David , Brown Eric C. , Briney John S. , Oesterle Sabrina , Abbott Robert D.
Published: February 2010

Although advances in prevention science over the past two decades have produced a growing list of tested and effective programs and policies for preventing adolescent delinquency and drug use, widespread dissemination and high‐quality implementation of effective programs and policies in communities has not been achieved. The Community Youth Development Study (CYDS) is a randomized, community‐level trial of the Communities That Care (CTC) system for promoting science‐based prevention in communities. This study compares 12 community prevention coalitions implementing the CTC system in 12 intervention communities as part of the CYDS to prevention coalitions located in the 12 control communities. As hypothesized, the CYDS coalitions implemented significantly more of the CTC core intervention elements, and also implemented significantly greater numbers of tested, effective prevention programs than the prevention coalitions in the control communities. Implications of the findings for efforts to achieve widespread dissemination of effective prevention programs, policies, and practices are discussed. © 2010 Wiley Periodicals, Inc.
Profilin‐1 overexpression inhibits proliferation of MDA‐MB‐231 breast cancer cells partly through p27kip1 upregulation
Authors: Zou Li , Ding Zhijie , Roy Partha
Published: February 2010

Profilin‐1 (Pfn1), a ubiquitously expressed actin‐binding protein, has gained interest in epithelial‐derived cancer because of its downregulation in expression in various adenocarcinoma. Pfn1 overexpression impairs tumorigenic ability of human breast cancer xenografts thus suggesting that Pfn1 could be a tumor‐suppressor protein. The objective of the present study was to determine how Pfn1 overexpression affects cell‐cycle progression of breast cancer cells. We show that Pfn1 overexpression in MDA‐MB‐231 breast cancer cells causes cell‐cycle arrest in G1 phase and dramatically reduced proliferation in culture. Pfn1 overexpression results in increased protein stability of p27kip1 (p27—a major cyclin‐dependent kinase inhibitor) and marked elevation in the overall cellular level of p27. Proliferation defect of Pfn1 overexpressers can be partly rescued by silencing p27 expression thus suggesting a critical role of p27 in Pfn1‐induced growth inhibition of MDA‐MB‐231 cells. Finally, Pfn1 overexpression was found to sensitize MDA‐MB‐231 cells to apoptosis in response to cytotoxic stimulus thus suggesting for the first time that survival of breast cancer cells can also be negatively influenced by Pfn1 upregulation. These findings may provide novel insights underlying Pfn1's tumor‐suppressive action. J. Cell. Physiol. 223:623–629, 2010. © 2010 Wiley‐Liss, Inc.
Both superficial and deep zone articular chondrocyte subpopulations exhibit the crabtree effect but have different basal oxygen consumption rates
Authors: Heywood Hannah K. , Knight Martin M. , Lee David A.
Published: February 2010

In the absence of in vivo measurements, the oxygen concentration within articular cartilage is calculated from the balance between cellular oxygen consumption and mass transfer. Current estimates of the oxygen tension within articular cartilage are based on oxygen consumption data from full‐depth tissue samples. However, superficial and deep cell subpopulations of articular cartilage express intrinsic metabolic differences. We test the hypothesis that the subpopulations differ with respect to their intrinsic oxygen consumption rate. Chondrocytes from the full cartilage thickness demonstrate enhanced oxygen consumption when deprived of glucose, consistent with the Crabtree phenomena. Chondrocyte subpopulations differ in the prevailing availability of oxygen and glucose, which decrease with distance from the cartilage–synovial fluid interface. Thus, we tested the hypothesis that the oxygen consumption of each subpopulation is modulated by nutrient availability, by examining the expression of the Crabtree effect. The deep cells had a greater oxygen consumption than the superficial cells (Vmax of 6.6 compared to 3.2 fmol/cell/h), consistent with our observations of mitochondrial volume (mean values 52.0 vs. 36.4 µm3/cell). Both populations expressed the Crabtree phenomena, with oxygen consumption increasing ∼2.5‐fold in response to glycolytic inhibition by glucose deprivation or 2‐deoxyglucose. Over 90% of this increase was oligomycin‐sensitive and thus accounted for by oxidative phosphorylation. The data contributes towards our understanding of chondrocyte energy metabolism and provides information valuable for the accurate calculation of the oxygen concentration that the cells experience in vivo. The work has further application to the optimisation of bioreactor design and engineered tissues. J. Cell. Physiol. 223:630–639, 2010. © 2010 Wiley‐Liss, Inc.
BMP‐3 promotes mesenchymal stem cell proliferation through the TGF‐β/activin signaling pathway
Authors: Stewart Aaron , Guan Haiyan , Yang Kaiping
Published: February 2010

Adipogenesis plays a key role in the pathogenesis of obesity. It begins with the commitment of mesenchymal stem cells (MSCs) to the adipocyte lineage, followed by terminal differentiation of preadipocytes to mature adipocytes. A critical, but poorly understood, component of adipogenesis involves proliferation of MSCs and preadipocytes. The present study was undertaken to examine the hypothesis that bone morphogenetic protein‐3 (BMP‐3) promotes adipogenesis using C3H10T1/2 MSCs and 3T3‐L1 preadipocytes as in vitro model systems. We demonstrated that although it did not promote the commitment of MSCs to the adipocyte lineage or the differentiation of preadipocytes to adipocytes, BMP‐3‐stimulated proliferation by threefold in both cell types. Owing to a lack of information on MSC proliferation, we then delineated the molecular mechanisms underlying BMP‐3‐stimulated MSC proliferation. We showed that BMP‐3 activated the transforming growth factor‐β (TGF‐β)/activin but not ERK1/2, p38 MAPK, or JNK signaling pathways in C3H10T1/2 cells. Furthermore, the TGF‐β/activin receptor kinase inhibitor SB‐431542 blocked BMP‐3‐stimulated proliferation. Importantly, siRNA‐mediated knockdown of the key TGF‐β/activin signaling pathway components, ActRIIB, ALK4, or Smad2, abrogated the mitogenic effects of BMP‐3 on MSCs. Together, these results demonstrate that BMP‐3 stimulates MSC proliferation via the TGF‐β/activin signaling pathway, thus revealing a novel role for this divergent and poorly understood member of the TGF‐β superfamily in regulating MSC proliferation. J. Cell. Physiol. 223:658–666, 2010. © 2010 Wiley‐Liss, Inc.
Non‐genomic rapid inhibition of Na+/H+‐exchange 1 and apoptotic immunosuppression in human T cells by glucocorticoids
Authors: Chang Ching‐Pang , Wang Shyi‐Wu , Huang Zih‐Ling , Wang Olivia Ya‐Hsuan , Huang Michael I‐Ta , Lu Li‐Ming , Tarng Der‐Cherng , Chien Chau‐Heng , Chien Eileen Jea
Published: February 2010

Glucocorticoids (GCs) have been employed as immunosuppressive agents for many years. However, it is still unclear how GCs instantly uncouple T cells from acute stressful inflammatory. In terms of time scale, the genomic activity of the classic GC receptor cannot fulfill this role under crisis; but a rapid non‐genomic response can. In a previous study, intracellular acidification was found to be due to a rapid non‐genomic inhibition of Na+/H+‐exchange 1 (NHE1) and this event led to the immunosuppression of T cell proliferation by progesterone. The aim of this study was to examine whether there is a rapid acidification response caused by an inhibition of NHE1 activity and to explore the differential non‐genomic effect on immunosuppression of hydrocortisone and dexamethasone. The IC50 values for NHE1‐dependent pHi recovery by hydrocortisone and dexamethasone are 250 and 1 nM, respectively. Co‐stimulation of GCs with phytohemagglutinin (PHA) is able to inhibit PHA‐induced IL‐2 secretion, IL‐4 secretion, and T‐cell proliferation. Furthermore, apoptosis in PHA‐activated T cells is not induced by hydrocortisone but by dexamethasone. The mechanism of immunosuppression on proliferation by dexamethasone was found to be different of hydrocortisone and seems to involve cytotoxicity against T cells. Moreover, apoptosis induced by dexamethasone and impermeable dexamethasone–bovine serum albumin suggests that the apoptotic immunosuppression occurs through both the plasma membrane and cytoplasmic sites. The rapid inhibitory responses triggered by GCs would seem to release T cells instantly when an acute stress‐related response is needed. Nonetheless, the apoptotic immunosuppression by dexamethasone is attributable to its severe cytotoxicity. J. Cell. Physiol. 223:679–686, 2010. © 2010 Wiley‐Liss, Inc.
Competitiveness for the niche and mutual dependence of the germline and somatic stem cells in the Drosophila testis are regulated by the JAK/STAT signaling
Authors: Singh Shree Ram , Zheng Zhiyu , Wang Hong , Oh Su‐Wan , Chen Xiu , Hou Steven X.
Published: February 2010

In many tissues, two or more types of stem cells share a niche, and how the stem cells coordinate their self‐renewal and differentiation is poorly understood. In the Drosophila testis, germ line stem cells (GSCs) and somatic cyst progenitor cells (CPCs) contact each other and share a niche (the hub). The hub expresses a growth factor unpaired (Upd) that activates the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway in GSCs to regulate the stem cell self‐renewal. Here, we demonstrate that the JAK/STAT signaling also regulates CPCs self‐renewal. We also show that a negative regulator, the suppressor of cytokine signaling 36E (SOCS36E), suppresses JAK/STAT signaling in somatic cells, preventing them from out‐competing the GSCs. Furthermore, through selectively manipulating the JAK/STAT signaling level in either CPCs or GSCs, we demonstrate that the somatic JAK/STAT signaling is essential for self‐renewal and maintenance of both CPCs and GSCs. These data suggest that a single JAK/STAT signal from the niche orchestrate the competitive and dependent co‐existence of GSCs and CPCs in the Drosophila testis niche. J. Cell. Physiol. 223: 500–510, 2010. © 2010 Wiley‐Liss, Inc.
Fine structure of the branchial epithelium in the teleost Oreochromis niloticus
Authors: Monteiro Sandra Mariza , Oliveira Elsa , Fontaínhas‐Fernandes António , Sousa Mário
Published: February 2010

We have studied the gill epithelium of Oreochromis niloticus using transmission electron microscopy with the particular interested relationship between cell morphology and osmotic, immunoregulatory, or other non‐regulatory functions of the gill. Pavement cells covered the filament epithelium and lamellae of gills, with filament pavement cells showing distinct features from lamellar pavement cells. The superficial layer of the filament epithelium was formed by osmoregulatory elements, the columnar mitochondria‐rich, mucous and support cells, as well as by their precursors. Light mitochondria‐rich cells were located next to lamellae. They exhibited an apical crypt with microvilli and horizontal small dense rod‐like vesicles, sealed by tight junctions to pavement cells. Dark mitochondria‐rich cells had long dense rod‐like vesicles and a small apical opening sealed by tight junctions to pavement cells. The deep layer of the filament epithelium was formed by a network of undifferentiated cells, containing neuroepithelial and myoepithelial cells, macrophage and eosinophil‐like cells and their precursors, as well as precursors of mucous cells. The lateral‐basal surface was coated by myoepithelial cells and a basal lamina. The lamellar blood lacunae was lined by pillar cells and surrounded by a basal lamina and pericytes. The data presented here support the existence of two distinct types of pavement cells, mitochondria‐rich cells, and mitochondria‐rich cells precursors, a structural role for support cells, a common origin for pavement cells and support cells, a paracrine function for neuroepithelial cells in the superficial layer, and the control of the lamellar capillary base by endocrine and contractile cells. Data further suggest that the filament superficial layer is involved in gill osmoregulation, that may interact, through pale mitochondria‐rich cells, with the deep layer and lamellae, whereas the deep layer, through immune and neuroendocrine systems, acts in the regeneration and defense of the tissue. J. Morphol. 2010. © 2010 Wiley‐Liss, Inc.
Laser desorption postionization for imaging MS of biological material
Authors: Akhmetov Artem , Moore Jerry F. , Gasper Gerald L. , Koin Peter J. , Hanley Luke
Published: February 2010

Vacuum ultraviolet single photon ionization (VUV SPI) is a soft ionization technique that has the potential to address many of the limitations of matrix‐assisted laser desorption/ionization (MALDI) for imaging MS. Laser desorption postionization (LDPI) uses VUV SPI for postionization and is experimentally analogous to a MALDI instrument with the addition of a pulsed VUV light source. This review discusses progress in LDPI‐MS over the last decade, with an emphasis on imaging MS of bacterial biofilms, analytes whose high salt environment make them particularly resistant to imaging by MALDI‐MS. This review first considers fundamental aspects of VUV SPI including ionization mechanisms, cross sections, quantum yields of ionization, dissociation and potential mass limits. The most common sources of pulsed VUV radiation are then described along with a newly constructed LDPI‐MS instrument with imaging capabilities. Next, the detection and imaging of small molecules within intact biofilms is demonstrated by LDPI‐MS using 7.87 eV (157.6 nm) VUV photons from a molecular fluorine excimer laser, followed by the use of aromatic tags for detection of selected species within the biofilm. The final section considers the future prospects for imaging intact biological samples by LDPI‐MS. Copyright © 2010 John Wiley & Sons, Ltd.
Nerve growth factor‐evoked nociceptor sensitization in pig skin in vivo
Authors: Rukwied Roman , Schley Marcus , Forsch Elmar , Obreja Otilia , Dusch Martin , Schmelz Martin
Published: February 2010

Peripheral sensitization of skin nociceptors by nerve growth factor (NGF) was explored in pig skin in vivo. As an objective output measure, the area of axon‐reflex‐mediated erythema was assessed upon mechanical, thermal, chemical, and electrical stimuli delivered at 1, 3, and 7 days after i.d. injection of 1 μg NGF into the pig's back skin (n = 8). Pretreatment with NGF provoked a sensitization to mechanical (600 mN), thermal (10 sec 49°C) and chemical (15 μl, pH 3) stimuli that lasted for 7 days. No sensitization, however, was found in response to weak mechanical (100 mN), weak thermal (10 sec 45°C), or electrical stimuli. Irrespective of the skin pretreatment (NGF or PBS vehicle control), the area of electrically induced erythema decreased upon repetition (days 1–7) by 70% (P < 0.05). Sensitization of sensory endings by NGF upon mechanical, heat, and chemical stimuli suggests recruitment of sensory transducer molecules [e.g., TRPV1, acid‐sensing ion channels (ASICs)]. In contrast, the gradual decrease in electrically induced erythema over 7 days might be attributable to axonal desensitization and possibly activity‐dependent down‐regulation of sodium channels. Thus, long‐lasting sensitization processes of nociceptor endings or axonal sodium channel desensitization mechanisms can be explored in the pig as a translational experimental animal model. © 2010 Wiley‐Liss, Inc.
Hexokinase II gene transfer protects against neurodegeneration in the rotenone and MPTP mouse models of Parkinson's disease
Authors: Corona Juan Carlos , Gimenez‐Cassina Alfredo , Lim Filip , Díaz‐Nido Javier
Published: February 2010

A typical feature of Parkinson's disease is the progressive loss of dopaminergic neurons in the substantia nigra, in which inhibition of mitochondrial complex I activity may play an important role. Rotenone or 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) inhibit the mitochondrial complex I and they cause the death of substantia nigra dopaminergic neurons, thereby providing acute murine models of Parkinson's disease. We have found that increasing mitochondrial hexokinase II activity can prevent cell death in neuronal cultures treated with rotenone. As a result, we have studied the effects of hexokinase II gene transfer in vivo using a herpes simplex virus type 1 (HSV‐1) amplicon vector. The placHK2 amplicon vector was injected into substantia nigra of mice that were subsequently administered rotenone or MPTP. Overexpression of hexokinase II prevented both rotenone and MPTP‐induced dopaminergic neuronal cell death, as well as reducing the associated motor defects. Our results provide the first proof‐of‐principle that hexokinase II protects against dopaminergic neurodegeneration in vivo, emphasizing the role of this enzyme in promoting neuronal survival. Thus, the increase of hexokinase II expression by gene transfer or other means represents a promising approach to treat Parkinson's and other neurodegenerative diseases. © 2010 Wiley‐Liss, Inc.
Oxygen resuscitation does not ameliorate neonatal hypoxia/ischemia‐induced cerebral edema
Authors: Ferrari Diana Carolina , Nesic Olivera B. , Perez‐Polo J. Regino
Published: February 2010

Neonatal hypoxia/ischemia (HI) is a common cause of cognitive and behavioral deficits in children with hyperoxia treatment (HHI) being the current therapy for newborn resuscitation. HI induces cerebral edema that is associated with poor neurological outcomes. Our objective was to characterize cerebral edema after HI and determine the consequences of HHI (40% or 100% O2). Dry weight analyses showed cerebral edema 1 to 21 days after HI in the ipsilateral cortex; and 3 to 21 days after HI in the contralateral cortex. Furthermore, HI increased blood‐brain barrier (BBB) permeability 1 to 7 days after HI, leading to bilateral cortical vasogenic edema. HHI failed to prevent HI‐induced increase in BBB permeability and edema development. At the molecular level, HI increased ipsilateral, but not contralateral, AQP4 cortical levels at 3 and up to 21 days after HI. HHI treatment did not further affect HI‐induced changes in AQP4. In addition, we observed developmental increases of AQP4 accompanied by significant reduction in water content and increase permeability of the BBB. Our results suggest that the ipsilateral HI‐induced increase in AQP4 may be beneficial and that its absence in the contralateral cortex may account for edema formation after HI. Finally, we showed that HI induced impaired motor coordination 21 days after the insult and HHI did not ameliorate this behavioral outcome. We conclude that HHI treatment is effective as a resuscitating therapy, but does not ameliorate HI‐induced cerebral edema and impaired motor coordination. © 2010 Wiley‐Liss, Inc.
Transgenic mouse and cell culture models demonstrate a lack of mechanistic connection between endoplasmic reticulum stress and tau dysfunction
Authors: Spatara M.L. , Robinson A.S.
Published: February 2010

In vivo aggregation of tau protein is a hallmark of many neurodegenerative disorders, including Alzheimer's disease (AD). Recent evidence has also demonstrated activation of the unfolded protein response (UPR), a cellular response to endoplasmic reticulum (ER) stress, in AD, although the role of the UPR in disease pathogenesis is not known. Here, three model systems were used to determine whether a direct mechanistic link could be demonstrated between tau aggregation and the UPR. The first model system used was SH‐SY5Y cells, a neuronal cultured cell line that endogenously expresses tau. In this system, the UPR was activated using chemical stressors, tunicamycin and thapsigargin, but no changes in tau expression levels, solubility, or phosphorylation were observed. In the second model system, wild‐type 4R tau and P301L tau, a variant with increased aggregation propensity, were heterologously overexpressed in HEK 293 cells. This overexpression did not activate the UPR. The last model system examined here was the PS19 transgenic mouse model. Although PS19 mice, which express the P301S variant of tau, display severe neurodegeneration and formation of tau aggregates, brain tissue samples did not show any activation of the UPR. Taken together, the results from these three model systems suggest that a direct mechanistic link does not exist between tau aggregation and the UPR. © 2010 Wiley‐Liss, Inc.
Adenosine A2A receptor but not HIF‐1 mediates Tyrosine hydroxylase induction in hypoxic PC12 cells
Authors: Gammella Elena , Cairo Gaetano , Tacchini Lorenza
Published: February 2010

Tyrosine hydroxylase (TH) is the rate‐limiting enzyme in the biosynthesis of catecholamines released by oxygen‐sensitive cells in response to hypoxic conditions. Adenosine is released in response to hypoxia in the central nervous system and CGS21680, an adenosine A2A receptor agonist, induces TH transcription. As we have previously demonstrated the A2A receptor‐mediated induction of HIF‐1 in macrophages and hepatocytes, we investigated the involvement of HIF‐1 in the adenosine‐mediated activation of TH expression. Exposure to adenosine or CGS21680 increased TH mRNA and protein levels in PC12 cells. Transcription of a reporter gene under the control of the wild type rat TH promoter was induced 3.5‐fold in CGS21680‐treated cells, but neither the mutation of the hypoxia responsive element in the TH promoter nor the co‐transfection of a dominant negative of the HIF‐1 β subunit prevented the increase in transcription; furthermore, CGS21680 increased CREB binding activity but did not induce HIF‐1 DNA binding activity or protein levels. To investigate whether HIF‐1 was involved in the hypoxia‐mediated induction of TH, PC12 cells were exposed to hypoxia in the presence of the A2A receptor antagonist ZM241385, which prevented hypoxia‐dependent TH induction despite HIF‐1 activation; in line with this finding, the inhibition of HIF‐1 did not abolish TH induction in hypoxic PC12 cells. These results indicate that, under hypoxic conditions, TH (a key factor in systemic adaptation to reduced oxygen availability) is not regulated by HIF‐1, the primary modulator of the response to hypoxia, but by the adenosine A2A receptor‐mediated signalling pathway. © 2010 Wiley‐Liss, Inc.
Salivary gland extract from engorged Ixodes ricinus (Acari: Ixodidae) stimulates in vitro growth of Borrelia burgdorferi sensu lato
Authors: Rudolf Ivo , Šikutová Silvie , Kopecký Jan , Hubálek Zdeněk
Published: February 2010

In vitro effect of salivary gland extract from fed Ixodes ricinus, the competent vector of Lyme borreliosis in Europe, on the growth of Borrelia burgdorferi sensu lato (B. garinii, B. afzelii and B. burgdorferi sensu stricto) was examined in BSK‐H medium. Motility rate, concentration of motile spirochetes and their morphology were estimated at intervals of 0, 2, 4, 6 and 8 days using darkfield microscopy. Salivary gland extract derived from I. ricinus stimulated markedly the growth of three genomic species of borreliae. The results confirm a substantial role of salivary glands in the mechanism of pathogen transmission to vertebrate host. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Practical procedures for genetic manipulation systems for medermycin‐producing Streptomyces sp. AM‐7161
Authors: Huiqun Deng , Xiaofeng Cai , Jianxin Peng , Huazhu Hong , Koji Ichinose , Aiying Li
Published: February 2010

Streptomyces sp. AM‐7161 is a producer of an aromatic polyketide medermycin with strong antibacterial and antitumor activity. It has been inefficient to perform genetic manipulations in this strain, which heavily hinders the genetic analysis of some interesting problems concerning tailoring modifications in medermycin biosynthesis. Here, condition optimizations of sporulation and mycelium growth of this strain as well as protoplast preparation and regeneration were conducted systematically. Based on these results, protoplast transformation system was established, and two types of foreign plasmids (integrative and auto‐replicating) were efficiently introduced into this strain. Additionally, a conjugation system to mediate plasmid transfer between Escherichia coli and Streptomyces sp. AM‐7161 was also optimized and established. These results provide the practical procedures to perform more convenient genetic analysis of medermycin biosynthesis in this strain. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Expression of hcp in freshwater Synechococcus spp., a gene encoding a hyperconserved protein in picocyanobacteria
Authors: Kutovaya Olga A. , McKay R. Michael L. , Bullerjahn George S.
Published: February 2010

Marine picoplankton of the genus Synechococcus and Prochlorococcus spp. are widely studied members of the picocyanobacterial clade, composed of unicellular cyanobacteria that dominate pelagic regions of the ocean. Less studied are the related freshwater Synechococcus spp. that similarly dominate the euphotic zone of oligotrophic lakes. Previous work has shown that marine picocyanobacteria harbor a small gene, hcp, that encodes a 62 amino acid protein 100% conserved among all strains examined. The gene is restricted exclusively to the picocyanobacterial lineage. The current study reveals that hcp is also 100% conserved in four freshwater Synechococcus spp. strains isolated from the Laurentian Great Lakes, and that the gene constitutively expressed with genes encoding a ribosomal protein and two tRNA genes. The synteny of the hcp region is also conserved between the marine and freshwater strains. Last, the hcp gene and the organization of the surrounding genetic region has been retained in the reduced genome of a picocyanobacterial endosymbiont of the amoeba Paulinella sp. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)