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1

Expression of growth differentiation factor 9, bone morphogenetic protein 15, and anti-Mullerian hormone in cultured mouse primary follicles  

Microsoft Academic Search

Growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), and anti-Mullerian hormone (AMH) play an important role in the primary to secondary follicle transition and follicle activation in vivo. In organ culture of neonatal mouse ovaries, it was observed that significantly fewer primary follicles develop to the secondary stage. The objectives of this study were: (1) to compare ovarian

J C Sadeu; T Adriaenssens; J Smitz

2008-01-01

2

Expression of growth differentiation factor 9, bone morphogenetic protein 15, and anti-Müllerian hormone in cultured mouse primary follicles.  

PubMed

Growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), and anti-Müllerian hormone (AMH) play an important role in the primary to secondary follicle transition and follicle activation in vivo. In organ culture of neonatal mouse ovaries, it was observed that significantly fewer primary follicles develop to the secondary stage. The objectives of this study were: (1) to compare ovarian follicular populations between organ-cultured neonatal mouse ovaries and freshly isolated age-matched control ovaries; (2) to quantify RNA levels of Gdf9, Bmp15, and Amh in cultured primary follicles; and (3) to immunolocalize GDF9 and AMH in cultured ovaries. Ovaries from 3-day-old (PND 3) mice were cultured for 7 or 10 days in the absence or presence of FSH. Follicular populations were counted in freshly isolated 13-day-old (PND 13) ovaries and organ-cultured ovaries. Transcripts were quantified in isolated primary follicles using real-time RT-PCR, and protein expressions were localized using immunohistochemistry. The number of secondary follicles in organ-cultured ovaries was significantly lower than in vivo controls. Gdf9 and Bmp15 mRNA expression levels were similar as in controls. Amh mRNA levels were significantly (P<0.05) lower after day 10 of culture in the absence of FSH. GDF9 and AMH proteins were respectively detected in the oocytes and the granulosa cells (GC) beginning at the primary and primordial stages onward. GDF9 and BMP15 production in cultured primary follicles are not different from in vivo controls; hence abnormal early follicular growth was not related to a deficient transcription of these factors. PMID:18469040

Sadeu, J C; Adriaenssens, T; Smitz, J

2008-05-09

3

Primary lymphomas of bone.  

PubMed

Primary lymphomas of bone are uncommon malignancies. The vast majority of them are non-Hodgkin lymphoma (NHL), whereas primary Hodgkin lymphoma (HL) of bone is extremely rare. Patients with primary NHL of bone commonly present with local bone pain, soft tissue swelling, and a mass or a pathological fracture. There is a slight male preponderance, and most patients are over 45-50 years of age. Primary NHL of bone can arise in any part of the skeleton, but long bones (femurs, tibia) are the most common sites of presentation. Comprehensive immunohistochemical studies are required to establish an accurate histological diagnosis of primary NHL of bone. Most cases of primary NHL of bone are classified as diffuse large B-cell lymphomas (DLBCL) in the World Health Organisation (WHO) classification of hematological malignancies. On full staging evaluation, most patients have disease of stage IE or IIE according to the Ann Arbor system. Several studies indicate that patients with primary NHL of bone have a favorable outcome, especially when treated by combined modality therapy. A number of studies reported that clinical stage is the most important prognostic variable in predicting overall survival. Interestingly, the rare occurrence of primary lymphoma of bone is in contrast with the frequency of plasma cell tumors in bone. This could be due to the fact that, during normal B-cell differentiation, the bone marrow is the normal site of homing of plasma cells which are terminally-differentiated, immunoglobulin-secreting post-germinal center B-cells. In this respect, there is circumstancial evidence that primary NHL of bone may represent tumors of post-germinal center B-cells. The present review summarizes data on the histogenesis of primary NHL of bone in view of the recent histogenetic classification of DLBCL on the basis of the B-cell differentiation gene expression profiles (germinal center vs. post-germinal center B-cell differentiation). PMID:16475714

Kitsoulis, Panagiotis; Vlychou, Marianna; Papoudou-Bai, Alexandra; Karatzias, Georgios; Charchanti, Antonia; Agnantis, Niki John; Bai, Maria

4

[Primary malignant bone tumors].  

PubMed

Among human neoplasms, primary malignant bone tumors are fairly rare. They present an incidence rate of roughly 10 cases per 1 million inhabitants per year. During childhood (<15 years), the percentage of malignant bone tumors amounts to 6% of all infantile malignancies. Only leukemia and lymphoma show a higher incidence in adolescence. Of all primary malignant bone tumors, 60% affect patients younger than 45 years and the peak incidence of all bone tumors occurs between 15 and 19 years. The most common primary malignant bone tumors are osteosarcoma (35%), chondrosarcoma (25%), and Ewing's sarcoma (16%). Less frequently (??5%) occurring tumors are chordoma, malignant fibrous histiocytoma of bone, and fibrosarcoma of bone. Vascular primary malignant tumors of bone and adamantinoma are very rare. Staging of the lesion is essential for systemic therapeutic decision-making and includes complete imaging and histo-pathological confirmation of the suspected entity. In most cases, this is established by open- or image-guided biopsy. Based on this information, an interdisciplinary tumor board will determine the individual therapeutic approach. Endoprosthetic or biological reconstruction following wide tumor resection is the most common surgical therapy for primary malignant bone tumors. There is vital importance in a thorough postoperative follow-up and continous after-care by a competent tumor center which is permanentely in charge of therapy. PMID:22130624

von Eisenhart-Rothe, R; Toepfer, A; Salzmann, M; Schauwecker, J; Gollwitzer, H; Rechl, H

2011-12-01

5

Primary leiomyosarcoma of bone  

Microsoft Academic Search

Sixteen cases of primary leiomyosarcoma of bone are described. The patients, 11 males and 5 females, ranged in age from 9 to 74 years. The annual incidence of this tumor in Sweden was calculated to be 0.09 cases per million. This figure was obtained by reviewing a Swedish series of spindle cell sarcomas of bone of which one quarter (11\\/44)

Lennart Angervall; Lars-Gunnar Kindblom; Isabelita C. Berlin; Bertil Stener

1987-01-01

6

Estrogen enhances differentiation of osteoblasts in mouse bone marrow culture  

Microsoft Academic Search

The effects of estrogen on bone are possibly mediated by several cell types. In the present study, the effect of 17?-estradiol (E2) on osteoblast-like cells was investigated by using mouse bone marrow cultures. Bone marrow cells were harvested from the shafts of femurs of 10-week-old NMRI mice and cultured. On day 6, confluent primary cultures were trypsinized and subcultured. Under

Q. Qu; M. Perälä-Heape; A. Kapanen; J. Dahllund; J. Salo; H. K. Väänänen; P. Härkönen

1998-01-01

7

Bone disease in primary hyperparathyrodism.  

PubMed

Nowadays, primary hyperparathyroidism (PHPT) is mostly a mild disease. Overt skeletal manifestations are rare but decreased bone mineral density (BMD) can still be demonstrated. Even in mild cases, excess parathyroid hormone (PTH) increases bone turnover leading to bone loss particularly at cortical sites. Conversely, a relative preservation of cancellous bone has been shown by histomorphometric analyses and advanced imaging techniques. An increased fracture rate has been demonstrated in untreated patients with PHPT at peripheral sites and in the spine. Parathyroidectomy (PTx) is the definitive cure for PHPT. With the restoration of normal PTH, bone resorption is quickly tapered down, while bone formation proceeds at the level of bone multicellular units, which were activated prior to PTx. The rapid refilling of the enlarged remodeling space and the subsequent matrix mineralization will result in an increase in BMD at sites rich in trabecular bone, such as lumbar spine and hip, which mainly occurs during the first 6-12 months after PTx. Cortical bone is less responsive to PTX because of the low rate of bone turnover, but sensible increases in BMD at the distal third of the radius can be observed in the long term. PTx seems to decrease the risk of fractures but more data are needed before a definitive conclusion on this important matter can be reached. Treatment with bisphosphonates can be considered for patients with low BMD who do not undergo PTx. Two-year treatment with alendronate has been shown to decrease bone turnover markers and increase BMD at the lumbar spine and hip, but not at the distal radius. Cinacalcet stably decreased serum calcium levels across a broad range of PHPT severity, but no change in BMD occurred in patients treated for up to 5.5 years. PMID:23024712

Marcocci, Claudio; Cianferotti, Luisella; Cetani, Filomena

2012-10-01

8

Bone disease in primary hyperparathyrodism  

PubMed Central

Nowadays, primary hyperparathyroidism (PHPT) is mostly a mild disease. Overt skeletal manifestations are rare but decreased bone mineral density (BMD) can still be demonstrated. Even in mild cases, excess parathyroid hormone (PTH) increases bone turnover leading to bone loss particularly at cortical sites. Conversely, a relative preservation of cancellous bone has been shown by histomorphometric analyses and advanced imaging techniques. An increased fracture rate has been demonstrated in untreated patients with PHPT at peripheral sites and in the spine. Parathyroidectomy (PTx) is the definitive cure for PHPT. With the restoration of normal PTH, bone resorption is quickly tapered down, while bone formation proceeds at the level of bone multicellular units, which were activated prior to PTx. The rapid refilling of the enlarged remodeling space and the subsequent matrix mineralization will result in an increase in BMD at sites rich in trabecular bone, such as lumbar spine and hip, which mainly occurs during the first 6–12 months after PTx. Cortical bone is less responsive to PTX because of the low rate of bone turnover, but sensible increases in BMD at the distal third of the radius can be observed in the long term. PTx seems to decrease the risk of fractures but more data are needed before a definitive conclusion on this important matter can be reached. Treatment with bisphosphonates can be considered for patients with low BMD who do not undergo PTx. Two-year treatment with alendronate has been shown to decrease bone turnover markers and increase BMD at the lumbar spine and hip, but not at the distal radius. Cinacalcet stably decreased serum calcium levels across a broad range of PHPT severity, but no change in BMD occurred in patients treated for up to 5.5 years.

Cianferotti, Luisella; Cetani, Filomena

2012-01-01

9

Effect of allogeneic bone marrow–derived mesenchymal stem cells transplantation in a polyI:C-induced primary biliary cirrhosis mouse model  

Microsoft Academic Search

Primary biliary cirrhosis (PBC) is a slowly progressive autoimmune disease of unknown mechanism. We established a PBC animal\\u000a model by injecting C57BL\\/6 mice with polyinosinic–polycytidylic acid sodium (polyI:C) to investigate the therapeutic effect\\u000a of bone marrow–derived mesenchymal stem cells (BM-MSC) on this model. After 6 weeks of MSC infusion, serum aminotransferase\\u000a and autoimmune antibodies declined, and histological examination by hematoxylin and

Dandan WangHuayong; Huayong Zhang; Jun Liang; Zhifeng Gu; Xiaolei Ma; Jing Huang; Jing Lin; Yayi Hou; Liwei Lu; Lingyun Sun

2011-01-01

10

Number of stromal precursors in mouse bone marrow and expression of cytokine genes in primary cultures of mouse bone marrow cells during various periods after immunization with S. typhimurium antigens.  

PubMed

Administration of S. typhimurium microbial mass to mice was followed by a significant increase (by 3-4 times) in the efficiency of cloning and number of stromal precursors in the femoral bone marrow. These parameters were maximum on days 1-3, but returned to normal by the 8th-15th day after immunization. As differentiated from intact animals, the expression of genes for proinflammatory cytokines IL-1? (day 1 after immunization), IL-6 (days 1-3), TNF-? (days 1, 3, and 6), and IFN-? (days 1-3) was detected in bone marrow cultures from immunized mice. The expression of genes for IFN-?, IL-18, and IFN-? was decreased on days 1, 3, and 6 after immunization of animals, respectively. Gene expression for the anti-inflammatory cytokine IL-4 was observed on day 6 after immunization. Therefore, this system was not characterized by a decrease in the immune response of stromal cells. The stromal component of hemopoietic and lymphoid organs has the vector of influences in response to bacterial antigens. This vector is directed to the stimulation and progression, but not to the suppression of immune reactions. Our results indicate that resident stromal cells play a role in the immune response of the body. PMID:21234434

Gorskaya, U F; Danilova, T A; Mesentzeva, M V; Shapoval, I M; Narovlyansky, A N; Nesterenko, V G

2010-10-01

11

Bone turnover markers in primary hyperparathyroidism.  

PubMed

Primary hyperparathyroidism is an endocrine disorder characterized by elevated or inappropriate normal levels of parathyroid hormone in a setting of hypercalcemia. The inclusion of calcium on the basic metabolic bone panel has allowed this disorder to be diagnosed even in the absence of symptoms. Nevertheless, the skeleton can be a target of excess parathyroid hormone activity even during its asymptomatic presentation. Bone turnover markers a surrogate index of the process of the remodeling process at the level of bone, and thus can be useful to monitor skeleton involvement in primary hyperparathyroidism. PMID:23374737

Costa, Aline G; Bilezikian, John P

12

[Surgical management of primary bone cancer].  

PubMed

Patients with primary bone malignancies must be treated by specialized multidisciplinary teams composed of pathologists, surgeons, orthopedists, oncologists, radiologists and radiotherapists, all with experience in the diagnosis and treatment of these tumors. If a malignancy is suspected, the biopsy must also be performed in such a center. Biopsy is part of the treatment and must be done by a senior surgeon, before starting specific treatment. Indeed, inappropriate biopsy can compromise the patient's functional prognosis and sometimes the vital outcome. The biopsy can be done percutaneously under radiological control with a True-cut needle or a trocart to obtain cores of pathological tissue. The pathologist must be well-versed in bone disorders. Open surgical biopsy is preferable for primary bone tumors, especially when a cartilaginous tumor is suspected. A short incision is used, situated on the same approach as that which will be used for surgical resection of the tumor, so that the biopsy scar is excised along with the tumor, in a single block. Surgical treatment of primary bone malignancies requires extensive resection, i.e. excision of the affected bone segment and any invaded soft tissues, as a single block, without breaching the tumor, and preserving a peripheral margin of healthy tissue. In most cases, reconstruction is necessary to preserve the function of the resected region. It is based on standard orthopedic techniques, namely osteosynthesis, bone grafts (autografts and allografts), prostheses of variable size, or a combination of prostheses and allografts (composite reconstruction). Amputation is only indicated if conservative resection is impossible. It has been shown that conservative resection, now possible in about 80% of cases, does not reduce the survival chances of patients with osteosarcoma. The indications for amputation include massive tumors invading vessels and nerves, resection of which would leave the limb non functional, as sell as tumor infection (often secondary to biopsy), inappropriate biopsy (infection of vessels or periarticular muscles, etc.), and local relapse. Amputation must respect the same oncologic principles as conservative resection. PMID:19718984

Anract, Philippe

2009-01-01

13

THE GROWTH OF MOUSE BONE MARROW CELLS IN VITRO  

Microsoft Academic Search

A simple in vitro technique is described for the growth of colonies from single cell suspensions of mouse bone marrow. The system involves the plating of marrow cells in agar on feeder layers of other cells, those from 8-day-old mouse kidney and 17th day mouse embryo being shown to be the most efficient types of feeder layers.Approximalely 400 colonies per

TR Bradley; D Metcalf

1966-01-01

14

Spatial integration in mouse primary visual cortex.  

PubMed

Responses of many neurons in primary visual cortex (V1) are suppressed by stimuli exceeding the classical receptive field (RF), an important property that might underlie the computation of visual saliency. Traditionally, it has proven difficult to disentangle the underlying neural circuits, including feedforward, horizontal intracortical, and feedback connectivity. Since circuit-level analysis is particularly feasible in the mouse, we asked whether neural signatures of spatial integration in mouse V1 are similar to those of higher-order mammals and investigated the role of parvalbumin-expressing (PV+) inhibitory interneurons. Analogous to what is known from primates and carnivores, we demonstrate that, in awake mice, surround suppression is present in the majority of V1 neurons and is strongest in superficial cortical layers. Anesthesia with isoflurane-urethane, however, profoundly affects spatial integration: it reduces the laminar dependency, decreases overall suppression strength, and alters the temporal dynamics of responses. We show that these effects of brain state can be parsimoniously explained by assuming that anesthesia affects contrast normalization. Hence, the full impact of suppressive influences in mouse V1 cannot be studied under anesthesia with isoflurane-urethane. To assess the neural circuits of spatial integration, we targeted PV+ interneurons using optogenetics. Optogenetic depolarization of PV+ interneurons was associated with increased RF size and decreased suppression in the recorded population, similar to effects of lowering stimulus contrast, suggesting that PV+ interneurons contribute to spatial integration by affecting overall stimulus drive. We conclude that the mouse is a promising model for circuit-level mechanisms of spatial integration, which relies on the combined activity of different types of inhibitory interneurons. PMID:23719206

Vaiceliunaite, Agne; Erisken, Sinem; Franzen, Florian; Katzner, Steffen; Busse, Laura

2013-05-29

15

The Primary Cilium as a Novel Extracellular Sensor in Bone  

PubMed Central

Mechanically induced adaptation of bone is required to maintain a healthy skeleton and defects in this process can lead to dramatic changes in bone mass, resulting in bone diseases such as osteoporosis. Therefore, understanding how this process occurs could yield novel therapeutics to treat diseases of excessive bone loss or formation. Over the past decade the primary cilium has emerged as a novel extracellular sensor in bone, being required to transduce changes in the extracellular mechanical environment into biochemical responses regulating bone adaptation. In this review, we introduce the primary cilium as a novel extracellular sensor in bone; discuss the in vitro and in vivo findings of primary cilia based sensing in bone; explore the role of the primary cilium in regulating stem cell osteogenic fate commitment and finish with future directions of research and possible development of cilia targeting therapeutics to treat bone diseases.

Hoey, David A.; Chen, Julia C.; Jacobs, Christopher R.

2012-01-01

16

Morphogenesis and bone integration of the mouse mandibular third molar.  

PubMed

The mouse third molar (M3) develops postnatally and is thus a unique model for studying the integration of a non-mineralized tooth with mineralized bone. This study assessed the morphogenesis of the mouse M3, related to the alveolar bone, comparing M3 development with that of the first molar (M1), the most common model in odontogenesis. The mandibular M3 was evaluated from initiation to eruption by morphology and by assessing patterns of proliferation, apoptosis, osteoclast distribution, and gene expression. Three-dimensional reconstruction and explant cultures were also used. Initiation of M3 occurred perinatally, as an extension of the second molar (M2) which grew into a region of soft mesenchymal tissue above the M2, still far away from the alveolar bone. The bone-free M3 bud gradually became encapsulated by bone at the cap stage at postnatal day 3. Osteoclasts were first visible at postnatal day 4 when the M3 came into close contact with the bone. The number of osteoclasts increased from postnatal day 8 to postnatal day 12 to form a space for the growing tooth. The M3 had erupted by postnatal day 26. The M3, although smaller than the M1, passed through the same developmental stages over a similar time span but showed differences in initiation and in the timing of bone encapsulation. PMID:21726286

Chlastakova, Ivana; Lungova, Vlasta; Wells, Kirsty; Tucker, Abigail S; Radlanski, Ralf J; Misek, Ivan; Matalova, Eva

2011-08-01

17

The bone diagnostic instrument III: Testing mouse femora  

NASA Astrophysics Data System (ADS)

Here we describe modifications that allow the bone diagnostic instrument (BDI) [P. Hansma et al., Rev. Sci. Instrum. 79, 064303 (2008); Rev. Sci. Instrum. 77, 075105 (2006)], developed to test human bone, to test the femora of mice. These modifications include reducing the effective weight of the instrument on the bone, designing and fabricating new probe assemblies to minimize damage to the small bone, developing new testing protocols that involve smaller testing forces, and fabricating a jig for securing the smaller bones for testing. With these modifications, the BDI was used to test the hypothesis that short-term running has greater benefit on the mechanical properties of the femur for young growing mice compared to older, skeletally mature mice. We measured elastic modulus, hardness, and indentation distance increase (IDI), which had previously been shown to be the best discriminators in model systems known to exhibit differences in mechanical properties at the whole bone level. In the young exercised murine femora, the IDI was significantly lower than in young control femora. Since IDI has a relation to postyield properties, these results suggest that exercise during bone development increases post yield mechanical competence. We were also able to measure effects of aging on bone properties with the BDI. There was a significant increase in the IDI, and a significant decrease in the elastic modulus and hardness between the young and old groups. Thus, with the modifications described here, the BDI can take measurements on mouse bones and obtain statistically significant results.

Randall, Connor; Mathews, Phillip; Yurtsev, Eugene; Sahar, Nadder; Kohn, David; Hansma, Paul

2009-06-01

18

New mouse models for metabolic bone diseases generated by genome-wide ENU mutagenesis.  

PubMed

Metabolic bone disorders arise as primary diseases or may be secondary due to a multitude of organ malfunctions. Animal models are required to understand the molecular mechanisms responsible for the imbalances of bone metabolism in disturbed bone mineralization diseases. Here we present the isolation of mutant mouse models for metabolic bone diseases by phenotyping blood parameters that target bone turnover within the large-scale genome-wide Munich ENU Mutagenesis Project. A screening panel of three clinical parameters, also commonly used as biochemical markers in patients with metabolic bone diseases, was chosen. Total alkaline phosphatase activity and total calcium and inorganic phosphate levels in plasma samples of F1 offspring produced from ENU-mutagenized C3HeB/FeJ male mice were measured. Screening of 9,540 mice led to the identification of 257 phenodeviants of which 190 were tested by genetic confirmation crosses. Seventy-one new dominant mutant lines showing alterations of at least one of the biochemical parameters of interest were confirmed. Fifteen mutations among three genes (Phex, Casr, and Alpl) have been identified by positional-candidate gene approaches and one mutation of the Asgr1 gene, which was identified by next-generation sequencing. All new mutant mouse lines are offered as a resource for the scientific community. PMID:22527485

Sabrautzki, Sibylle; Rubio-Aliaga, Isabel; Hans, Wolfgang; Fuchs, Helmut; Rathkolb, Birgit; Calzada-Wack, Julia; Cohrs, Christian M; Klaften, Matthias; Seedorf, Hartwig; Eck, Sebastian; Benet-Pagès, Ana; Favor, Jack; Esposito, Irene; Strom, Tim M; Wolf, Eckhard; Lorenz-Depiereux, Bettina; Hrab? de Angelis, Martin

2012-04-21

19

The terminator mouse: salvation for primary cell culture.  

PubMed

The Terminator had to come back from the future already several times in an effort to bring salvation to mankind. In the present issue of Kidney International, Guo et al. brought us a novel transgenic mouse model: the terminator mouse. This highly elegant mouse may facilitate significantly the derivation of primary cultures of a specific cell type from a tissue containing multiple cell populations. PMID:24172731

Kabgani, Nazanin; Moeller, Marcus J

2013-11-01

20

Estrogen deficiency stimulates B lymphopoiesis in mouse bone marrow.  

PubMed Central

We have found that an estrogen deficiency causes a marked increase in bone marrow cells. To examine the effect of estrogen on hemopoiesis, we characterized the increased population of bone marrow cells after ovariectomy (OVX). In OVX mice, the percentage of myeloid cells and granulocytes was decreased, whereas that of B220-positive B lymphocytes was selectively increased 2-4 wk after surgery. The total number of myeloid cells and granulocytes did not change appreciably, but that of B220-positive cells was greatly increased by OVX. When OVX mice were treated with estrogen, the increased B lymphopoiesis returned to normal. B220-positive cells were classified into two subpopulations, B220low and B220high. The majority of the B220low cells were negative for the IgM mu chain, whereas most of the B220high cells were mu-positive. OVX selectively increased the precursors of B lymphocytes identified by B220low. mu-negative phenotype, suggesting that an estrogen deficiency stimulates accumulation of B lymphocyte precursors. When bone marrow-derived stromal cells (ST2) were pretreated with estrogen then co-cultured with bone marrow cells in the presence of estrogen, the stromal cell-dependent B lymphopoiesis was greatly inhibited. The present study suggests that estrogen plays an important role in the regulation of B lymphocyte development in mouse bone marrow.

Masuzawa, T; Miyaura, C; Onoe, Y; Kusano, K; Ohta, H; Nozawa, S; Suda, T

1994-01-01

21

The effect of diet on bone shape in the mouse.  

PubMed Central

The effects of four deficient diets (oats, barley, wheat, buckwheat) on the shape of first and second cervical vertebrae and scapulae in C57BL mice have been measured using Fourier analysis. Bone shape was found to be robust, and only minimally affected by dietary change. The significance of this lack of change is discussed in the light of changes induced by diet in non-metrical variants in the skeleton. The study further emphasises the dangers of using certain non-metrical characters in taxonomic studies and indicates that the shapes of mouse bones are affected to a lesser degree by dietary influences than are the incidences of certain non-metrical character states.

Johnson, D R; O'Higgins, P; McAndrew, T J

1990-01-01

22

Effects of bleomycin on mouse bone-marrow stem cells.  

PubMed

The mouse hematopoietic stem-cell population was tested by the spleen colony technique for effects of the antineoplastic agent bleomycin (BLM). The time response of normal bone marrow was investigated by a single dose of BLM (400 mg/kg) between 0 and 72 hours. The dose response was studied over a wide range of doses (from 40 to 1,600 mg/kg) at a 4-hour exposure. Additional experiments concerned 1) the fraction of colony-forming units in the S phase after BLM administration (by means of pulse hydroxyurea treatment), 2) the response of bone marrow stimulated by endotoxin, and 3) the effects of split-dose treatments. The relatively low toxicity of BLM on both the differentiated and stem-cell populations of unstimulated bone marrow was confirmed and detailed. This drug exhibited peculiar, proliferation-dependent cell inactivation kinetics. Furthermore, BLM induced parasynchronous behavior in the unstimulated stem-cell population. The various aspects of BLM action are discussed with regard to its use in cancer chemotherapy. PMID:51084

Briganti, G; Galloni, L; Levi, G; Spalletta, V; Mauro, F

1975-07-01

23

Toxic effects of cobalt in primary cultures of mouse astrocytes  

Microsoft Academic Search

Cobalt is suspected to cause memory deficit in humans and was reported to induce neurotoxicity in animal models. We have studied the effects of cobalt in primary cultures of mouse astrocytes. CoCl2 (0.2–0.8mM) caused dose-dependent ATP depletion, apoptosis (cell shrinkage, phosphatidylserine externalization and chromatin rearrangements) and secondary necrosis. The mitochondria appeared to be a main target of cobalt toxicity, as

Olga Karovic; Ilaria Tonazzini; Nelson Rebola; Erik Edström; Cecilia Lövdahl; Bertil B. Fredholm; Elisabetta Daré

2007-01-01

24

Binocular integration and disparity selectivity in mouse primary visual cortex.  

PubMed

Signals from the two eyes are first integrated in primary visual cortex (V1). In many mammals, this binocular integration is an important first step in the development of stereopsis, the perception of depth from disparity. Neurons in the binocular zone of mouse V1 receive inputs from both eyes, but it is unclear how that binocular information is integrated and whether this integration has a function similar to that found in other mammals. Using extracellular recordings, we demonstrate that mouse V1 neurons are tuned for binocular disparities, or spatial differences, between the inputs from each eye, thus extracting signals potentially useful for estimating depth. The disparities encoded by mouse V1 are significantly larger than those encoded by cat and primate. Interestingly, these larger disparities correspond to distances that are likely to be ecologically relevant in natural viewing, given the stereo-geometry of the mouse visual system. Across mammalian species, it appears that binocular integration is a common cortical computation used to extract information relevant for estimating depth. As such, it is a prime example of how the integration of multiple sensory signals is used to generate accurate estimates of properties in our environment. PMID:23515794

Scholl, Benjamin; Burge, Johannes; Priebe, Nicholas J

2013-03-20

25

Bone Densitometry: Patients with Asymptomatic Primary Hyperparathyroidism. Health Technology Assessment Number 6, 1996.  

National Technical Information Service (NTIS)

Bone mass loss and osteoporosis are caused by various conditions, such as asymptomatic primary hyperparathyroidism (APHPT), and treatments, such as prolonged steroid therapy. Bone densitometry is used to measure bone mass density to determine the degree o...

M. Erlichman T. V. Holohan

1995-01-01

26

Primary Mesenchymal Stem and Progenitor Cells from Bone Marrow Lack Expression of CD44 Protein*  

PubMed Central

Despite significant progress in our understanding of mesenchymal stem cell (MSC) biology during recent years, much of the information is based on experiments using in vitro culture-selected stromal progenitor cells. Therefore, the natural cellular identity of MSCs remains poorly defined. Numerous studies have reported that CD44 expression is one of the characteristics of MSCs in both humans and mice; however, we here have prospectively isolated bone marrow stromal cell subsets from both human and mouse bone marrow by flow cytometry and characterized them by gene expression analysis and function assays. Our data provide functional and molecular evidence suggesting that primary mesenchymal stem and progenitor cells of bone marrow reside in the CD44? cell fraction in both mice and humans. The finding that these CD44? cells acquire CD44 expression after in vitro culture provides an explanation for the previous misconceptions concerning CD44 expression on MSCs. In addition, the other previous reported MSC markers, including CD73, CD146, CD271, and CD106/VCAM1, are also differentially expressed on those two cell types. Our microarray data revealed a distinct gene expression profile of the freshly isolated CD44? cells and the cultured MSCs generated from these cells. Thus, we conclude that bone marrow MSCs physiologically lack expression of CD44, highlighting the natural phenotype of MSCs and opening new possibilities to prospectively isolate MSCs from the bone marrow.

Qian, Hong; Le Blanc, Katarina; Sigvardsson, Mikael

2012-01-01

27

Mesenchymal Stem Cells Expressing Osteogenic and Angiogenic Factors Synergistically Enhance Bone Formation in a Mouse Model of Segmental Bone Defect  

PubMed Central

The potential of mesenchymal stem cells (MSC) in tissue regeneration is increasingly gaining attention. There is now accumulating evidence that MSC make an important contribution to postnatal vasculogenesis. During bone development and fracture healing, vascularization is observed before bone formation. The present study determined the potential of MSC, transduced ex vivo with a recombinant adeno-associated virus 6 (rAAV6) encoding bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor (VEGF) in a mouse model of segmental bone defect created in the tibiae of athymic nude mice. Mouse MSC that were mock-transduced or transduced with rAAV6-BMP2:VEGF were systemically transplanted following radiographic confirmation of the osteotomy. Effects of the therapy were determined by enzyme-linked immunosorbent assay measurements for BMP2 and VEGF, dual-energy X-ray absorptiometry (DXA) for bone density, three-dimensional microcomputed tomography (µCT) for bone and capillary architecture, and histomorphometry for bone remodeling. Results of these analyses indicated enhanced bone formation in the group that received BMP2+VEGF-expressing MSC compared to other groups. The therapeutic effects were accompanied by increased vascularity and osteoblastogenesis, indicating its potential for effective use while treating difficult nonunion bone defects in humans.

Kumar, Sanjay; Wan, Chao; Ramaswamy, Girish; Clemens, Thomas L; Ponnazhagan, Selvarangan

2010-01-01

28

Effect of electromagnetic fields on proliferation and differentiation of cultured mouse bone marrow mesenchymal stem cells  

Microsoft Academic Search

Summary  In order to study the effects of electromagnetic fields (EMFs) on proliferation, differentiation and intercellular cyclic\\u000a AMP (cAMP) in mouse bone marrow mesenchymal stem cells (MSCs)in vitro, the mouse bone MSCs were isolated and culturedin vitro. The third passage MSCs were divided into 4 groups and stimulated with EMFs. The cellular proliferation (MTT), the cellular\\u000a differentiation (alkaline phosphatase activity, ALP),

Wu Hua; Ren Kai; Zhao Wenchun; Ge Baojian; Peng Songlin

2005-01-01

29

Unusual primary malignant reticulosis of the bone marrow.  

PubMed

An unusual case of primary reticulosis of the bone marrow in a 67-year-old female has been described. Pathological-anatomical characteristics of the present case are as follows; 1) Primary non-destructive proliferation of neoplastic reticulum cells or their variants, confined to the systemic bone marrow. Metastatic foci were noted in several areas where extramedullary hematopoiesis occurred. 2) Cytologically, neoplastic cells resemble atypical plasmacytic cells. No active phagocytosis was noted in neoplastic cells. 3) Histologically a significant increase in reticulin fibers was observed closely associated with proliferating cells. 4) Electron-microscopically these cells showed developed endoplasmic reticula but were distinguishable from myeloma cells by their irregular cytoplasmic border, which became frequently obscure, and by the presence of a desmosome-like structure. 5) These cells showed a negative immunofluorescent reaction to all types of human immunoglobulin classes. The morphological characteristics of these cells distinguished the disorder from poorly differentiated myelomatosis. PMID:685694

Inoue, T; Kimula, Y; Tanaka, Y; Shirakura, T; Hayashi-Nakao, J

1978-05-01

30

Isolation of primary myofibroblasts from mouse and human colon tissue.  

PubMed

The myofibroblast is a stromal cell of the gastrointestinal (GI) tract that has been gaining considerable attention for its critical role in many GI functions. While several myofibroblast cell lines are commercially available to study these cells in vitro, research results from a cell line exposed to experimental cell culture conditions have inherent limitations due to the overly reductionist nature of the work. Use of primary myofibroblasts offers a great advantage in terms of confirming experimental findings identified in a cell line. Isolation of primary myofibroblasts from an animal model allows for the study of myofibroblasts under conditions that more closely mimic the disease state being studied. Isolation of primary myofibroblasts from human colon tissue provides arguably the most relevant experimental data, since the cells come directly from patients with the underlying disease. We describe a well-established technique that can be utilized to isolate primary myofibroblasts from both mouse and human colon tissue. These isolated cells have been characterized to be alpha-smooth muscle actin and vimentin-positive, and desmin-negative, consistent with subepithelial intestinal myofibroblasts. Primary myofibroblast cells can be grown in cell culture and used for experimental purposes over a limited number of passages. PMID:24145735

Khalil, Hassan; Nie, Wenxian; Edwards, Robert A; Yoo, James

2013-10-12

31

MRI detection of early bone metastases in B16 mouse melanoma models  

PubMed Central

Bone metastasis causes significant morbidity in cancer patients, including bone pain, pathologic fractures, nerve compression syndrome, and hypercalcemia. Animal models are utilized to study the pathogenesis of skeletal metastases and to evaluate potential therapeutic agents. Previously published methods for imaging bone metastasis in rodent models have focused on identifying advanced stage metastasis using simple X-rays. Here we report MRI as a method for detecting early bone metastases in mouse models in vivo. B16 mouse melanoma cells were injected into the left cardiac ventricle of C57BL/6 mice and magnetic resonance (MR) images were obtained of the left leg following the development of metastatic disease, when tumor associated bone destruction was histologically present but not visible by X-ray. T1 and T2 relaxation times of bone marrow were measured in healthy control mice and B16 melanoma tumor-bearing mice. Mean T2 values for normal marrow were 28 ms (SD 5) and for diseased bone marrow were 41 ms (SD 3). T2 relaxation time of diseased bone marrow is significantly longer than that of normal bone marrow (P < 0.0001) and can be used as a marker of early bone metastases. These studies demonstrate that MR imaging can detect bone marrow metastases in small animals prior to development of cortical bone loss identified by X-ray.

Gauvain, Karen M.; Garbow, Joel R.; Song, Sheng-Kwei; Hirbe, Angela C.; Weilbaecher, Katherine

2009-01-01

32

Modulation of Doxorubicin-Induced Genotoxicity by Aegle marmelos in Mouse Bone Marrow: A Micronucleus Study  

Microsoft Academic Search

The effect of various concentrations of Aegle marmelos (AME) on the doxorubicin (DOX)-induced genotoxic effects in mice bone marrow was studied. Treatment of mice with different concentrations of DOX resulted in a dose-dependent elevation in the frequency of micronucleated polychromatic (MPCE) as well as normochromatic (MNCE) erythrocytes in mouse bone marrow. The frequencies of MPCE and MNCE increased with scoring

Ponemone Venkatesh; Bellary Shantala; Ganesh Chandra Jagetia; K. Koteshwer Rao; Manjeshwar Shrinath Baliga

2007-01-01

33

The relevance of mouse models for investigating age-related bone loss in humans.  

PubMed

Mice are increasingly used for investigation of the pathophysiology of osteoporosis because their genome is easily manipulated, and their skeleton is similar to that of humans. Unlike the human skeleton, however, the murine skeleton continues to grow slowly after puberty and lacks osteonal remodeling of cortical bone. Yet, like humans, mice exhibit loss of cancellous bone, thinning of cortical bone, and increased cortical porosity with advancing age. Histologic evidence in mice and humans alike indicates that inadequate osteoblast-mediated refilling of resorption cavities created during bone remodeling is responsible. Mouse models of progeria also show bone loss and skeletal defects associated with senescence of early osteoblast progenitors. Additionally, mouse models of atherosclerosis, which often occurs in osteoporotic participants, also suffer bone loss, suggesting that common diseases of aging share pathophysiological pathways. Knowledge of the causes of skeletal fragility in mice should therefore be applicable to humans if inherent limitations are recognized. PMID:23689830

Jilka, Robert L

2013-05-20

34

Orientation specificity of contrast adaptation in mouse primary visual cortex.  

PubMed

Contrast adaptation is a commonly studied phenomenon in vision, where prolonged exposure to spatial contrast alters perceived stimulus contrast and produces characteristic shifts in the contrast response functions of primary visual cortex neurons in cats and primates. In this study we investigated contrast adaptation in mouse primary visual cortex with two goals in mind. First, we sought to establish a quantitative description of contrast adaptation in an animal model, where genetic tools are more readily applicable to this phenomenon. Second, the orientation specificity of contrast adaptation was studied to comparatively assess the possible role of local cortical networks in contrast adaptation. In cats and primates, predictable differences in visual processing across the cortical surface are thought to be caused by inhomogeneous local network membership that arises from the pinwheel organization of orientation columns. Because mice lack this pinwheel organization, we predicted that local cortical networks would have access to a broad spectrum of orientation signals, and contrast adaptation in mice would not be specific to the recorded cell's preferred orientation. We found that most mouse V1 neurons showed contrast adaptation that was robust regardless of whether the adapting stimulus matched the cell's preferred orientation or was orthogonal to it. PMID:22696541

Stroud, Aaron C; Ledue, Emily E; Crowder, Nathan A

2012-06-13

35

Isolation of primary mouse trophoblast cells and trophoblast invasion assay.  

PubMed

The placenta is responsible for the transport of nutrients, gasses and growth factors to the fetus, as well as the elimination of wastes. Thus, defects in placental development have important consequences for the fetus and mother, and are a major cause of embryonic lethality. The major cell type of the fetal portion of the placenta is the trophoblast. Primary mouse placental trophoblast cells are a useful tool for studying normal and abnormal placental development, and unlike cell lines, may be isolated and used to study trophoblast at specific stages of pregnancy. In addition, primary cultures of trophoblast from transgenic mice may be used to study the role of particular genes in placental cells. The protocol presented here is based on the description by Thordarson et al., in which a percoll gradient is used to obtain a relatively pure trophoblast cell population from isolated mouse placentas. It is similar to the more widely used methods for human trophoblast cell isolation. Purity may be assessed by immunocytochemical staining of the isolated cells for cytokeratin 7. Here, the isolated cells are then analyzed using a matrigel invasion assay to assess trophoblast invasiveness in vitro. The invaded cells are analyzed by immunocytochemistry and stained for counting. PMID:22257865

Pennington, Kathleen A; Schlitt, Jessica M; Schulz, Laura C

2012-01-08

36

Resveratrol Inhibited Hydroquinone-Induced Cytotoxicity in Mouse Primary Hepatocytes  

PubMed Central

Hydroquinone (1,4-benzenediol) has been widely used in clinical situations and the cosmetic industry because of its depigmenting effects. Most skin-lightening hydroquinone creams contain 4%–5% hydroquinone. We have investigated the role of resveratrol in prevention of hydroquinone induced cytotoxicity in mouse primary hepatocytes. We found that 400 µM hydroquinone exposure alone induced apoptosis of the cells and also resulted in a significant drop of cell viability compared with the control, and pretreatment of resveratrol to a final concentration of 0.5 mM 1 h before hydroquinone exposure did not show a significant improvement in the survival rate of the hepatocytes, however, relatively higher concentrations of resveratrol (?1 mM) inhibited apoptosis of the mouse primary hepatocytes and increased cell viability in a dose-dependent manner, and in particular the survival rate of the hepatocytes was recovered from 28% to near 100% by 5 mM resveratrol. Interestingly, pretreatment with resveratrol for longer time (24 h), even in very low concentrations (50 µM, 100 µM), blocked the damage of hydroquinone to the cells. We also observed that resveratrol pretreatment suppressed hydroquinone-induced expression of cytochrome P450 2E1 mRNA dose-dependently. The present study suggests that resveratrol protected the cells against hydroquinone-induced toxicity through its antioxidant function and possibly suppressive effect on the expression of cytochrome P450 2E1.

Wang, Da-Hong; Ootsuki, Yoshie; Fujita, Hirofumi; Miyazaki, Masahiro; Yie, Qinxia; Tsutsui, Ken; Sano, Kuniaki; Masuoka, Noriyoshi; Ogino, Keiki

2012-01-01

37

Mechanical loading, damping, and load-driven bone formation in mouse tibiae  

PubMed Central

Mechanical loads play a pivotal role in the growth and maintenance of bone and joints. Although loading can activate anabolic genes and induce bone remodeling, damping is essential for preventing traumatic bone injury and fracture. In this study we investigated the damping capacity of bone, joint tissue, muscle, and skin using a mouse hindlimb model of enhanced loading in conjunction with finite element modeling to model bone curvature. Our hypothesis was that loads were primarily absorbed by the joints and muscle tissue, but that bone also contributed to damping through its compression and natural bending. To test this hypothesis, fresh mouse distal lower limb segments were cyclically loaded in axial compression in sequential bouts, with each subsequent bout having less surrounding tissue. A finite element model was generated to model effects of bone curvature in silico. Two damping-related parameters (phase shift angle and energy loss) were determined from the output of the loading experiments. Interestingly, the experimental results revealed that the knee joint contributed to the largest portion of the damping capacity of the limb, and bone itself accounted for approximately 38% of the total phase shift angle. Computational results showed that normal bone curvature enhanced the damping capacity of the bone by approximately 40%, and the damping effect grew at an accelerated pace as curvature was increased. Although structural curvature reduces critical loads for buckling in beam theory, evolution apparently favors maintaining curvature in the tibia. Histomorphometric analysis of the tibia revealed that in response to axial loading, bone formation was significantly enhanced in the regions that were predicted to receive a curvature-induced bending moment. These results suggest that in addition to bone’s compressive damping capacity, surrounding tissues, as well as naturally-occurring bone curvature, also contribute to mechanical damping, which may ultimately affect bone remodeling and bone quality.

Dodge, Todd; Wanis, Mina; Ayoub, Ramez; Zhao, Liming; Watts, Nelson B.; Bhattacharya, Amit; Akkus, Ozan; Robling, Alexander; Yokota, Hiroki

2012-01-01

38

Clinical characteristics and outcomes of primary bone lymphoma in Korea  

PubMed Central

Background This study evaluates the effectiveness of immunochemotherapy and radiation therapy in the treatment of patients with primary bone lymphoma (PBL). Methods We retrospectively reviewed the medical records of 33 patients with PBL who were treated at 6 medical centers in Korea from 1992 to 2010. Clinicopathological features and treatment outcomes were analyzed. Results The median age of the patients participating in our study was 40 years. The most common sites of involvement were the pelvis (12.36%) and femur (11.33%). CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisolone) or CHOP-like regimens were administered to 20 patients (61%), and R-CHOP (rituximab plus CHOP) was administered to the remaining 13 patients (39%). The overall response rate was 89% (complete response, 76%; partial response, 12%). The overall survival (OS) of patients with solitary bone lesions was longer than that of patients with multiple bone lesions (median OS: not reached vs. 166 months, respectively; P=0.089). Addition of rituximab to CHOP did not significantly affect either OS or progression-free survival (P=0.53 and P=0.23, respectively). Combining radiation therapy with chemotherapy also did not improve the OS or progression-free survival of patients with solitary bone lesions. Conclusion Conventional cytotoxic chemotherapy remains an effective treatment option for patients with PBL. Additional benefits of supplementing chemotherapy with either rituximab or radiation therapy were not observed in this study. Further investigation is needed to characterize the role of immunochemotherapy in treating patients with PBL.

Kim, So Yeon; Shin, Dong-Yeop; Lee, Seung-Sook; Suh, Cheolwon; Kwak, Jae-Yong; Kim, Hoon-Gu; Lee, Jae Hoon; Lee, Soon Il; Lee, Ye Rim; Kang, Seung Hwa; Mun, Se Kwon; Lee, Min Jae; Lee, Hyo-Rak; Yang, Sung Hyun

2012-01-01

39

Spatiotemporal specificity of contrast adaptation in mouse primary visual cortex  

PubMed Central

Prolonged viewing of high contrast gratings alters perceived stimulus contrast, and produces characteristic changes in the contrast response functions of neurons in the primary visual cortex (V1). This is referred to as contrast adaptation. Although contrast adaptation has been well-studied, its underlying neural mechanisms are not well-understood. Therefore, we investigated contrast adaptation in mouse V1 with the goal of establishing a quantitative description of this phenomenon in a genetically manipulable animal model. One interesting aspect of contrast adaptation that has been observed both perceptually and in single unit studies is its specificity for the spatial and temporal characteristics of the stimulus. Therefore, in the present work we determined if the magnitude of contrast adaptation in mouse V1 neurons was dependent on the spatial frequency and temporal frequency of the adapting grating. We used protocols that were readily comparable with previous studies in cats and primates, and also a novel contrast ramp stimulus that characterized the spatial and temporal specificity of contrast adaptation simultaneously. Similar to previous work in higher mammals, we found that contrast adaptation was strongest when the spatial frequency and temporal frequency of the adapting grating matched the test stimulus. This suggests similar mechanisms underlying contrast adaptation across animal models and indicates that the rapidly advancing genetic tools available in mice could be used to provide insights into this phenomenon.

LeDue, Emily E.; King, Jillian L.; Stover, Kurt R.; Crowder, Nathan A.

2013-01-01

40

Spatiotemporal specificity of contrast adaptation in mouse primary visual cortex.  

PubMed

Prolonged viewing of high contrast gratings alters perceived stimulus contrast, and produces characteristic changes in the contrast response functions of neurons in the primary visual cortex (V1). This is referred to as contrast adaptation. Although contrast adaptation has been well-studied, its underlying neural mechanisms are not well-understood. Therefore, we investigated contrast adaptation in mouse V1 with the goal of establishing a quantitative description of this phenomenon in a genetically manipulable animal model. One interesting aspect of contrast adaptation that has been observed both perceptually and in single unit studies is its specificity for the spatial and temporal characteristics of the stimulus. Therefore, in the present work we determined if the magnitude of contrast adaptation in mouse V1 neurons was dependent on the spatial frequency and temporal frequency of the adapting grating. We used protocols that were readily comparable with previous studies in cats and primates, and also a novel contrast ramp stimulus that characterized the spatial and temporal specificity of contrast adaptation simultaneously. Similar to previous work in higher mammals, we found that contrast adaptation was strongest when the spatial frequency and temporal frequency of the adapting grating matched the test stimulus. This suggests similar mechanisms underlying contrast adaptation across animal models and indicates that the rapidly advancing genetic tools available in mice could be used to provide insights into this phenomenon. PMID:24106461

Ledue, Emily E; King, Jillian L; Stover, Kurt R; Crowder, Nathan A

2013-10-03

41

Primary homologies of the circumorbital bones of snakes.  

PubMed

Some snakes have two circumorbital ossifications that in the current literature are usually referred to as the postorbital and supraorbital. We review the arguments that have been proposed to justify this interpretation and provide counter-arguments that reject those conjectures of primary homology based on the observation of 32 species of lizards and 81 species of snakes (both extant and fossil). We present similarity arguments, both topological and structural, for reinterpretation of the primary homologies of the dorsal and posterior orbital ossifications of snakes. Applying the test of similarity, we conclude that the posterior orbital ossification of snakes is topologically consistent as the homolog of the lacertilian jugal, and that the dorsal orbital ossification present in some snakes (e.g., pythons, Loxocemus, and Calabaria) is the homolog of the lacertilian postfrontal. We therefore propose that the terms postorbital and supraorbital should be abandoned as reference language for the circumorbital bones of snakes, and be replaced with the terms jugal and postfrontal, respectively. The primary homology claim for the snake "postorbital" fails the test of similarity, while the term "supraorbital" is an unnecessary and inaccurate application of the concept of a neomorphic ossification, for an element that passes the test of similarity as a postfrontal. This reinterpretation of the circumorbital bones of snakes is bound to have important repercussions for future phylogenetic analyses and consequently for our understanding of the origin and evolution of snakes. J. Morphol. 274:973-986, 2013. © 2013 Wiley Periodicals, Inc. PMID:23630161

Palci, Alessandro; Caldwell, Michael W

2013-04-30

42

Magnetic resonance imaging of primary tumours and tumour-like lesions of bone  

Microsoft Academic Search

The experience with magnetic resonance imaging (MRI) of 81 patients with primary bone tumours and tumour-like lesions is reported. MRI proved to be a sensitive method of detecting primary bone tumours. Intramedullary and extraosseous parts of bone tumours were, delineated better than by plain films and computed tomography (CT). Surgical clips and Harrington rods did not appreciably limit the estimation

K. Bohndorf; M. Reiser; B. Lochner; W. Féaux de Lacroix; W. Steinbrich

1986-01-01

43

Normocalcemic versus Hypercalcemic Primary Hyperparathyroidism: More Stone than Bone?  

PubMed Central

Introduction. Normocalcemic primary hyperparathyroidism (NPHPT) is considered a variant of the more frequent form of the disease characterized by normal serum calcium levels with high PTH. The higher prevalence of renal stones in patients with HPTP and the well established association with bone disorders show the importance of studies on how to manage asymptomatic patients. Objective. To compare the clinical and laboratory data between the normocalcemic and mild hypercalcemic forms of PHPT. Methods. We retrospectively evaluated 70 patients with PHPT, 33 normocalcemic and 37 mild hypercalcemic. Results. The frequency of nephrolithiasis was 18.2% in normocalcemic patients and 18.9% in the hypercalcemic ones (P = 0.937). Fifteen percent of normocalcemic patients had a previous history of fractures compared to 10.8% of hypercalcemic patients, although there was no statistically significant difference (P = 0.726). Conclusion. Our data confirms a high prevalence of urolithiasis in normocalcemic primary hyperparathyroidism, but with the preservation of cortical bone. This finding supports the hypothesis that this disease is not an idle condition and needs treatment.

Amaral, L. M.; Queiroz, D. C.; Marques, T. F.; Mendes, M.; Bandeira, F.

2012-01-01

44

Alveolar bone loss and restorative dentistry in the primary molars.  

PubMed

This study examined the relationship between the status of the interproximal alveolar bone and the nature of the adjacent proximal surfaces in the primary molar area. In bite wing radiographs of 354 children, aged 6 to 9 years, 5091 sites were examined; 72.7% of the sites were adjacent to intact proximal surfaces, 14.2% to untreated proximal carious surfaces, and 13.0% to restored proximal surfaces. Marginal alveolar bone loss (ABL) was evident in 26.8% of the children, at 4.0% of the sites. Two thirds of the children with ABL had bone defects in more than 1 site. Males had a significantly higher mean number of sites affected with marginal ABL, per child, than females (mean = 2.4, SE = 0.2 and mean = 1.8, SE = 0.2 respectively). ABL was found adjacent to: 0.8% of the intact surfaces; 16.9% of the carious surfaces; 7.8% of the restored surfaces; 1.8% and 53.8% of the sites without or with proximal contact loss respectively; 3.8% and 30.8% of the sites with or without an adequate amalgam restoration respectively; 4.9% and 25.8% of the sites with an adequate or inadequate crown restoration respectively. The differences in distribution of marginal alveolar bone loss were highly significant (Chi square analysis, p = < 0.0001) for sites with intact, carious or restored sites, and for the presence or absence of contact loss, adequate amalgam or adequate crown. PMID:9161207

Bimstein, E; Zaidenberg, R; Soskolne, A W

1996-01-01

45

Cadmium stimulates osteoclast-like multinucleated cell formation in mouse bone marrow cell cultures  

SciTech Connect

Most of cadmium (Cd)-treated animals have been reported to show osteoporosis-like changes in bones. This suggests that Cd may promote bone loss by a direct action on bone. It was found that Cd stimulated prostaglandin E{sub 2}(PGE{sub 2}) production in the osteoblast-like cell, MC3T3-E1. Therefore, Cd stimulates bone resorption by increasing PGE{sub 2} production. Recently, several bone marrow cell culture systems have been developed for examining the formation of osteoclast-like multinucleated cells in vitro. As osteoblasts produce PGE{sub 2} by Cd-induced cyclooxygenase and may play an important role in osteoclast formation, the present study was undertaken to clarify the possibility that Cd might stimulate osteoclast formation in a mouse bone marrow culture system.

Miyahara, Tatsuro; Takata, Masakazu; Miyata, Masaki; Nagai, Miyuki; Sugure, Akemi; Kozuka, Hiroshi; Kuze, Shougo (Toyama Medical and Pharmaceutical Univ. (Japan))

1991-08-01

46

Tumor-induced injury of primary afferent sensory nerve fibers in bone cancer pain  

Microsoft Academic Search

Bone is the most common site of chronic pain in patients with metastatic cancer. What remains unclear are the mechanisms that generate this pain and why bone cancer pain can be so severe and refractory to treatment with opioids. Here we show that following injection and confinement of NCTC 2472 osteolytic tumor cells within the mouse femur, tumor cells sensitize

Christopher M. Peters; Joseph R. Ghilardi; Cathy P. Keyser; Kazufumi Kubota; Theodore H. Lindsay; Nancy M. Luger; David B. Mach; Matthew J. Schwei; Molly A. Sevcik; Patrick W. Mantyh

2005-01-01

47

Sex differences in bone resorption in the mouse femur  

Microsoft Academic Search

In male and female dd-mice at 4, 7, and 14 weeks of age and in 7 and 14-week-old mice gonadectomized at 4 weeks of age, the number of osteoclasts and the number and size of bone resorption areas along the surface of bone trabeculae in the distal metaphysis of the femur were determined. Osteoclasts were counted at the light-microscopic level

K. Abe; Y. Aoki

1989-01-01

48

Periodontal Disease: Linking the Primary Inflammation to Bone Loss  

PubMed Central

Periodontal disease (PD), or periodontitis, is defined as a bacterially induced disease of the tooth-supporting (periodontal) tissues. It is characterized by inflammation and bone loss; therefore understanding how they are linked would help to address the most efficacious therapeutic approach. Bacterial infection is the primary etiology but is not sufficient to induce the disease initiation or progression. Indeed, bacteria-derived factors stimulate a local inflammatory reaction and activation of the innate immune system. The innate response involves the recognition of microbial components by host cells, and this event is mediated by toll-like receptors (TLRs) expressed by resident cells and leukocytes. Activation of these cells leads to the release of proinflammatory cytokines and recruitment of phagocytes and lymphocytes. Activation of T and B cells initiates the adaptive immunity with Th1 Th2 Th17 Treg response and antibodies production respectively. In this inflammatory scenario, cytokines involved in bone regulation and maintenance have considerable relevance because tissue destruction is believed to be the consequence of host inflammatory response to the bacterial challenge. In the present review, we summarize host factors including cell populations, cytokines, and mechanisms involved in the destruction of the supporting tissues of the tooth and discuss treatment perspectives based on this knowledge.

Di Benedetto, Adriana; Gigante, Isabella; Colucci, Silvia; Grano, Maria

2013-01-01

49

Pathological features of bone marrow transplantation-related toxicity in a mouse  

Microsoft Academic Search

In this case report, we present a mock-transduced bone marrow (BM) transplantation in a mouse, which was found moribund and autopsied to evaluate pathogenesis. Macroscopically, red discoloration of systemic organs was observed. Hematological values revealed a decrease in white blood cells, red blood cells, hematocrit, hemoglobin, and platelets, but an increase in reticulocytes. In BM cytology, hematopoietic cell lines were

Yong-Hoon Kim; Chang-Su Ha; Hyun-Sook Lee; Sun-Hwa Lim; Kyoung-Sik Moon; Moon-Koo Chung; Hwa-Young Son

2009-01-01

50

Activity of the human carcinogen MeCCNU in the mouse bone marrow micronucleus assay  

SciTech Connect

The nitrosourea mustard MeCCNU is the most recent organic chemical to be classified as a human carcinogen by IARC. MeCCNU gave a strong positive response when tested in the mouse bone marrow micronucleus assay. Activity was evident using either ip injection or oral gavage of the test chemical. These results further support the correlation between human carcinogens and their genotoxicity.

Tinwell, H.; Ashby, J. (ICI Central Toxicology Lab, Cheshire (England))

1991-01-01

51

Primary cilia act as mechanosensors during bone healing around an implant.  

PubMed

The primary cilium is an organelle that senses cues in a cell's local environment. Some of these cues constitute molecular signals; here, we investigate the extent to which primary cilia can also sense mechanical stimuli. We used a conditional approach to delete Kif3a in pre-osteoblasts and then employed a motion device that generated a spatial distribution of strain around an intra-osseous implant positioned in the mouse tibia. We correlated interfacial strain fields with cell behaviors ranging from proliferation through all stages of osteogenic differentiation. We found that peri-implant cells in the Col1Cre;Kif3a(fl/fl) mice were unable to proliferate in response to a mechanical stimulus, failed to deposit and then orient collagen fibers to the strain fields caused by implant displacement, and failed to differentiate into bone-forming osteoblasts. Collectively, these data demonstrate that the lack of a functioning primary cilium blunts the normal response of a cell to a defined mechanical stimulus. The ability to manipulate the genetic background of peri-implant cells within the context of a whole, living tissue provides a rare opportunity to explore mechanotransduction from a multi-scale perspective. PMID:22784673

Leucht, P; Monica, S D; Temiyasathit, S; Lenton, K; Manu, A; Longaker, M T; Jacobs, C R; Spilker, R L; Guo, H; Brunski, J B; Helms, J A

2012-07-10

52

Bone morphogenetic protein signaling is impaired in an Hfe knockout mouse model of hemochromatosis  

PubMed Central

Background and Aims Mutations in HFE are the most common cause of the iron-overload disorder hereditary hemochromatosis (HH). Levels of the main iron regulatory hormone, hepcidin, are inappropriately low in HH mouse models and patients with HFE mutations, indicating that HFE regulates hepcidin. The bone morphogenetic protein 6 (BMP6)-SMAD signaling pathway is an important endogenous regulator of hepcidin expression. We investigated whether HFE is involved in BMP6-SMAD regulation of hepcidin expression. Methods The BMP6-SMAD pathway was examined in Hfe knockout (KO) mice and in wild-type (WT) mice as controls. Mice were placed on diets of varying iron content. Hepcidin induction by BMP6 was examined in primary hepatocytes from Hfe KO mice; data were compared with those of WT mice. Results Liver levels of Bmp6 mRNA were higher in Hfe KO mice; these were appropriate for the increased hepatic levels of iron in these mice, compared with WT mice. However, levels of hepatic phosphorylated Smad 1/5/8 protein (an intracellular mediator of Bmp6 signaling) and Id1 mRNA (a target gene of Bmp6) were inappropriately low for the body iron burden and Bmp6 mRNA levels in Hfe KO, compared with WT mice. BMP6 induction of hepcidin expression was reduced in Hfe KO hepatocytes compared with WT hepatocytes. Conclusions HFE is not involved in regulation of BMP6 by iron, but does regulate the downstream signals of BMP6 that are triggered by iron.

Corradini, Elena; Garuti, Cinzia; Montosi, Giuliana; Ventura, Paolo; Andriopoulos, Billy; Lin, Herbert Y.; Pietrangelo, Antonello; Babitt, Jodie L.

2009-01-01

53

Bone tumor  

MedlinePLUS

Tumor - bone; Bone cancer; Primary bone tumor; Secondard bone tumor ... malignant) bone tumors include: Chondrosarcoma Ewing's sarcoma Fibrosarcoma Osteosarcomas The cancers that most often spread to the ...

54

Function and Malfunction of Hematopoietic Stem Cells in Primary Bone Marrow Failure Syndromes  

Microsoft Academic Search

Hematopoietic stem cells (HSCs) are responsible for the production of mature blood cells in bone marrow; peripheral pancytopenia is a common clinical presentation resulting from several different conditions, including hematological or extra-hematol ogical diseases (mostly cancers) affecting the marrow function, as well as primary failure of hematopoiesis. Primary bone marrow failure syndromes are a heterogeneous group of diseases with specific

Antonio M. Risitano; Jaroslaw P. Maciejewski; Carmine Selleri; Bruno Rotoli

2007-01-01

55

Neovascular Niche for Human Myeloma Cells in Immunodeficient Mouse Bone  

PubMed Central

The interaction with bone marrow (BM) plays a crucial role in pathophysiological features of multiple myeloma (MM), including cell proliferation, chemoresistance, and bone lesion progression. To characterize the MM-BM interactions, we utilized an in vivo experimental model for human MM in which a GFP-expressing human MM cell line is transplanted into NOG mice (the NOG-hMM model). Transplanted MM cells preferentially engrafted at the metaphyseal region of the BM endosteum and formed a complex with osteoblasts and osteoclasts. A subpopulation of MM cells expressed VE-cadherin after transplantation and formed endothelial-like structures in the BM. CD138+ myeloma cells in the BM were reduced by p53-dependent apoptosis following administration of the nitrogen mustard derivative bendamustine to mice in the NOG-hMM model. Bendamustine maintained the osteoblast lining on the bone surface and protected extracellular matrix structures. Furthermore, bendamustine suppressed the growth of osteoclasts and mesenchymal cells in the NOG-hMM model. Since VE-cadherin+ MM cells were chemoresistant, hypoxic, and HIF-2?-positive compared to the VE-cadherin? population, VE-cadherin induction might depend on the oxygenation status. The NOG-hMM model described here is a useful system to analyze the dynamics of MM pathophysiology, interactions of MM cells with other cellular compartments, and the utility of novel anti-MM therapies.

Miyakawa, Yoshitaka; Nakamura-Ishizu, Ayako; Miyauchi, Yoshiteru; Fujita, Nobuyuki; Miyamoto, Kana; Miyamoto, Takeshi; Ikeda, Eiji; Kizaki, Masahiro; Nojima, Yoshihisa; Suda, Toshio

2012-01-01

56

Effect of cyclophosphamide and electromagnetic fields on mouse bone marrow  

SciTech Connect

The authors have previously shown that the exposure to low frequency pulsing electromagnetic fields (PEMF) of mice X-ray irradiated resulted in an increased damage to the bone marrow. The series of experiments here reported were designed to investigate the effect of PEMF exposure after intraperitoneum injection of 200mg/kg of cyclophosphamide (CY). Control mice were CY injected only; experimental mice were CY injected and then exposed to PEMF. Exposure to PEMF (24 hours/day) increased the rate of decline of white blood cells in peripheral blood. Spleen weight was statistically higher among control mice than among mice exposed to PEMF at day 6, 8 and 10 after CY injection. Spleen autoradiography proved to be higher among PEMF exposed mice than among controls at day 8 and 9 after CY injection. The grafting efficiency of the bone marrow obtained from control mice was higher than the grafting efficiency of the bone marrow recovered from mice exposed to PEMF. All these data indicate that the exposure to PEMF increases the cytotoxic effect of CY.

Cadossi, R.; Zucchini, P.; Emilia, G.; Torelli, G. (Univ. di Modena (Italy))

1990-02-26

57

Bisphosphonate treatment in the oim mouse model alters bone modeling during growth  

PubMed Central

Osetogenesis Imperfecta (OI) is a heritable disease, which results from an abnormal amount or structure of Type I collagen. Bisphosphonates, a class of synthetic antiresorptive drugs used in osteoporosis management, are also used to decrease fracture incidence and improve quality of life in children with OI. In this study we used the oim mouse to test the hypotheses that pamidronate treatment during active growth 1. produces larger, stronger, stiffer long bone diaphyses without altering bone material properties, and 2. negatively impacts longitudinal bone growth. Our results indicate that femoral cross-sectional moment of inertia in the distal metaphysis tended to increase with pamidronate treatment and that the treated bones are thicker and structurally stiffer, but shorter than their control-dose counterpar

Rao, S.H.; Evans, K.D.; Oberbauer, A.M.; Martin, R.B.

2009-01-01

58

DNA-Mediated gene transfection into primary cultures of adult mouse keratinocytes  

Microsoft Academic Search

Summary  An efficient and reproducible technique for the transfection of primary cultures of adult mouse keratinocytes has been developed.\\u000a The procedure involves culturing the primary adult mouse epidermal cells at 32° C in an enriched media until they reach 70\\u000a to 95% confluency, followed by transfection with exogenous DNA in a low potassium environment. Using chloramphenicol acetyl\\u000a transferase (CAT) transient gene

Natalie A. Betz; Ken J. Wolterman; John J. Reiners; Jill C. Pelling

1992-01-01

59

Altered plasma membrane dynamics of bone morphogenetic protein receptor type Ia in a low bone mass mouse model.  

PubMed

Bone morphogenetic proteins (BMPs) are growth factors that initiate differentiation of bone marrow stromal cells to osteoblasts and adipocytes, yet the mechanism that decides which lineage the cell will follow is unknown. BMP2 is linked to the development of osteoporosis and variants of BMP2 gene have been reported to increase the development of osteoporosis. Intracellular signaling is transduced by BMP receptors (BMPRs) of type I and type II that are serine/threonine kinase receptors. The BMP type I a receptor (BMPRIa) is linked to osteogenesis and bone mineral density (BMD). BMPRs are localized to caveolae enriched with Caveolin1 alpha/beta and Caveolin beta isoforms to facilitate signaling. BMP2 binding to caveolae was recently found to be crucial for the initiation of the Smad signaling pathway. Here we determined the role of BMP receptor localization within caveolae isoforms and aggregation of caveolae as well as BMPRIa in bone marrow stromal cells (BMSCs) on bone mineral density using the B6.C3H-6T as a model system. The B6.C3H-6T is a congenic mouse with decreased bone mineral density (BMD) with increased marrow adipocytes and decreased osteoprogenitor proliferation. C57BL/6J mice served as controls since only a segment of Chr6 from the C3H/HeJ mouse was backcrossed to a C57BL/6J background. Family of image correlation spectroscopy was used to analyze receptor cluster density and co-localization of BMPRIa and caveolae. It was previously shown that BMP2 stimulation results in an aggregation of caveolae and BMPRIa. Additionally, BMSCs isolated from the B6.C3H-6T mice showed a dispersion of caveolae domains compared to C57BL/6J. The aggregation of BMPRIa that is necessary for signaling to occur was inhibited in BMSCs isolated from B6.C3H-6T. Additionally, we analyzed the co-localization of BMPRIa with caveolin-1 isoforms. There was increased percentage of BMPRIa co-localization with caveolae compared to C57BL/6J. BMP2 stimulation had no effect on the colocalization of BMPRIa with caveolin-1. Disrupting caveolae initiated Smad signaling in the isolated BMSCs from B6.C3H-6T. These data suggest that in congenic 6T mice BMP receptors aggregation is inhibited causing an inhibition of signaling and reduced bone mass. PMID:22036911

Bragdon, Beth; D'Angelo, Alex; Gurski, Lauren; Bonor, Jeremy; Schultz, Kathryn L; Beamer, Wesley G; Rosen, Clifford J; Nohe, Anja

2011-10-22

60

Amelioration of cognitive ability in senescence-accelerated mouse prone 8 (SAMP8) by intra-bone marrow-bone marrow transplantation  

Microsoft Academic Search

Bone marrow cells (BMCs) can increase the number of activated microglias, which play a central role in the inflammatory response in Alzheimer's disease (AD). Senescence-accelerated mouse (SAM) prone 8 (SAMP8) are widely used in various experiments because of cognitive deficits observed with age. In the present study, 4-month-old SAMP8 were reconstituted with BMCs of C57BL\\/6 mice by intra-bone marrow-bone marrow

Ming Li; Muneo Inaba; Kequan Guo; Nader G. Abraham; Susumu Ikehara

2009-01-01

61

[Comparative investigation of mesenchymal stem cells isolated from the bone marrow and embryonic liver of mouse and rat].  

PubMed

We studied the properties of cells forming fibroblast colonies from the bone marrow and embryonic liver of mouse and rat. Bone marrow and embryonic liver cells formed colonies in vitro including fibroblasts as well as a considerable proportion of macrophages. The colonies formed from bone marrow and hepatic cells of rat differed from the murine ones by a higher proportion of fibroblasts. Most colonies derived from the bone marrow of both mouse and rat included a proportion of cells expressing acid phosphatase, and hence, capable of osteogenic differentiation; the colonies derived from the embryonic liver included low proportions of such cells. Cell layers derived from the colony-forming fibroblasts of both the bone marrow and embryonic liver of mouse maintained hematopoiesis in the peritoneal cavity of irradiated mice, which indicates that these precursor cells can form hematopoietic environment. PMID:15615444

Paniushina, O V; Bueverova, é I; Satdykova, G P; Starostin, V I; Domaratskaia, E I; Khrushchev, N G

62

Protective effects of indole-3-carbinol on cyclophosphamide-induced clastogenecity in mouse bone marrow cells  

Microsoft Academic Search

Indole-3-carbinol (I3C) is present in many cruciferous vegetables and is known to possess protective properties against chemically induced toxicity and carcinogenesis. In the present study, the antimutagenic potential of I3C has been evaluated using in vivo chromosomal aberration (CA) assay as a cytogenetic end point. Chromosomal analysis was carried out in mouse bone marrow cells following administration of I3C (5

Yogeshwer Shukla; Bhawna Srivastava; Annu Arora; L KS Chauhan

2004-01-01

63

Prevention of chromosomal aberration in mouse bone marrow by indole-3-carbinol  

Microsoft Academic Search

In this study, we report the protective effect of indole-3-carbinol (I3C), one of the glucobrassicin derivative isolated from cruciferous vegetables against cyclophosphamide induced chromosomal aberrations in mouse bone marrow cells. The three test doses namely 1000,500 and 250 mg\\/kg b.wt. of I3C provided protection when given 48 h prior to the single i.p. administration of cyclophosphamide (50 mg\\/kg). I3C alone

R. C. Agrawal; S. Kumar

1999-01-01

64

Characterization of Murine Dendritic Cell Line JAWS II and Primary Bone Marrow-Derived Dendritic Cells in Chlamydia muridarum Antigen Presentation and Induction of Protective Immunity  

Microsoft Academic Search

Dendritic cells (DCs) appear to orchestrate much of the immunobiology of Chlamydia infection, but most studies of Chlamydia-DC interaction have been limited by the availability and heterogeneity of primary bone marrow-derived DCs (BMDCs). We therefore evaluated the immunobiology of Chlamydia muridarum infection in an immortal DC line termed JAWS II derived from BMDCs of a C57BL\\/6 p53-knockout mouse. JAWS II

Xiaozhou Jiang; Caixia Shen; Jose Rey-Ladino; Hong Yu; Robert C. Brunham

2008-01-01

65

Use of a calcium sulfate-calcium phosphate synthetic bone graft composite in the surgical management of primary bone tumors.  

PubMed

Benign primary bone tumors are commonly treated with intralesional curettage with or without the use of surgical adjuvants. The reconstructive approach to the resulting contained bone defects is controversial, and clinical practice is varied. Synthetic bone substitutes may provide early mechanical support while minimizing the risks of disease transmission, nonunion, infection, and donor-site morbidity. Limited data exists regarding the use of calcium sulfate-calcium phosphate composite bone substitute for this purpose. The authors retrospectively reviewed the clinical outcomes of 24 patients with benign primary bone tumors who underwent intralesional curettage followed by reconstruction with a calcium sulfate-calcium phosphate composite bone substitute. Mean follow-up was 23 months. The most common diagnosis was giant cell tumor of bone. Six patients had upper-extremity tumors and 18 had lower-extremity tumors. Mean preoperative radiographic tumor volume was 41.0 cm(3). Mean volume of PRO-DENSE (Wright Medical Technology, Arlington, Tennessee) used in each patient was 15.6 cm(3). Mean time to full weight bearing for all patients was 7.3 weeks. Two patients sustained local tumor recurrences. No postoperative fractures occurred, and no complications occurred related to the use of the calcium sulfate-calcium phosphate composite. One case of deep infection occurred secondary to wound breakdown. The use of a calcium sulfate-calcium phosphate composite was associated with rapid biological integration and an early return to activities of daily living, with no composite-related complications. This technique is a viable option in the reconstruction of cavitary bone defects following intralesional curettage of primary benign bone tumors. PMID:23380017

Evaniew, Nathan; Tan, Victoria; Parasu, Naveen; Jurriaans, Erik; Finlay, Karen; Deheshi, Benjamin; Ghert, Michelle

2013-02-01

66

Abnormal Parathyroid Cell Proliferation Precedes Biochemical Abnormalities in a Mouse Model of Primary Hyperparathyroidism  

Microsoft Academic Search

The properties of neoplastic proliferation and hor- monal dysregulation are tightly linked in primary hyperparathyroidism (HPT). However, whether ab- normal parathyroid proliferation is the cause or result of a shift in calcium-sensitive parathyroid hormonal regulation has been controversial. We addressed this issue by analyzing the temporal se- quence of these fundamental abnormalities in a mouse model of primary HPT. These

Sanjay M. Mallya; James J. Gallagher; Yvette K. Wild; Olga Kifor; Jessica Costa-Guda; Kirsten Saucier; Edward M. Brown; Andrew Arnold

2005-01-01

67

Adipocytes regulate the bone marrow microenvironment in a mouse model of obesity.  

PubMed

Obesity is markedly associated with abnormal bone density indicating the importance of adipocytes in bone metabolism. However, the specific function of adipocytes remains unclear, with marked discrepancies in observations of previous studies. In the present study, the effect of adipocytes on osteoblasts/osteoclasts was analyzed. A mouse model of obesity was established and an in vitro co?culture system was utilized containing adipocyte and MC3T3/RAW 264.7 cells in a Transwell plate. Compared with control mice, obese mice exhibited low body weight and bone mineral density of the tibia and fat cells were observed to accumulate in bone marrow. MC3T3/RAW 264.7 cells were co?cultured with adipocytes and the mRNA and protein expression of alkaline phosphatase and osteocalcin was found to be decreased in MC3T3?E1 cells and mRNA and protein expression of tartrate-resistant acid phosphatase and cathepsin K was significantly increased in RAW 264.7 cells. In addition, the effect of adipocytes on the osteoprotegerin (OPG)/receptor activator of nuclear factor ?B ligand (RANKL)/RANK system indicated that the RANKL/OPG ratio secreted by osteoblasts increased and RANK expression by osteoclasts increased, leading to increased osteoclastogenesis. These results indicate that bone metabolism is impaired in obese mice leading to decreased osteoblastogenesis and marked increases in osteoclastogenesis and low bone mass. PMID:23835909

Xu, Fei; Du, Yu; Hang, Shilong; Chen, Anmin; Guo, Fengjin; Xu, Tao

2013-07-05

68

Accretion of Bone Quantity and Quality in the Developing Mouse Skeleton  

SciTech Connect

To meet the mechanical challenges during early development, the skeleton requires the rapid accretion of bone quality and bone quantity. Here, we describe early bone development in the mouse skeleton and test the hypothesis that specific compositional properties determine the stiffness of the tissue. Tibias of female BALB mice were harvested at eight time-points (n = 4 each) distributed between 1 and 40 days of age and subjected to morphometric ({mu}CT), chemical (Fourier transform infrared microscpectroscopy), and mechanical (nanoindentation) analyses. Tibias of 450-day-old mice served as fully mineralized control specimens. In this work, we found that bone mineral formation proceeded very rapidly in mice by 1 day of age, where the degree of mineralization, the tissue mineral density, and the mineral crystallinity reached 36%, 51%, and 87% of the adult values, respectively. However, even though significant mineralization had occurred, the elastic modulus of 1-day-old bone was only 14% of its adult value, indicating that the intrinsic stiffening of the bone lags considerably behind the initial mineral formation.

Miller,L.; Little, W.; Schirmer, A.; Sheik, F.; Busa, B.; Judex, S.

2007-01-01

69

Interferon regulatory factor-2 induces megakaryopoiesis in mouse bone marrow hematopoietic cells.  

PubMed

Megakaryopoiesis is associated with inflammatory reactions. To investigate the role of interferon regulatory factors (IRFs) in inflammation-associated megakaryopoiesis, mouse bone marrow hematopoietic stem cells (HSCs) were analyzed. IFN-gamma treatment induced IRF-2 expression as well as the expression of CD41 and IRF-1 in HSCs. An in vitro clonogenic assay showed that IRF-2- but not IRF-1-overexpressing cells increased the number of megakaryocytic colonies. IRF-2 transfection up-regulated CD41 promoter activity in hematopoietic cell lines. The number of CD41-positive bone marrow cells increased in mice injected with IRF-2-expressing bone marrow cells. These findings suggest that IRF-2 plays an important role in megakaryopoiesis in inflammatory states. PMID:19818776

Masumi, Atsuko; Hamaguchi, Isao; Kuramitsu, Madoka; Mizukami, Takuo; Takizawa, Kazuya; Momose, Haruka; Naito, Seishiro; Yamaguchi, Kazunari

2009-10-08

70

?-Actinin-3 deficiency is associated with reduced bone mass in human and mouse.  

PubMed

Bone mineral density (BMD) is a complex trait that is the single best predictor of the risk of osteoporotic fractures. Candidate gene and genome-wide association studies have identified genetic variations in approximately 30 genetic loci associated with BMD variation in humans. ?-Actinin-3 (ACTN3) is highly expressed in fast skeletal muscle fibres. There is a common null-polymorphism R577X in human ACTN3 that results in complete deficiency of the ?-actinin-3 protein in approximately 20% of Eurasians. Absence of ?-actinin-3 does not cause any disease phenotypes in muscle because of compensation by ?-actinin-2. However, ?-actinin-3 deficiency has been shown to be detrimental to athletic sprint/power performance. In this report we reveal additional functions for ?-actinin-3 in bone. ?-Actinin-3 but not ?-actinin-2 is expressed in osteoblasts. The Actn3(-/-) mouse displays significantly reduced bone mass, with reduced cortical bone volume (-14%) and trabecular number (-61%) seen by microCT. Dynamic histomorphometry indicated this was due to a reduction in bone formation. In a cohort of postmenopausal Australian women, ACTN3 577XX genotype was associated with lower BMD in an additive genetic model, with the R577X genotype contributing 1.1% of the variance in BMD. Microarray analysis of cultured osteoprogenitors from Actn3(-/-) mice showed alterations in expression of several genes regulating bone mass and osteoblast/osteoclast activity, including Enpp1, Opg and Wnt7b. Our studies suggest that ACTN3 likely contributes to the regulation of bone mass through alterations in bone turnover. Given the high frequency of R577X in the general population, the potential role of ACTN3 R577X as a factor influencing variations in BMD in elderly humans warrants further study. PMID:21784188

Yang, Nan; Schindeler, Aaron; McDonald, Michelle M; Seto, Jane T; Houweling, Peter J; Lek, Monkol; Hogarth, Marshall; Morse, Alyson R; Raftery, Joanna M; Balasuriya, Dominic; MacArthur, Daniel G; Berman, Yemima; Quinlan, Kate G R; Eisman, John A; Nguyen, Tuan V; Center, Jacqueline R; Prince, Richard L; Wilson, Scott G; Zhu, Kathy; Little, David G; North, Kathryn N

2011-07-19

71

Purification of Mouse Primary Myoblasts Based on ?7 Integrin Expression  

Microsoft Academic Search

Fundamental insights have come from the study of myogenesis. Primary myoblasts isolated directly from muscle tissue more closely approximate myogenesis than established cell lines. However, contamination of primary muscle cultures with nonmyogenic cells can complicate the results. To overcome this problem, we previously described a method for myoblast purification based on novel culture conditions (T. A. Rando and H. M.

William E. Blanco-Bose; Chung-Chen Yao; Randall H. Kramer; Helen M. Blau

2001-01-01

72

Determining the elastic modulus of mouse cortical bone using electronic speckle pattern interferometry (ESPI) and micro computed tomography: a new approach for characterizing small-bone material properties.  

PubMed

Mice phenotypes are invaluable for understanding bone formation and function, as well as bone disease. The elastic modulus is an important property of bones that can provide insights into bone quality. The determination of the elastic modulus of mouse cortical bone is complicated by the small dimensions of the bones. Whole bone bending tests are known to under estimate the elastic modulus compared to nanoindentation tests. The latter however provides information on extremely localized areas that do not necessarily correspond to the bulk elastic modulus under compression. This study presents a novel method for determining the bulk or effective elastic modulus of mouse cortical bone using the femur. We use Electronic Speckle Pattern Interferometry (ESPI), an optical method that enables the measurement of displacements on the bone surface, as it is compressed under water. This data is combined with geometric information obtained from micro-CT to calculate the elastic modulus. Roughly tubular cortical bone segments (2 mm) were cut from the diaphyses of femora of four week old C57BL/6 (B6) female mice and compressed axially using a mechanical tension-compression device. Displacements in the loading direction were mapped on the bone surface after loading the specimen. A linear regression of the displacement vs. axial-position enabled the calculation of the effective strain. Effective stress was calculated using force (N) data from the system's load cell and the mean cross-sectional area of the sample as determined by micro-CT. The effective elastic modulus (E) was calculated from the stress to strain ratio. The method was shown to be accurate and precise using a standard material machined to similar dimensions as those of the mouse femoral segments. Diaphyses of mouse femora were shown to have mean elastic moduli of 10.4+/-0.9 GPa for femora frozen for eight months, 8.6+/-1.4 GPa for femora frozen for two weeks and 8.9+/-1.1 GPa for the fresh femora. These values are much higher than those measured using three-point bending, and lower than values reported in the literature based on nanoindentation tests from mice bones of the same age. We show that this method can be used to accurately and precisely measure the effective elastic modulus of mouse cortical bone. PMID:19332167

Chattah, Netta Lev-Tov; Sharir, Amnon; Weiner, Steve; Shahar, Ron

2009-03-28

73

Primary vascular tumors of bone: a spectrum of entities?  

PubMed Central

Vascular tumors of bone are a heterogeneous group. Numerous terms have been introduced as well as different classification systems. None of the classification schemes have been accepted due to lack of consistent terminology, accepted histologic criteria, and limited correlation with clinical outcome. It is acknowledged that vascular tumors of bone originate from endothelial cells, resulting in variable expression of endothelial markers. None of these markers are useful to discriminate between benign and malignant lesions. Although radiologic appearance is not specific, radiologic multifocality should trigger to include a vascular neoplasm in the differential diagnosis. This review gives an overview of current literature by describing all different histologic subtypes in correspondence with clinical, radiologic and genetic data. We propose the classification of vascular tumors of bone according to the three-tiered World Health Organization classification scheme for soft tissue tumors dividing entities into a benign, intermediate and malignant category. Hemangioma is the most often and commonly recognized benign lesion. Epithelioid hemangioma has been better defined over the past few years. Based on its locally aggressive behavior and occurrence of lymph node metastases, classification within the intermediate category could be considered. Angiosarcoma is the only accepted term for high-grade malignant vascular tumor of bone and so far, epithelioid hemangioendo-thelioma is the only accepted low-grade malignant vascular tumor of bone. It is still unclear whether other low-grade malignant vascular tumors of bone (e.g. hemangioendothelioma) truly exist. Unfortunately, molecular / genetic studies of vascular tumors of bone which might support the proposed classification are very sparse.

Verbeke, Sofie LJ; Bovee, Judith VMG

2011-01-01

74

Phenotypic integration among trabecular and cortical bone traits establishes mechanical functionality of inbred mouse vertebrae.  

PubMed

Conventional approaches to identifying quantitative trait loci (QTLs) regulating bone mass and fragility are limited because they examine cortical and trabecular traits independently. Prior work examining long bones from young adult mice and humans indicated that skeletal traits are functionally related and that compensatory interactions among morphological and compositional traits are critical for establishing mechanical function. However, it is not known whether trait covariation (i.e., phenotypic integration) also is important for establishing mechanical function in more complex, corticocancellous structures. Covariation among trabecular, cortical, and compositional bone traits was examined in the context of mechanical functionality for L(4) vertebral bodies across a panel of 16-wk-old female AXB/BXA recombinant inbred (RI) mouse strains. The unique pattern of randomization of the A/J and C57BL/6J (B6) genome among the RI panel provides a powerful tool that can be used to measure the tendency for different traits to covary and to study the biology of complex traits. We tested the hypothesis that genetic variants affecting vertebral size and mass are buffered by changes in the relative amounts of cortical and trabecular bone and overall mineralization. Despite inheriting random sets of A/J and B6 genomes, the RI strains inherited nonrandom sets of cortical and trabecular bone traits. Path analysis, which is a multivariate analysis that shows how multiple traits covary simultaneously when confounding variables like body size are taken into consideration, showed that RI strains that tended to have smaller vertebrae relative to body size achieved mechanical functionality by increasing mineralization and the relative amounts of cortical and trabecular bone. The interdependence among corticocancellous traits in the vertebral body indicated that variation in trabecular bone traits among inbred mouse strains, which is often thought to arise from genetic factors, is also determined in part by the adaptive response to variation in traits describing the cortical shell. The covariation among corticocancellous traits has important implications for genetic analyses and for interpreting the response of bone to genetic and environmental perturbations. PMID:19063678

Tommasini, Steven M; Hu, Bin; Nadeau, Joseph H; Jepsen, Karl J

2009-04-01

75

Altered canonical hedgehog-gli signalling axis in pesticide-induced bone marrow aplasia mouse model.  

PubMed

The mechanistic interplay between pesticide exposure and development of marrow aplasia is not yet well established but there are indices that chronic pesticide exposure in some instances causes marrow aplasia like haematopoietic degenerative condition in human beings. Canonical Hedgehog (Hh) signalling has multiple roles in a wide range of developmental processes, including haematopoiesis. The present study was designed to explore the status of four important components of the canonical Hedgehog signalling cascade, the Sonic Hedgehog (Shh), Ptch1, Smo, and Gli1, in a mouse model of chronic pesticide-induced bone marrow aplasia. We used 5 % aqueous mixture of pesticides (chlorpyriphos, prophenophos, cypermethrin, alpha-methrin, and hexaconazole) for inhalation and dermal exposure of 6 hours per day and 5 days a week up to 90 days. Murine bone marrow aplasia related to chronic pesticide treatment was confirmed primarily by haemogram, bone marrow cellularity, short term bone marrow explant culture for cellular kinetics, bone marrow smear, and fl ow cytometric Lin-Sca-1+C-kit+ extracellular receptor expression pattern. Later, components of hedgehog signalling were analysed in the bone marrow of both control and pesticide-treated aplastic groups of animals. The results depicted pancytopenic feature of peripheral blood, developmental anomaly of neutrophils, depression of primitive stem and progenitor population along with Shh, Ptch1, Smo and Gli1 expression in aplasia group. This investigation suggests that pesticide-induced downregulation of two critically important proteins--Ptch1 and Gli1--inside the haematopoietic stem and progenitor cell population impairs haematopoietic homeostasis and regeneration mechanism in vivo concurrent with bone marrow aplasia. PMID:23152377

Chaklader, Malay; Das, Prosun; Pereira, Jacintha Archana; Chaudhuri, Samaresh; Law, Sujata

2012-09-01

76

Nasal Bone Shape Is under Complex Epistatic Genetic Control in Mouse Interspecific Recombinant Congenic Strains  

PubMed Central

Background Genetic determinism of cranial morphology in the mouse is still largely unknown, despite the localization of putative QTLs and the identification of genes associated with Mendelian skull malformations. To approach the dissection of this multigenic control, we have used a set of interspecific recombinant congenic strains (IRCS) produced between C57BL/6 and mice of the distant species Mus spretus (SEG/Pas). Each strain has inherited 1.3% of its genome from SEG/Pas under the form of few, small-sized, chromosomal segments. Results The shape of the nasal bone was studied using outline analysis combined with Fourier descriptors, and differential features were identified between IRCS BcG-66H and C57BL/6. An F2 cross between BcG-66H and C57BL/6 revealed that, out of the three SEG/Pas-derived chromosomal regions present in BcG-66H, two were involved. Segments on chromosomes 1 (?32 Mb) and 18 (?13 Mb) showed additive effect on nasal bone shape. The three chromosomal regions present in BcG-66H were isolated in congenic strains to study their individual effect. Epistatic interactions were assessed in bicongenic strains. Conclusions Our results show that, besides a strong individual effect, the QTL on chromosome 1 interacts with genes on chromosomes 13 and 18. This study demonstrates that nasal bone shape is under complex genetic control but can be efficiently dissected in the mouse using appropriate genetic tools and shape descriptors.

Burgio, Gaetan; Baylac, Michel; Heyer, Evelyne; Montagutelli, Xavier

2012-01-01

77

Composite treatment for primary long-bone hydatidosis.  

PubMed

Hydatid disease is a parasitic tapeworm infection caused by the Echinococcus species. Involvement of the long tubular bones is rare in hydatid bone disease. Patients are initially asymptomatic and usually present at a later stage of the disease when the bony lesions are extensive. Diagnosing bone hydatid disease is challenging, even in endemic regions, and a high index of suspicion is required because the radiologic findings often mimic other bone pathologies. Recurrence following treatment can occur after a long period of quiescence.This article describes a case of hydatid disease in a 62-year-old woman with extensive diaphyseal tibial involvement. She was treated with initial chemotherapy followed by extended curettage, polymethylmethacrylate cementation, and intramedullary fixation. Functional outcome was excellent, with no recurrence at 60-month follow-up. She was fully weight bearing with no pain or discomfort and had full hip, knee, and ankle range of motion.This case was important due to its rarity, the diagnostic challenge it presented, and the composite nature of the treatment used to avoid recurrence. Diaphyseal bone hydatidosis can be initially treated like a low-grade malignant tumor with curettage and high-speed burring, followed by filling the defect with polymethylmethacrylate cement. The composite treatment of chemotherapy with the surgical protocol described offers a reasonable chance of long-term disease suppression. Recurrent disease can be treated with repeat curettage and cementation. Wide excision with reconstruction of the resulting defect should only be considered for recalcitrant diaphyseal hydatid disease. PMID:23218646

Banerjee, Samik; Sabui, Kanchan Kumar; Mondal, Jayanta; Nath, Chinmoy; Pal, Dilip Kumar

2012-12-01

78

The cellular and molecular toxicity of lead in primary and clonal osteoblastic bone cells  

SciTech Connect

First, steady state kinetic models of lead metabolism and calcium homeostasis were developed in both primary and clonal osteoblastic bone cells. Secondly, the effect of lead on cellular calcium homeostasis was determined. Finally, the effect of lead on 1,25 (OH){sub 2}D{sub 3} induced production of osteocalcin, a protein synthesized and secreted by osteoblasts, was investigated. Lead metabolism in osteoblastic bone cells was characterized by three intracellular pools. The largest of these, S{sub 3}, included mitochondrial lead and accounted for 70 percent of total cell lead in primary osteoblastic bone cells and 85 percent of total lead in clonal osteoblastic bone cells. None of the kinetic pools were saturated at lead concentrations up to 100 {mu}M lead. Calcium homeostasis in osteoblastic bone cells was also described by a three compartment, intracellular kinetic model.

Long, G.J.

1989-01-01

79

Newly developed multiple myeloma in a patient with primary T-cell lymphoma of bone.  

PubMed

Primary non-Hodgkin's lymphoma of bone (PLB) is rare, and generally presents as a single extensive and destructive bone lesion. Histopathologically, most cases present as diffuse large B-cell lymphoma, and T-cell lymphoma is rare. By contrast, multiple myeloma is a disease defined as the neoplastic proliferation of a single clone of plasma cells producing a monoclonal immunoglobulin. We report a case of multiple myeloma that developed during treatment of PLB in a type of T-cell. A 48-yr-old man was diagnosed as T-cell PLB, stage IE, 18 months ago. The patient received the chemoradiotherapy and salvage chemotherapy for PLB. However, the lymphoma progressed with generalized bone pain, and laboratory findings showed bicytopenia and acute renal failure. On bone marrow biopsy, the patient was diagnosed as having multiple myeloma newly developed with primary T-cell lymphoma of bone. In spite of chemotherapy, the patient died of renal failure. PMID:18583898

Hwang, Jun-Eul; Cho, Sang-Hee; Kim, Ok-Ki; Shim, Hyun-Jeong; Lee, Se-Ryeon; Ahn, Jae-Sook; Yang, Duk-Hwan; Kim, Yeo-Kyeoung; Lee, Je-Jung; Kim, Hyeoung-Joon; Chung, Ik-Joo

2008-06-01

80

Sl/Sld mouse bone marrow stroma in vitro contains an active radiation-sensitive inhibitor of normal hemopoiesis  

SciTech Connect

Sl/Sld mice have a defective hemopoietic microenvironment. It has been assumed, based upon previous studies, that the primary abnormality in these mice is simply lack of a necessary supportive or inductive material within the hemopoietic stroma. We used in vitro long-term bone marrow cultures to characterize further the nature of the hemopoietic microenvironmental defect in Sl/Sld mice. Sl/Sld mouse bone marrow cells consistently produced less than 10% of the total hemopoietic cells and multipotent and unipotent hemopoietic progenitor cells produced in cultures of marrow from normal, congenic +/+ mice. If fresh Sl/Sld and +/+ marrow cells were mixed prior to establishing long-term marrow cultures, there was a direct correlation between number of Sl/Sld cells added and degree of inhibition of +/+ hemopoiesis. A pre-established, confluent Sl/Sld adherent stromal layer inhibited hemopoiesis by fresh +/+ marrow cells by nearly 70%, as compared with dishes with irradiated +/+ or no stroma. This inhibitory effect was abrogated by irradiation of the Sl/Sld stroma prior to addition of the fresh +/+ marrow cells. Similarly, unirradiated, but not 9 to 200 Gy irradiated Sl/Sld stroma inhibited proliferation of the factor-dependent FDC-P1 hemopoietic progenitor cell line. Thus, the Sl/Sld hemopoietic microenvironment actively inhibits hemopoiesis in vitro, and this inhibition can be at least partially eliminated by irradiation of the Sl/Sld stroma.

Zuckerman, K.S.; Prince, C.W.; Ribadeneira, M.

1986-12-01

81

Strain measurement of a mouse bone by 3D-electronic speckle pattern interferometry (3D-ESPI)  

NASA Astrophysics Data System (ADS)

Bone is a mechanosensitive tissue that adapts its mass, architecture and mechanical properties to mechanical loading. Appropriate mechanical loads provide an effective means to stimulate bone remodeling and prevent from bone loss. It is controversial whether in situ strain in bone is a critical determinant in enhancement of bone formation, and it is therefore important to evaluate load-driven strain in bone. Using electronic speckle pattern interferometry, we determined high-resolution three-dimensional strains on the mouse femur in response to two loading modalities: an axial loading modality (ALM) and a knee loading modality (KLM). We demonstrated that these two loading modalities induced a different pattern of strain distributions. ALM generated strain in the midshaft of cortical bone, while strains with KLM were concentrated on the distal epiphysis of the mouse femur. Since KLM is capable of enhancing bone formation in cortical bone distant from the knee, the current results indicate that in situ strain is not always necessary for load-driven bone formation.

Samala, Praveen R.; Su, Min; Liu, Sheng; Jiang, Hui H.; Yokota, Hiroki; Yang, Lianxiang

2005-08-01

82

Intra–Bone Marrow Cotransplantation of Donor Mesenchymal Stem Cells in Pig-to–NOD\\/SCID Mouse Bone Marrow Transplantation Facilitates Short-Term Xenogeneic Hematopoietic Engraftment  

Microsoft Academic Search

We directly injected porcine donor mesenchymal stem cells (MSC) into murine bone marrow (BM) cavities to examine the effects of intra-BM cotransplantation of MSC in pig-to–NOD\\/SCID mouse bone marrow transplantation (BMT) on xenogeneic engraftment. Porcine MSC prepared by aspiration of iliac BM of miniature swine were identified as CD90+CD29+CD45?CD31- and shown to differentiate into osteoblastocytes and adipocytes. A few weeks

H. Eguchi; Y. Kuroiwa; A. Matsui; M. Sada; N. Nagaya; S. Kawano

2008-01-01

83

Pathological features of bone marrow transplantation-related toxicity in a mouse.  

PubMed

In this case report, we present a mock-transduced bone marrow (BM) transplantation in a mouse, which was found moribund and autopsied to evaluate pathogenesis. Macroscopically, red discoloration of systemic organs was observed. Hematological values revealed a decrease in white blood cells, red blood cells, hematocrit, hemoglobin, and platelets, but an increase in reticulocytes. In BM cytology, hematopoietic cell lines were severely depleted. Histopathologically, hemorrhage in the cerebellar parenchyma, hemosiderin deposition and hemorrhage in the heart, necrosis and telangiectasia in liver, pulmonary parenchymal cysts, spermatogenic germ cells necrosis, atrophy and hemorrhage in testis, oligospermia and hemorrhage in the epididymis, and atrophy of BM, thymus and spleen were observed. In conclusion, autoimmune-like complications such as hematological value change, BM dysplasia and systemic hemorrhage appear to be the lethal cause of the mouse transplanted with mock-transduced BM. PMID:19934605

Kim, Yong Hoon; Ha, Chang Su; Lee, Hyun Sook; Lim, Sun Hwa; Moon, Kyoung Sik; Chung, Moon Koo; Son, Hwa Young

2009-12-01

84

Pathological features of bone marrow transplantation-related toxicity in a mouse  

PubMed Central

In this case report, we present a mock-transduced bone marrow (BM) transplantation in a mouse, which was found moribund and autopsied to evaluate pathogenesis. Macroscopically, red discoloration of systemic organs was observed. Hematological values revealed a decrease in white blood cells, red blood cells, hematocrit, hemoglobin, and platelets, but an increase in reticulocytes. In BM cytology, hematopoietic cell lines were severely depleted. Histopathologically, hemorrhage in the cerebellar parenchyma, hemosiderin deposition and hemorrhage in the heart, necrosis and telangiectasia in liver, pulmonary parenchymal cysts, spermatogenic germ cells necrosis, atrophy and hemorrhage in testis, oligospermia and hemorrhage in the epididymis, and atrophy of BM, thymus and spleen were observed. In conclusion, autoimmune-like complications such as hematological value change, BM dysplasia and systemic hemorrhage appear to be the lethal cause of the mouse transplanted with mock-transduced BM.

Kim, Yong-Hoon; Ha, Chang-Su; Lee, Hyun-Sook; Lim, Sun-Hwa; Moon, Kyoung-Sik; Chung, Moon-Koo

2009-01-01

85

Improved culture-based isolation of differentiating endothelial progenitor cells from mouse bone marrow mononuclear cells.  

PubMed

Numerous endothelial progenitor cell (EPC)-related investigations have been performed in mouse experiments. However, defined characteristics of mouse cultured EPC have not been examined. We focused on fast versus slow adherent cell population in bone marrow mononuclear cells (BMMNCs) in culture and examined their characteristics. After 24 h-culture of BMMNCs, attached (AT) cells and floating (FL) cells were further cultured in endothelial differentiation medium separately. Immunological and molecular analyses exhibited more endothelial-like and less monocyte/macrophage-like characteristics in FL cells compared with AT cells. FL cells formed thick/stable tube and hypoxia or shear stress overload further enhanced these endothelial-like features with increased angiogenic cytokine/growth factor mRNA expressions. Finally, FL cells exhibited therapeutic potential in a mouse myocardial infarction model showing the specific local recruitment to ischemic border zone and tissue preservation. These findings suggest that slow adherent (FL) but not fast attached (AT) BMMNCs in culture are EPC-rich population in mouse. PMID:22216102

Sekiguchi, Haruki; Ii, Masaaki; Jujo, Kentaro; Yokoyama, Ayumi; Hagiwara, Nobuhisa; Asahara, Takayuki

2011-12-28

86

Radioprotection by superoxide dismutase of macrophage progenitor cells from mouse bone marrow  

SciTech Connect

X-ray survival of cultured macrophage progenitor cells from mouse bone marrow was represented by a two component curve, both in the absence and presence of superoxide dismutase. Protection by the enzyme was limited to the radiosensitive fraction, for which a dose modifying factor of 2.8 +/- 0.7 was obtained. Catalase did not protect. Survival of the radiosensitive fraction, with and without exogenous superoxide dismutase, was temperature-dependent, whereas that of the radioresistant fraction was not. In the former case, the energy required for the enzyme-treated cells was approx. 13kJ/mol. 15 references, 2 figures, 1 table.

Petkau, A.; Chelack, W.S.

1984-03-30

87

Mouse basophils reside in extracellular matrix-enriched bone marrow niches which control their motility.  

PubMed

Basophils co-express Fc?RI? and CD49b, the ?-2 chain of integrin-type receptor VLA-2 (?2?1), which recognizes type-1 collagen as a major natural ligand. The physiological relevance of this integrin for interactions with extracellular bone marrow matrix remains unknown. Herein, we examined the expression of several receptors of this family by bone marrow-derived basophils sorted either ex-vivo or after culture with IL-3. Having established that both populations display CD49d, CD49e and CD49f (?-4, ?-5 and ?-6 integrins subunits, respectively), we addressed receptor functions by measuring migration, adhesion, proliferation and survival after interacting with matched natural ligands. Type I collagen, laminin and fibronectin promoted basophil migration/adhesion, the former being the most effective. None of these ligands affected basophil viability and expansion. Interactions between basophils and extracellular matrix are likely to play a role in situ, as supported by confocal 3D cell imaging of femoral bone marrow sections, which revealed basophils exclusively in type-1 collagen-enriched niches that contained likewise laminin and fibronectin. This is the first evidence for a structure/function relationship between basophils and extracellular matrix proteins inside the mouse bone marrow. PMID:24086246

Smaniotto, Salete; Schneider, Elke; Goudin, Nicolas; Bricard-Rignault, Rachel; Machavoine, François; Dardenne, Mireille; Dy, Michel; Savino, Wilson

2013-09-27

88

Mouse Basophils Reside in Extracellular Matrix-Enriched Bone Marrow Niches Which Control Their Motility  

PubMed Central

Basophils co-express Fc?RI? and CD49b, the ?-2 chain of integrin-type receptor VLA-2 (?2?1), which recognizes type-1 collagen as a major natural ligand. The physiological relevance of this integrin for interactions with extracellular bone marrow matrix remains unknown. Herein, we examined the expression of several receptors of this family by bone marrow-derived basophils sorted either ex-vivo or after culture with IL-3. Having established that both populations display CD49d, CD49e and CD49f (?-4, ?-5 and ?-6 integrins subunits, respectively), we addressed receptor functions by measuring migration, adhesion, proliferation and survival after interacting with matched natural ligands. Type I collagen, laminin and fibronectin promoted basophil migration/adhesion, the former being the most effective. None of these ligands affected basophil viability and expansion. Interactions between basophils and extracellular matrix are likely to play a role in situ, as supported by confocal 3D cell imaging of femoral bone marrow sections, which revealed basophils exclusively in type-1 collagen-enriched niches that contained likewise laminin and fibronectin. This is the first evidence for a structure/function relationship between basophils and extracellular matrix proteins inside the mouse bone marrow.

Smaniotto, Salete; Schneider, Elke; Goudin, Nicolas; Bricard-Rignault, Rachel; Machavoine, Francois; Dardenne, Mireille

2013-01-01

89

Reduction in cement-bone interface shear strength between primary and revision arthroplasty.  

PubMed

This study quantified changes in the cement-bone interface shear strength between primary and first- and second-revision arthroplasties as a function of mechanical interlock between the cement and bone. There were 128 segments obtained from four pairs of fresh human femora that were prepared sequentially as for primary and first and second revisions, taking care to maintain original canal morphology. Cement was pressurized into the cavity of the anatomic specimens, and the maximum interface shear strength between the cement plug and the bone was experimentally determined for each revision. First-revision interface shear strength was reduced to 20.6% of primary strength, and second revision strength to 6.8% of primary strength. PMID:3180573

Dohmae, Y; Bechtold, J E; Sherman, R E; Puno, R M; Gustilo, R B

1988-11-01

90

Sodium current properties of primary skeletal myocytes and cardiomyocytes derived from different mouse strains  

PubMed Central

The mouse has become the preferred animal for genetic manipulations. Because of the diverse genetic backgrounds of various mouse strains, these can manifest strikingly different characteristics. Here, we studied the functional properties of currents through voltage-gated sodium channels in primary cultures of skeletal myocytes and cardiomyocytes derived from the three commonly used mouse strains BL6, 129/Sv, and FVB, by using the whole-cell patch-clamp technique. We found strain-specific sodium current function in skeletal myocytes, which could partly be explained by differences in sodium channel isoform expression. In addition, we found significant effects of cell source (neonatal or adult animal-derived) and variation of the differentiation time period. In contrast to skeletal myocytes, sodium current function in cardiomyocytes was similar in all strains. Our findings are relevant for the design and proper interpretation of electrophysiological studies, which use excitable cells in primary culture as a model system.

Mille, M.; Koenig, X.; Zebedin, E.; Uhrin, P.; Cervenka, R.; Todt, H.; Hilber, K.

2010-01-01

91

Cell wounding activates phospholipase D in primary mouse keratinocytes.  

PubMed

Plasma membrane disruptions occur in mechanically active tissues such as the epidermis and can lead to cell death if the damage remains unrepaired. Repair occurs through fusion of vesicle patches to the damaged membrane region. The enzyme phospholipase D (PLD) is involved in membrane traffickiing; therefore, the role of PLD in membrane repair was investigated. Generation of membrane disruptions by lifting epidermal keratinocytes from the substratum induced PLD activation, whereas removal of cells from the substratum via trypsinization had no effect. Pretreatment with 1,25-dihydroxyvitamin D?, previously shown to increase PLD1 expression and activity, had no effect on, and a PLD2-selective (but not a PLD1-selective) inhibitor decreased, cell lifting-induced PLD activation, suggesting PLD2 as the isoform activated. PLD2 interacts functionally with the glycerol channel aquaporin-3 (AQP3) to produce phosphatidylglycerol (PG); however, wounding resulted in decreased PG production, suggesting a potential PG deficiency in wounded cells. Cell lifting-induced PLD activation was transient, consistent with a possible role in membrane repair, and PLD inhibitors inhibited membrane resealing upon laser injury. In an in vivo full-thickness mouse skin wound model, PG accelerated wound healing. These results suggest that PLD and the PLD2/AQP3 signaling module may be involved in membrane repair and wound healing. PMID:23288946

Arun, Senthil N; Xie, Ding; Howard, Amber C; Zhong, Quincy; Zhong, Xiaofeng; McNeil, Paul L; Bollag, Wendy B

2013-01-02

92

Mouse primary spermatocytes can complete two meiotic divisions within the oocyte cytoplasm  

Microsoft Academic Search

This study was undertaken to determine whether primary spermatocyte nuclei can complete meiosis after transfer into maturing or mature oocytes and can participate in normal embryogenesis. When injected into maturing mouse oocytes at prometaphase of the first meiotic division, spermatocyte chromosomes became arranged on a first meiotic metaphase (Met-I) spindle. Thus, oocytes contained two sets of Met-I chromosomes. When these

I. Sasagawa; S Kuretake; J J Eppig; R Yanagimachi

1998-01-01

93

Activation of protein kinase C isozymes in primary mouse myotubes by carbachol  

Microsoft Academic Search

The activation of muscle PKC isozymes following treatment with carbachol, an acetylcholine receptor agonist, has been investigated. Primary mouse myotubes were treated with carbachol, and protein extracts from the cytosol and membrane fractions of the myotubes were subjected to Western blot analyses. Carbachol treatment resulted in a rapid translocation of PKC-? to the membrane. This effect was dependent on both

Sunghee Kim; Tzvetanka Bondeva; Phillip G Nelson

2002-01-01

94

Vitamin D, hydroxyapatite, and calcium gluconate in treatment of cortical bone thinning in postmenopausal women with primary biliary  

Microsoft Academic Search

Women with primary bihiary cirrhosis malabsorb calcium, phosphate and vitamin D, and develop accelerated cortical bone thinning. We have assessed the value of parenteral vitamin D, oral hydroxyapatite (HA), and calcium gluconate (CG) in the treatment of cortical bone thinning in primary biliary cirrhosis. Sixty-four postmenopausal women with primary biliary cirrhosis were assigned randomly into three groups: one group receiving

Owen Epstein; Yasuhiro Kato; Robert Dick; Sheila Sherlock

95

Tenofovir treatment of primary osteoblasts alters gene expression profiles: implications for bone mineral density loss  

PubMed Central

There is strong clinical evidence that implicates tenofovir in the loss of bone mineral density during treatment of human immunodeficiency virus infection. In this study, we sought to test the hypothesis that tenofovir treatment of osteoblasts causes changes in the gene expression profile that would impact osteoblast function during bone formation. Primary osteoblasts were isolated and then treated with the tenofovir prodrug, tenofovir disoproxil fumarate (TDF). Total RNA from TDF-treated and untreated osteoblasts were extracted and used for microarray analysis to assess TDF-associated changes in the gene expression profile. Strikingly, the changes in gene expression profiles involved in cell signaling, cell cycle and amino acid metabolism, which would likely impact osteoblast function in bone formation. Our findings demonstrate for the first time that tenofovir treatment of primary osteoblasts results in gene expression changes that implicate loss of osteoblast function in tenofovir-associated bone mineral density loss.

Grigsby, Iwen F.; Pham, Lan; Mansky, Louis M.; Gopalakrishnan, Raj; Carlson, Ann E.; Mansky, Kim C.

2010-01-01

96

An RNA-seq protocol to identify mRNA expression changes in mouse diaphyseal bone: Applications in mice with bone property altering Lrp5 mutations.  

PubMed

Loss-of-function and certain missense mutations in the Wnt coreceptor low-density lipoprotein receptor-related protein 5 (LRP5) significantly decrease or increase bone mass, respectively. These human skeletal phenotypes have been recapitulated in mice harboring Lrp5 knockout and knock-in mutations. We hypothesized that measuring mRNA expression in diaphyseal bone from mice with Lrp5 wild-type (Lrp5(+/+) ), knockout (Lrp5(-/-) ), and high bone mass (HBM)-causing (Lrp5(p.A214V/+) ) knock-in alleles could identify genes and pathways that regulate or are regulated by LRP5 activity. We performed RNA-seq on pairs of tibial diaphyseal bones from four 16-week-old mice with each of the aforementioned genotypes. We then evaluated different methods for controlling for contaminating nonskeletal tissue (ie, blood, bone marrow, and skeletal muscle) in our data. These methods included predigestion of diaphyseal bone with collagenase and separate transcriptional profiling of blood, skeletal muscle, and bone marrow. We found that collagenase digestion reduced contamination, but also altered gene expression in the remaining cells. In contrast, in silico filtering of the diaphyseal bone RNA-seq data for highly expressed blood, skeletal muscle, and bone marrow transcripts significantly increased the correlation between RNA-seq data from an animal's right and left tibias and from animals with the same Lrp5 genotype. We conclude that reliable and reproducible RNA-seq data can be obtained from mouse diaphyseal bone and that lack of LRP5 has a more pronounced effect on gene expression than the HBM-causing LRP5 missense mutation. We identified 84 differentially expressed protein-coding transcripts between LRP5 "sufficient" (ie, Lrp5(+/+) and Lrp5(p.A214V/+) ) and "insufficient" (Lrp5(-/-) ) diaphyseal bone, and far fewer differentially expressed genes between Lrp5(p.A214V/+) and Lrp5(+/+) diaphyseal bone. © 2013 American Society for Bone and Mineral Research. PMID:23553928

Ayturk, Ugur M; Jacobsen, Christina M; Christodoulou, Danos C; Gorham, Joshua; Seidman, Jonathan G; Seidman, Christine E; Robling, Alexander G; Warman, Matthew L

2013-10-01

97

Capsaicin promotes the development of burst-forming units-erythroid (BFU-E) from mouse bone marrow cells  

Microsoft Academic Search

Capsaicin, the pungent component of chilli peppers, is known to induce mediators of hematopoiesis. We investigated the effect of capsaicin on hemato - poiesis in mouse progenitor cells. Treatment of mouse bone marrow cells with capsaicin induced the formation of colony of burst-forming units-erythroid (BFU-E). We also found that the number of ery - thropoietin receptor (EpoR)-positive cells was increased

Seong-Ae Lee; Young-Shin Ryu; Hyung-Im Choi; In-Seob Han

98

Multifocal primary bone lymphoma: durable complete remission after R-CHOP chemotherapy.  

PubMed

Primary bone lymphoma (PBL) is a type of non-Hodgkin's lymphoma predominantly affecting the skeletal system. PBL is an extremely rare cancer in adults affecting mainly the axial skeleton. The extent of bone involvement in these patients is variable. Most of the cases reported had single or a few skeletal lesions. We report a patient who had extensive multifocal lymphoma involving the axial skeleton and a very good and durable response to R-CHOP chemotherapy. PMID:23704456

Mohamed, Muhajir; Brain, Terry; Sharma, Sharad

2013-05-22

99

Analysis of human primary bone cells by fluorescence activated cell scanning: methodological problems and preliminary results  

Microsoft Academic Search

We describe the development of flowcytometrical methods to analyse human primary osteoblast-like cultures obtained from trabecular bone explants in comparison to the human osteosarcoma cell line HOS 58. Two antigens typical of osteoblasts were studied: bone alkaline phosphatase and collagen\\/procollagen I; the non-specific attachment protein fibronectin served as control. The morphology of all different antigens is shown by immunocytochemistry before

Heide Siggelkow; Dirk Hilmes; Katja Rebenstorff; Wiebke Kurre; Iris Engel; Michael Hüfner

1998-01-01

100

Fusobacterium nucleatum and Tannerella forsythia induce synergistic alveolar bone loss in a mouse periodontitis model.  

PubMed

Tannerella forsythia is strongly associated with chronic periodontitis, an inflammatory disease of the tooth-supporting tissues, leading to tooth loss. Fusobacterium nucleatum, an opportunistic pathogen, is thought to promote dental plaque formation by serving as a bridge bacterium between early- and late-colonizing species of the oral cavity. Previous studies have shown that F. nucleatum species synergize with T. forsythia during biofilm formation and pathogenesis. In the present study, we showed that coinfection of F. nucleatum and T. forsythia is more potent than infection with either species alone in inducing NF-?B activity and proinflammatory cytokine secretion in monocytic cells and primary murine macrophages. Moreover, in a murine model of periodontitis, mixed infection with the two species induces synergistic alveolar bone loss, characterized by bone loss which is greater than the additive alveolar bone losses induced by each species alone. Further, in comparison to the single-species infection, mixed infection caused significantly increased inflammatory cell infiltration in the gingivae and osteoclastic activity in the jaw bones. These data show that F. nucleatum subspecies and T. forsythia synergistically stimulate the host immune response and induce alveolar bone loss in a murine experimental periodontitis model. PMID:22547549

Settem, Rajendra P; El-Hassan, Ahmed Taher; Honma, Kiyonobu; Stafford, Graham P; Sharma, Ashu

2012-04-30

101

Impaired cholesterol esterification in primary brain cultures of the lysosomal cholesterol storage disorder (LCSD) mouse mutant  

SciTech Connect

Esterification of cholesterol was investigated in primary neuroglial cultures obtained from newborn lysosomal cholesterol storage disorder (LCSD) mouse mutants. An impairment in /sup 3/H-oleic acid incorporation into cholesteryl esters was demonstrated in cultures of homozygous LCSD brain. Primary cultures derived from other phenotypically normal pups of the carrier breeders esterified cholesterol at normal levels or at levels which were intermediary between normal and deficient indicating a phenotypic expression of the LCSD heterozygote genotype. These observations on LCSD mutant brain cells indicate that the defect in cholesterol esterification is closely related to the primary genetic defect and is expressed in neuroglial cells in culture.

Patel, S.C.; Suresh, S.; Weintroub, H.; Brady, R.O.; Pentchev, P.G.

1987-02-27

102

Effect of MPC11 myeloma and MPC11 + IL1 receptor antagonist treatment on mouse bone properties  

Microsoft Academic Search

This study examines the effects of an IL-6-producing murine multiple myeloma cell line on trabecular and cortical mouse bone, and evaluates the efficacy of interleukin-1 receptor antagonist (IL-1ra) in mitigating bone destruction. Six-week-old BALB\\/c mice were assigned to two groups: normal controls and myeloma animals (5 × 107 MPC-11 cells on day 0). Myeloma animals were further assigned to three

V. L Ferguson; S. J Simske; R. A Ayers; T. A Bateman; H. T Wang; A Bendele; B Rich; D Collins; J Scherrer; R Sennello; D. B Colagiovanni

2002-01-01

103

Antibiotic bone cements: their use in routine primary total joint arthroplasty is justified.  

PubMed

The use of antibiotic bone cement in total hip arthroplasty (THA) is the standard of care in countries with long-standing national registries, as data from the registries demonstrates an improvement in survivorship by reducing the incidence of both septic and aseptic failures. There is reluctance in North America to embrace antibiotic bone cement, although the percentage of use is increasing. Reasons cited for not using antibiotic cement include lack of efficacy, adverse effects on mechanical properties, increased costs, bacterial resistance, and systemic toxicity. Little to no compelling data in the literature support these claims, and significant evidence refutes them, specifically: registry data and randomized controlled trials demonstrate a clear reduction in deep joint infections with the use of antibiotic bone cement; antibiotic bone cement has lower incidence of both septic and aseptic loosening, indicating no negative effect on mechanical properties; antibiotic bone cement is cost-effective, given its proven ability to reduce revision rates and prevent poor patient outcomes; there is no evidence to support bacterial resistance, and antibiotic bone cement may reduce the incidence of resistance; and there are no reported cases of systemic toxicity from manufacturer-prepared antibiotic cement in primary THA or total knee arthroplasty. Based on the strong evidence supporting the benefits of antibiotic bone cement and the lack of evidence against its use, it is difficult to justify why antibiotic cement is not the standard of care for primary cemented THA in North America. PMID:19751021

Dunbar, Michael J

2009-09-01

104

Lentiviral-mediated integrin ?5 expression in human adult mesenchymal stromal cells promotes bone repair in mouse cranial and long-bone defects.  

PubMed

Abstract Adult human mesenchymal stromal cells (hMSCs) are an important source for tissue repair in regenerative medicine. Notably, targeted gene therapy in hMSCs to promote osteogenic differentiation may help in the development of novel therapeutic approaches for bone repair. We recently showed that ?5 integrin (ITGA5) promotes osteoblast differentiation in bone marrow-derived hMSCs. Here, we determined whether lentiviral (LV)-mediated expression of ITGA5 in hMSCs derived from the bone-marrow stroma of healthy individuals may promote bone repair in vivo in two relevant critical-size bone defects in the mouse. In a first series of experiments, control or LV-ITGA5-transduced hMSCs were seeded on collagen-based gelatin sponge and transplanted in a cranial critical-size defect (5?mm) in Nude-Foxn1nu mice. Microcomputed tomography and quantitative histological analyses after 8 weeks showed no or little de novo bone formation in defects implanted with collagen sponge alone or with hMSCs, respectively. In contrast, implantation of collagen sponge with LV-ITGA5-transduced hMSCs showed greater bone formation compared with control hMSCs. We also tested the bone-repair potential of LV-mediated ITGA5 expression in hMSCs in a critical-size long-bone defect (2?mm) in femur in Nude-Foxn1nu mice. Bone remnants were stabilized with external fixation, and control or LV-ITGA5-transduced hMSCs mixed with coral/hydroxyapatite particles were transplanted into the critical-size long-bone defect. Histological analysis after 8 weeks showed that LV-ITGA5-transduced hMSCs implanted with particles induced 85% bone regeneration and repair. These results demonstrate that repair of critical-size mouse cranial and long-bone defects can be induced using LV-mediated ITGA5 gene expression in hMSCs, which provides a novel gene therapy for bone regeneration. PMID:21958321

Srouji, Samer; Ben-David, Dror; Fromigué, Olivia; Vaudin, Pascal; Kuhn, Gisela; Müller, Ralph; Livne, Erella; Marie, Pierre J

2011-12-02

105

Inhibition of RANKL blocks skeletal tumor progression and improves survival in a mouse model of breast cancer bone metastasis.  

PubMed

Bone metastases cause severe skeletal morbidity including fractures and hypercalcemia. Tumor cells in bone induce activation of osteoclasts, which mediate bone resorption and release of growth factors from bone matrix, resulting in a "vicious cycle" of bone breakdown and tumor proliferation. Receptor activator of NF-kappaB ligand (RANKL) is an essential mediator of osteoclast formation, function, and survival, and is blocked by a soluble decoy receptor, osteoprotegerin (OPG). In human malignancies that metastasize to bone, dysregulation of the RANK/RANKL/OPG pathway can increase the RANKL:OPG ratio, a condition which favors excessive osteolysis. In a mouse model of bone metastasis, RANKL protein levels in MDA-MB-231 (MDA-231) tumor-bearing bones were significantly higher than tumor-free bones. The resulting tumor-induced osteoclastogenesis and osteolysis was dose-dependently inhibited by recombinant OPG-Fc treatment, supporting the essential role for RANKL in this process. Using bioluminescence imaging in a mouse model of metastasis, we monitored the anti-tumor efficacy of RANKL inhibition on MDA-231 human breast cancer cells in a temporal manner. Treatment with OPG-Fc in vivo inhibited growth of MDA-231 tumor cells in bony sites when given both as a preventative (dosed day 0) and as a therapeutic agent for established bone metastases (dosed day 7). One mechanism by which RANKL inhibition reduced tumor burden appears to be indirect through inhibition of the "vicious cycle" and involved an increase in tumor cell apoptosis, as measured by active caspase-3. Here, we demonstrate for the first time that OPG-Fc treatment of mice with established bone metastases resulted in an overall improvement in survival. PMID:18064531

Canon, Jude R; Roudier, Martine; Bryant, Rebecca; Morony, Sean; Stolina, Marina; Kostenuik, Paul J; Dougall, William C

2007-12-05

106

Mouse preproacrosin: cDNA sequence, primary structure and postmeiotic expression in spermatogenesis.  

PubMed

The primary structure of mouse preproacrosin was deduced by nucleotide sequencing of cDNA clones isolated from a mouse testis cDNA library. The largest cDNA, with 1373 bp, consists of a 11-bp 5'untranslated sequence, a 1254-bp open reading frame terminated by a TGA triplet and a 105-bp 3' untranslated end, including one potential polyadenylation signal. The NH2-terminus of the polypeptide contains a hydrophobic 15-amino acid signal peptide. This cleavable signal sequence is followed by 403 amino acids, representing the acrosin light and the heavy chain of 23 and 380 amino acid residues, respectively. The proteolytic active site segments His, Asp and Ser are part of the heavy chain, as well as a proline-rich COOH-terminus, which is not present in any other serine proteinase studied so far. Furthermore the postmeiotic expression of the preproacrosin gene during mouse spermatogenesis was studied. PMID:2111255

Klemm, U; Maier, W M; Tsaousidou, S; Adham, I M; Willison, K; Engel, W

1990-02-01

107

Mouse Genome-Wide Association and Systems Genetics Identify Asxl2 As a Regulator of Bone Mineral Density and Osteoclastogenesis  

Microsoft Academic Search

Significant advances have been made in the discovery of genes affecting bone mineral density (BMD); however, our understanding of its genetic basis remains incomplete. In the current study, genome-wide association (GWA) and co-expression network analysis were used in the recently described Hybrid Mouse Diversity Panel (HMDP) to identify and functionally characterize novel BMD genes. In the HMDP, a GWA of

Charles R. Farber; Brian J. Bennett; Luz Orozco; Wei Zou; Ana Lira; Emrah Kostem; Hyun Min Kang; Nicholas Furlotte; Ani Berberyan; Anatole Ghazalpour; Jaijam Suwanwela; Thomas A. Drake; Eleazar Eskin; Q. Tian Wang; Steven L. Teitelbaum; Aldons J. Lusis

2011-01-01

108

Histology of the bone marrow antibody response. An immunocytochemical study during the secondary response in mouse and rat.  

PubMed

The histology of the specific and non-specific antibody response in mouse and rat bone marrow was studied after subcutaneous priming and intravenous boosting with horseradish peroxidase (HRP). Cells producing specific antibody against HRP were found only occasionally in the bone marrow after subcutaneous priming. After the intravenous boost injection their number gradually increased. These anti-HRP forming cells were found as single cells, randomly dispersed throughout the bone marrow. Such a random distribution was also found for cytoplasmic (non-specific) immunoglobulin containing cells. At no time point after immunization could lymphoid aggregates or trapping of immune complexes be observed in the bone marrow of either species. On the basis of these observations it is concluded that the bone marrow forms a suitable microenvironment for immigrating antibody-forming cells but does not contribute actively to the induction of the immune response. PMID:6138895

Geldof, A A; Rimmelzwaan, G F; Langevoort, H L

1983-01-01

109

The effect of implant shape and bone preparation on primary stability  

PubMed Central

Purpose The purpose of this study was to evaluate the effects of implant shape and bone preparation on the primary stability of the implants using resonance frequency analysis. Methods Sixty bovine rib blocks were used for soft and hard bone models. Each rib block received two types of dental implant fixtures; a straight-screw type and tapered-screw type. Final drilling was done at three different depths for each implant type; 1 mm under-preparation, standard preparation, and 1 mm over-preparation. Immediately after fixture insertion, the implant stability quotient (ISQ) was measured for each implant. Results Regardless of the bone type, the ISQ values of the straight-screw type and tapered-screw type implants were not significantly different (P > 0.05). Depth of bone preparation had no significant effect on the ISQ value of straight-screw type implants (P > 0.05). For the tapered-screw type implants, under-preparation significantly increased the ISQ value (P < 0.05), whereas overpreparation significantly decreased the ISQ value (P < 0.05). Conclusions Within the limitations of this study, it is concluded that bone density seemed to have a prevailing effect over implant shape on primary stability. The primary stability of the tapered-screw type implants might be enhanced by delicate surgical techniques.

Moon, Sang-Hyun; Lee, Jae-Kwan; Chang, Beom-Seok; Lee, Min-Ku

2010-01-01

110

Role of aquaporin 9 in cellular accumulation of arsenic and its cytotoxicity in primary mouse hepatocytes  

SciTech Connect

Aquaporin (AQP) 9 is a member of the aquaglyceroporin subfamily of AQPs in the transfer of water and small solutes such as glycerol and arsenite. It is well recognized that arsenic toxicity is associated with intracellular accumulation of this metalloid. In the present study, we examined the contribution of AQP9 to the uptake of inorganic arsenite, thereby increasing arsenic-induced cytotoxicity in primary mouse hepatocytes. Pretreatment with sorbitol as a competitive inhibitor of AQP9 and siRNA-mediated knockdown of AQP9 resulted in a significant decrease of arsenite uptake in the cell and its cytotoxicity. Furthermore, overexpression of AQP9 in HEK293 cells led to the enhancement of intracellular arsenic concentration, resulting in enhanced cytotoxicity after arsenite exposure. These results suggest that AQP9 is a channel to define arsenite sensitivity in primary mouse hepatocytes.

Shinkai, Yasuhiro [Organization for Frontier Research in Preventive Pharmaceutical Sciences, Hokuriku University, Ho-3 Kanagawa-machi, Kanazawa, Ishikawa 920-1181 (Japan); Sumi, Daigo; Toyama, Takashi [Department of Environmental Medicine, Doctoral Programs in Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575 (Japan); Kaji, Toshiyuki [Organization for Frontier Research in Preventive Pharmaceutical Sciences, Hokuriku University, Ho-3 Kanagawa-machi, Kanazawa, Ishikawa 920-1181 (Japan); Kumagai, Yoshito [Department of Environmental Medicine, Doctoral Programs in Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575 (Japan)], E-mail: yk-em-tu@md.tsukuba.ac.jp

2009-06-01

111

Role of protein kinase C in growth stimulation of primary mouse colonic epithelial cells  

Microsoft Academic Search

Summary  The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (160 nM) and the secondary bile acid, deoxycholic acid (50 µM) stimulated DNA synthesis in quiescent primary epithelial cells from the normal mouse colon as measured by autoradiographic\\u000a analysis of [3H]thymidine incorporation. The purpose of the present study was to investigate the involvement of protein kinase C in the\\u000a proliferative response of the normal colonic cells.

Christina Branting; Rune Toftgård; Inger Porsch Hällström; Joseph Rafter

1995-01-01

112

HMG-CoA reductase inhibitors induce apoptosis in mouse proximal tubular cells in primary culture  

Microsoft Academic Search

HMG-CoA reductase inhibitors induce apoptosis in mouse proximal tubular cells in primary culture. Renal cyst formation in polycystic diseases or after nephron reduction is attributed to enhanced tubular cell proliferation with unbalanced cell death. The induction of tubular cell death could be effective to reduce renal cyst formation. In this study, we examined the effects of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA)

Osamu Iimura; François Vrtovsnik; Fabiola Terzi; Gérard Friedlander

1997-01-01

113

Mechanism of suppression of nitric oxide synthase expression by interleukin-4 in primary mouse macrophages  

Microsoft Academic Search

Nitric oxide (NO) contributes to the anti- tumor, antimicrobial, and immunosuppressive activity of macrophages. An inducible form of NO synthase (iNOS) is responsible for high output generation of nitric oxide by macrophages after stimulation with cytokines and\\/or lipopolysaccharide (LPS). In the present study, we demonstrate that interleukin 4 (IL-4) suppressed produc- tion of NO by primary mouse peritoneal macrophages cx-

Christian Bogdan; Yoram Vodovotz; John Paik; Qiao-wen Xie; Carl Nathan

1994-01-01

114

Primary bone marrow T-cell/histiocyte-rich large B-cell lymphoma: a diagnostic challenge.  

PubMed

We report a primary bone marrow diffuse large B-cell lymphoma in a 52-year-old post-menopausal woman that evaded definitive diagnosis initially due to deceptive clinical features, non-contributory radiological findings, and an extensive reactive lymphoid infiltrate masking the relatively few neoplastic B-cells on bone marrow biopsy. The correct diagnosis was apparent only after a repeat bone marrow procedure and review of the previous histology and immunohistochemistry. The case illustrates the pitfall of assuming mixed B- and T-cell infiltrates in the bone marrow to be invariably benign. A high index of suspicion regardless of the clinical and/or radiological absence of lymphadenopathy or organomegaly and critical examination of each individual cell population in immunohistochemically stained material are essential for correct identification of this rare entity. PMID:23562349

Sharma, Prashant; Ahluwalia, Jasmina; Sachdeva, Man Updesh Singh; Varma, Neelam; Malhotra, Pankaj; Varma, Subhash

2013-03-01

115

Primary bone neoplasms in beagle dogs exposed by inhalation to aerosols of plutonium-238 dioxide  

SciTech Connect

Primary bone neoplasms developed in beagle dogs briefly exposed by inhalation to aerosols of /sup 238/PuO/sub 2/. /sup 238/PuO/sub 2/ was initially deposited in the respiratory tract where it was retained with a half time greater than 100 days. A portion of the /sup 238/Pu was solubilized and translocated to the liver and skeleton. Five years after exposure, 46 osteosarcomas developed in 35 of 144 exposed dogs. The cumulative absorbed radiation doses to skeleton for these dogs ranged from 210 to 830 rad. Of the 46 bone tumors, 22 originated in the vertebrae, 12 in the humeri, 6 in the pelves, and 6 in miscellaneous long and flat bones. Most of the tumors were well-differentiated sarcomas. Only 10 of the tumors metastasized; the lung was the organ most often invaded. Bone tumors were associated with lesions of radiation osteodysplasia. The number of bone tumors found in this study indicated that inhaled /sup 238/PuO/sub 2/ was an effective skeletal carcinogen. The rate of solubilization in the lung and translocation to bone may be a factor in the radiation dose pattern and type and location of bone tumors that developed after inhalation of /sup 238/PuO/sub 2/.

Hahn, F.F.; Mewhinneyk, J.A.; Merickel, B.S.; Guilmette, R.A.; Boecker, B.B.; McClellan, R.D.

1981-10-01

116

Primary bone neoplasms in beagle dogs exposed by inhalation to aerosols of plutonium-238 dioxide  

SciTech Connect

Primary bone neoplasms developed in beagle dogs briefly exposed by inhalation to aerosols of 238PuO2. 238PuO2 was initially deposited in the respiratory tract where it was retained with a half time greater than 100 days. A portion of the 238Pu was solubilized and translocated to the liver and skeleton. Five years after exposure, 46 osteosarcomas developed in 35 of 144 exposed dogs. The cumulative absorbed radiation doses to skeleton for these dogs ranged from 210 to 830 rad. Of the 46 bone tumors, 22 originated in the vertebrae, 12 in the humeri, 6 in the pelves, and 6 in miscellaneous long and flat bones. Most of the tumors were well-differentiated sarcomas. Only 10 of the tumors metastasized; the lung was the organ most often invaded. Bone tumors were associated with lesions of radiation osteodysplasia. The number of bone tumors found in this study indicated that inhaled 238PuO2 was an effective skeletal carcinogen. The rate of solubilization in the lung and translocation to bone may be a factor in the radiation dose pattern and type and location of bone tumors that developed after inhalation of 238PuO2.

Hahn, F.F.; Mewhinney, J.A.; Merickel, B.S.; Guilmette, R.A.; Boecker, B.B.; McClellan, R.O.

1981-10-01

117

Malignant primary neoplasms of the ear and temporal bone studied by high-resolution computed tomography  

SciTech Connect

Ten patients with malignant primary neoplasms of the ear and temporal bone were examined with high-resolution computed tomography (CT) and the results correlated with the operative findings. CT was found to be highly accurate in establishing the presence and extent of tumor and is recommended as the procedure of choice for preoperative treatment planning.

Bird, C.R.; Hasso, A.N.; Stewart, C.E.; Hinshaw, D.B. Jr.; Thompson, J.R.

1983-10-01

118

Bone conducted vibration selectively activates irregular primary otolithic vestibular neurons in the guinea pig  

Microsoft Academic Search

The main objective of this study was to determine whether bone-conducted vibration (BCV) is equally effective in activating both semicircular canal and otolith afferents in the guinea pig or whether there is preferential activation of one of these classes of vestibular afferents. To answer this question a large number (346) of single primary vestibular neurons were recorded extracellularly in anesthetized

Ian S. Curthoys; Juno Kim; Samara K. McPhedran; Aaron J. Camp

2006-01-01

119

Cancer Cell Expression of Autotaxin Controls Bone Metastasis Formation in Mouse through Lysophosphatidic Acid-Dependent Activation of Osteoclasts  

PubMed Central

Background Bone metastases are highly frequent complications of breast cancers. Current bone metastasis treatments using powerful anti-resorbtive agents are only palliative indicating that factors independent of bone resorption control bone metastasis progression. Autotaxin (ATX/NPP2) is a secreted protein with both oncogenic and pro-metastatic properties. Through its lysosphospholipase D (lysoPLD) activity, ATX controls the level of lysophosphatidic acid (LPA) in the blood. Platelet-derived LPA promotes the progression of osteolytic bone metastases of breast cancer cells. We asked whether ATX was involved in the bone metastasis process. We characterized the role of ATX in osteolytic bone metastasis formation by using genetically modified breast cancer cells exploited on different osteolytic bone metastasis mouse models. Methodology/Principal Findings Intravenous injection of human breast cancer MDA-B02 cells with forced expression of ATX (MDA-B02/ATX) to inmmunodeficiency BALB/C nude mice enhanced osteolytic bone metastasis formation, as judged by increased bone loss, tumor burden, and a higher number of active osteoclasts at the metastatic site. Mouse breast cancer 4T1 cells induced the formation of osteolytic bone metastases after intracardiac injection in immunocompetent BALB/C mice. These cells expressed active ATX and silencing ATX expression inhibited the extent of osteolytic bone lesions and decreased the number of active osteoclasts at the bone metastatic site. In vitro, osteoclast differentiation was enhanced in presence of MDA-B02/ATX cell conditioned media or recombinant autotaxin that was blocked by the autotaxin inhibitor vpc8a202. In vitro, addition of LPA to active charcoal-treated serum restored the capacity of the serum to support RANK-L/MCSF-induced osteoclastogenesis. Conclusion/Significance Expression of autotaxin by cancer cells controls osteolytic bone metastasis formation. This work demonstrates a new role for LPA as a factor that stimulates directly cancer growth and metastasis, and osteoclast differentiation. Therefore, targeting the autotaxin/LPA track emerges as a potential new therapeutic approach to improve the outcome of patients with bone metastases.

David, Marion; Wannecq, Estelle; Descotes, Francoise; Jansen, Silvia; Deux, Blandine; Ribeiro, Johnny; Serre, Claire-Marie; Gres, Sandra; Bendriss-Vermare, Nathalie; Bollen, Mathieu; Saez, Simone; Aoki, Junken; Saulnier-Blache, Jean-Sebastien; Clezardin, Philippe; Peyruchaud, Olivier

2010-01-01

120

The impact of pathological fractures on therapy outcome in patients with primary malignant bone tumours.  

PubMed

The primary objective of this study was to investigate the implications of pathological fractures on therapy outcome in patients with primary malignant bone tumours and to determine whether limb salvage can be safely performed. A retrospective analysis of 447 patients with primary malignant bone tumours, treated between 1985 and 2005, was performed. Multivariate Cox regression analysis was used to investigate the influence of pathological fractures and further independent variables on survival rate. In 52 of the 447 patients, the primary malignant bone tumour was complicated by a pathological fracture. These fractures were more common in malignant fibrous histiocytoma (MFH) of the bone and in the tumour stages IIa/b and III. Ablative surgery was performed in ten patients and limb salvage surgery in 42 patients. The mortality risk for patients with pathological fractures was significantly increased by a factor of 1.82 (p = 0.015), and overall duration of survival was significantly lower in the fracture group, with a median of 6.2 years (p < 0.00001). In univariate and multivariate analysis, fracture, higher tumour stages and resection margins remained a significant predictor of worse survival. Overall survival, rate of local recurrence and distant metastases were not affected by the type of surgical treatment selected; there was no difference between the patients who underwent limb salvage and those who underwent an amputation. Pathological fracture in patients with primary malignant bone tumours is a predictor of worse survival and significantly increases mortality risk. Reconstructive surgery did not influence the survival rate, showing that limb salvage therapy is safe when adequate resection margins are achieved. PMID:20012861

Moradi, Babak; Zahlten-Hinguranage, Anita; Lehner, Burkhard; Zeifang, Felix

2009-12-13

121

Low Oxygen Tension Maintains Multipotency, Whereas Normoxia Increases Differentiation of Mouse Bone Marrow Stromal Cells  

PubMed Central

Optimization of mesenchymal stem cells (MSC) culture conditions is of great importance for their more successful application in regenerative medicine. O2 regulates various aspects of cellular biology and, in vivo, MSC are exposed to different O2 concentrations spanning from very low tension in the bone marrow niche, to higher amounts in wounds. In our present work, we isolated mouse bone marrow stromal cells (BMSC) and showed that they contained a population meeting requirements for MSC definition. In order to establish the effect of low O2 on cellular properties, we examined BSMC cultured under hypoxic (3% O2) conditions. Our results demonstrate that 3% O2 augmented proliferation of BMSC, as well as the formation of colonies in the colony-forming unit assay (CFU-A), the percentage of quiescent cells, and the expression of stemness markers Rex-1 and Oct-4, thereby suggesting an increase in the stemness of culture when exposed to hypoxia. In contrast, intrinsic differentiation processes were inhibited by 3% O2. Overall yield of differentiation was dependent on the adjustment of O2 tension to the specific stage of BMSC culture. Thus, we established a strategy for efficient BMSC in vitro differentiation using an initial phase of cell propagation at 3% O2, followed by differentiation stage at 21% O2. We also demonstrated that 3% O2 affected BMSC differentiation in p53 and reactive oxygen species (ROS) independent pathways. Our findings can significantly contribute to the obtaining of high-quality MSC for effective cell therapy.

Berniakovich, Ina; Giorgio, Marco

2013-01-01

122

Raman spectroscopy detects deterioration in biomechanical properties of bone in a glucocorticoid-treated mouse model of rheumatoid arthritis  

PubMed Central

Although glucocorticoids are frequently prescribed for the symptomatic management of inflammatory disorders such as rheumatoid arthritis, extended glucocorticoid exposure is the leading cause of physician-induced osteoporosis and leaves patients at a high risk of fracture. To study the biochemical effects of glucocorticoid exposure and how they might affect biomechanical properties of the bone, Raman spectra were acquired from ex vivo tibiae of glucocorticoid- and placebo-treated wild-type mice and a transgenic mouse model of rheumatoid arthritis. Statistically significant spectral differences were observed due to both treatment regimen and mouse genotype. These differences are attributed to changes in the overall bone mineral composition, as well as the degree of phosphate mineralization in tibial cortical bone. In addition, partial least squares regression was used to generate a Raman-based prediction of each tibia's biomechanical strength as quantified by a torsion test. The Raman-based predictions were as accurate as those produced by microcomputed tomography derived parameters, and more accurate than the clinically-used parameter of bone mineral density. These results suggest that Raman spectroscopy could be a valuable tool for monitoring bone biochemistry in studies of bone diseases such as osteoporosis, including tests of drugs being developed to combat these diseases.

Maher, Jason R.; Takahata, Masahiko; Awad, Hani A.; Berger, Andrew J.

2011-01-01

123

Raman spectroscopy detects deterioration in biomechanical properties of bone in a glucocorticoid-treated mouse model of rheumatoid arthritis  

NASA Astrophysics Data System (ADS)

Although glucocorticoids are frequently prescribed for the symptomatic management of inflammatory disorders such as rheumatoid arthritis, extended glucocorticoid exposure is the leading cause of physician-induced osteoporosis and leaves patients at a high risk of fracture. To study the biochemical effects of glucocorticoid exposure and how they might affect biomechanical properties of the bone, Raman spectra were acquired from ex vivo tibiae of glucocorticoid- and placebo-treated wild-type mice and a transgenic mouse model of rheumatoid arthritis. Statistically significant spectral differences were observed due to both treatment regimen and mouse genotype. These differences are attributed to changes in the overall bone mineral composition, as well as the degree of phosphate mineralization in tibial cortical bone. In addition, partial least squares regression was used to generate a Raman-based prediction of each tibia's biomechanical strength as quantified by a torsion test. The Raman-based predictions were as accurate as those produced by microcomputed tomography derived parameters, and more accurate than the clinically-used parameter of bone mineral density. These results suggest that Raman spectroscopy could be a valuable tool for monitoring bone biochemistry in studies of bone diseases such as osteoporosis, including tests of drugs being developed to combat these diseases.

Maher, Jason R.; Takahata, Masahiko; Awad, Hani A.; Berger, Andrew J.

2011-08-01

124

Recombinant Entactin Promotes Mouse Primary Trophoblast Cell Adhesion and Migration Through the Arg-Gly-Asp (RGD) Recognition Sequence  

Microsoft Academic Search

In vitro culture of mouse blastocysts during the period coinciding with implantation has revealed that primary trophoblast cells can adhere and migrate in serum-free medium when provided with certain ex- tracellular matrix components, including fibronectin and laminin. Tightly associated with laminin is the glycoprotein, entactin, that may play an important role in basement membrane assembly and cell attachment. Mouse blastocysts

Frank D. Yelian; Nancy A. Edgeworth; Li-Jin Dong; Albert E. Chung; D. Randall Armant

125

Orientation Selectivity of Synaptic Input to Neurons in Mouse and Cat Primary Visual Cortex  

PubMed Central

Primary visual cortex (V1) is the site at which orientation selectivity emerges in mammals: visual thalamus afferents to V1 respond equally to all stimulus orientations whereas their target V1 neurons respond selectively to stimulus orientation. The emergence of orientation selectivity in V1 has long served as a model for investigating cortical computation. Recent evidence for orientation selectivity in mouse V1 opens cortical computation to dissection by genetic and imaging tools, but also raises two essential questions: 1) how does orientation selectivity in mouse V1 neurons compare with that in previously described species? 2) what is the synaptic basis for orientation selectivity in mouse V1? A comparison of orientation selectivity in mouse and in cat, where such measures have traditionally been made, reveals that orientation selectivity in mouse V1 is weaker than in cat V1, but that spike threshold plays a similar role in narrowing selectivity between membrane potential and spike rate. To uncover the synaptic basis for orientation selectivity, we made whole-cell recordings in vivo from mouse V1 neurons, comparing neuronal input selectivity - based on membrane potential, synaptic excitation, and synaptic inhibition - to output selectivity based on spiking. We found that a neuron's excitatory and inhibitory inputs are selective for the same stimulus orientations as is its membrane potential response, and that inhibitory selectivity is not broader than excitatory selectivity. Inhibition has different dynamics than excitation, adapting more rapidly. In neurons with temporally modulated responses, the timing of excitation and inhibition was different in mice and cats.

Tan (???), Andrew Y. Y.; Brown, Brandon D.; Scholl, Benjamin; Mohanty, Deepankar; Priebe, Nicholas J.

2011-01-01

126

Tooth-bone morphogenesis during postnatal stages of mouse first molar development  

PubMed Central

The first mouse molar (M1) is the most common model for odontogenesis, with research particularly focused on prenatal development. However, the functional dentition forms postnatally, when the histogenesis and morphogenesis of the tooth is completed, the roots form and the tooth physically anchors into the jaw. In this work, M1 was studied from birth to eruption, assessing morphogenesis, proliferation and apoptosis, and correlating these with remodeling of the surrounding bony tissue. The M1 completed crown formation between postnatal (P) days 0–2, and the development of the tooth root was initiated at P4. From P2 until P12, cell proliferation in the dental epithelium reduced and shifted downward to the apical region of the forming root. In contrast, proliferation was maintained or increased in the mesenchymal cells of the dental follicle. At later stages, before tooth eruption (P20), cell proliferation suddenly ceased. This withdrawal from the cell cycle correlated with tooth mineralization and mesenchymal differentiation. Apoptosis was observed during all stages of M1 postnatal morphogenesis, playing a role in the removal of cells such as osteoblasts in the mandibular region and working together with osteoclasts to remodel the bone around the developing tooth. At more advanced developmental stages, apoptotic cells and bodies accumulated in the cell layers above the tooth cusps, in the path of eruption. Three-dimensional reconstruction of the developing postnatal tooth and bone indicates that the alveolar crypts form by resorption underneath the primordia, whereas the ridges form by active bone growth between the teeth and roots to form a functional complex.

Lungova, Vlasta; Radlanski, Ralf J; Tucker, Abigail S; Renz, Herbert; Misek, Ivan; Matalova, Eva

2011-01-01

127

Clinical utility of bone SPECT scintigraphy in renal metastases from primary osteosarcoma.  

PubMed

Renal metastases from primary osteosarcomas are rather uncommon and rarely diagnosed early because the patients are asymptomatic and frequently die from other metastatic involvement before renal symptoms develop. The authors present a patient with two clinically silent renal metastases from primary osteosarcoma of the right femur 2 years after surgery of the primary lesion that was first detected on radionuclide bone imaging. Subsequently, a CT scan and a CT guided needle biopsy were performed for confirmation. The patient underwent a left nephrectomy and two separate lesions were proven to be metastatic osteosarcoma. The clinical importance of the nuclear bone scan in the initial management, as well as in the subsequent follow-up of patients after surgery, cannot be overemphasized. PMID:7874811

Balingit, A G; Rudd, S; Williams, S

1994-12-01

128

Influence of the surgical technique and surface roughness on the primary stability of an implant in artificial bone with a density equivalent to maxillary bone: a laboratory study  

Microsoft Academic Search

OBJECTIVE: The aim of this biomechanical study was to assess the effect of surgical technique and surface roughness on primary implant stability in low-density bone. MATERIAL AND METHODS: Eighty screw-shaped (Biocomp) implants with machined or etched surface topography were inserted into a low-density bone equivalent. Solid rigid polyurethane blocks (Sawbones) with two different bone densities (group A=0.32 g\\/cm(3); group B=0.48

Afsheen Tabassum; Gert J. Meijer; Johannes G. C. Wolke; John A. Jansen

2009-01-01

129

Parathyroid hormone related peptide and receptor expression in paired primary prostate cancer and bone metastases  

PubMed Central

Parathyroid hormone-related peptide is a regulatory protein implicated in the pathogenesis of bone metastases, particularly in breast carcinoma. Parathyroid hormone-related peptide is widely expressed in primary prostate cancers but there are few reports of its expression in prostatic metastases. The aim of this study was to examine the expression of parathyroid hormone-related peptide and its receptor in matched primary and in bone metastatic tissue from patients with untreated adenocarcinoma of the prostate. Eight-millimetre trephine iliac crest bone biopsies containing metastatic prostate cancer were obtained from 14 patients from whom matched primary tumour tissue was also available. Histological grading was performed by an independent pathologist. The cellular location of mRNA for parathyroid hormone-related peptide and parathyroid hormone-related peptide receptor was identified using in situ hybridization with 35S-labelled probe. Expression of parathyroid hormone-related peptide and its receptor was described as uniform, heterogenous or negative within the tumour cell population. Parathyroid hormone-related peptide expression was positive in 13 out of 14 primary tumours and in all 14 metastases. Receptor expression was evident in all 14 primaries and 12 out of 14 metastases. Co-expression of parathyroid hormone-related peptide and parathyroid hormone-related peptide receptor was common (13 primary tumours, 12 metastases). The co-expression of parathyroid hormone-related peptide and its receptor suggest that autocrine parathyroid hormone-related peptide mediated stimulation may be a mechanism of escape from normal growth regulatory pathways. The high frequency of parathyroid hormone-related peptide expression in metastases is consistent with a role in the pathogenesis of bone metastases. British Journal of Cancer (2002) 86, 322–325. DOI: 10.1038/sj/bjc/6600115 www.bjcancer.com © 2002 The Cancer Research Campaign

Bryden, A A G; Hoyland, J A; Freemont, A J; Clarke, N W; George, N J R

2002-01-01

130

Homeobox genes d11-d13 and a13 control mouse autopod cortical bone and joint formation  

PubMed Central

The molecular mechanisms that govern bone and joint formation are complex, involving an integrated network of signaling pathways and gene regulators. We investigated the role of Hox genes, which are known to specify individual segments of the skeleton, in the formation of autopod limb bones (i.e., the hands and feet) using the mouse mutant synpolydactyly homolog (spdh), which encodes a polyalanine expansion in Hoxd13. We found that no cortical bone was formed in the autopod in spdh/spdh mice; instead, these bones underwent trabecular ossification after birth. Spdh/spdh metacarpals acquired an ovoid shape and developed ectopic joints, indicating a loss of long bone characteristics and thus a transformation of metacarpals into carpal bones. The perichondrium of spdh/spdh mice showed abnormal morphology and decreased expression of Runt-related transcription factor 2 (Runx2), which was identified as a direct Hoxd13 transcriptional target. Hoxd11–/–Hoxd12–/–Hoxd13–/– triple-knockout mice and Hoxd13–/–Hoxa13+/– mice exhibited similar but less severe defects, suggesting that these Hox genes have similar and complementary functions and that the spdh allele acts as a dominant negative. This effect was shown to be due to sequestration of other polyalanine-containing transcription factors by the mutant Hoxd13 in the cytoplasm, leading to their degradation. These data indicate that Hox genes not only regulate patterning but also directly influence bone formation and the ossification pattern of bones, in part via Runx2.

Villavicencio-Lorini, Pablo; Kuss, Pia; Friedrich, Julia; Haupt, Julia; Farooq, Muhammed; Turkmen, Seval; Duboule, Denis; Hecht, Jochen; Mundlos, Stefan

2010-01-01

131

Bone Metastasis from Primary Hepatocellular Carcinoma: Characteristics of Soft Tissue Formation  

PubMed Central

Purpose To assess the characteristics of bone metastasis from hepatocellular carcinoma and the radiation field arrangement based on imaging studies. Materials and Methods Fifty-three patients (84 lesions) with bone metastasis from a primary hepatocellular carcinoma completed palliative radiation therapy. All patients underwent one of following imaging studies prior to the initiation of radiation therapy: a bone scan, computed tomography or magnetic resonance imaging. The median radiation dose was 30 Gy (7~40 Gy). We evaluated retrospectively the presence of soft tissue formation and the adjustment of the radiation field based on the imaging studies. Results Soft tissue formation at the site of bony disease was identified from either a CT/MRI scan (41 lesions) or from a symptomatic palpable mass (5 lesions). The adjustment of the radiation field size based on a bone scan was necessary for 31 of 41 soft tissue forming lesions (75.6%), after a review of the CT/MRI scan. The median survival from the initial indication of a hepatoma diagnosis was 8 months (2 to 71 months), with a 2-year survival rate of 38.6%. The median survival from the detection of a bone metastasis was 5 months (1 to 38 months) and the 1-year overall survival rate was 8.7%. Conclusion It was again identified that bone metastasis from a primary hepatocellular carcinoma is accompanied by soft tissue formation. From this finding, an adjustment of the radiation field size based on imaging studies is required. It is advisable to obtain a CT or MRI scan of suspected bone metastasis for better tumor volume coverage prior to the initiation of radiation therapy.

Kim, Sangwon; Wang, Heejung; Cho, Sungwon; Oh, Young-Taek; Kang, Seung-Hee; Yang, Juno

2007-01-01

132

Defective endochondral ossification-derived matrix and bone cells alter the lymphopoietic niche in collagen x mouse models.  

PubMed

Despite the appreciated interdependence of skeletal and hematopoietic development, the cell and matrix components of the hematopoietic niche remain to be fully defined. Utilizing mice with disrupted function of collagen X (ColX), a major hypertrophic cartilage matrix protein associated with endochondral ossification, our data identified a cytokine defect in trabecular bone cells at the chondro-osseous hematopoietic niche as a cause for aberrant B lymphopoiesis in these mice. Specifically, analysis of ColX transgenic and null mouse chondro-osseous regions via micro-computed tomography revealed an altered trabecular bone environment. Additionally, cocultures with hematopoietic and chondro-osseous cell types highlighted impaired hematopoietic support by ColX transgenic and null mouse derived trabecular bone cells. Further, cytokine arrays with conditioned media from the trabecular osteoblast cocultures suggested an aberrant hematopoietic cytokine milieu within the chondro-osseous niche of the ColX deficient mice. Accordingly, B lymphopoiesis was rescued in the ColX mouse derived trabecular osteoblast cocultures with interlukin-7, stem cell factor, and stromal derived factor-1 supplementation. Moreover, B cell development was restored in vivo after injections of interlukin-7. These data support our hypothesis that endrochondrally-derived trabecular bone cells and matrix constituents provide cytokine-rich niches for hematopoiesis. Furthermore, this study contributes to the emerging concept that niche defects may underlie certain immuno-osseous and hematopoietic disorders. PMID:23656481

Sweeney, Elizabeth; Roberts, Douglas; Lin, Angela; Guldberg, Robert; Jacenko, Olena

2013-06-18

133

Defective Endochondral Ossification-Derived Matrix and Bone Cells Alter the Lymphopoietic Niche in Collagen X Mouse Models  

PubMed Central

Despite the appreciated interdependence of skeletal and hematopoietic development, the cell and matrix components of the hematopoietic niche remain to be fully defined. Utilizing mice with disrupted function of collagen X (ColX), a major hypertrophic cartilage matrix protein associated with endochondral ossification, our data identified a cytokine defect in trabecular bone cells at the chondro-osseous hematopoietic niche as a cause for aberrant B lymphopoiesis in these mice. Specifically, analysis of ColX transgenic and null mouse chondro-osseous regions via micro-computed tomography revealed an altered trabecular bone environment. Additionally, cocultures with hematopoietic and chondro-osseous cell types highlighted impaired hematopoietic support by ColX transgenic and null mouse derived trabecular bone cells. Further, cytokine arrays with conditioned media from the trabecular osteoblast cocultures suggested an aberrant hematopoietic cytokine milieu within the chondro-osseous niche of the ColX deficient mice. Accordingly, B lymphopoiesis was rescued in the ColX mouse derived trabecular osteoblast cocultures with interlukin-7, stem cell factor, and stromal derived factor-1 supplementation. Moreover, B cell development was restored in vivo after injections of interlukin-7. These data support our hypothesis that endrochondrally-derived trabecular bone cells and matrix constituents provide cytokine-rich niches for hematopoiesis. Furthermore, this study contributes to the emerging concept that niche defects may underlie certain immuno-osseous and hematopoietic disorders.

Sweeney, Elizabeth; Roberts, Douglas; Lin, Angela; Guldberg, Robert

2013-01-01

134

Elimination of an accessory population involved in proliferation of pleuripotent hematopoietic stem cells from bone marrow by mouse antibrain serum  

SciTech Connect

The authors study changes in the number of splenic exocolonies and also postradiation repopulation of hematopoietic organs in recipients after receiving an injection of bone marrow treated with mouse antibrain serum (MABS), with or without the addition of thymocytes in the later stages. The recipient mice were irradiated with /sup 60/Co gamma rays in a dose of 8.5-8.0 Gy, on the Luch-1 radiotherapeutic apparatus 18 h before receiving the injection of bone marrow and thymocytes. It is postulated that MABS inactivates a population of cells in the bone marrow which are invloved in the control of colony-forming-units (CFUs) proliferation. This population is restored in the animal by the 14th day after irradiation and of injection of MABS-treated bone marrow.

Semina, O.V.; Man'ko, V.M.; Poverennyi, A.M.; Semenets, T.N.

1986-04-01

135

Effect of macrophage migration inhibition factor on the content of stromal precursor cells in mouse bone marrow and efficiency of bone marrow precursor cell cloning in vitro.  

PubMed

The content of stromal precursor cells in the bone marrow of mice decreased 2-5.7 times 24 h after injection of macrophage migration inhibition factor in doses of 0.1-50 ng/kg, this reduction depending on the dose of inhibition factor. The content of precursor cells in the bone marrow of mice increased 2-fold 24 h after injection of S. typhimurium bacterial mass. One day after injection of S. typhimurium bacterial mass, the count of precursor cells in mouse spleen was 7-fold higher than 24 h after injection of macrophage migration inhibition factor. The efficiency of cloning of mouse bone marrow stromal precursor cells in vitro was suppressed 1.7-2.8 times in the presence of macrophage migration inhibition factor in doses of 0.1 to 50 ng/ml culture medium. The effect of cloning inhibition was preserved, if macrophage migration inhibition factor was added to the culture medium after 2 days of bone marrow cell culturing. In general, macrophage migration inhibition factor inhibits stromal precursor cells in vivo and in vitro. The data also indicate that macrophage migration inhibition factor is not responsible for rapid and sharp increase in the count of stromal precursor cells after immunization of animals. PMID:19513377

Gorskaya, U F; Tretyakov, O U; Suslov, A P; Nesterenko, V G

2008-12-01

136

Oocyte-specific overexpression of mouse bone morphogenetic protein-15 leads to accelerated folliculogenesis and an early onset of acyclicity in transgenic mice.  

PubMed

Whereas mutations in the bmp15 gene cause infertility in ewes and women due to defects in folliculogenesis, most defects in female mice lacking bone morphogenetic protein (BMP)-15 are confined to the ovulation process, supportive of the observation that functional mouse BMP-15 is barely detected in oocytes in vivo until after the LH surge. In addition, the mouse BMP-15 proprotein is not processed into the functional mature protein in transfected cells. However, a chimeric protein consisting of the human proregion, human cleavage site, and mouse mature region (termed hhmBMP-15) is processed and the mature protein secreted. To study the role of BMP-15 in folliculogenesis, we generated transgenic mice overexpressing hhmBMP-15, exclusively in oocytes during folliculogenesis and confirmed the overexpression of mouse BMP-15 mature protein. Immature transgenic mice exhibited accelerated follicle growth with decreased primary follicles and an increase in secondary follicles. Granulosa cells of immature mice displayed an increased mitotic index and decreased FSH receptor mRNA expression. Adult mice had normal litter sizes but an increased number of atretic antral follicles. Interestingly, aging mice exhibited an early onset of acyclicity marked by increased diestrus length and early occurrence of constant diestrus. These findings indicate the role of BMP-15 in vivo in promoting follicle growth and preventing follicle maturation, resulting in an early decline in the ovarian reserve of transgenic mice. Therefore, the lack of mouse BMP-15 during early folliculogenesis in the wild-type mice may be relevant to their polyovulatory nature as well as the preservation of ovarian function as the mice age. PMID:18308851

McMahon, Heather E; Hashimoto, Osamu; Mellon, Pamela L; Shimasaki, Shunichi

2008-02-28

137

Primary structure of the mouse sperm receptor polypeptide determined by genomic cloning.  

PubMed Central

The mouse sperm receptor, a glycoprotein called ZP3, is synthesized and secreted by growing oocytes. It is present in more than a billion copies in the unfertilized egg's extracellular coat, or zona pellucida. We have cloned and characterized a region of the mouse (CD-1) genome that spans 10 kilobases of the ZP3 locus. The genomic clones described encompass the entire ZP3 coding region, which contains eight exons. The exons were identified, mapped, and sequenced, yielding the entire primary structure of the ZP3 polypeptide chain (424 amino acids; Mr, 46,300), which includes a 22-amino acid signal sequence. In addition, sequencing of genomic clones has revealed some unusual features of ZP3 mRNA and a region just downstream of the ZP3 gene. Images

Kinloch, R A; Roller, R J; Fimiani, C M; Wassarman, D A; Wassarman, P M

1988-01-01

138

Functional analysis of the mouse alpha-fetoprotein enhancers and their subfragments in primary mouse hepatocyte cultures.  

PubMed

We have compared the activities of mouse alpha-fetoprotein (AFP) enhancers I, II, and III with their minimal enhancer fragments (Mers) I, II, and III and with the entire 7-kilobase pair enhancer domain by transient expression assay in primary fetal mouse liver cells. The level of expression directed by the AFP promoter [p(-1009)AFPcat] alone is stimulated at least 10-fold by the entire AFP enhancer domain (-1009 to -6983). Enhancer I can drive the level of chloramphenicol acetyltransferase activity equivalent to that of the entire enhancer domain, whereas the increase in activity by enhancers II and III is significantly lower (1.5-fold). MersI, II, and III all mediate a greater increase in activity than their corresponding enhancer regions. The increase with MerI is 16-fold. Using DNase I protection analyses we identified 3 protein-binding regions in MerI; site Ia binds liver and brain nuclear proteins; site Ib binds liver, kidney, and brain nuclear proteins as well as purified C/EBP; site Ic binds liver and kidney nuclear proteins. Site-specific mutation of Ia, Ib, or Ic showed a 10-25% reduction in chloramphenicol acetyltransferase expression; deletion of the C/EBP-binding site in Ib showed a 45% reduction in activity and mutation of all 3 sites (Ia, Ib, and Ic) resulted in a 75% reduction in activity. Our studies indicate no single trans-acting factor is absolutely essential for enhancer activity, and that the enhancer activity of MerI is mediated via a combinatorial and additive mechanism. PMID:1375227

Zhang, D E; Rabek, J P; Hsieh, C C; Torres-Ramos, C; Papaconstantinou, J

1992-05-25

139

The relief of bone pain in primary biliary cirrhosis with calcium infusions  

PubMed Central

Intravenous calcium infusions produced subjective relief of bone pain in 14 patients with primary biliary cirrhosis. The bone pain had developed despite long-term parenteral vitamin D therapy. The pain returned after two to three months, but a subsequent course of infusions again brought relief. Before treatment satisfactory iliac crest bone biopsies were obtained in 11 of the patients and were normal in seven; two patients had biopsies indicating osteomalacia and two osteoporosis. After treatment a repeat biopsy in one of the patients with osteomalacia showed marked reduction in osteoid. The infusion treatment produced no change in plasma calcium concentration, serum phosphate, or serum alkaline phosphatase. Absorption of oral calcium was also unchanged.

Ajdukiewicz, A. B.; Agnew, J. E.; Byers, P. D.; Wills, M. R.; Sherlock, Sheila

1974-01-01

140

Controlled release of BMP-2 from a sintered polymer scaffold enhances bone repair in a mouse calvarial defect model.  

PubMed

Sustained and controlled delivery of growth factors, such as bone morphogenetic protein 2 (BMP-2), from polymer scaffolds has excellent potential for enhancing bone regeneration. The present study investigated the use of novel sintered polymer scaffolds prepared using temperature-sensitive PLGA/PEG particles. Growth factors can be incorporated into these scaffolds by mixing the reconstituted growth factor with the particles prior to sintering. The ability of the PLGA/PEG scaffolds to deliver BMP-2 in a controlled and sustained manner was assessed and the osteogenic potential of these scaffolds was determined in a mouse calvarial defect model. BMP-2 was released from the scaffolds in vitro over 3 weeks. On average, ca. 70% of the BMP-2 loaded into the scaffolds was released by the end of this time period. The released BMP-2 was shown to be active and to induce osteogenesis when used in a cell culture assay. A substantial increase in new bone volume of 55% was observed in a mouse calvarial defect model for BMP-2-loaded PLGA/PEG scaffolds compared to empty defect controls. An increase in new bone volume of 31% was observed for PLGA/PEG scaffolds without BMP-2, compared to empty defect controls. These results demonstrate the potential of novel PLGA/PEG scaffolds for sustained BMP-2 delivery for bone-regeneration applications. Copyright © 2012 John Wiley & Sons, Ltd. PMID:22678704

Rahman, Cheryl V; Ben-David, Dror; Dhillon, Amritpaul; Kuhn, Gisela; Gould, Toby W A; Müller, Ralph; Rose, Felicity R A J; Shakesheff, Kevin M; Livne, Erella

2012-06-01

141

Bone Quality determined by Fourier Transform Infrared Imaging Analysis in Mild Primary Hyperparathyroidism  

PubMed Central

Context: Mild primary hyperparathyroidism (PHPT) is characterized by asymptomatic hypercalcemia, most commonly in the absence of classical signs and symptoms. Hence, there is need to characterize this disorder with particular attention to the skeleton. Design: We determined the ratio of pyridinium and dehydrodihydroxylysinonorleucine collagen cross-links in 46 iliac crest bone biopsies from patients with PHPT (14 men, aged 28–68 yr; 32 women, aged 26–74 yr) by Fourier transform infrared imaging. The results were compared with previously reported collagen cross-links ratio determined in iliac crest biopsies from normal subjects. Results: PHPT patients exhibited significantly lower pyridinium to dehydrodihydroxylysinonorleucine collagen cross-links ratio, compared with normal controls. Parathyroidectomy restored values to those comparable with normal controls. Moreover, the differences among PHPT subjects were gender dependent, with female PHPT patients having a statistically significant lower ratio, compared with either male PHPT patients or normal controls. Comparison of the obtained outcomes with histomorphometry showed that the collagen cross-link ratio was strongly correlated with rate of bone formation, and mineralizing surface, in individual patients. This ratio was also correlated with bone mineralization density distribution parameters obtained in the same patients. The strongest correlations were with bone mineralization density distribution variables reflecting heterogeneity of mineralization and primary mineralization parameters. Conclusions: The results are consistent with the high turnover state manifested in PHPT patients. Reduced collagen cross-link ratio in patients with PHPT would be expected to reduce the stiffness of bone tissue. These observations provide a more complete assessment of bone material properties in this disorder.

Zoehrer, Ruth; Dempster, David W.; Bilezikian, John P.; Zhou, Hua; Silverberg, Shonni J.; Shane, Elizabeth; Roschger, Paul; Paschalis, Eleftherios P.; Klaushofer, Klaus

2008-01-01

142

Retroviral-mediated gene transfer and expression of human phenylalanine hydroxylase in primary mouse hepatocytes  

SciTech Connect

Genetic therapy for phenylketonuria (severe phenylalanine hydroxylase deficiency) may require introduction of a normal phenylalanine hydroxylase gene into hepatic cells of patients. The authors report development of a recombinant retrovirus based on the N2 vector for gene transfer and expression of human phenylalanine hydroxylase cDNA in primary mouse hepatocytes. This construct contains an internal promoter of the human {alpha}{sub 1}-antitrypsin gene driving transcription of the phenylalanine hydroxylase cDNA. Primary mouse hepatocytes were isolated from newborn mice, infected with the recombinant virus, and selected for expression of the neomycin-resistance gene. Hepatocytes transformed with the recombinant virus contained high levels of human phenylalanine hydroxylase mRNA transcripts originating from the retroviral and internal promoters. These results demonstrate that the transcriptional regulatory elements of the {alpha}{sub 1} antitrypsin gene retain their tissue-specific function in the recombinant provirus and establish a method for efficient transfer and high-level expression of human phenylalanine hydroxylase in primary hepatocytes.

Peng, H.; Armentano, D.; Mackenzie-Graham, L.; Shen, R.F.; Darlington, G.; Ledley, F.D.; Woo, S.L.C. (Baylor College of Medicine, Houston, TX (USA))

1988-11-01

143

Construction of ectopic xenogeneic bone marrow structure associated with persistent multi-lineage mixed chimerism by engraftment of rat bone marrow plugs into mouse kidney capsules.  

PubMed

Poor bone marrow (BM) engraftment in a xenogeneic combination results at least in part from the limited engraftment capacity of BM-derived stromal cells, which support hematopoietic repopulation in a species-specific fashion. We attempted to construct a BM stromal microenvironment by engraftment of BM plug fragments into kidney capsules in a rat-to-mouse combination. BM plugs from F344/N Jcl-rnu/rnu (F344 nu) rats were transplanted into the kidney capsules of C.B-17 scid/scid (C.B-17 scid) mice treated with rabbit anti-asialo-GM1 serum to deplete natural killer (NK) cells and then with 3 Gy of whole body irradiation. As a conventional control, an equivalent amount of F344 nu bone marrow cells (BMCs) was intravenously injected into C.B-17 scid mice treated with a similar conditioning regimen. In both mouse recipients of rat BM plug engraftment in the kidney capsules and recipients of intravenous injection of rat BMC suspension, comparable extents of donor rat class I+ cells were persistently detected in the peripheral blood. However, the differentiation of rat-derived B cells in the mouse recipients of rat BM plugs was more rapid than that in the recipients of rat BMC suspension. In the late phase (10 weeks after BM transplantation), the percentage of rat-derived T cells (CD4+ cells) in the mouse recipients of rat BM plugs was significantly higher than that in the recipients of rat BMC suspension. At this time point, ectopic BM structure consisting of bone, mesenchymal cells, and hematopoietic progenitors was constructed in the kidney capsules of mice that received rat BM plugs. Most of the cells in the ectopic BM were derived from the donor rat. Thus, engraftment of BM plugs into the kidney capsules results in the construction of a donor-derived BM microenvironment, facilitating multilineage mixed xenogeneic chimerism. PMID:12694546

Hara, Hidetaka; Ohdan, Hideki; Tokita, Daisuke; Onoe, Takashi; Zhou, Wendy; Asahara, Toshimasa

2003-05-01

144

Isolation and characterization of mouse bone marrow-derived Lin(-)/VEGF-R2 (+) progenitor cells.  

PubMed

Circulating endothelial progenitor cells (EPCs) in the peripheral blood (PB) have physiological roles in the maintenance of the existing vascular beds and rescue of vascular injury. In this study, we have evaluated the properties of Lin(-)/VEGF-R2(+) progenitor cells isolated from the mouse bone marrow (BM) and further studied their distribution and integration in an animal model of laser-induced retinal vascular injury. Lin(-)/VEGF-R2(+) cells were enriched from C57BL/6 mice BM using magnetic cell sorting with hematopoietic lineage (Lin) depletion followed by VEGF-R2 positive selection. Lin(-)/VEGF-R2(+) BM cells were characterized using flow cytometry and immunocytochemistry and further tested for colony formation during culture and tube formation on Matrigel®. Lin(-)/VEGF-R2(+) BM cells possessed typical EPC properties such as forming cobble-stone shaped colonies after 3 to 4 weeks of culture, CD34(+) expression, take up of Dil-acLDL and binding to Ulex europaeus agglutinin. However, they did not form tube-like structures on Matrigel®. The progenitor cells retained their phenotype over extended period of culture. After intravitreal transplantation in eyes subjected to the laser-induced retinal vascular injury, some Lin(-)/VEGF-R2(+) cells were able to integrate into the damaged retinal vasculature but the level of cell integration seemed less efficient when compared with previous reports in which EPCs from the human PB were employed. Our results indicate that Lin(-)/VEGF-R2(+) cells isolated from the mouse BM share some similarities to EPCs from the human PB but most of them are at a very early stage of maturation and remain quiescent during culture and after intravitreal transplantation. PMID:23771478

Barthelmes, Daniel; Irhimeh, Mohammad R; Gillies, Mark C; Zhu, Ling; Shen, Weiyong

2013-06-16

145

Proton-gated ion channels in mouse bone marrow stromal cells.  

PubMed

A variety of ion channels like acid sensing ion channels (ASICs) and several members of the transient receptor potential (TRP) cation channel family are known to be activated by protons. The present study describes proton-gated current in mouse bone marrow stromal cells (BMSCs), by using whole cell patch clamp. Rapid application of extracellular solution of pH ? 6.5, evoked slow inactivating current with mean peak value of 328 ± 31pA, (n = 25) at pH 5.0. The reversal potential was close to the theoretical Na(+) equilibrium potential, indicating that majority of the current is mediated by Na(+) and partially carried by Ca(2+) as revealed by ion substitution experiments and Ca(2+) imaging. ASICs blocker amiloride (1mM) and nonselective cation channel blocker flufenamic acid (0.3mM) reduced the current amplitudes by 36 ± 5% (n = 10) and 39 ± 7% (n = 14) respectively. Co-application of flufenamic acid and amiloride further decreased the current by 70 ± 7% (n = 7). However, capsazepine, SKF 96365 and ruthenium red had no effect. 10mM of Ca(2+) and 2mM of La(3+) inhibited the current by 39 ± 6% (n = 5) and 46 ± 6% (n = 4) respectively. Zn(2+) (300 ?M) and Gd(3+) (500 ?M) had no effect on the current amplitude. Low pH mediated cell death was completely inhibited by co-application of La(3+) and amiloride. Reverse Transcriptase-PCR detected expression of mRNAs of ASICs and TRP family. In summary, our results demonstrate the functional expression of low pH-activated ion channels in mouse BMSCs. PMID:22677706

Swain, Sandip Madhusudan; Parameswaran, Sreejit; Sahu, Giriraj; Verma, Rama Shanker; Bera, Amal Kanti

2012-05-09

146

Effect of Coadministration of Vancomycin and BMP-2 on Cocultured Staphylococcus aureus and W-20-17 Mouse Bone Marrow Stromal Cells In Vitro  

PubMed Central

In this study, we aimed to establish an in vitro bacterium/bone cell coculture model system and to use this model for dose dependence studies of dual administration of antibiotics and growth factors in vitro. We examined the effect of single or dual administration of the antibiotic vancomycin (VAN) at 0 to 16 ?g/ml and bone morphogenetic protein-2 (BMP-2) at 0 or 100 ng/ml on both methicillin-sensitive Staphylococcus aureus and mouse bone marrow stromal cells (W-20-17) under both mono- and coculture conditions. Cell metabolic activity, Live/Dead staining, double-stranded DNA (dsDNA) amounts, and alkaline phosphatase activity were measured to assess cell viability, proliferation, and differentiation. An interleukin-6 (IL-6) enzyme-linked immunosorbent assay (ELISA) kit was used to test the bone cell inflammation response in the presence of bacteria. Our results suggest that, when delivered together in coculture, VAN and BMP-2 maintain their primary functions as an antibiotic and a growth factor, respectively. Most interestingly, this dual-delivery type of approach has shown itself to be effective at lower concentrations of VAN than those required for an approach relying strictly on the antibiotic. It may be that BMP-2 enhances cell proliferation and differentiation before the cells become infected. In coculture, a dosage of VAN higher than that used for treatment in monoculture may be necessary to effectively inhibit growth of Staphylococcus aureus. This could mean that the coculture environment may be limiting the efficacy of VAN, possibly by way of bacterial invasion of the bone cells. This report of a coculture study demonstrates a potential beneficial effect of the coadministration of antibiotics and growth factors compared to treatment with antibiotic alone.

Nguyen, A. H.; Kim, S.; Maloney, W. J.; Wenke, J. C.

2012-01-01

147

Bones  

MedlinePLUS

Sections Bone, Joint, and Muscle Disorders Chapters Biology of the Musculoskeletal System Bones Bone, although strong, is a constantly changing tissue that has several functions. Bones serve as rigid structures to ...

148

Bone marrow endothelial progenitors augment atherosclerotic plaque regression in a mouse model of plasma lipid lowering.  

PubMed

The major event initiating atherosclerosis is hypercholesterolemia-induced disruption of vascular endothelium integrity. In settings of endothelial damage, endothelial progenitor cells (EPCs) are mobilized from bone marrow into circulation and home to sites of vascular injury where they aid endothelial regeneration. Given the beneficial effects of EPCs in vascular repair, we hypothesized that these cells play a pivotal role in atherosclerosis regression. We tested our hypothesis in the atherosclerosis-prone mouse model in which hypercholesterolemia, one of the main factors affecting EPC homeostasis, is reversible (Reversa mice). In these mice, normalization of plasma lipids decreased atherosclerotic burden; however, plaque regression was incomplete. To explore whether endothelial progenitors contribute to atherosclerosis regression, bone marrow EPCs from a transgenic strain expressing green fluorescent protein (GFP) under the control of endothelial cell-specific Tie2 promoter (Tie2-GFP(+)) were isolated. These cells were then adoptively transferred into atheroregressing Reversa recipients where they augmented plaque regression induced by reversal of hypercholesterolemia. Advanced plaque regression correlated with engraftment of Tie2-GFP(+) EPCs into endothelium and resulted in an increase in atheroprotective nitric oxide and improved vascular relaxation. Similarly augmented plaque regression was also detected in regressing Reversa mice treated with the stem cell mobilizer AMD3100 which also mobilizes EPCs to peripheral blood. We conclude that correction of hypercholesterolemia in Reversa mice leads to partial plaque regression that can be augmented by AMD3100 treatment or by adoptive transfer of EPCs. This suggests that direct cell therapy or indirect progenitor cell mobilization therapy may be used in combination with statins to treat atherosclerosis. PMID:23081735

Yao, Longbiao; Heuser-Baker, Janet; Herlea-Pana, Oana; Iida, Ryuji; Wang, Qilong; Zou, Ming-Hui; Barlic-Dicen, Jana

2012-12-01

149

Long-term results of combined modality therapy in primary bone lymphomas  

Microsoft Academic Search

Purpose: To report the Massachusetts General Hospital experience in the management of patients with primary bone lymphoma (PBL) treated with combined modality therapy (CMT).Methods and Materials: Records from 37 eligible patients were reviewed. Two patients were treated with complete resection of the tumor, while 35 patients underwent radiation therapy with a median total dose of 54 Gy (range 38.35–66.5). All

Panos Fidias; Ira Spiro; Mark L Sobczak; Gunnlaugur P Nielsen; Eugene F Ruffolo; Henry Mankin; Herman D Suit; David C Harmon

1999-01-01

150

Primary bone tumors in birds: a review and description of two new cases.  

PubMed

Primary bone tumors are only occasionally reported in avian species. This paper presents the cases of an osteosarcoma in a 6-yr-old free-range chicken and a chondrosarcoma in a 3-yr-old barred Plymouth Rock chicken. The well-differentiated, moderately productive osteoblastic osteosarcoma arose from the synsacral vertebrae and had metastasized to the liver. The chondrosarcoma was well differentiated and firmly attached to the left side of the keel. There was no evidence of metastasis. PMID:22856206

Dittmer, K E; French, A F; Thompson, D J; Buckle, K N; Thompson, K G

2012-06-01

151

Growth characteristics of bone marrow cells from beige mutant, the mouse homologue of the Chediak-Highshi syndrome of man, propagated in semisolid agar cultures  

Microsoft Academic Search

Summary  Suspensions of bone marrow cells from the beige (bg\\/bg) mouse, a homologue of the Chediak-Higashi syndrome (C-HS) of man, and normal mouse bone marrow cells, when stimulated by\\u000a colony-stimulating factor (CSF) from different sources, proliferate in semisolid agar cultures and produce colonies composed\\u000a of granulocytic and\\/or mononuclear cells. Studies with CSF from various sources (embryo and kidney feeder monolayers, conditioned

Harland W. Renshaw; William C. Davis

1975-01-01

152

Multiple Biological Activities are Expressed by a Mouse Interleukin 6 cDNA Clone Isolated from Bone Marrow Stromal Cells  

Microsoft Academic Search

Interleukin 6 (IL-6) refers to the gene product that was characterized initially as beta 2 interferon\\/26-kDa protein produced by human fibroblasts and later was found to be identical to B-cell stimulatory factor 2, hybridoma\\/plasmacytoma growth factor, and probably hepatocyte-stimulating factor. Using the human IL-6 cDNA as a probe, we have isolated functional cDNA clones from mouse bone marrow stromal cell

Choy-Pik Chiu; Courtney Moulds; Robert L. Coffman; Donna Rennick; Frank Lee

1988-01-01

153

Fifty-hertz magnetic fields induce free radical formation in mouse bone marrow-derived promonocytes and macrophages  

Microsoft Academic Search

Our findings show a significant increase of free radical production after exposure to 50 Hz electromagnetic fields at a flux density of 1 mT to mouse bone marrow-derived (MBM) promonocytes and macrophages, indicating the cell-activating capacity of extremely low frequency magnetic fields (ELF-MF). We demonstrate that after exposure to ELF-MF mainly superoxide anion radicals were produced, both in MBM macrophages

Jana Rollwitz; Madeleine Lupke; Myrtill Simkó

2004-01-01

154

Identification of clonogenic common Flt3+M-CSFR+ plasmacytoid and conventional dendritic cell progenitors in mouse bone marrow  

Microsoft Academic Search

Lymphoid tissue plasmacytoid and conventional dendritic cells (DCs) are continuously regenerated from hematopoietic stem cells. The cytokine dependence and biology of plasmacytoid and conventional DCs suggest that regeneration might proceed through common DC-restricted developmental intermediates. By selecting for cytokine receptor expression relevant to DC development, we identify here highly cycling Lin?c-KitintFlt3+M-CSFR+ cells with a distinct gene-expression profile in mouse bone

Nobuyuki Onai; Aya Obata-Onai; Michael A Schmid; Toshiaki Ohteki; David Jarrossay; Markus G Manz

2007-01-01

155

An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow  

Microsoft Academic Search

As dendritic cells (DC) are rare populations in all organs, their generation from hematopoietic precursors in large quantities has proven critical to study their biology. From murine bone marrow about 5×106 cells at 70% purity are obtained per mouse after 8 days of culture with GM-CSF. We have improved this standard method and routinely achieve a 50-fold higher yield, i.e.,

Manfred B Lutz; Nicole Kukutsch; Alexandra L. J Ogilvie; Susanne Rößner; Franz Koch; Nikolaus Romani; Gerold Schuler

1999-01-01

156

Evaluation and validation of multiple cell lines and primary mouse macrophages to predict phospholipidosis potential.  

PubMed

Phospholipidosis (PLD) in preclinical species can lead to regulatory delays thereby creating incentives to screen for PLD during drug discovery. The objective of this work was to compare, optimize, and validate in vitro PLD assays in primary mouse macrophages and hepatocyte- (HepG2, HuH7) or macrophage-derived cells lines (I.13.35, RAW264.7) and to evaluate whether primary cells were better at predicting PLD. Assay precision, determined by a measure of signal to noise window (Z'), within assay variability, and day-to-day variability, using amiodarone, was generally acceptable for all cell types; however, precision limits for HepG2 and HuH7 were slightly below assay acceptance criteria. Up to 66 known PLD inducers and non-inducers were subsequently tested to validate the assays. The concordance for predicting PLD in primary macrophages, I-13.35, RAW264.7, HuH7, and HepG2 cells was 91%, 74%, 73%, 62%, and 62% respectively using a decision limit of EC50?125 ?M as a positive finding. Increasing the number of negative controls tested in RAW264.7 cells and changing the decision limit to ?4-fold increase in PLD, improved the specificity and overall concordance to 88%. RAW264.7 cells were selected as the primary screen for predicting PLD, and together with the primary macrophages, were integrated into an overall testing paradigm proposed for use in PLD risk identification. PMID:21767630

LeCureux, Lloyd; Cheng, Charles S; Herbst, John; Reilly, Timothy P; Lehman-McKeeman, Lois; Otieno, Monicah

2011-07-13

157

Peptidomic Analyses of Mouse Astrocytic Cell Lines and Rat Primary Cultured Astrocytes  

PubMed Central

Astrocytes play an active role in the modulation of synaptic transmission by releasing cell-cell signaling molecules in response to various stimuli that evoke a Ca2+ increase. We expand on recent studies of astrocyte intracellular and secreted proteins by examining the astrocyte peptidome in mouse astrocytic cell lines and rat primary cultured astrocytes, as well as those peptides secreted from mouse astrocytic cell lines in response to Ca2+-dependent stimulations. We identified 57 peptides derived from 24 proteins with LC–MS/MS and CE–MS/MS in the astrocytes. Among the secreted peptides, four peptides derived from elongation factor 1, macrophage migration inhibitory factor, peroxiredoxin-5, and galectin-1, were putatively identified by mass-matching to peptides confirmed to be found in astrocytes. Other peptides in the secretion study were mass-matched to those found in prior peptidomics analyses on mouse brain tissue. Complex peptide profiles were observed after stimulation, suggesting that astrocytes are actively involved in peptide secretion. Twenty-six peptides were observed in multiple stimulation experiments but not in controls and thus appear to be released in a Ca2+-dependent manner. These results can be used in future investigations to better understand stimulus-dependent mechanisms of astrocyte peptide secretion.

Yin, Ping; Knolhoff, Ann M.; Rosenberg, Harry J.; Millet, Larry J.; Gillette, Martha U.; Sweedler, Jonathan V.

2012-01-01

158

Antibodies against ClC7 inhibit extracellular acidification-induced Cl? currents and bone resorption activity in mouse osteoclasts.  

PubMed

The Cl? channel/transporter ClC7 is crucial for osteoclastic bone resorption and might become a therapeutic target for osteoporosis. In this study, we raised anti-ClC7 polyclonal antibodies against three different peptide sequences, including G215, P249, and R286, which are the mutation regions found in autosomal dominant osteopetrosis type II patients and examined the effects of these antibodies on the ClC7 Cl? current induced by extracellular acidification (acid-activated Cl? current) using the whole-cell patch clamp technique and bone resorption activity in mouse osteoclasts. Intracellular dialysis of osteoclasts with antibodies to intracellular G215 (Ab-G215) and extracellular application of antibodies to extracellular P249 (Ab-P249) or R286 (Ab-R286) inhibited the acid-activated Cl? current. These antibodies also suppressed the acid-activated Cl? current in ClC7 overexpressing Raw264.7 cells; however, Cl? currents evoked by hypotonic stimulation and the inherent inwardly rectifying K+ currents in mouse osteoclasts were unaffected by these antibodies. Furthermore, extracellularly applied Ab-P249 and Ab-R286 also reduced bone resorption activity. Our results demonstrate that these antibodies specifically block ClC7 in mouse osteoclasts. Thus, anti-ClC7 antibodies have potential promise for treatment of osteoporosis. PMID:21061117

Ohgi, Kimiko; Okamoto, Fujio; Kajiya, Hiroshi; Sakagami, Ryuji; Okabe, Koji

2011-01-01

159

Remodeling of Actin Cytoskeleton in Mouse Periosteal Cells under Mechanical Loading Induces Periosteal Cell Proliferation during Bone Formation  

PubMed Central

Background The adaptive nature of bone formation under mechanical loading is well known; however, the molecular and cellular mechanisms in vivo of mechanical loading in bone formation are not fully understood. To investigate both mechanisms at the early response against mechanotransduction in vivo, we employed a noninvasive 3-point bone bending method for mouse tibiae. It is important to investigate periosteal woven bone formation to elucidate the adaptive nature against mechanical stress. We hypothesize that cell morphological alteration at the early stage of mechanical loading is essential for bone formation in vivo. Principal Findings We found the significant bone formation on the bone surface subjected to change of the stress toward compression by this method. The histological analysis revealed the proliferation of periosteal cells, and we successively observed the appearance of ALP-positive osteoblasts and increase of mature BMP-2, resulting in woven bone formation in the hypertrophic area. To investigate the mechanism underlying the response to mechanical loading at the molecular level, we established an in-situ immunofluorescence imaging method to visualize molecules in these periosteal cells, and with it examined their cytoskeletal actin and nuclei and the extracellular matrix proteins produced by them. The results demonstrated that the actin cytoskeleton of the periosteal cells was disorganized, and the shapes of their nuclei were drastically changed, under the mechanical loading. Moreover, the disorganized actin cytoskeleton was reorganized after release from the load. Further, inhibition of onset of the actin remodeling blocked the proliferation of the periosteal cells. Conclusions These results suggest that the structural change in cell shape via disorganization and remodeling of the actin cytoskeleton played an important role in the mechanical loading-dependent proliferation of cells in the periosteum during bone formation.

Sakai, Daisuke; Kii, Isao; Nakagawa, Kazuki; Matsumoto, Hiroko N.; Takahashi, Masateru; Yoshida, Suguru; Hosoya, Takamitsu; Takakuda, Kazuo; Kudo, Akira

2011-01-01

160

Adenosine A2A receptors play an active role in mouse bone marrow-derived mesenchymal stem cell development  

PubMed Central

Bone marrow-derived mesenchymal stem cells (BM-MSCs) play a role in wound healing and tissue repair and may also be useful for organ regeneration. As we have demonstrated previously that A2A adenosine receptors (A2AR) promote tissue repair and wound healing by stimulating local repair mechanisms and enhancing accumulation of endothelial progenitor cells, we investigated whether A2AR activation modulates BM-MSC proliferation and differentiation. BM-MSCs were isolated and cultured from A2A-deficient and ecto-5?nucleotidase (CD73)-deficient female mice; the MSCs were identified and quantified by a CFU-fibroblast (CFU-F) assay. Procollagen ?2 type I expression was determined by Western blotting and immunocytochemistry. MSC-specific markers were examined in primary cells and third-passage cells by cytofluorography. PCR and real time-PCR were used to quantitate adenosine receptor and CD73 expression. There were significantly fewer CFU-Fs in cultures of BM-MSCs from A2AR knockout (KO) mice or BM-MSCs treated with the A2AR antagonist ZM241385, 1 ?M. Similarly, there were significantly fewer procollagen ?2 type I-positive MSCs in cultures from A2AR KO and antagonist-treated cultures as well. In late passage cells, there were significantly fewer MSCs from A2A KO mice expressing CD90, CD105, and procollagen type I (P<0.05 for all; n=3). These findings indicate that adenosine and adenosine A2AR play a critical role in promoting the proliferation and differentiation of mouse BM-MSCs.

Katebi, Majid; Soleimani, Mansooreh; Cronstein, Bruce N.

2009-01-01

161

EVIDENCE THAT THE BONE RESORPTION-STIMULATING FACTOR PRODUCED BY MOUSE FIBROSARCOMA CELLS IS PROSTAGLANDIN E2  

PubMed Central

A transplantable mouse fibrosarcoma, HSDM1, produces a potent bone resorption-stimulating factor. The factor can be extracted from the tumor tissue and harvested from the medium of clonal strains of HSDM1 tumor cells growing in monolayer culture. It has several chemical and biological properties of a prostaglandin. Using radioimmunoassay techniques, we have shown that HSDM1 cells synthesize and secrete large quantities of prostaglandin E2 (PGE2). The specific bone resorption-stimulating activity of the HSDM1 factor extracted from the tumor is high and approximately equal to that of PGE2 as measured in a bone tissue culture system in vitro. Indomethacin, a potent inhibitor of PGE2 synthesis in HSDM1 cells, also inhibits production by the cells of the bone resorption-stimulating factor, and has no detectable nonspecific effects on the bone culture assay system. Mice bearing the HSDM1 tumor have higher levels of both calcium and PGE2 in serum than control mice. We conclude that PGE2 is the bone resorption-stimulating factor produced by HSDM1 tumor cells, and that secretion of PGE2 by the tumor in vivo accounts for the relative hypercalcemia observed in tumor-bearing animals. The HSDM1 tumor cell system constitutes a new model for studying the pathogenesis of hypercalcemia associated with certain malignant tumors.

Tashjian, Armen H.; Voelkel, Edward F.; Levine, Lawrence; Goldhaber, Paul

1972-01-01

162

Minor histocompatibility antigens on transfused leukoreduced units of red blood cells induce bone marrow transplant rejection in a mouse model  

PubMed Central

When successful, human leukocyte antigen (HLA)–matched bone marrow transplantation with reduced-intensity conditioning is a cure for several nonmalignant hematologic disorders that require chronic transfusion, such as sickle cell disease and aplastic anemia. However, there are unusually high bone marrow transplant (BMT) rejection rates in these patients. Rejection correlates with the number of transfusions before bone marrow transplantation, and it has been hypothesized that preimmunization to antigens on transfused blood may prime BMT rejection. Using a novel mouse model of red blood cell (RBC) transfusion and major histocompatibility complex–matched bone marrow transplantation, we report that transfusion of RBC products induced BMT rejection across minor histocompatibility antigen (mHA) barriers. It has been proposed that contaminating leukocytes are responsible for transfusion-induced BMT rejection; however, filter leukoreduction did not prevent rejection in the current studies. Moreover, we generated a novel transgenic mouse with RBC-specific expression of a model mHA and demonstrated that transfusion of RBCs induced a CD8+ T-cell response. Together, these data suggest that mHAs on RBCs themselves are capable of inducing BMT rejection. Cellular immunization to mHAs is neither monitored nor managed by current transfusion medicine practice; however, the current data suggest that mHAs on RBCs may represent an unappreciated and significant consequence of RBC transfusion.

Desmarets, Maxime; Cadwell, Chantel M.; Peterson, Kenneth R.; Neades, Renee

2009-01-01

163

Establishment of primary cultures for mouse ameloblasts as a model of their lifetime  

SciTech Connect

To understand how the properties of ameloblasts are spatiotemporally regulated during amelogenesis, two primary cultures of ameloblasts in different stages of differentiation were established from mouse enamel epithelium. Mouse primary ameloblasts (MPAs) prepared from immature enamel epithelium (MPA-I) could proliferate, whereas those from mature enamel epithelium (MPA-M) could not. MPA-M but not MPA-I caused apoptosis during culture. The mRNA expression of amelogenin, a marker of immature ameloblasts, was down-regulated, and that of enamel matrix serine proteiase-1, a marker of mature ameloblasts, was induced in MPA-I during culture. Using green fluorescence protein as a reporter, a visualized reporter system was established to analyze the promoter activity of the amelogenin gene. The region between -1102 bp and -261 bp was required for the reporter expression in MPA-I. These results suggest that MPAs are valuable in vitro models for investigation of ameloblast biology, and that the visualized system is useful for promoter analysis in MPAs.

Suzawa, Tetsuo [Department of Biochemistry, School of Dentistry, Showa University, Tokyo 142-8555 (Japan)]. E-mail: suzawa@dent.showa-u.ac.jp; Itoh, Nao [Department of Biochemistry, School of Dentistry, Showa University, Tokyo 142-8555 (Japan); Department of Clinical Pharmacy, School of Pharmaceutical Science, Showa University (Japan); Takahashi, Naoyuki [Institute for Oral Science, Matsumoto Dental University, Shiojiri 399-0781 (Japan); Katagiri, Takenobu [Division of Pathophysiology, Research Center for Genomic Medicine, Saitama Medical School, Hidaka 350-1241 (Japan); Morimura, Naoko [Department of Biochemistry, School of Dentistry, Showa University, Tokyo 142-8555 (Japan); Kobayashi, Yasuna [Department of Clinical Pharmacy, School of Pharmaceutical Science, Showa University (Japan); Yamamoto, Toshinori [Department of Clinical Pharmacy, School of Pharmaceutical Science, Showa University (Japan); Kamijo, Ryutaro [Department of Biochemistry, School of Dentistry, Showa University, Tokyo 142-8555 (Japan)

2006-07-07

164

Benzene-induced micronuclei formation in mouse fetal liver blood, peripheral blood, and maternal bone marrow cells  

SciTech Connect

The transplacental cytogenetic effects of benzene were studied by using the micronucleus test of polychromatic erythrocytes (PCE) found in both fetal liver and fetal peripheral blood, and were compared with PCE from maternal bone marrow. Timed-pregnant mice received single intraperitoneal doses of benzene on the 14th day of gestation and were sacrificed 21 hours after injection. Benzene elicited a significant increase in the frequency of micronucleated polychromatic erythrocytes (MNPCE) in fetal liver blood cells at doses of 219 to 874 mg/kg, and in fetal peripheral blood cells and maternal bone marrow cells at doses of 437 and 874 mg/kg. The data demonstrate that benzene is a moderate transplacental clastogenic agent, and that the mouse transplacental micronucleus test using fetal liver blood cells is a potentially more sensitive indicator of the genotoxicity of benzene than either fetal peripheral blood or maternal bone marrow cells.

Ning, Hansun (Univ. of California, Davis (United States) Ministry of Railways, Beijing (China)); Kado, N.Y. (Univ. of California, Davis (United States) California Air Resources Board, Sacramento (United States)); Kuzmicky, P.A.; Hsieh, D.P.H. (Univ. of California, Davis (United States))

1991-01-01

165

Evaluation of mouse whole body bone marrow cellularity and distribution of hematopoietic progenitors  

Microsoft Academic Search

In order to perform murine transplant experiments, marrow cells are usually harvested from hind legs bones either by bone marrow flushing or bone crushing. We examined the feasibility of harvesting whole skeleton marrow in regards to cell numbers and progenitor potential. Bones of BALB\\/c mice were dissected and flushed with cold HBSS or crushed in a mortar with cold HBSS

J.-F Lambert; J. E Carlson; G. A Colvin; P. J Quesenberry

2000-01-01

166

Effect of rhBMP-2 on the osteogenic potential of bone marrow stromal cells from an osteogenesis imperfecta mouse ( oim)  

Microsoft Academic Search

To understand whether osteogenesis imperfecta (OI) could result from defective differentiation of osteoprogenitor cells, we investigated the osteogenic potential of bone marrow stromal cells from a mouse model of human OI (oim). Bone marrow was flushed from the femurs and tibias of oim and normal littermates using a syringe with Dulbecco's modified Eagle's medium, and cells were allowed to adhere

M. L. Balk; J. Bray; C. Day; M. Epperly; J. Greenberger; C. H. Evans; C. Niyibizi

1997-01-01

167

Metastatic malignant melanoma in bone marrow with occult primary site - a case report with review of literature  

PubMed Central

Background Metastases of malignant melanoma to the bone marrow are very rare. A few case reports are published in the literature with a known primary site. Case presentation Herein we present a case of metastatic malignant melanoma in bone marrow with occult primary site in a 22- year-old-male. Diagnosis was confirmed by morphology and immunohistochemistry. A pertinent review of literature is also presented by using relevant articles indexed in PubMed (National Library of Medicine) database. The search was based on the following terms: metastasis or metastases, malignant melanoma and bone marrow. Conclusion In this report we discuss a rare case of metastatic malignant melanoma to the bone marrow with an unknown primary. Clinicians must be aware of the varied clinical manifestations of disseminated malignant melanoma even if the primary site is not evident.

Jain, Deepali; singh, Tejindar; Kumar, Naresh; Daga, Mradul K

2007-01-01

168

Abnormal parathyroid cell proliferation precedes biochemical abnormalities in a mouse model of primary hyperparathyroidism.  

PubMed

The properties of neoplastic proliferation and hormonal dysregulation are tightly linked in primary hyperparathyroidism (HPT). However, whether abnormal parathyroid proliferation is the cause or result of a shift in calcium-sensitive parathyroid hormonal regulation has been controversial. We addressed this issue by analyzing the temporal sequence of these fundamental abnormalities in a mouse model of primary HPT. These transgenic mice (PTH-D1) harbor a transgene that targets overexpression of the cyclin D1 oncogene to parathyroid cells, resulting in parathyroid hypercellularity with a phenotype of chronic biochemical HPT and, notably, an abnormal in vivo PTH-calcium set point. We examined parathyroid cell proliferation and biochemical alterations in PTH-D1 and control wild-type mice from ages 1-14 months. Strikingly, abnormal parathyroid proliferation regularly preceded dysregulation of the calcium-PTH axis, supporting the concept that disturbed parathyroid proliferation is the crucial primary initiator leading to the development of the biochemical phenotype of HPT. Furthermore, we observed that decreased expression of the calcium-sensing receptor in the parathyroid glands occurs several months before development of biochemical HPT, suggesting that decreased calcium-sensing receptor may not be sufficient to cause PTH dysregulation in this animal model of primary HPT. PMID:15928311

Mallya, Sanjay M; Gallagher, James J; Wild, Yvette K; Kifor, Olga; Costa-Guda, Jessica; Saucier, Kirsten; Brown, Edward M; Arnold, Andrew

2005-05-31

169

Primary cultures and the levels of cytochrome P 450 in hepatocytes from mouse, rat, hamster, and rabbit liver  

Microsoft Academic Search

Summary  Hepatocyte primary cultures (HPC) derived from rat, mouse, hamster, and rabbit liver were characterized for a variety of parameters.\\u000a The conditions that maximized recovery, attachment, and survival varied between species. Hepatocytes from all four species\\u000a were capable of attaching in serum-free Williams’ medium E (WME), but optimal attachment as monolayer cultures was achieved\\u000a for mouse and hamster HPC in medium

Carol J. Maslansky; Gary M. Williams

1982-01-01

170

The influence of mechanical stimulation on osteoclast localization in the mouse maxilla: bone histomorphometry and finite element analysis.  

PubMed

The mechanism of traumatic bone resorption in the denture-bearing bone has not yet been established with regard to the osteoclastic activity in relation to the mechanical stimulus. The purpose of this study was to clarify whether osteoclast appearance in maxilla depends on the strain intensity, using the murine loading model. The maxillary palate of thirteen-week-old male C57BL/6 mice was subjected to continuous pressure of 2 kPa (low stimulation, n = 4) or 7 kPa (high stimulation, n = 4) for 30 min/day for 7 consecutive days, and the mice were sacrificed after the last loading. The control group underwent the same protocol without load (n = 4). An animal-specific finite element model was constructed based on morphology and characteristics obtained from the micro-CT data and used to calculate the strain intensity of the bone. The bone histomorphometric technique revealed significant reduction of cortical bone volume and significant increase of bone resorption parameters such as osteoclast number in the bone tissue under the loading contact in comparison to the control (p < 0.05). The osteoclasts were observed in the subsurface region adjacent to the loading contact and the peripheral region of the marrow space in the intracortical region of the cortical bone in the mouse maxilla in both stimulation groups. An average of more than 90 % of the osteoclasts was observed in the areas with strain intensity higher than 85.0? strain for the high stimulation group. The result suggests that the osteoclastic resorption is location-dependent and is also sensitive to the local strain intensity. PMID:22584607

Fujiki, K; Aoki, K; Marcián, P; Borák, L; Hudieb, M; Ohya, K; Igarashi, Y; Wakabayashi, N

2012-05-15

171

Examination of ER? Signaling Pathways in Bone of Mutant Mouse Models Reveals the Importance of ERE-Dependent Signaling  

PubMed Central

The mechanisms of estrogen receptor (ER)-? activity can be categorized into those involving direct (classical) or indirect (nonclassical) DNA binding. Although various mouse models have demonstrated the importance of ER? in bone, the specific gene expression patterns affected by these modes of ER? action are unknown. In this report, the gene expression patterns of ER?-deficient (ERKO) mice and nonclassical ER knock-in (NERKI) mice, which can function only by nonclassical means, were analyzed. Three-month-old mice were ovariectomized and implanted with estrogen pellets for 1 month to normalize estrogen levels. Microarray analysis of flushed cortical bone revealed 28% (210 of 763) of the genes differentially expressed in ERKO mice were altered in NERKI mice, suggesting estrogen response element-dependent regulation of these genes in bone. Pathway analysis revealed alterations in genes involved in focal adhesion and extracellular matrix interactions. However, the majority of genes regulated in ERKO mice (72%) were unique (i.e. not altered in NERKI mice), suggesting these are regulated by nonclassical mechanisms. To further explore the pathways affected in ERKO mice, we performed focused quantitative PCR arrays for genes involved in various aspects of bone physiology. Genes involved in bone formation, senescence, apoptosis, and autophagy were significantly regulated. Overall, the majority of the genes regulated by ER? in bone are via nonclassical pathways. However, because NERKI mice display an osteoporotic phenotype, it can be deduced that the minority of the estrogen response element-dependent genes/pathways play critical roles in the regulation of bone physiology. These data demonstrate the importance of classical ER? signaling in regulating bone metabolism.

Chokalingam, Kumar; Roforth, Matthew M.; Nicks, Kristy M.; McGregor, Ulrike; Fraser, Daniel; Khosla, Sundeep

2012-01-01

172

The role of bone marrow-derived cells in bone fracture repair in a green fluorescent protein chimeric mouse model  

SciTech Connect

We investigated the role of bone marrow cells in bone fracture repair using green fluorescent protein (GFP) chimeric model mice. First, the chimeric model mice were created: bone marrow cells from GFP-transgenic C57BL/6 mice were injected into the tail veins of recipient wild-type C57BL/6 mice that had been irradiated with a lethal dose of 10 Gy from a cesium source. Next, bone fracture models were created from these mice: closed transverse fractures of the left femur were produced using a specially designed device. One, three, and five weeks later, fracture lesions were extirpated for histological and immunohistochemical analyses. In the specimens collected 3 and 5 weeks after operation, we confirmed calluses showing intramembranous ossification peripheral to the fracture site. The calluses consisted of GFP- and osteocalcin-positive cells at the same site, although the femur consisted of only osteocalcin-positive cells. We suggest that bone marrow cells migrated outside of the bone marrow and differentiated into osteoblasts to make up the calluses.

Taguchi, Kazuhiro [Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo (Japan) and Department of Orthopedic Surgery, Nippon Medical School, Tokyo (Japan)]. E-mail: s3061@nms.ac.jp; Ogawa, Rei [Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo (Japan); Department of Plastic and Reconstructive Surgery, Nippon Medical School, Tokyo (Japan); Migita, Makoto [Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo (Japan); Department of Pediatrics, Nippon Medical School, Tokyo (Japan); Hanawa, Hideki [Department of Plastic and Reconstructive Surgery, Nippon Medical School, Tokyo (Japan); Ito, Hiromoto [Department of Orthopedic Surgery, Nippon Medical School, Tokyo (Japan); Orimo, Hideo [Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo (Japan)

2005-05-27

173

Constructing a multi-scan synchrotron X-ray microscope to study the function of osteocyte canaliculi in mouse bone  

SciTech Connect

Formulating a multi-scan method applied to an X-ray microscope CT with synchrotron radiation, we attempted to analyze the 3D functional structure of osteocyte canaliculi inside the cortical bone of a mouse tibia. We employed a two-method combination to scan the same position of the specimen. To extract the internal bone canalicular structure, we first combined a Talbot interferometer with an X-ray microscope, and applied a differential phase imaging method to measure the absolute value of bone mineral around the canaliculi. Next, we used the X-ray microscope without the Talbot interferometer under a defocus condition, moving the specimen toward the zone plate by 6 mm. This defocus contrast method visualizes the canaliculi by emphasizing the edges of the bone. We performed CT scans by the two configurations and precisely aligned resultant 3D images so that the same position in the specimen is compared. We could extract the osteocyte canaliculi and evaluate the mineral density of their surroundings. The degree of mineralization varied for each osteocyte lacuna and canaliculus. The multi-scan microscopic X-ray CT is a powerful tool for analyzing bone mineralization.

Nango, Nobuhito; Kubota, Shogo; Yashiro, Wataru; Momose, Atsushi; Takada, Yasunari; Matsuo, Koichi [Ratoc System Engineering Co., Ltd, Toho Edogawabashi Bldg. 4F, 1-24-8 Sekiguchi, Bunkyo-ku, Tokyo 112-0014 (Japan); Dept. of Advanced Materials Science, Graduate School of Frontier Sciences, The University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa-shi, Chiba 277-8561 (Japan); Lab. of Cell and Tissue Biology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan)

2012-07-31

174

Enumeration of the colony-forming units-fibroblast from mouse and human bone marrow in normal and pathological conditions  

PubMed Central

Bone marrow stromal cell populations, containing a subset of multipotential skeletal stem cells, are increasingly contemplated for use in tissue engineering and stem cell therapy, whereas their involvement in the pathogenetic mechanisms of skeletal disorders is far less recognized. We compared the concentrations of stromal clonogenic cells, colony forming units–fibroblast (CFU-Fs), in norm and pathology. Initially, culture conditions were optimized by demonstrating that fetal bovine serum heat inactivation could significantly repress colony formation. Using non-heat-inactivated fetal bovine serum, the concentration of CFU-Fs (colony-forming efficiency, CFE) ranged from 3.5 ± 1.0 to 11.5 ± 4.0 per 1 × 105 nucleated cells in five inbred mouse strains. In four transgenic lines with profound bone involvement, CFE was either significantly reduced or increased compared to wild-type littermates. In normal human donors, CFE decreased slightly with age and averaged 52.2 ± 4.1 for children and 32.3 ± 3.0 for adults. CFE was significantly altered in patients with several skeletal, metabolic, and hematological disorders: reduced in congenital generalized lipodystrophy, achondroplasia (SADDAN), pseudoachondroplasia, and Paget disease of bone and elevated in alcaptonuria and sickle cell anemia. Our findings indicate that under appropriate culture conditions, CFE values may provide useful insights into bone/bone marrow pathophysiology.

Kuznetsov, Sergei A.; Mankani, Mahesh H.; Bianco, Paolo; Robey, Pamela G.

2009-01-01

175

Regulation of STAT signaling in mouse bone marrow derived dendritic cells by respiratory syncytial virus  

PubMed Central

Background/aims Dendritic cells (DCs) act as a portal for virus invasion as well as potent antigen-presenting cells (APCs) involved in the antiviral host response. Interferons (IFNs) are produced in response to bacterial and viral infection and activate innate immune responses to efficiently counteract and remove pathogenic invaders. Respiratory syncytial virus (RSV) could inhibit IFN-mediated signaling pathway in epithelial cells; however, the effects of RSV on IFN signaling in the dendritic cells (DCs) are still unknown. Methods Mouse bone marrow derived DCs (BMDCs) were mock or infected with RSV at different multiplicity of infection (MOI) for 24 h, and then treated with different cytokines such as interferon-? (IFN-?), IFN-? or interleukin-10 (IL-10). The mRNA expression of RSV nonstructural protein-1 (NS-1) and NS-2 was detected by RT-PCR. The expression of Janus family kinase-signal transducer and activator of transcription (JAK/STAT) signaling proteins was assessed by immunoblotting assays. The nuclear localization of specific signaling proteins was determined by immunofluorescence assay. Results Increasing amounts of NS-1 or NS-2 mRNA expression in BMDCs were observed with infected RSV at increasing MOI, suggesting BMDCs were permissive for viral gene expression. Further examination of the IFN-? signaling cascade showed RSV infection increased the total cellular levels of STAT1 and STAT2 in BMDCs, but impaired the IFN-?-dependent phosphorylation and nuclear localization of STAT1 and STAT2. The inhibitory effects of RSV on STAT1 and STAT2 phosphorylation and translocation were abolished by UV inactivation. In contrast, RSV did not inhibit the IFN-?-stimulated STAT1 phosphorylation and nuclear localization. IL-10-stimulated STAT3 phosphorylation was also unaffected by RSV. Conclusions As well as RSV inhibiting STAT protein levels through degradation mechanisms in epithelial cells, these findings demonstrate that RSV also can specifically inhibit the type I interferon response in BMDCs through regulation of STAT1 and STAT2 phosphorylation and nuclear translocation.

Jie, Zhijun; Dinwiddie, Darrell L.; Senft, Albert P.; Harrod, Kevin S.

2013-01-01

176

Does Bone Morphogenetic Protein 6 (BMP6) Affect Female Fertility in the Mouse?1  

PubMed Central

Bone morphogenetic protein 6 (BMP6) is a transforming growth factor beta superfamily member produced by mammalian oocytes as well as other cell types. Despite well-characterized effects of recombinant BMP6 on granulosa cells in vitro, the function of BMP6 in vivo has been ill-defined. Therefore, the effects of genetic deletion of the Bmp6 gene on female mouse fertility were assessed. The mean litter size of Bmp6?/? females was reduced by 22% (P < 0.05) compared to Bmp6+/+ controls. Not only did Bmp6?/? females naturally ovulate 24% fewer eggs, but competence of in vitro-matured oocytes to complete preimplantation development after fertilization in vitro was decreased by 50%. No apparent effect of Bmp6 deletion on either the morphology or the dynamics of follicular development was apparent. Nevertheless, levels of luteinizing hormone (LH)/human chorionic gonadotropin (hCG)-induced transcripts, which encode proteins required for cumulus expansion (HAS2, PTGS2, PTX3, and TNFAIP6), and of epidermal growth factor-like peptides (AREG, BTC, and EREG) were lower in Bmp6?/? mice than in controls after administration of a reduced dose of hCG (1 IU) in vivo. LH receptor (Lhcgr) transcript levels were not significantly lower in Bmp6?/? granulosa cells, suggesting that BMP6 is required for processes downstream of LH receptors. To assess whether another oocyte-derived BMP, BMP15, could have BMP6-redundant functions in vivo, the fertility of Bmp15/Bmp6 double mutants was assessed. Fertility was not significantly reduced in double-homozygous mutants compared with that in double-heterozygous controls. Therefore, BMP6 promotes normal fertility in female mice, at least in part, by enabling appropriate responses to LH and normal oocyte quality. Thus, Bmp6 probably is part of the complex genetic network that determines female fertility.

Sugiura, Koji; Su, You-Qiang; Eppig, John J.

2010-01-01

177

Positive selection in bone morphogenetic protein 15 targets a natural mutation associated with primary ovarian insufficiency in human.  

PubMed

Bone Morphogenetic Protein 15 (BMP15) is a TGF?-like oocyte-derived growth factor involved in ovarian folliculogenesis as a critical regulator of many granulosa cell processes. Alterations of the BMP15 gene have been found associated with different ovarian phenotypic effects depending on the species, from sterility to increased prolificacy in sheep, slight subfertility in mouse or associated with primary ovarian insufficiency (POI) in women. To investigate the evolving role of BMP15, a phylogenetic analysis of this particular TGF? family member was performed. A maximum likelihood phylogenetic tree of several TGF?/BMP family members expressed by the ovary showed that BMP15 has a very strong divergence and a rapid evolution compared to others. Moreover, among 24 mammalian species, we detected signals of positive selection in the hominidae clade corresponding to F146, L189 and Y235 residues in human BMP15. The biological importance of these residues was tested functionally after site directed-mutagenesis in a COV434 cells luciferase assay. By replacing the positively selected amino acid either by alanine or the most represented residue in other studied species, only L189A, Y235A and Y235C mutants showed a significant increase of BMP15 signaling when compared to wild type. Additionally, the Y235C mutant was more potent than wild type in inhibiting progesterone secretion of ovine granulosa cells in primary culture. Interestingly, the Y235C mutation was previously identified in association with POI in women. In conclusion, this study evidences that the BMP15 gene has evolved faster than other members of the TGFß family and was submitted to a positive selection pressure in the hominidae clade. Some residues under positive selection are of great importance for the normal function of the protein and thus for female fertility. Y235 represents a critical residue in the determination of BMP15 biological activity, thus indirectly confirming its role in the onset of POI in women. PMID:24147118

Auclair, Sylvain; Rossetti, Raffaella; Meslin, Camille; Monestier, Olivier; Di Pasquale, Elisa; Pascal, Géraldine; Persani, Luca; Fabre, Stéphane

2013-10-16

178

Positive Selection in Bone Morphogenetic Protein 15 Targets a Natural Mutation Associated with Primary Ovarian Insufficiency in Human  

PubMed Central

Bone Morphogenetic Protein 15 (BMP15) is a TGF?-like oocyte-derived growth factor involved in ovarian folliculogenesis as a critical regulator of many granulosa cell processes. Alterations of the BMP15 gene have been found associated with different ovarian phenotypic effects depending on the species, from sterility to increased prolificacy in sheep, slight subfertility in mouse or associated with primary ovarian insufficiency (POI) in women. To investigate the evolving role of BMP15, a phylogenetic analysis of this particular TGF? family member was performed. A maximum likelihood phylogenetic tree of several TGF?/BMP family members expressed by the ovary showed that BMP15 has a very strong divergence and a rapid evolution compared to others. Moreover, among 24 mammalian species, we detected signals of positive selection in the hominidae clade corresponding to F146, L189 and Y235 residues in human BMP15. The biological importance of these residues was tested functionally after site directed-mutagenesis in a COV434 cells luciferase assay. By replacing the positively selected amino acid either by alanine or the most represented residue in other studied species, only L189A, Y235A and Y235C mutants showed a significant increase of BMP15 signaling when compared to wild type. Additionally, the Y235C mutant was more potent than wild type in inhibiting progesterone secretion of ovine granulosa cells in primary culture. Interestingly, the Y235C mutation was previously identified in association with POI in women. In conclusion, this study evidences that the BMP15 gene has evolved faster than other members of the TGFß family and was submitted to a positive selection pressure in the hominidae clade. Some residues under positive selection are of great importance for the normal function of the protein and thus for female fertility. Y235 represents a critical residue in the determination of BMP15 biological activity, thus indirectly confirming its role in the onset of POI in women.

Meslin, Camille; Monestier, Olivier; Di Pasquale, Elisa; Pascal, Geraldine; Persani, Luca; Fabre, Stephane

2013-01-01

179

ACTH stimulates insulin secretion from MIN6 cells and primary mouse and human islets of Langerhans.  

PubMed

It has previously been suggested that ACTH and ACTH-related peptides may act as paracrine modulators of insulin secretion in the islets of Langerhans. We have, therefore, examined the expression and function of the ACTH receptor (the melanocortin 2 receptor, MC2-R) in human and mouse primary islet tIssue and in the MIN6 mouse insulinoma cell line. Mouse MC2-R mRNA was detected in both MIN6 cells and mouse islet tIssue by PCR amplification of cDNA. In perifusion experiments with MIN6 pseudo-islets, a small, transient increase in insulin secretion was obtained when ACTH(1-24) (1 nM) was added to medium containing 2 mM glucose (control) but not when the medium glucose content was increased to 8 mM. Further investigations were performed using static incubations of MIN6 cell monolayers; ACTH(1-24) (1 pM-10 nM) provoked a concentration-dependent increase in insulin secretion from MIN6 monolayer cells that achieved statistical significance at concentrations of 1 and 10 nM (150 +/- 13.6% basal secretion; 187 +/- 14.9% basal secretion, P<0.01). Similar responses were obtained with ACTH(1-39). The phosphodiesterase inhibitor IBMX (100 microM) potentiated the responses to sub-maximal doses of ACTH(1-24). Two inhibitors of the protein kinase A (PKA) signaling pathway, Rp-cAMPS (500 microM) and H-89 (10 microM), abolished the insulin secretory response to ACTH(1-24) (0.5-10 nM). Treatment with 1 nM ACTH(1-24) caused a small, statistically significant increase in intracellular cAMP levels. Secretory responses of MIN6 cells to ACTH(1-24) were also influenced by changes in extracellular Ca2+ levels. Incubation in Ca2+-free buffer supplemented with 0.1 mM EGTA blocked the MIN6 cells' secretory response to 1 and 10 nM ACTH(1-24). Similar results were obtained when a Ca2+ channel blocker (nitrendipine, 10 microM) was added to the Ca2+-containing buffer. ACTH(1-24) also evoked an insulin secretory response from primary tIssues. The addition of ACTH(1-24) (0.5 nM) to perifusions of mouse islets induced a transient increase in insulin secretion at 8 mM glucose. Perifused human primary islets also showed a secretory response to ACTH(1-24) at basal glucose concentration (2 mM) with a rapid initial spike in insulin secretion followed by a decline to basal levels. Overall the results demonstrate that the MC2-R is expressed in beta-cells and suggest that activation of the receptor by ACTH initiates insulin secretion through the activation of PKA in association with Ca2+ influx into beta-cells. PMID:14709154

Al-Majed, H T; Jones, P M; Persaud, S J; Sugden, D; Huang, G C; Amiel, S; Whitehouse, B J

2004-01-01

180

Differential requirement for B-memory and T-memory cells in adoptive antibody formation in mouse bone marrow.  

PubMed Central

During the secondary response of mice to T-dependent antigens, antibody-producing plaque-forming cells (PFC) appear not only in peripheral lymphoid organs, but also in the bone marrow. This bone marrow antibody formation is feeble after primary immunization. The capacity of bone marrow antibody formation is dependent on the presence of antigen-specific memory cells at the moment of secondary immunization. We investigated whether hapten-primed B memory, carrier-primed T memory or both B-memory and T-memory cells are required for the adoptive PFC response in the bone marrow to T-dependent hapten-carrier conjugates. Adoptive antibody formation in the bone marrow was found after transfer of hapten-primed spleen cells, but not after transfer of carrier-primed spleen cells or virgin spleen cells. Thus, B-memory cells are obligatory for adoptive antibody formation in the bone marrow, in contrast to T-memory cells. However, T-memory cells did facilitate the bone marrow PFC response mediated by the infused B-memory cells.

Koch, G; Benner, R

1982-01-01

181

The Effects of RANKL Inhibition on Fracture Healing and Bone Strength in a Mouse Model of Osteogenesis Imperfecta  

PubMed Central

Summary Currently, the standard treatment for osteogenesis imperfecta (OI) is bisphosphonate therapy. Recent studies, however, have shown delayed healing of osteotomies in a subset of OI patients treated with such agents. The current study sought to determine the effects of another therapy, RANKL inhibition, on bone healing and bone strength in the growing oim/oim mouse, a model of moderate-to-severe OI. Mice (73 oim/oim and 69 wildtype (WT)) were injected twice weekly with either soluble murine RANK (RANK-Fc) (1.5mg/kg) or saline beginning at 6 weeks of age. At 8 weeks of age, the animals underwent transverse mid-diaphyseal osteotomies of the right femur. Therapy was continued until sacrifice at 2, 3, 4 or 6 weeks post-fracture. At 6 weeks post-fracture, greater callus area (6.59±3.78mm2 vs 2.67±2.05mm2, p=0.003) and increased radiographic intensity (mineral density) (0.48 ± 0.14 vs. 0.30 ± 0.80, p=0.005) were found in the RANK-Fc vs saline oim/oim group, indicating a delay in callus remodeling. Despite this delay, mechanical tests at 6 weeks post-fracture revealed no significant differences in whole bone properties of stiffness and failure moment. Further, RANKL inhibition resulted in a greater failure moment and greater work to failure for the non-fractured contralateral WT bones compared to the non-fractured saline WT bones. Together, these results demonstrate that RANKL-inhibition does not adversely affect the mechanical properties of healing bone in the oim/oim mice, and is associated with increased strength in intact bone in the WT mice.

Delos, D.; Yang, X.; Ricciardi, B.F.; Myers, E.R.; Bostrom, M.P.G.; Pleshko Camacho, N.

2009-01-01

182

Proteases as Modulators of Tumor-Stromal Interaction: Primary Tumors to Bone Metastases  

PubMed Central

Summary As cells undergo oncogenic transformation and as transformed cells arrive at metastatic sites, a complex interplay occurs with the surrounding stroma. This dialogue between tumor and stroma ultimately dictates the success of the tumor cells in the given microenvironment. As a result, understanding the molecular mechanisms at work is important for developing new therapeutic modalities. Proteases are major players in the interaction between tumor and stroma. This review will focus on the role of proteases in modulating tumor-stromal interactions of both primary breast and prostate tumors as well as at bone metastatic sites in a way that favors tumor growth.

Wilson, Thomas J.; Singh, Rakesh K.

2008-01-01

183

Primary Ewing’s sarcoma of cranial bones: analysis of ten patients  

Microsoft Academic Search

Objective  Ewing’s sarcomas are the second most common bone tumors in children and primary involvement of the cranium is uncommon. We\\u000a analyzed retrospectively the data of ten patients with this rare subset of disease, who had been treated at our institute\\u000a since 2005. Our aim was to assess the outcomes, recurrence rates and the selection of appropriate treatment methods.\\u000a \\u000a \\u000a \\u000a \\u000a Methods  The patients

Pravin Shashikant Salunke; Kirti Gupta; Vinod Malik; Narendra Kumar; Lauren E. Henke; Chunyu Cai; Wei-Shen Chen; John D. Pfeifer

2011-01-01

184

Sensorimotor mismatch signals in primary visual cortex of the behaving mouse.  

PubMed

Studies in anesthetized animals have suggested that activity in early visual cortex is mainly driven by visual input and is well described by a feedforward processing hierarchy. However, evidence from experiments on awake animals has shown that both eye movements and behavioral state can strongly modulate responses of neurons in visual cortex; the functional significance of this modulation, however, remains elusive. Using visual-flow feedback manipulations during locomotion in a virtual reality environment, we found that responses in layer 2/3 of mouse primary visual cortex are strongly driven by locomotion and by mismatch between actual and expected visual feedback. These data suggest that processing in visual cortex may be based on predictive coding strategies that use motor-related and visual input to detect mismatches between predicted and actual visual feedback. PMID:22681686

Keller, Georg B; Bonhoeffer, Tobias; Hübener, Mark

2012-06-01

185

Calcitonin (but not calcitonin gene-related peptide) increases mouse bone cell proliferation in a dose-dependent manner, and increases mouse bone formation, alone and in combination with fluoride  

Microsoft Academic Search

Summary  Previousin vitro studies have shown that salmon calcitonin had direct effects to increase parameters associated with embryonic chicken bone\\u000a formation and to increase mouse and chicken osteoblast-line cell proliferation. The current studies demonstrate increased\\u000a cell proliferation (i.e., [3H]-thymidine incorporation into DNA and tetrazolium salt reduction\\/deposition) in the osteoblastic murine cell line MC-3T3-E1\\u000a in response to salmon calcitonin (PP<0.005), but not

John R. Farley; Susan L. Hall; Nanine M. Tarbaux

1989-01-01

186

Cytotoxic effects of propiconazole and its metabolites in mouse and human hepatoma cells and primary mouse hepatocytes  

EPA Science Inventory

Abstract: Propiconazole is a triazole-containing fungicide that is used agriculturally on grasses, fruits, grains, seeds, hardwoods, and conifers. Propiconazole is a mouse liver hepatotoxicant and a hepatocarcinogen and has adverse reproductive and developmental toxicities in exp...

187

Gene expression of bone morphogenic protein 8B in the primary site, peripheral blood and bone marrow of patients with gastric cancer  

PubMed Central

The prognosis for individuals that are diagnosed with gastric cancer remains poor due to the high frequency of metastatic disease. In response to tumor-derived secreted factors, the bone marrow generates a suitable microenvironment for the development of metastasis. However, it is largely unknown whether secreted factors in bone marrow associated with metastatic disease of patients with gastric cancer are present. Secreted factors from the bone marrow of patients with metastatic gastric cancer were identified using a DNA microarray analysis and the mRNA expression levels were investigated in 355 bone marrow, 295 peripheral blood and 144 primary site samples using quantitative PCR (qPCR). Using DNA microarray analysis, the present study identified bone morphogenetic protein 8B (BMP8B) as a secreted signaling molecule in the bone marrow that was associated with the metastatic disease of human gastric cancer. The expression levels of BMP8B in the bone marrow of 355 gastric cancer patients were increased with metastatic disease. A significant correlation was demonstrated between BMP8B mRNA expression in the bone marrow and in the peripheral blood. High BMP8B expression in the bone marrow was associated with the diffuse type of gastric cancer (P=0.009), lymph node metastasis (P=0.009), liver metastasis (P=0.044) and peritoneal dissemination (P<0.001). In the primary site, a multivariate analysis revealed BMP8B mRNA expression as one of the independent prognostic factors of gastric cancer [hazard ratio (HR), 2.066; 95% CI, 1.132–3.772]. This study suggests that BMP8B, a previously unknown secreted factor in cancer progression, has the potential to be used as a prognostic biomarker. The present study may provide insight into a new mechanism that underlies the dissemination of gastric cancer cells.

MIMA, KOSUKE; FUKAGAWA, TAKEO; KURASHIGE, JUNJI; TAKANO, YUKI; UCHI, RYUTARO; UEO, HIROKI; MATSUMURA, TAE; ISHIBASHI, MASAHISA; SAWADA, GENTA; TAKAHASHI, YUSUKE; AKIYOSHI, SAYURI; EGUCHI, HIDETOSHI; SUDO, TOMOYA; SUGIMACHI, KEISHI; WATANABE, MASAYUKI; ISHII, HIDESHI; MORI, MASAKI; BABA, HIDEO; SASAKO, MITSURU; MIMORI, KOSHI

2013-01-01

188

MOUSE  

NSDL National Science Digital Library

Based in New York City, the MOUSE organization works to empower "underserved students to provide technology support and leadership in their schools, supporting their academic and career success." On their homepage, visitors can learn about their programs, learn about supporting the MOUSE organization, and read up on their resources. In the "Resources" area, visitors can learn about their outreach activities in New York City, Chicago, and California. Visitors working in educational outreach will appreciate the information offered here, including materials on how different groups can receive assistance from the MOUSE organization. Also, visitors can look over the "News" updates to learn about their new programs, their educational seminars, and their outreach activities.

189

New bone formation in nude mouse calvaria induced by canine prostate tissue.  

PubMed

Osteoblastic metastases are common in patients with advanced prostate cancer. The pathophysiology of the new bone formation at metastatic sites is not currently known, but it is hypothesized that growth factors secreted by the prostate may be involved. Unfortunately, most rodent models of prostate cancer with metastasis to bone are osteolytic and not osteoblastic. Significant osteolysis by tumor cells at metastatic sites also may lead to fractures or bone instability. Misinterpretation of new periosteal bone due to bone instability as tumor-cell osteo-induction is another disadvantage of the osteolytic models. To circumvent these problems, we have developed a model system of new bone formation in the calvaria of nude mice stimulated by normal canine prostate tissue. Collagenase-digested normal prostate tissue was implanted adjacent to the calvaria of nude mice. Calvaria were examined at 2 weeks post-implantation for changes in the bone microenvironment by histology, calcein uptake at sites of bone mineralization, and tartrate-resistant acid phosphatase staining for osteoclasts. The prostate tissue remained viable and induced abundant new woven bone formation on the adjacent periosteal surface. In some cases new bone formation also was induced on the distant or concave calvarial periosteum. The new bone stained intensely with calcein, which demonstrated mineralization of the bone matrix. The new bone formation on prostate-implanted calvaria significantly increased (1.7-fold) the thickness of the calvaria compared with control calvaria. New bone formation was not induced in calvaria of mice implanted with normal canine kidney, urinary bladder, spleen, or skeletal muscle tissue, or mice with surgically-induced disruption of the periosteum. Osteoclast numbers in the medullary spaces and periosteum of calvaria were mildly increased (61%) in mice with implanted prostate tissue. In conclusion, this animal model will be useful for investigating the roles of prostate-derived growth factors on new bone formation in vivo. PMID:12431820

LeRoy, Bruce E; Bahnson, Robert R; Rosol, Thomas J

2002-11-29

190

Late malignant transformation of giant cell tumor of bone 41 years after primary surgery.  

PubMed

Giant cell tumor of bone is an uncommon benign tumor that frequently recurs locally. Spontaneous malignant transformation of conventional giant cell tumor of bone is rare and usually occurs with irradiation.This article describes a case of malignant transformation of a giant cell tumor 41 years after initial curettage and subsequent resection. A 68-year-old man presented with a 6-month history of left hip pain. He had been diagnosed 41 years previously with giant cell tumor in the left femoral neck treated by simple curettage and bone grafting, followed by resection of the femoral head 1 year later for local recurrence. On presentation, radiographs revealed a destructive lesion in the left proximal femur. Incisional biopsy revealed recurrence of giant cell tumor with suspected malignant transformation. The patient underwent en bloc resection of the proximal femur with adequately wide margins and reconstruction of the hip joint with a prosthesis. Pathological findings showed malignant transformation of a giant cell tumor to osteosarcoma and leiomyosarcoma. No recurrence or metastasis developed during 2-year follow-up. Benign local recurrences usually arise in the first 3 postoperative years, whereas malignant transformation tends to take longer than 3 years. To the authors' knowledge, the 41-year interval from primary surgery to diagnosis of malignancy for the current patient is the longest interval reported among cases in which patients received no radiation therapy. PMID:23027500

Kadowaki, Masaru; Yamamoto, Soichiro; Uchio, Yuji

2012-10-01

191

Primary culture of cellular subtypes from postnatal mouse for in vitro studies of oxygen glucose deprivation.  

PubMed

One of the most widely utilized in vitro models of ischemia or oxygen glucose deprivation (OGD) is the hippocampal organotypical culture (HOTC). The HOTC is used not only for the study of the mechanisms of cell death, but also has been the cornerstone of synaptic physiology. Although the intact nature of the HOTC is one of its primary advantages, some studies require a dissociated preparation in order to distinguish cell type specific responses. Typically, primary dissociated neuronal cultures are prepared from embryonic tissue. Since the HOTC is prepared from postnatal pups, we wanted to establish a primary culture of hippocampus from postnatal pups to parallel our studies in the HOTC preparation. Mixed cultures were prepared by enzymatic dissociation of hippocampus from 7-day-old mouse pups. These cultures responded to OGD with a time course of delayed cell death that was similar to that reported in HOTC. Dual label immunocytochemical staining revealed that neurons, but not astrocytes, were dying from apoptosis following OGD. To examine this vulnerability further, we also prepared neuronal enriched cultures by treating mixed cultures with cytosine-?-d-arabinofuranoside (CBA). These neuronal cultures appear to be even more sensitive to OGD. In addition, we have established primary astrocyte-enriched cultures from the same age pups to examine the vulnerability of astrocytes to OGD. These three culture preparations are useful for comparison of the responses of the two major cell types in the same culture, and the enriched cultures will allow biochemical, electrophysiological and molecular studies of homogenous cell populations. PMID:21620892

Jones, Susan M; Novak, Alicia E; Elliott, J Paul

2011-05-19

192

Radionuclide bone scanning in neuroblastoma: skeletal metastases and primary tumor localization of /sup 99m/Tc-MDP  

SciTech Connect

Of 42 radionuclide bone scans in 35 children with neuroblastoma, 21 were abnormal for the presence of skeletal metastases. Of the 21 abnormal scans, 16 were corroborated by positive bone-marrow biopsy or clinical data. The false-negative and false-positive rates for bone scanning were 4.8% and 9.5%, respectively. Calcification of the primary tumor was seen on pretreatment computed tomographic (CT) scans in 24 (89%) of 27 cases, while only 13 (48%) of 27 were detectable by plain radiographs. Uptake of /sup 99m/Tc methylene diphosphate /sup 99m/Tc-MDP) by the primary tumor occurred in 20 of 27 cases, but correlation between tumor uptake and calcification was not statistically significant. All children with markedly elevated urinary vanillylmandelic acid exhibited primary tumor uptake. Survival was not affected independently by primary tumor uptake.

Podrasky, A.E.; Stark, D.D.; Hattner, R.S.; Gooding, C.A.; Moss, A.A.

1983-09-01

193

Accuracy of Peripheral Quantitative Computed Tomography (pQCT) For Assessing Area and Density of Mouse Cortical Bone  

Microsoft Academic Search

Peripheral quantitative computed tomography (pQCT) is increasingly used for measurement of cortical bone geometry and density in mice. We evaluated the accuracy of pQCT for area and density measurements of thin-walled aluminum phantoms and mouse femora. Aluminum tubes with varying wall thicknesses and femora from 1- to 6-month-old C3H\\/HeJ (C3H) and C57B1\\/6J (B6) mice (average cortical thickness 0.14–0.29 mm) were

M. D. Brodt; G. B. Pelz; J. Taniguchi; M. J. Silva

2003-01-01

194

Effects of phorbol esters and pertussis toxin on calcitonin-stimulated accumulation of cyclic AMP in neonatal mouse calvarial bones  

Microsoft Academic Search

Summary  Calcitonin (CT) is a well-known inhibitor of osteoclastic bone resorption bothin vivo andin vitro. The effect is mediated by activation of adenylate cyclase and subsequent increased levels of cyclic AMP (cAMP). We report\\u000a here that CT-induced (30 nmol\\/liter) accumulation of cAMP in cultured neonatal mouse calvaria is enhanced two-fold by 12-O-tetradecanoylphorbol-13-acetate\\u000a (TPA; 100 nmol\\/liter) and phorbol 12,13-dibutyrate (PDBU; 100 nmol\\/liter),

Maria Ransjö; Ulf H. Lerner

1991-01-01

195

Dura mater stimulates human adipose-derived stromal cells to undergo bone formation in mouse calvarial defects.  

PubMed

Human adipose-derived stromal cells (hASCs) have a proven capacity to aid in osseous repair of calvarial defects. However, the bone defect microenvironment necessary for osseous healing is not fully understood. In this study, we postulated that the cell-cell interaction between engrafted ASCs and host dura mater (DM) cells is critical for the healing of calvarial defects. hASCs were engrafted into critical sized calvarial mouse defects. The DM-hASC interaction was manipulated surgically by DM removal or by insertion of a semipermeable or nonpermeable membrane between DM and hASCs. Radiographic, histologic, and gene expression analyses were performed. Next, the hASC-DM interaction is assessed by conditioned media (CM) and coculture assays. Finally, bone morphogenetic protein (BMP) signaling from DM was investigated in vivo using novel BMP-2 and anti-BMP-2/4 slow releasing scaffolds. With intact DM, osseous healing occurs both from host DM and engrafted hASCs. Interference with the DM-hASC interaction dramatically reduced calvarial healing with abrogated BMP-2-Smad-1/5 signaling. Using CM and coculture assays, mouse DM cells stimulated hASC osteogenesis via BMP signaling. Through in vivo manipulation of the BMP-2 pathway, we found that BMP-2 plays an important role in DM stimulation of hASC osteogenesis in the context of calvarial bone healing. BMP-2 supplementation to a defect with disrupted DM allowed for bone formation in a nonhealing defect. DM is an osteogenic cell type that both participates in and stimulates osseous healing in a hASC-engrafted calvarial defect. Furthermore, DM-derived BMP-2 paracrine stimulation appears to play a key role for hASC mediated repair. PMID:21656608

Levi, Benjamin; Nelson, Emily R; Li, Shuli; James, Aaron W; Hyun, Jeong S; Montoro, Daniel T; Lee, Min; Glotzbach, Jason P; Commons, George W; Longaker, Michael T

2011-08-01

196

Effect of taxol and okadaic acid on microtubule dynamics in thimerosal-arrested primary mouse oocytes: a confocal study  

Microsoft Academic Search

A pulse of thimerosal (TMS), a sulfhydryl reagent, induces an instantaneous, complete and long-lasting microtubule interphasic network disassembly in mouse primary oocytes, correlated with the irreversible inhibition of meiosis reinitiation This inhibition is bypassed by dithiothreitol (DTT) while thiosalicylic acid, an analog of TMS, does induce neither microtubules depolymerisation nor inhibition of reinitiation and resumption of meiosis. This strongly suggests

H. Alexandre; V. Delsinne; J.-J. Goval; A. Van Cauwenberge

2003-01-01

197

Hydrocortisone effect of arylsulfatase A in primary mouse brain cell cultures  

SciTech Connect

The primary goal of this study was to study the mechanism of action of hydrocortisone (HC) on arylsulfatase A (ASA) in primary cultures of cells that were dissociated from the brains of embryonic mice. Cells were cultured in a defined medium in the absence or in the presence of 3 ..mu..M HC. The specific activity of ASA in nontreated cells was 1.297 U/mg (U = ..mu..mol/hr) while the value for the HC-treated cells was 0.783 U/mg. The authors data shows that HC inhibits ASA activity in these cultures cells (p < 0.001). The determination of the ASA enzyme activity was assayed primarily with the artificial substrate p-nitrocatechol sulfate. However, the natural substrate (cerebroside /sup 35/S-sulfate) also as active and correlated linearly with the activity of p-nitrocatechol sulfate. Purified ASA was isolated from calf brains and used to generate an antibody (Ab) against ASA. The specificity of the Ab for the ASA protein of cell cultures was tested in Ouchterlony double immunodiffusion studies. The Ab was used in a competitive enzyme-linked immunosorbent assay to quantify the number of ASA molecules in the cell extracts from the embryonic mouse cell cultures. Preliminary data suggest that HC decreases the number of ASA molecules.

Marcelo, A.; Pieringer, R.A.

1986-05-01

198

Primary culture of adult mouse lung fibroblasts in serum-free medium: responses to growth factors.  

PubMed

We report a completely serum-free system for primary culture of fibroblasts from explants of adult mouse lung tissue which permits bioassays for cytokine activity to be performed using unselected populations of cells at low passage number, without interference by serum binding proteins or interacting growth factors. Cultures were established on collagen-coated surfaces in medium MCDB 201 containing albumin, transferrin, epidermal growth factor, lipids, prostaglandin E1, vitamin E, and reducing agents. The cells were morphologically and ultrastructurally typical of fibroblasts in culture and demonstrated expression of vimentin and induction of expression of desmin in culture. Proliferation of the cells was reproducible between different primary cultures and was growth factor dependent. Both cycling and growth-arrested cells exhibited increased DNA synthesis when stimulated with epidermal growth factor, platelet-derived growth factor, or basic fibroblast growth factor, which functioned as complete mitogens, but did not respond to insulin, tumor necrosis factor or interleukin-1 beta. Maximal induction of DNA synthesis by epidermal growth factor required the continued presence of the mitogen in the culture medium. These results cannot be satisfactorily explained by the competence-progression model of responses to mitogenic stimuli but support and extend the findings of other studies using diploid fibroblasts. PMID:2004652

Kumar, R K; O'Grady, R; Li, W; Smith, L W; Rhodes, G C

1991-04-01

199

Role of PTH1R internalization in osteoblasts and bone mass using a phosphorylation-deficient knock-in mouse model  

PubMed Central

Phosphorylation, internalization, and desensitization of G protein-coupled receptors, such as the parathyroid hormone (PTH) and PTH-related peptide (PTHrP) receptor (PTH1R), are well characterized and known to regulate the cellular responsiveness in vitro. However, the role of PTH1R receptor phosphorylation in bone formation and osteoblast functions has not yet been elucidated. In previous studies, we demonstrated impaired internalization and sustained cAMP stimulation of a phosphorylation-deficient (pd) PTH1R in vitro, and exaggerated cAMP and calcemic responses to s.c. PTH infusion in pdPTH1R knock-in mouse model. In this study, we examined the impact of impaired PTH1R phosphorylation on the skeletal phenotype of mice maintained on normal, low, and high calcium diets. The low calcium diet moderately reduced (P<0.05) bone volume and trabecular number, and increased trabecular spacing in both wild-type (WT) and pd mice. The effects, however, seem to be less pronounced in the female pd compared to WT mice. In primary calvarial osteoblasts isolated from 2-week-old pd or WT mice, PTH and PTHrP decreased phosphorylated extracellular signal-regulated kinases 1/2 (pERK1/2), a member of mitogen-activated protein kinase, and cyclin D1, a G1/S phase cyclin, in vitro. In contrast to WT osteoblasts, down-regulation of cyclin D1 was sustained for longer periods of time in osteoblasts isolated from the pd mice. Our results suggest that adaptive responses of intracellular signaling pathways in the pd mice may be important for maintaining bone homeostasis.

Datta, Nabanita S; Samra, Tareq A; Mahalingam, Chandrika D; Datta, Tanuka; Abou-Samra, Abdul B

2013-01-01

200

Effects and differentiation activity of IGF-I, IGF-II, insulin and preptin on human primary bone cells.  

PubMed

The importance of the complex interrelated regulatory pathways involving IGF factors and pancreatic hormones can be observed in several metabolic diseases, where the deregulation of these factors has a wide impact on bone health. These findings have stimulated us to compare the effect of IGF-I, IGF-II, insulin and preptin on human bone cells. The effect on cell differentiation and cell activity of osteoblasts and osteoclasts has been analysed. We have observed a significant effect by IGF-I, a modest effect by IGF-II and preptin and no effect after insulin administration on human primary osteoblast-like cells. All studied factors have shown an induction on human primary osteoclast differentiation and bone resorption activity, with IGF-I being the most potent factor. We hypothesize that these findings may be on the basis of decreased bone mass density observed in several diseases. PMID:23410103

Bosetti, Michela; Sabbatini, Maurizio; Nicolì, Elena; Fusaro, Luca; Cannas, Mario

2013-02-15

201

Therapeutic impact of low amplitude high frequency whole body vibrations on the osteogenesis imperfecta mouse bone?  

PubMed Central

Osteogenesis imperfecta (OI) is characterized by extremely brittle bone. Currently, bisphosphonate drugs allow a decrease of fracture by inhibiting bone resorption and increasing bone mass but with possible long term side effects. Whole body mechanical vibrations (WBV) treatment may offer a promising route to stimulate bone formation in OI patients as it has exhibited health benefits on both muscle and bone mass in human and animal models. The present study has investigated the effects of WBV (45 Hz, 0.3 g, 15 minutes/days, 5 days/week) in young OI (oim) and wild type female mice from 3 to 8 weeks of age. Vibration therapy resulted in a significant increase in the cortical bone area and cortical thickness in the femur and tibia diaphysis of both vibrated oim and wild type mice compared to sham controls. Trabecular bone was not affected by vibration in the wild type mice; vibrated oim mice, however, exhibited significantly higher trabecular bone volume fraction in the proximal tibia. Femoral stiffness and yield load in three point bending were greater in the vibrated wild type mice than in sham controls, most likely attributed to the increase in femur cortical cross sectional area observed in the ?CT morphology analyses. The vibrated oim mice showed a trend toward improved mechanical properties, but bending data had large standard deviations and there was no significant difference between vibrated and non-vibrated oim mice. No significant difference of the bone apposition was observed in the tibial metaphyseal trabecular bone for both the oim and wild type vibrated mice by histomorphometry analyses of calcein labels. At the mid diaphysis, the cortical bone apposition was not significantly influenced by the WBV treatment in both the endosteum and periosteum of the oim vibrated mice while a significant change is observed in the endosteum of the vibrated wild type mice. As only a weak impact in bone apposition between the vibrated and sham groups is observed in the histological sections, it is possible that WBV reduced bone resorption, resulting in a relative increase in cortical thickness. Whole body vibration appears as a potential effective and innocuous means for increasing bone formation and strength, which is particularly attractive for treating the growing skeleton of children suffering from brittle bone disease or low bone density pathologies without the long term disadvantages of current pharmacological therapies.

Vanleene, Maximilien; Shefelbine, Sandra J.

2013-01-01

202

Therapeutic impact of low amplitude high frequency whole body vibrations on the osteogenesis imperfecta mouse bone.  

PubMed

Osteogenesis imperfecta (OI) is characterized by extremely brittle bone. Currently, bisphosphonate drugs allow a decrease of fracture by inhibiting bone resorption and increasing bone mass but with possible long term side effects. Whole body mechanical vibrations (WBV) treatment may offer a promising route to stimulate bone formation in OI patients as it has exhibited health benefits on both muscle and bone mass in human and animal models. The present study has investigated the effects of WBV (45Hz, 0.3g, 15minutes/days, 5days/week) in young OI (oim) and wild type female mice from 3 to 8weeks of age. Vibration therapy resulted in a significant increase in the cortical bone area and cortical thickness in the femur and tibia diaphysis of both vibrated oim and wild type mice compared to sham controls. Trabecular bone was not affected by vibration in the wild type mice; vibrated oim mice, however, exhibited significantly higher trabecular bone volume fraction in the proximal tibia. Femoral stiffness and yield load in three point bending were greater in the vibrated wild type mice than in sham controls, most likely attributed to the increase in femur cortical cross sectional area observed in the ?CT morphology analyses. The vibrated oim mice showed a trend toward improved mechanical properties, but bending data had large standard deviations and there was no significant difference between vibrated and non-vibrated oim mice. No significant difference of the bone apposition was observed in the tibial metaphyseal trabecular bone for both the oim and wild type vibrated mice by histomorphometry analyses of calcein labels. At the mid diaphysis, the cortical bone apposition was not significantly influenced by the WBV treatment in both the endosteum and periosteum of the oim vibrated mice while a significant change is observed in the endosteum of the vibrated wild type mice. As only a weak impact in bone apposition between the vibrated and sham groups is observed in the histological sections, it is possible that WBV reduced bone resorption, resulting in a relative increase in cortical thickness. Whole body vibration appears as a potential effective and innocuous means for increasing bone formation and strength, which is particularly attractive for treating the growing skeleton of children suffering from brittle bone disease or low bone density pathologies without the long term disadvantages of current pharmacological therapies. PMID:23352925

Vanleene, Maximilien; Shefelbine, Sandra J

2013-01-22

203

Characterization of phenobarbital-inducible mouse Cyp2b10 gene transcription in primary hepatocytes.  

PubMed

The mouse phenobarbital (PB)-inducible Cyp2b10 gene promoter has been isolated and sequenced, and control of its expression has been characterized. The 1405-base pair (bp) Cyp2bl0 promoter sequence is 83% identical to the corresponding region from the rat CYP2B2 gene. In addition to the lack of CA repeats, differences include insertion of 42 base pairs (-123/-82 bp) into the middle of a consensus sequence to the so-called "Barbie box." In this report, we have developed a primary mouse hepatocyte culture system in which endogenous 2B10 mRNA as well as Cyp2b10-driven CAT activity were induced by PB and 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP), but not by the 3-chloro derivative of TCPOBOP. Deletion analysis of the Cyp2b10 promoter identified a basal transcription element at -64/-34 bp and a negative element at -971/-775 bp. Sequences contained within the -1404/-971 bp region are responsible for the induced CAT activity. DNase I protection and gel shift assays detected five major protein binding sites within the -1404/-971 bp fragment, one of which shared high sequence identity with a portion of a regulatory element in CYP2B2 gene (Trottier, E., Belzil, A., Stoltz, C., and Anderson, A. (1995) Gene 158, 263-268). Our results indicate that sequences important for PB-induced transcription of Cyp2b10 gene are located in the distal promoter. PMID:8621653

Honkakoski, P; Moore, R; Gynther, J; Negishi, M

1996-04-19

204

Cinacalcet hydrochloride in combination with alendronate normalizes hypercalcemia and improves bone mineral density in patients with primary hyperparathyroidism  

Microsoft Academic Search

Cinacalcet is effective in controlling the biochemical abnormalities in patients with primary hyperparathyroidism (PHPT) but\\u000a it seems to be less effective on bone mineral density (BMD). In the same patients, bisphosphonates are reported to be effective\\u000a on bone resorption but less effective on calcium and PTH excess. In this study, the efficacy of cinacalcet in combination\\u000a with alendronate has been

A. Faggiano; C. Di Somma; V. Ramundo; R. Severino; L. Vuolo; A. Coppola; F. Panico; S. Savastano; G. Lombardi; A. Colao; M. Gasperi

2011-01-01

205

Co-culture of canine mesenchymal stem cells with primary bone-derived osteoblasts promotes osteogenic differentiation  

Microsoft Academic Search

Tissue engineering of bone grafts with osteogenic progenitor cells such as adult mesenchymal stem cells (MSC) represents a\\u000a promising strategy for the treatment of large bone defects. The aim of this experimental study was to evaluate the osteogenic\\u000a potential of primary osteoblasts on MSCs in co-culture at different ratios. The co-cultures were treated with or without a\\u000a specific osteogenic induction

C. Csaki; U. Matis; A. Mobasheri; M. Shakibaei

2009-01-01

206

Vitamin D Deficiency Influences Histomorphometric Features of Bone in Primary Hyperparathyroidism  

PubMed Central

Introduction Vitamin D deficiency is common in patients with primary hyperparathyroidism (PHPT). The presence of low levels of vitamin D may affect the skeletal consequences of PHPT. Methods In this cross-sectional study, transiliac crest bone biopsies were performed after double tetracycline labeling in patients with mild PHPT and analyzed according to serum levels of 25 hydroxyvitamin D (25OHD). Results We studied 30 patients with mild PHPT (age 53±11 years; 67% women; calcium 11.1±1.0 mg/dl; PTH 149±129 pg/ml). Serum 25OHD levels were low in the majority of subjects (mean 21±11 ng/ml) and inversely associated with PTH (r=-0.69; p<0.01). 25OHD levels were directly associated with cortical width (Ct.Wi; r=0.46, p<0.03) and trabecular separation (Tb.Sp; r=0.41; p<0.04), but inversely associated with cancellous bone volume (BV/TV; r=-0.39, p<0.04). Subjects with 25OHD levels <20 ng/ml (n=14) and ?20 ng/ml (n=16) were compared. Groups did not differ by age, sex, menopausal status, serum calcium, creatinine, or 1,25(OH)2D. PTH was 1.8-fold higher in subjects with 25OHD <20 (265±166 pg/ml vs. 95±50 pg/ml; p <0.01). On histomorphometric analysis, those with low 25OHD had lower Ct.Wi (541±167 ?m vs. 712±200 ?m; p<0.03). Conversely, measures of trabecular microarchitecture were better in those with lower 25OHD, with higher BV/TV (26.1±6.1 % vs. 20.4±6.4 %; p<0.03), greater trabecular number (Tb.N: 2.0±0.4 mm -1 vs. 1.8±0.4 mm -1; p<0.04) and lower Tb.Sp (371±90 ?m vs. 472±137 ?m; p<0.04). There were no differences between the groups in bone remodeling indices. Conclusions Low levels of 25OHD in patients with PHPT are associated with higher concentrations of PTH, greater catabolic effects in cortical bone and greater anabolic effects in trabecular bone.

Stein, Emily M.; Dempster, David W.; Udesky, Julia; Zhou, Hua; Bilezikian, John P.; Shane, Elizabeth; Silverberg, Shonni J.

2010-01-01

207

Cytotoxic effects of propiconazole and its metabolites in mouse and human hepatoma cells and primary mouse hepatocytes  

Microsoft Academic Search

Propiconazole is a triazole-containing fungicide that is used agriculturally on grasses, fruits, grains, seeds, hardwoods, and conifers. Propiconazole is a mouse liver hepatotoxicant and a hepatocarcinogen that has adverse reproductive and developmental toxicities in experimental animals. The goal of this study was to investigate the cytotoxic responses of propiconazole and its metabolites to determine if metabolism of this agent differentially

Pei-Jen Chen; Tanya Moore; Stephen Nesnow

2008-01-01

208

Primary hyperparathyroidism diagnosed after surgical ablation of a costal mass mistaken for giant-cell bone tumor: a case report  

PubMed Central

Introduction Primary hyperparathyroidism is a common endocrine disorder characterized by elevated parathyroid hormone levels, which cause continuous osteoclastic bone resorption. Giant cell tumor of bone is an expansile osteolytic tumor that contains numerous osteoclast-like giant cells. There are many similarities in the radiological and histological features of giant cell tumor of bone and brown tumor. This is a rare benign focal osteolytic process most commonly caused by hyperparathyroidism. Case presentation We report the unusual case of a 40-year-old Caucasian woman in which primary hyperparathyroidism was diagnosed after surgical ablation of a costal mass. The mass was suspected of being neoplastic and histopathology was compatible with a giant cell tumor of bone. On the basis of the biochemical results (including serum calcium, phosphorous and intact parathyroid hormone levels) primary hyperparathyroidism was suspected and a brown tumor secondary to refractory hyperparathyroidism was diagnosed. Conclusions Since giant cell tumor is a bone neoplasm that has major implications for the patient, the standard laboratory tests in patients with bone lesions are important for a correct diagnosis.

2011-01-01

209

Characterization of xenogeneic mouse-to-rat bone marrow chimeras. I. Examination of hematologic and immunologic function  

SciTech Connect

Eighteen xenogeneic chimeric rats (survival: greater than 100 days) were established by transplanting bone marrow cells from femurs of 10 gnotobiotic CFW mice into each germfree Sprague-Dawley or Wistar rat. The erythrocytes circulating in the rats were of mouse origin as determined by hemagglutination. Hemoglobin electrophoresis, radial immunodiffusion for IgG, and assay of granulocytic neutrophils for leukocyte alkaline phosphatase verified that true chimerism was achieved. The extent of hematological and immunological reconstitution varied. In general, hematocrit levels were low to normal, white blood cell counts and differentials were within normal limits, and serum protein levels were normal. Levels of circulating IgG of each species were comparable to those of germfree rat and mouse controls. Natural killer (NK) activity was depressed, a phenomenon that may be attributable to the radiation treatment of recipients, or to failure to transfer NK cells or precursors. Mitogenic stimulation reactions were varied, but most chimeric rats demonstrated moderately depressed responses. Reactions as a whole suggested that gnotobiotic rats with xenogeneic bone marrow are incompletely reconstituted, both hematologically and immunologically. No acute graft-versus-host reaction was seen.

Wade, A.C.; Luckert, P.H.; Tazume, S.; Niedbalski, J.L.; Pollard, M.

1987-07-01

210

Relief of Preintegration Inhibition and Characterization of Additional Blocks for HIV Replication in Primary Mouse T Cells  

PubMed Central

Development of a small animal model to study HIV replication and pathogenesis has been hampered by the failure of the virus to replicate in non-primate cells. Most studies aimed at achieving replication in murine cells have been limited to fibroblast cell lines, but generating an appropriate model requires overcoming blocks to viral replication in primary T cells. We have studied HIV-1 replication in CD4+ T cells from human CD4/ CCR5/Cyclin T1 transgenic mice. Expression of hCD4 and hCCR5 in mouse CD4+ T cells enabled efficient entry of R5 strain HIV-1. In mouse T cells, HIV-1 underwent reverse transcription and nuclear import as efficiently as in human T cells. In contrast, chromosomal integration of HIV-1 proviral DNA was inefficient in activated mouse T cells. This process was greatly enhanced by providing a secondary T cell receptor (TCR) signal after HIV-1 infection, especially between 12 to 24 h post infection. This effect was specific for primary mouse T cells. The pathways involved in HIV replication appear to be PKC??, CARMA1-, and WASp-independent. Treatment with Cyclosporin A (CsA) further relieved the pre-integration block. However, transcription of HIV-1 RNA was still reduced in mouse CD4+ T cells despite expression of the hCyclin T1 transgene. Additional post-transcriptional defects were observed at the levels of Gag expression, Gag processing, Gag release and virus infectivity. Together, these post-integration defects resulted in a dramatically reduced yield of infectious virus (300–500 fold) after a single cycle of HIV-1 replication. This study implies the existence of host factors, in addition to those already identified, that are critical for HIV-1 replication in mouse cells. This study also highlights the differences between primary T cells and cell lines regarding pre-integration steps in the HIV-1 replication cycle.

Zhang, Jing-xin; Diehl, Gretchen E.; Littman, Dan R.

2008-01-01

211

INHIBITION OF DIRECT BONE FORMATION ASSOCIATED WITH CHRONIC ETHANOL EXPOSURE IN A MOUSE MODEL OF DISTRACTIONOSTEOGENESIS  

Technology Transfer Automated Retrieval System (TEKTRAN)

Excessive alcohol consumption has been reported to interfere with human bone homeostasis and repair in multiple ways. Previous studies have demonstrated that chronic ethanol (EtOH) exposure in the rat inhibits direct bone formation during distraction osteogenesis (DO, limb lengthening). The opportun...

212

Upregulating CXCR4 in Human Fetal Mesenchymal Stem Cells Enhances Engraftment and Bone Mechanics in a Mouse Model of Osteogenesis Imperfecta  

PubMed Central

Stem cells have considerable potential to repair damaged organs and tissues. We previously showed that prenatal transplantation of human first trimester fetal blood mesenchymal stem cells (hfMSCs) in a mouse model of osteogenesis imperfecta (oim mice) led to a phenotypic improvement, with a marked decrease in fracture rate. Donor cells differentiated into mature osteoblasts, producing bone proteins and minerals, including collagen type I?2, which is absent in nontransplanted mice. This led to modifications of the bone matrix and subsequent decrease of bone brittleness, indicating that grafted cells directly contribute to improvement of bone mechanical properties. Nevertheless, the therapeutic effect was incomplete, attributing to the limited level of engraftment in bone. In this study, we show that although migration of hfMSCs to bone and bone marrow is CXCR4-SDF1 (SDF1 is stromal-derived factor) dependent, only a small number of cells present CXCR4 on the cell surface despite high levels of internal CXCR4. Priming with SDF1, however, upregulates CXCR4 to increase the CXCR4+ cell fraction, improving chemotaxis in vitro and enhancing engraftment in vivo at least threefold in both oim and wild-type bone and bone marrow. Higher engraftment in oim bones was associated with decreased bone brittleness. This strategy represents a step to improve the therapeutic benefits of fetal cell therapy toward being curative.

Jones, Gemma N.; Moschidou, Dafni; Lay, Kenneth; Abdulrazzak, Hassan; Vanleene, Maximilien; Shefelbine, Sandra J.; Polak, Julia; de Coppi, Paolo; Fisk, Nicholas M.

2012-01-01

213

Activation and Anergy in Bone Marrow B Cells of a Novel Immunoglobulin Transgenic Mouse that Is Both Hapten Specific and Autoreactive  

Microsoft Academic Search

Available evidence indicates that B cell tolerance is attained by receptor editing, anergy, or clonal deletion. Here, we describe a p-azophenylarsonate (Ars)-specific immunoglobulin transgenic mouse in which B cells become anergic as a consequence of cross-reaction with autoantigen in the bone marrow. Developing bone marrow B cells show no evidence of receptor editing but transiently upregulate activation markers and appear

Robert J Benschop; Katja Aviszus; Xianghua Zhang; Tim Manser; John C Cambier; Lawrence J Wysocki

2001-01-01

214

Chitosan-poly(butylene succinate) scaffolds and human bone marrow stromal cells induce bone repair in a mouse calvaria model  

Microsoft Academic Search

Tissue engineering sustains the need of a three-dimensional (3D)\\u000d\\u000a scaffold to promote the regeneration of tissues in volume. Usually,\\u000d\\u000a scaffolds are seeded with an adequate cell population, allowing their\\u000d\\u000a growth and maturation upon implantation in vivo. Previous studies\\u000d\\u000a obtained by our group evidenced significant growth patterns and\\u000d\\u000a osteogenic differentiation of human bone marrow mesenchymal stem cells\\u000d\\u000a (hBMSCs) when seeded and

A. R. Costa-Pinto; V. M. Correlo; P. C. Sol; M. Bhattacharya; S. Srouji; E. Livne; R. L. Reis; N. M. Neves

2012-01-01

215

Evaluation of the correlation between insertion torque and primary stability of dental implants using a block bone test  

PubMed Central

Purpose Implant stability at the time of surgery is crucial for the long-term success of dental implants. Primary stability is considered of paramount importance to achieve osseointegration. The purpose of the present study was to investigate the correlation between the insertion torque and primary stability of dental implants using artificial bone blocks with different bone densities and compositions to mimic different circumstances that are encountered in routine daily clinical settings. Methods In order to validate the objectives, various sized holes were made in bone blocks with different bone densities (#10, #20, #30, #40, and #50) using a surgical drill and insertion torque together with implant stability quotient (ISQ) values that were measured using the Osstell Mentor. The experimental groups under evaluation were subdivided into 5 subgroups according to the circumstances. Results In group 1, the mean insertion torque and ISQ values increased as the density of the bone blocks increased. For group 2, the mean insertion torque values decreased as the final drill size expanded, but this was not the case for the ISQ values. The mean insertion torque values in group 3 increased with the thickness of the cortical bone, and the same was true for the ISQ values. For group 4, the mean insertion torque values increased as the cancellous bone density increased, but the correlation with the ISQ values was weak. Finally, in group 5, the mean insertion torque decreased as the final drill size increased, but the correlation with the ISQ value was weak. Conclusions Within the limitations of the study, it was concluded that primary stability does not simply depend on the insertion torque, but also on the bone quality.

Bayarchimeg, Dorjpalam; Namgoong, Hee; Kim, Byung Kook; Kim, Myung Duk; Kim, Sungtae; Kim, Tae-Il; Seol, Yang Jo; Lee, Yong Moo; Ku, Young; Rhyu, In-Chul; Lee, Eun Hee

2013-01-01

216

Differential gene expression in mouse primary hepatocytes exposed to the peroxisome proliferator-activated receptor ? agonists  

PubMed Central

Background Fibrates are a unique hypolipidemic drugs that lower plasma triglyceride and cholesterol levels through their action as peroxisome proliferator-activated receptor alpha (PPAR?) agonists. The activation of PPAR? leads to a cascade of events that result in the pharmacological (hypolipidemic) and adverse (carcinogenic) effects in rodent liver. Results To understand the molecular mechanisms responsible for the pleiotropic effects of PPAR? agonists, we treated mouse primary hepatocytes with three PPAR? agonists (bezafibrate, fenofibrate, and WY-14,643) at multiple concentrations (0, 10, 30, and 100 ?M) for 24 hours. When primary hepatocytes were exposed to these agents, transactivation of PPAR? was elevated as measured by luciferase assay. Global gene expression profiles in response to PPAR? agonists were obtained by microarray analysis. Among differentially expressed genes (DEGs), there were 4, 8, and 21 genes commonly regulated by bezafibrate, fenofibrate, and WY-14,643 treatments across 3 doses, respectively, in a dose-dependent manner. Treatments with 100 ?M of bezafibrate, fenofibrate, and WY-14,643 resulted in 151, 149, and 145 genes altered, respectively. Among them, 121 genes were commonly regulated by at least two drugs. Many genes are involved in fatty acid metabolism including oxidative reaction. Some of the gene changes were associated with production of reactive oxygen species, cell proliferation of peroxisomes, and hepatic disorders. In addition, 11 genes related to the development of liver cancer were observed. Conclusion Our results suggest that treatment of PPAR? agonists results in the production of oxidative stress and increased peroxisome proliferation, thus providing a better understanding of mechanisms underlying PPAR? agonist-induced hepatic disorders and hepatocarcinomas.

Guo, Lei; Fang, Hong; Collins, Jim; Fan, Xiao-hui; Dial, Stacey; Wong, Alex; Mehta, Kshama; Blann, Ernice; Shi, Leming; Tong, Weida; Dragan, Yvonne P

2006-01-01

217

myo-inositol transport in mouse astroglia-rich primary cultures.  

PubMed

Uptake of radiolabeled myo-inositol was studied in astroglia-rich primary cultures derived from neonatal mouse brains. The uptake was saturable in the presence of Na+ with a Km of 25 microM and a Vmax of 60 pmol.min-1.(mg protein)-1, suggesting a high-affinity transport system for myo-inositol in astroglial cells. In addition, a Na(+)-independent, nonsaturable component was found. Carrier-mediated uptake was not inhibited by cytochalasin B (50 microM), but was reduced by depolarizing concentrations of K+ and, to different extents, in the presence of phloretin, ouabain, or amiloride (1 mM each). scyllo-Inositol, glucose, and galactose also reduced myo-inositol uptake; inhibition by the two hexoses was not reversed in the presence of 0.4 mM sorbinil. On the other hand, uptake of 2-deoxyglucose was not inhibited by high concentrations of myo-inositol. Preincubation of the cells with glucose-free or inositol-free medium stimulated uptake of myo-inositol and preincubation with 25 mM glucose in the presence of 0.4 mM sorbinil had no effect on the rate of uptake. The results suggest that myo-inositol is taken up into the astroglial cells by a transport mechanism that is distinct from that of glucose and probably is an active one. Sorbitol pathway activity does not interfere with myo-inositol uptake. PMID:2013761

Wiesinger, H

1991-05-01

218

Biocompatibility effects of biologically synthesized graphene in primary mouse embryonic fibroblast cells  

NASA Astrophysics Data System (ADS)

Due to unique properties and unlimited possible applications, graphene has attracted abundant interest in the areas of nanobiotechnology. Recently, much work has focused on the synthesis and properties of graphene. Here we show that a successful reduction of graphene oxide (GO) using spinach leaf extract (SLE) as a simultaneous reducing and stabilizing agent. The as-prepared SLE-reduced graphene oxide (S-rGO) was characterized by ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy. Dynamic light scattering technique was used to determine the average size of GO and S-rGO. Scanning electron microscopy and atomic force microscopy images provide clear surface morphological evidence for the formation of graphene. The resulting S-rGO has a mostly single-layer structure, is stable, and has significant water solubility. In addition, the biocompatibility of graphene was investigated using cell viability, leakage of lactate dehydrogenase and alkaline phosphatase activity in primary mouse embryonic fibroblast (PMEFs) cells. The results suggest that the biologically synthesized graphene has significant biocompatibility with PMEF cells, even at a higher concentration of 100 ?g/mL. This method uses a `green', natural reductant and is free of additional stabilizing reagents; therefore, it is an environmentally friendly, simple, and cost-effective method for the fabrication of soluble graphene. This study could open up a promising view for substitution of hydrazine by a safe, biocompatible, and powerful reduction for the efficient deoxygenation of GO, especially in large-scale production and potential biomedical applications.

Gurunathan, Sangiliyandi; Han, Jae Woong; Eppakayala, Vasuki; Dayem, Ahmed Abdal; Kwon, Deug-Nam; Kim, Jin-Hoi

2013-09-01

219

Primary biliary cirrhosis in the mouse: induction by human mycoplasma-like organisms.  

PubMed Central

Human intraocular and orbital chronic inflammatory disease with autoimmune features has been reported to be caused by mycoplasma-like organisms (MLO). MLO are intracellular cell-wall deficient pathogenic bacteria, closely related to rickettsia, with a characteristic ultrastrural pleomorphic tubulo-spherical and filamentous appearance. No culture system has been developed for MLO and diagnosis of MLO disease is made by detecting these bacteria within infected cells using a transmission electron microscope. In human MLO ocular and orbital disease the organisms are found in parasitized leucocytes at the disease site. Inoculation of human MLO into mouse eyelids produces a high incidence of orbital and introcular disease. MLO disseminate to produce randomly distributed lethal systemic disease with infected leucocytes found in all disease sites and with similar histologic features in all disease sites. Microvasculitis is the initial lesion. Disease progression results in lysis of vascular and parenchymal structures, stromal lymphocytic infiltrates, granulomas, and fibrosis. This report describes the hepatic portal chronic progressive inflammatory disease in 11 of 100 of those mice versus 0 in 200 controls. MLO parasitized portal leucocytes are present in all 11 inflamed livers versus 0 in 5 control livers (P less than 0.05). The resemblance of the animal liver disease induced by MLO to human primary biliary cirrhosis and rifampin treatment of MLO disease are discussed. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8

Johnson, L.; Wirostko, E.; Wirostko, W.

1990-01-01

220

Spatial distribution of mRNAs for myelin proteins in primary cultures of mouse brain.  

PubMed

A nonradioactive in situ hybridization procedure was employed to study the distribution of mRNAs for myelin basic protein (MBP), proteolipid protein (PLP), myelin-associated glycoprotein (MAG) and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) in oligodendrocytes in primary cultures of mouse brain. This procedure provided good cellular localization and allowed rapid detection of the mRNAs with low backgrounds. Gradual movement of MBP mRNAs from oligodendrocyte somas into processes was observed with time in culture. The MBP mRNAs were observed to be distributed in an asymmetric fashion within the somas and cell processes. Antigalactocerebroside staining indicated the presence of oligodendrocyte processes prior to movement of MBP mRNA, suggesting that the presence of processes alone was insufficient for translocation of MBP mRNAs. The mRNAs for PLP and MAG remained confined to the oligodendrocyte somas at all times in culture at least up to 28 days. While most of the CNP mRNAs were observed to be associated with the perikarya of oligodendrocytes, in less than 1% of these cells the presence of CNP mRNA in the processes was evident. This suggests that there may exist a subset of oligodendrocytes in which the translocation of these messages occurs. PMID:1705208

Amur-Umarjee, S G; Hall, L; Campagnoni, A T

1990-01-01

221

The excitatory neuronal network of the C2 barrel column in mouse primary somatosensory cortex.  

PubMed

Local microcircuits within neocortical columns form key determinants of sensory processing. Here, we investigate the excitatory synaptic neuronal network of an anatomically defined cortical column, the C2 barrel column of mouse primary somatosensory cortex. This cortical column is known to process tactile information related to the C2 whisker. Through multiple simultaneous whole-cell recordings, we quantify connectivity maps between individual excitatory neurons located across all cortical layers of the C2 barrel column. Synaptic connectivity depended strongly upon somatic laminar location of both presynaptic and postsynaptic neurons, providing definitive evidence for layer-specific signaling pathways. The strongest excitatory influence upon the cortical column was provided by presynaptic layer 4 neurons. In all layers we found rare large-amplitude synaptic connections, which are likely to contribute strongly to reliable information processing. Our data set provides the first functional description of the excitatory synaptic wiring diagram of a physiologically relevant and anatomically well-defined cortical column at single-cell resolution. PMID:19186171

Lefort, Sandrine; Tomm, Christian; Floyd Sarria, J-C; Petersen, Carl C H

2009-01-29

222

Efficacy of Phosphatidylinositol-3 Kinase Inhibitors in a Primary Mouse Model of Undifferentiated Pleomorphic Sarcoma  

PubMed Central

Recent advances in sarcoma genomics have identified novel mutations in the PI3K pathway in human sarcomas. Here, we use a mouse model of primary soft-tissue sarcoma for preclinical testing of doxorubicin and inhibitors of the PI3K pathway: BKM120 (PI3K inhibitor) and BEZ235 (a dual PI3K/mTOR inhibitor). Doxorubicin-treated tumors (n = 15) showed a partial response rate of 6.6%, just as the majority of human sarcomas do not respond to doxorubicin. Treatment with BKM120 elicited a partial response in 50% of tumors (n = 10), which was also seen in combination with doxorubicin (n = 10). Additionally, BKM120 treatment produced a robust delay in tumor growth kinetics. BEZ235-treated tumors (n = 9) showed a complete response rate of 11.1%. Combining BEZ235 with doxorubicin (n = 10) increased the complete response rate to 50% (P = 0.035). These studies demonstrate that PI3K pathway inhibition is a viable and attractive target for soft-tissue sarcomas.

Kim, Suzy; Dodd, Rebecca D.; Mito, Jeffrey K.; Ma, Yan; Kim, Yongbaek; Riedel, Richard F.; Kirsch, David G.

2012-01-01

223

Efficacy of phosphatidylinositol-3 kinase inhibitors in a primary mouse model of undifferentiated pleomorphic sarcoma.  

PubMed

Recent advances in sarcoma genomics have identified novel mutations in the PI3K pathway in human sarcomas. Here, we use a mouse model of primary soft-tissue sarcoma for preclinical testing of doxorubicin and inhibitors of the PI3K pathway: BKM120 (PI3K inhibitor) and BEZ235 (a dual PI3K/mTOR inhibitor). Doxorubicin-treated tumors (n = 15) showed a partial response rate of 6.6%, just as the majority of human sarcomas do not respond to doxorubicin. Treatment with BKM120 elicited a partial response in 50% of tumors (n = 10), which was also seen in combination with doxorubicin (n = 10). Additionally, BKM120 treatment produced a robust delay in tumor growth kinetics. BEZ235-treated tumors (n = 9) showed a complete response rate of 11.1%. Combining BEZ235 with doxorubicin (n = 10) increased the complete response rate to 50% (P = 0.035). These studies demonstrate that PI3K pathway inhibition is a viable and attractive target for soft-tissue sarcomas. PMID:22619567

Kim, Suzy; Dodd, Rebecca D; Mito, Jeffrey K; Ma, Yan; Kim, Yongbaek; Riedel, Richard F; Kirsch, David G

2012-04-29

224

Ovarian Abnormalities in a Mouse Model of Fragile X Primary Ovarian Insufficiency  

PubMed Central

FMR1 premutation (PM) alleles have 55–200 CGG·CCG-repeats in their 5? UTR. PM carriers are at risk of fragile X–associated tremor and ataxia syndrome (FXTAS). Females are also at risk for FX primary ovarian insufficiency (FXPOI). PM pathology is generally attributed to deleterious properties of transcripts with long CGG-tracts. For FXPOI, hormone changes suggest a reduced residual follicle pool. Whether this is due to a smaller than normal original follicle pool or an increased rate of follicle depletion is unclear. A FX-PM mouse the authors generated with 130 CGG·CCG-repeats in the endogenous Fmr1 gene recapitulates features of FXTAS. Here the authors demonstrate that the gross development of the ovary and the establishment of the primordial follicle pool is normal in these mice. However, these animals show a faster loss of follicles of all follicle classes, suggesting that the problem is intrinsic to the ovary. In addition, many oocytes show aberrant nuclear accumulation of FMRP and elevated levels of ubiquitination. Furthermore, PM follicles are smaller and have fewer granulosa cells (GCs) than normal. Thus, these animals have ovarian abnormalities involving both the oocytes and GCs that may shed light on the molecular basis of FXPOI in humans.

Le, Wei Wei; Entezam, Ali; Otsuka, Noriyuki; Tong, Zhi-Bin; Nelson, Lawrence; Flaws, Jodi A.; McDonald, John H.; Jafar, Sanjeeda; Usdin, Karen

2012-01-01

225

Similar effects of phospholipase C and phorbol ester tumor promoters on primary mouse epidermal cells  

SciTech Connect

Interaction of tumor promoting phorbol esters with specific high affinity receptors is probably essential for many of the biological responses elicited by these agents. Since diacylglycerols which can be produced enzymatically from phospholipids by phospholipase C are postulated to be the physiological ligands for the phorbol ester receptor, the authors have examined primary cultures of mouse epidermal basal cells exposed to phospholipase C (Clostridium perfringens) for several biological and biochemical responses characteristic of treatment with 12-O-tetradecanoyl-phorbol-13-acetate, the most potent phorbol ester tumor promoter. Formation of diacylglycerols by treatment with phospholipase C was demonstrated by the dose-dependent release of radioactive diacylglycerols in cells prelabeled with (TH)arachidonic acid. Treatment with phospholipase C led to the morphological changes and to the reduction in epidermal growth factor binding (90%) associated with 12-O-tetradecanoylphorbol-13-acetate treatment. Continuous treatment at the same dose led to the induction of the enzymes ornithine decarboxylase and transglutaminase with a time course and extent similar to the inductions by 12-O-tetradecanoylphorbol-13-acetate. Treatment with phospholipase C yielded substantial suppression of the binding affinity of phorbol-12,13-dibutyrate for its receptors without reduction in total number of binding sites, consistent with the production by phospholipase C of a competitive inhibitor of phorbol ester binding.

Jeng, A.Y.; Lichti, U.; Strickland, J.E.; Blumberg, P.M.

1985-11-01

226

CCR1 blockade reduces tumor burden and osteolysis in vivo in a mouse model of myeloma bone disease  

PubMed Central

The chemokine CCL3/MIP-1? is a risk factor in the outcome of multiple myeloma (MM), particularly in the development of osteolytic bone disease. This chemokine, highly overexpressed by MM cells, can signal mainly through 2 receptors, CCR1 and CCR5, only 1 of which (CCR1) is responsive to CCL3 in human and mouse osteoclast precursors. CCR1 activation leads to the formation of osteolytic lesions and facilitates tumor growth. Here we show that formation of mature osteoclasts is blocked by the highly potent and selective CCR1 antagonist CCX721, an analog of the clinical compound CCX354. We also show that doses of CCX721 selected to completely inhibit CCR1 produce a profound decrease in tumor burden and osteolytic damage in the murine 5TGM1 model of MM bone disease. Similar effects were observed when the antagonist was used prophylactically or therapeutically, with comparable efficacy to that of zoledronic acid. 5TGM1 cells were shown to express minimal levels of CCR1 while secreting high levels of CCL3, suggesting that the therapeutic effects of CCX721 result from CCR1 inhibition on non-MM cells, most likely osteoclasts and osteoclast precursors. These results provide a strong rationale for further development of CCR1 antagonists for the treatment of MM and associated osteolytic bone disease.

Oyajobi, Babatunde O.; Gupta, Anjana; McCluskey, Brandon; Miao, Shichang; Powers, Jay P.; Seitz, Lisa C.; Wang, Yu; Zeng, Yibin; Zhang, Penglie; Schall, Thomas J.; Jaen, Juan C.

2012-01-01

227

Transition Pattern and Mechanism of B-lymphocyte Precursors in Regenerated Mouse Bone Marrow after Subtotal Body Irradiation  

PubMed Central

Little is known about the effects of ionizing radiation on the transition and the related signal transduction of progenitor B cells in the bone marrow. Thus, using an NIH Swiss mouse model, we explored the impact of ionizing radiation on the early stage of B-cell development via an examination of the transition of CLP to pro-B to pre-B cells within bone marrow as a function of radiation doses and times. Our results showed that while the total number of bone marrow lymphoid cells at different stages were greatly reduced by subtotal body irradiation (sub-TBI), the surviving cells continued to transition from common lymphoid progenitors to pro-B and then to pre-B in a reproducible temporal pattern. The rearrangement of the immunoglobulin heavy chain increased significantly 1–2 weeks after irradiation, but no change occurred after 3–4 weeks. The rearrangement of the immunoglobulin light chain decreased significantly 1–2 weeks after sub-TBI but increased dramatically after 3–4 weeks. In addition, several key transcription factors and signaling pathways were involved in B-precursor transitions after sub-TBI. The data indicate that week 2 after irradiation is a critical time for the transition from pro-B cells to pre-B cells, reflecting that the functional processes for different B-cell stages are well preserved even after high-dose irradiation.

Han, Deping; Zhang, Mei; Ma, Jun; Hong, Jingshen; Chen, Chun; Zhang, Bingrong; Huang, Luqiang; Lv, Wenlong; Yin, Liangjie; Zhang, Amy; Zhang, Hengshan; Zhang, Zhenhuan; Vidyasagar, Sadasivan; Okunieff, Paul; Zhang, Lurong

2012-01-01

228

Miltefosine decreases the cytotoxic effect of epirubicine and cyclophosphamide on mouse spermatogenic, thymic and bone marrow cells.  

PubMed

A new class of potent anticancer drugs, alkylphosphocholines has been recognized lately. Miltefosine (Hexadecylphosphochlorine, HPC) has been found to express select antineoplastic effect on human breast cancer skin metastases with simultaneous preservation of bone marrow proliferative activity and low clastogenicity. In the current study, we present data about the specific effect of two widely used cytostatics Cyclophosphamide (CP) and Epirubicine (ERb) applied separately or in combination with Miltefosine. C57BL6 mice were treated per os or intraperitonieally in doses corresponding to that in clinical use. Morphological, autoradiographic, ultrastructural and cytogenetic studies on spermatogenic, thymic and bone marrow cells were performed. It is found that compared with separate application, combinations of ERb or CP with Miltefosine slightly decreases spermatogonial proliferation and exerts milder effect on the structure of germinal and thymic cells. In addition, a lot of plasmocytes showed signs of active protein (antibody) synthesis. A significant reduction of aberrant chromosomes (clastogenicity) without changes in proliferative activity of bone marrow cells were recorded. In conclusion, the combine application of Miltefosine with ERb and CP decreased the destructive cytotoxic effects of ERb and CP on mouse spermatogenic and hematopoietic cells. PMID:16079990

Martinova, Yordanka; Topashka-Ancheva, Margarita; Konstantinov, Spiro; Petkova, Svetlozara; Karaivanova, Margarita; Berger, Martin

2005-08-04

229

Detection of mineral density on the surface of mouse parietal bones: backscattered electron imaging of low accelerating voltage scanning electron microscopy.  

PubMed

Backscattered electron (BSE) imaging of scanning electron microscopy (SEM) was applied to a study on the mineral density of the bone surface. The neonatal and adult mouse parietal bones freed of the periosteum and covering cells were examined in a field emission scanning electron microscope equipped with a high sensitivity BSE detector at 1-30 kV accelerating voltages. The mineral density of the bone surface was observable in BSE images at 5 kV accelerating voltage while only the topographic structures of the surface were obtained under an accelerating voltage less than 5 kV. As the accelerating voltages increased from 5 kV, the bright areas were extended, probably due to the imaging of the calcified bone matrix under the uncalcified osteoid. The bone surface is usually divided into smooth and rough areas according to its irregularities. BSE images at 5 kV clearly showed that the smooth areas were further divided into dark and bright areas which apparently corresponded to the uncalcified osteoid and calcified bone matrix, respectively. Bright granules, about 1.0-3.0 microns in diameter, were sometimes observed at the border between the osteoid and calcified bone matrix; these granular calcified areas were regarded as the calcifying front forming the calcified bone matrix from the osteoid. The present study demonstrated that the distribution of the osteoid on the mouse parietal bone surface changes depending on age: the osteoid occupied a large area in the parietal bone surface in neonatal mice, but was small in adult mice. Thus, low accelerating voltage SEM using BSE provides new information on the distribution of the osteoid and the bone matrix calcification under both normal and pathological conditions. PMID:9232183

Hashizume, H; Abe, K; Ushiki, T

1997-06-01

230

Characterization of a new renal cell carcinoma bone metastasis mouse model  

Microsoft Academic Search

Metastatic bone disease caused by renal cell carcinoma (RCC) occurs frequently and becomes more and more prevalent presumably\\u000a because survival times among patients with disseminated cancers are increasing. Patients with bone metastases from renal cell\\u000a carcinoma suffer from severe pain, nerve compression syndromes and pathologic fractures. Very little is known about the mechanisms\\u000a of skeletal metastases of RCC. Thus, to

Anne Strube; Elizaveta Stepina; Dominik Mumberg; Arne Scholz; Peter Hauff; Sanna-Maria Käkönen

2010-01-01

231

Excessive bone formation in a mouse model of ankylosing spondylitis is associated with decreases in Wnt pathway inhibitors  

PubMed Central

Introduction Ankylosing spondylitis (AS) is unique in its pathology where inflammation commences at the entheses before progressing to an osteoproliferative phenotype generating excessive bone formation that can result in joint fusion. The underlying mechanisms of this progression are poorly understood. Recent work has suggested that changes in Wnt signalling, a key bone regulatory pathway, may contribute to joint ankylosis in AS. Using the proteoglycan-induced spondylitis (PGISp) mouse model which displays spondylitis and eventual joint fusion following an initial inflammatory stimulus, we have characterised the structural and molecular changes that underlie disease progression. Methods PGISp mice were characterised 12 weeks after initiation of inflammation using histology, immunohistochemistry (IHC) and expression profiling. Results Inflammation initiated at the periphery of the intervertebral discs progressing to disc destruction followed by massively excessive cartilage and bone matrix formation, as demonstrated by toluidine blue staining and IHC for collagen type I and osteocalcin, leading to syndesmophyte formation. Expression levels of DKK1 and SOST, Wnt signalling inhibitors highly expressed in joints, were reduced by 49% and 63% respectively in the spine PGISp compared with control mice (P < 0.05) with SOST inhibition confirmed by IHC. Microarray profiling showed genes involved in inflammation and immune-regulation were altered. Further, a number of genes specifically involved in bone regulation including other members of the Wnt pathway were also dysregulated. Conclusions This study implicates the Wnt pathway as a likely mediator of the mechanism by which inflammation induces bony ankylosis in spondyloarthritis, raising the potential that therapies targeting this pathway may be effective in preventing this process.

2012-01-01

232

Is bone marrow biopsy always indicated in patients with primary cutaneous marginal zone B-cell lymphoma?  

PubMed

Bone marrow involvement at the time of diagnosis is uncommon in patients with primary cutaneous marginal zone B-cell lymphoma (PCMZL). Moreover, in these patients such involvement is rarely found in isolation on diagnosis. Typically the few patients with PCMZL who have early bone marrow involvement also present secondary nodal or visceral involvement, which is detected by other staging studies (usually computed tomography). In recent years, this has given rise to some debate about whether a bone marrow biopsy should be routinely performed in patients diagnosed with PCMZL in view of the good prognosis and low incidence of bone marrow infiltration and/or extracutaneous involvement in this type of lymphoma. PMID:23954046

Muniesa, C; Hernández-Machín, B

2013-08-13

233

Genetic modification of mouse bone marrow by lentiviral vector-mediated delivery of hypoxanthine-Guanine phosphoribosyltransferase short hairpin RNA confers chemoprotection against 6-thioguanine cytotoxicity.  

PubMed

We have recently developed a novel and highly efficient strategy that exclusively uses the purine analog 6-thioguanine (6TG) for both pretransplantation conditioning and post-transplantation chemoselection of hypoxanthine-guanine phosphoribosyltransferase (HPRT)-deficient bone marrow (BM). In a mouse BM transplantation model, combined 6TG preconditioning and in vivo chemoselection consistently achieved >95% engraftment of HPRT-deficient donor BM and long-term reconstitution of histologically and immunophenotypically normal hematopoiesis in both primary and secondary recipients, without significant toxicity and in the absence of any other cytotoxic conditioning regimen. To translate this strategy for combined 6TG conditioning and chemoselection into a clinically feasible approach, it is necessary to develop methods for genetic modification of normal hematopoietic stem cells (HSC) to render them HPRT-deficient and thus 6TG-resistant. Here we investigated a strategy to reduce HPRT expression and thereby confer protection against 6TG myelotoxicity to primary murine BM cells by RNA interference (RNAi). Accordingly, we constructed and validated a lentiviral gene transfer vector expressing short-hairpin RNA (shRNA) that targets the murine HPRT gene. Our results showed that lentiviral vector-mediated delivery of HPRT-targeted shRNA could achieve effective and long-term reduction of HPRT expression. Furthermore, in both an established murine cell line as well as in primary murine BM cells, lentiviral transduction with HPRT-targeted shRNA was associated with enhanced resistance to 6TG cytotoxicity in vitro. Hence this represents a translationally feasible method to genetically engineer HSC for implementation of 6TG-mediated preconditioning and in vivo chemoselection. PMID:23769104

Hacke, K; Treger, J A; Bogan, B T; Schiestl, R H; Kasahara, N

2013-06-01

234

Primary aneurysmal bone cyst of the maxillary sinus in a child: case report and review of the literature  

Microsoft Academic Search

Our case report describes a primary aneurysmal bone cyst (ABC) of the maxillary sinus in a 12-year-old girl. The young patient presented with progressive diplopia, strabismus, and rapidly growing painless swelling of the left cheek. Imaging studies showed a heterogeneous contrast enhancing mass expanding the left maxillary sinus. The lesion was completely resected endoscopically and histological examination reported it as

G. Fyrmpas; J. Constantinidis; D. Televantou; I. Konstantinidis; J. Daniilidis

2006-01-01

235

Spontaneous but not experimental metastatic activities differentiate primary tumor-derived vs metastasis-derived mouse prostate cancer cell lines  

Microsoft Academic Search

We previously developed an in vivomouse prostate reconstitution (MPR) model of metastatic prostate cancer using p53 ‘knockout’ mouse urogenital sinus tissue for retroviral transduction of rasand myconcogenes (Thompson et al., Oncogene, 10, 869, 1995). We further demonstrated contrasting responses to transforming growth factor beta-1 (TGF-ß1) in three matched pairs of early passage cell lines derived from primary prostate tumors and

Simon J. Hall; Timothy C. Thompson

1997-01-01

236

Responses of L929 mouse fibroblasts, primary and immortalized bovine dental papilla-derived cell lines to dental resin components  

Microsoft Academic Search

Objective: The use of adequate target cells for cytotoxicity testing of dental restorative materials has often been experimentally assessed with respect to the clinical relevance of the test results. In the present study, the responses in primary bovine dental papilla-derived cells (pulp cells) were compared with those in transformed dental papilla-derived cell lines and L929 mouse fibroblasts after exposure to

B Thonemann; G Schmalz; K.-A Hiller; H Schweikl

2002-01-01

237

Effects of chronic exposure to ammonia on glutamate and glutamine interconversion and compartmentation in homogeneous primary cultures of mouse astrocytes  

Microsoft Academic Search

Accumulation of radioactivity was studied in primary cultures of mouse astrocytes as a function of time of exposure (4–60 min) to 50 µM glutamate and 200 µM glutamine (initial concentrations), of whicheither glutamateor glutamine was14C-labeled. Both the glutamate pool and the glutamine pool were compartmentalized. Initially, by far the major intracellular glutamate pool (=90%) was derived from extracellular glutamate and

Rong Huang; Geeta Kala; Ch. R. K. Murthy; Leif Hertz

1994-01-01

238

Ultrastructural and biochemical alterations induced in human, rat and mouse hepatocytes in primary culture exposed to selected carcinogens  

SciTech Connect

Aflatoxin B1 (AFB{sub 1}), dimethylnitrosamine (DMN), 2-acetylaminofluorene (2-AAF) and actinomycin D are all potential human liver carcinogens. In order to investigate carcinogenic susceptibility of human liver to these agents, primary cultures of normal human hepatocytes were exposed to the four carcinogens. In the first series of experiments, human, rat, and mouse hepatocytes in primary culture were exposed to actinomycin D, AFB{sub 1}, and DMN for 24 h and examined for ultrastructural alterations. In an effort to relate the ultrastructural effects with total covalent binding of the carcinogen to DNA, human, rat and mouse hepatocytes were exposed to 2.0 {times} 10{sup {minus}7} M ({sup 3}H)AFB{sub 1} for 24 h. Hepatocytes from male rats had the highest degree of ({sup 3}H)AFB{sub 1}-DNA binding. Human hepatocytes contained the next highest binding level, while hepatocytes from female rats bound 38 pmoles/mg DNA. The AFB{sub 1}-DNA binding level in mouse hepatocytes was 1.4 pmoles/mg DNA. In similar experiments, human, and male and female rat hepatocytes in primary culture were exposed to the carcinogen 2-acetylamino (9-{sup 14}C)fluorene for 24 h. It was determined that male rat hepatocytes had the highest amount of radiolabeled 2-AAF bound to their DNA, female rats contained 0.57 nmoles/mg DNA, while human hepatocytes contained 0.29 nmoles/mg DNA.

Cole, K.H.

1987-01-01

239

Antibiotic Loaded Bone Cement Reduces Deep Infection Rates For Primary Reverse Total Shoulder Arthroplasty. A Retrospective, Cohort Study of 501 Shoulders  

Microsoft Academic Search

BackgroundDeep infection following primary reverse total shoulder arthroplasty is a devastating event and has an increased incidence compared to anatomic total shoulder arthroplasty. Recent reports in the hip and knee arthroplasty literature suggest that antibiotic loaded bone cement may lower infection rates for primary arthroplasties. We conducted a retrospective cohort study to evaluate the effect of antibiotic loaded bone cement

Robert J. Nowinski; Robert J. Gillespie; Yousef Shishani; Brian Cohen; Gilles Walch; Reuben Gobezie

240

Proteolytic processing of osteopontin by PHEX and accumulation of osteopontin fragments in Hyp mouse bone, the murine model of X-linked hypophosphatemia.  

PubMed

X-linked hypophosphatemia (XLH/HYP)-with renal phosphate wasting, hypophosphatemia, osteomalacia, and tooth abscesses-is caused by mutations in the zinc-metallopeptidase PHEX gene (phosphate-regulating gene with homologies to endopeptidase on the X chromosome). PHEX is highly expressed by mineralized tissue cells. Inactivating mutations in PHEX lead to distal renal effects (implying accumulation of a secreted, circulating phosphaturic factor) and accumulation in bone and teeth of mineralization-inhibiting, acidic serine- and aspartate-rich motif (ASARM)-containing peptides, which are proteolytically derived from the mineral-binding matrix proteins of the SIBLING family (small, integrin-binding ligand N-linked glycoproteins). Although the latter observation suggests a local, direct matrix effect for PHEX, its physiologically relevant substrate protein(s) have not been identified. Here, we investigated two SIBLING proteins containing the ASARM motif-osteopontin (OPN) and bone sialoprotein (BSP)-as potential substrates for PHEX. Using cleavage assays, gel electrophoresis, and mass spectrometry, we report that OPN is a full-length protein substrate for PHEX. Degradation of OPN was essentially complete, including hydrolysis of the ASARM motif, resulting in only very small residual fragments. Western blotting of Hyp (the murine homolog of human XLH) mouse bone extracts having no PHEX activity clearly showed accumulation of an ?35?kDa OPN fragment that was not present in wild-type mouse bone. Immunohistochemistry and immunogold labeling (electron microscopy) for OPN in Hyp bone likewise showed an accumulation of OPN and/or its fragments compared with normal wild-type bone. Incubation of Hyp mouse bone extracts with PHEX resulted in the complete degradation of these fragments. In conclusion, these results identify full-length OPN and its fragments as novel, physiologically relevant substrates for PHEX, suggesting that accumulation of mineralization-inhibiting OPN fragments may contribute to the mineralization defect seen in the osteomalacic bone characteristic of XLH/HYP. PMID:22991293

Barros, Nilana M T; Hoac, Betty; Neves, Raquel L; Addison, William N; Assis, Diego M; Murshed, Monzur; Carmona, Adriana K; McKee, Marc D

2013-03-01

241

Development of adult bone marrow stem cells in H-2-compatible and -incompatible mouse fetuses.  

PubMed

Bone marrow of normal adult mice was found, after transplacental inoculation, to contain cells still able to seed the livers of early fetuses. The recipients' own hematopoietic stem cells, with a W-mutant defect, were at a selective disadvantage. Progression of donor strain cells to the bone marrow, long-term self-renewal, and differentiation into myeloid and lymphoid derivatives was consistent with the engraftment of totipotent hematopoietic stem cells (THSC) comparable to precursors previously identified (4) in normal fetal liver. More limited stem cells, specific for the myeloid or lymphoid cell lineages, were not detected in adult bone marrow. The bone marrow THSC, however, had a generally lower capacity for self-renewal than did fetal liver THSC. They had also embarked upon irreversible changes in gene expression, including partial histocompatibility restriction. While completely allogeneic fetal liver THSC were readily accepted by fetuses, H-2 incompatibility only occasionally resulted in engraftment of adult bone marrow cells and, in these cases, was often associated with sudden death at 3-5 mo. On the other hand, H-2 compatibility, even with histocompatibility differences at other loci, was sufficient to ensure long-term success as often as with fetal liver THSC. PMID:6607968

Fleischman, R A; Mintz, B

1984-03-01

242

Dose Response Effects of 810 nm Laser Light on Mouse Primary Cortical Neurons  

PubMed Central

Background and Objectives In the past four decades numerous studies have reported the efficacy of low level light (laser) therapy (LLLT) as a treatment for diverse diseases and injuries. Recent studies have shown that LLLT can biomodulate processes in the central nervous system and has been extensively studied as a stroke treatment. However there is still a lack of knowledge on the effects of LLLT at the cellular level in neurons. The present study aimed to study the effect of 810 nm laser on several cellular processes in primary cortical neurons cultured from embryonic mouse brains. Study Design/Materials and Methods Neurons were irradiated with fluences of 0.03, 0.3, 3, 10, or 30 J/cm2 of 810-nm laser delivered over varying times at 25 mW/cm2 and intracellular levels of reactive oxygen species (ROS), nitric oxide and calcium were measured using fluorescent probes within 5 minutes of the end of irradiation. The changes in mitochondrial function in response to light were studied in terms of adenosine triphosphate (ATP) and mitochondrial membrane potential (MMP). Results Light induced a significant increase in calcium, ATP and MMP at lower fluences and a decrease at higher fluences. ROS was significantly induced at low fluences, followed by a decrease and a second larger increase at 30 J/cm2. Nitric oxide levels showed a similar pattern of a double peak but values were less significant compared to ROS. Conclusions The results suggest that LLLT at lower fluences is capable of inducing mediators of cell signaling processes which in turn may be responsible for the beneficial stimulatory effects of the low level laser. At higher fluences beneficial mediators are reduced and high levels of Janus-type mediators such as ROS and NO (beneficial at low concentrations and harmful at high concentrations) may be responsible for the damaging effects of high-fluence light and the overall biphasic dose response.

Sharma, Sulbha K.; Kharkwal, Gitika B.; Sajo, Mari; Huang, Ying-Ying; De Taboada, Luis; McCarthy, Thomas; Hamblin, Michael R.

2011-01-01

243

The inhibitory action of aluminum on mouse bone marrow cell growth: evidence for an erythropoietin- and transferrin-mediated mechanism.  

PubMed

Aluminum (Al) has been associated with anemia in chronic renal failure patients under hemodialysis as well as in Al-overloaded animals. In an attempt to elucidate further the mechanism of Al toxicity we have investigated the effect of this ion on erythropoiesis in vitro. Mouse bone marrow cells were stimulated in vitro with erythropoietin (Epo) in the presence of Al3+ ion and erythroid colony-forming units were then determined. Results of this study indicate that Al compounds (chloride and citrate) at concentrations as low as 0.37 mumol Al/l inhibit erythropoiesis in vitro through a mechanism dependent upon the availability of transferrin to bind to aluminum. This process cannot be reversed by increasing Epo doses. This inhibition only occurs in the presence of Epo at early stages during the interaction of the hormone with its target cell. PMID:7816003

Garbossa, G; Gutnisky, A; Nesse, A

1994-01-01

244

Re-evaluation of the need for multiple sampling times in the mouse bone marrow micronucleus assay: results for DMBA  

SciTech Connect

7,12-dimethylbenzanthracene (DMBA) is confirmed as active in the mouse bone marrow micronucleus assay 24 hr after dosing as corn-oil homogenate via either oral gavage or intraperitoneal (ip) injection. These data are consistent with recent observations made by several investigators. However, when dosed via ip injection as a solution in DMSO, peak activity was evident 48 hr after dosing and a dramatic reduction in erythropoiesis was observed. It is suggested that a maximum of two sampling times is adequate and that, as a consequence, the number of animals employed in the conduct of the test could be reduced with no loss of sensitivity. The present data also suggest that the use of a corn-oil homogenate of insoluble test agents may provide an efficient replacement for the use of ground suspensions or solutions in DMSO.

Ashby, J.; Mirkova, E.

1987-01-01

245

Analysis of and prognostic information from disseminated tumour cells in bone marrow in primary breast cancer: a prospective observational study  

PubMed Central

Background Disseminated tumour cells (DTCs) in the bone marrow of patients with breast cancer have been identified as an independent predictor of poor prognosis in patients with non-metastatic disease. This prospective study aimed to evaluate the presence and prognostic value of DTCs in the bone marrow of female patients with primary breast cancer. Methods Between 1999 and 2003, bone marrow aspirates were obtained from patients at the time of surgery for primary invasive breast cancer. DTCs in bone marrow were identified using monoclonal antibodies against cytokeratins for detection of epithelial cells. The detection of DTCs was related to clinical follow-up with distant disease-free survival (DDFS) and breast cancer-specific survival as endpoints. Bone marrow aspirates from adult healthy bone marrow donors were analysed separately. Results DTCs were analysed in 401 patients, and cytokeratin-positive cells were found in 152 of these (38%). An immunofluorescence (IF) staining procedure was used in 327 patients, and immunocytochemistry (IC) was performed in 74 patients. The IF-based method resulted in 40% DTC-positive cases, whereas 30% were positive using IC (p?=?0.11). The presence of DTCs in bone marrow was not significantly related to patient or tumour characteristics. The presence of DTCs was not a prognostic factor for DDFS (IF: hazards ratio [HR], 2.2; 95% confidence interval [CI], 0.63–2.2; p?=?0.60; IC: HR, 0.84; 95% CI, 0.09–8.1; p?=?0.88). Significant prognostic factors were lymph node metastases, oestrogen receptor positivity, Nottingham histological grade, and tumour size using Cox univariate analysis. The analyses were positive for epithelial cells in bone marrow from adult healthy donors in 19 (25%) samples. Conclusions The detection of DTCs in bone marrow in primary breast cancer was previously shown to be a predictor of poor prognosis. We were not able to confirm these results in a prospective cohort including unselected patients before the standard procedure was established. Future studies with a standardised patient protocol and improved technique for isolating and detecting DTCs may reveal the clinical applications of DTC detection in patients with micrometastases in the bone marrow.

2012-01-01

246

The validity of decision rules for selecting women with primary osteoporosis for bone mineral density testing.  

PubMed

The purpose of this study was to determine the validity of the Osteoporosis Risk Assessment Instrument (ORAI), Osteoporosis Self-Assessment Tool (OST) chart and equation, and a criterion based on body weight for identifying women with asymptomatic primary osteoporosis. Prospective recruitment and chart abstractions from family practices of three University affiliated hospitals were completed for women aged 45 years or more with baseline bone mineral density (BMD) testing results by dual energy X-ray absorptiometry. Those taking bone active medication other than hormone therapy, with prior fragility fracture or with risk factors for secondary osteoporosis were excluded. Women were categorized as being normal, osteopenic or osteoporotic by lowest BMD T-score at either the femoral neck or lumbar spine (L1-L4). Sensitivity, specificity and area under the receiver operating characteristic (ROC) curve to identify those with osteoporosis were determined for each decision rule. The positive predictive value (PPV) for detecting osteoporosis after using a second cut point to convert each decision rule into a risk index (low, moderate or high risk) was also determined. The sensitivity of the decision rules to identify women with osteoporosis ranged from 92% to 95% and specificity from 35% to 46%. The area under the ROC curves were significantly better for the ORAI (0.80), OST chart (0.82) and OST equation (0.82) compared with the body weight criterion (0.73). PPV for detecting osteoporosis ranged from 30% to 58% among women deemed at high risk. These data confirm the validity of the ORAI, the OST chart and the OST equation as screening tools for BMD testing. Further evidence is required to confirm the validity of the body weight criterion. PMID:14730421

Cadarette, Suzanne M; McIsaac, Warren J; Hawker, Gillian A; Jaakkimainen, Liisa; Culbert, Alison; Zarifa, Gihane; Ola, Ebele; Jaglal, Susan B

2004-01-17

247

Three-dimensional culture of mouse bone marrow cells on stroma formed within a porous scaffold: influence of scaffold shape and cryopreservation of the stromal layer on expansion of haematopoietic progenitor cells.  

PubMed

This study's primary goal was to develop an effective ex vivo expansion method for haematopoietic cells. 3D culture of mouse bone marrow cells was performed in porous scaffolds using a sheet or cube shape. Bone marrow cells were cultured on bone marrow-derived stromal layers formed within the scaffolds and the effect of scaffold shape on the expansion of haematopoietic cells was examined. In some experiments, stromal layers within cubic scaffolds were frozen and then used to culture bone marrow cells after thawing. Results show that after comparison, total cell density and expansion of haematopoietic cells were greater in cultures using the cubic scaffold, suggesting that it was superior to the sheet-like scaffold for expanding haematopoietic cells. When cryopreserved stroma was used, it effectively supported the expansion of haematopoietic cells, and a greater expansion of haematopoietic cells [(erythroid and haematopoietic progenitor cells (HPCs)] was achieved than in cultures with stromal cells that had not been cryopreserved. Expansion of cells using cryopreserved stroma had several other advantages such as a shorter culture period than the conventional method, a stable supply of stromal cells, and ease of handling and scaling up. As a result, this is an attractive method for ex vivo expansion of haematopoietic stem cells (HSCs) and HPCs for clinical use. PMID:22081538

Miyoshi, Hirotoshi; Ohshima, Norio; Sato, Chiaki

2011-11-14

248

Intravenous Ibandronate Rapidly Reduces Pain, Neurochemical Indices of Central Sensitization, Tumor Burden, and Skeletal Destruction in a Mouse Model of Bone Cancer  

PubMed Central

Over half of all chronic cancer pain arises from metastases to bone and bone cancer pain is one of the most difficult of all persistent pain states to fully control. Currently, bone pain is treated primarily by opioid-based therapies, which are frequently accompanied by significant unwanted side effects. In an effort to develop non-opioid-based therapies that could rapidly attenuate tumor-induced bone pain, we examined the effect of intravenous administration of the bisphosphonate, ibandronate, in a mouse model of bone cancer pain. Following injection and confinement of green fluorescent protein-transfected murine osteolytic 2472 sarcoma cells into the marrow space of the femur of male C3H/HeJ mice, ibandronate was administered either as a single dose (300 µg/kg), at day 7 post-tumor injection, when tumor-induced bone destruction and pain were first evident, or in three consecutive doses (100 µg/kg/day) at day 7, 8 and 9 post-tumor injection. Intravenous ibandronate administered once or in three consecutive doses reduced ongoing and movement-evoked bone cancer pain-related behaviors, neurochemical markers of central sensitization, tumor burden and tumor-induced bone destruction. These results support limited clinical trials that suggest the potential of ibandronate to rapidly attenuate bone pain and illuminate the mechanisms that may be responsible for limiting pain and disease progression.

Halvorson, Kyle G.; Sevcik, Molly A.; Ghilardi, Joseph R.; Sullivan, Lucy J.; Koewler, Nathan J.; Bauss, Frieder; Mantyh, Patrick W.

2008-01-01

249

Intracellular cholesterol-binding proteins enhance HDL-mediated cholesterol uptake in cultured primary mouse hepatocytes.  

PubMed

A major gap in our knowledge of rapid hepatic HDL cholesterol clearance is the role of key intracellular factors that influence this process. Although the reverse cholesterol transport pathway targets HDL to the liver for net elimination of free cholesterol from the body, molecular details governing cholesterol uptake into hepatocytes are not completely understood. Therefore, the effects of sterol carrier protein (SCP)-2 and liver fatty acid-binding protein (L-FABP), high-affinity cholesterol-binding proteins present in hepatocyte cytosol, on HDL-mediated free cholesterol uptake were examined using gene-targeted mouse models, cultured primary hepatocytes, and 22-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-amino]-23,24-bisnor-5-cholen-3?-ol (NBD-cholesterol). While SCP-2 overexpression enhanced NBD-cholesterol uptake, counterintuitively, SCP-2/SCP-x gene ablation also 1) enhanced the rapid molecular phase of free sterol uptake detectable in <1 min and initial rate and maximal uptake of HDL free cholesterol and 2) differentially enhanced free cholesterol uptake mediated by the HDL3, rather than the HDL2, subfraction. The increased HDL free cholesterol uptake was not due to increased expression or distribution of the HDL receptor [scavenger receptor B1 (SRB1)], proteins regulating SRB1 [postsynaptic density protein (PSD-95)/Drosophila disk large tumor suppressor (dlg)/tight junction protein (ZO1) and 17-kDa membrane-associated protein], or other intracellular cholesterol trafficking proteins (steroidogenic acute response protein D, Niemann Pick C, and oxysterol-binding protein-related proteins). However, expression of L-FABP, the single most prevalent hepatic cytosolic protein that binds cholesterol, was upregulated twofold in SCP-2/SCP-x null hepatocytes. Double-immunogold electron microscopy detected L-FABP sufficiently close to SRB1 for direct interaction, similar to SCP-2. These data suggest a role for L-FABP in HDL cholesterol uptake, a finding confirmed with SCP-2/SCP-x/L-FABP null mice and hepatocytes. Taken together, these results suggest that L-FABP, particularly in the absence of SCP-2, plays a significant role in HDL-mediated cholesterol uptake in cultured primary hepatocytes. PMID:22241858

Storey, Stephen M; McIntosh, Avery L; Huang, Huan; Landrock, Kerstin K; Martin, Gregory G; Landrock, Danilo; Payne, H Ross; Atshaves, Barbara P; Kier, Ann B; Schroeder, Friedhelm

2012-01-12

250

Targeting A-type K(+) channels in primary sensory neurons for bone cancer pain in a rat model.  

PubMed

Cancer pain is one of the most severe types of chronic pain, and the most common cancer pain is bone cancer pain. The treatment of bone cancer pain remains a clinical challenge. Here, we report firstly that A-type K(+) channels in dorsal root ganglion (DRG) are involved in the neuropathy of rat bone cancer pain and are a new target for diclofenac, a nonsteroidal anti-inflammatory drug that can be used for therapy for this distinct pain. There are dynamically functional changes of the A-type K(+) channels in DRG neurons during bone cancer pain. The A-type K(+) currents that mainly express in isolectin B4-positive small DRG neurons are increased on post-tumor day 14 (PTD 14), then faded but still remained at a higher level on PTD 21. Correspondingly, the expression levels of A-type K(+) channel Kv1.4, Kv3.4, and Kv4.3 showed time-dependent changes during bone cancer pain. Diclofenac enhances A-type K(+) currents in the DRG neurons and attenuates bone cancer pain in a dose-dependent manner. The analgesic effect of diclofenac can be reversed or prevented by A-type K(+) channel blocker 4-AP or pandinotoxin-K?, also by siRNA targeted against rat Kv1.4 or Kv4.3. Repeated diclofenac administration decreased soft tissue swelling adjacent to the tumor and attenuated bone destruction. These results indicate that peripheral A-type K(+) channels were involved in the neuropathy of rat bone cancer pain. Targeting A-type K(+) channels in primary sensory neurons may provide a novel mechanism-based therapeutic strategy for bone cancer pain. PMID:22188869

Duan, Kai-Zheng; Xu, Qian; Zhang, Xiao-Meng; Zhao, Zhi-Qi; Mei, Yan-Ai; Zhang, Yu-Qiu

2011-12-19

251

Effects of magnetic resonance imaging (MRI) on the formation of mouse dentin and bone  

SciTech Connect

The effects of magnetic resonance imaging (MRI) on dentin and bone formation in mice were examined using standard autoradiographic and liquid scintillation procedures. It was observed that exposure to a standard 23.2 min clinical multislice MRI (0.15T) procedure caused a significant increase in the synthesis of the collagenous matrix of dentin in the incisors of mice. There were no significant effects on alveolar and tibial bone matrix synthesis. These results suggest that the magnetic fields associated with MRI can affect the activity of cells and/or tissues that are involved in rapid synthetic activity.

Kwong-Hing, A.; Sandhu, H.S.; Prato, F.S.; Frappier, J.R.; Kavaliers, M. (Univ. of Western Ontario, London (Canada))

1989-10-01

252

Pathological Fractures in Primary Non-Hodgkin's Lymphoma of the Bone: A Case Series with Review of the Literature  

PubMed Central

Primary non-Hodgkin’s lymphoma of bone (PLB) is a rare entity. Patients generally present with localized bone pain and, less frequently, soft-tissue swelling or a palpable mass. Pathological fracture of the proximal femur and proximal humerus secondary to soft-tissue tumours is well documented in the literature; however, lymphomas presenting primarily at these sites with pathological fracture is unusual. A review of the world literature shows that the incidence of skeletal manifestation from NHL is less than 5%, and in all these cases, bony involvement was reported many years after presentation of the primary cancer. Histopathologically, PLB usually represents diffuse large B-cell lymphoma. We report our experience with two cases of Primary non-Hodgkin’s lymphoma of proximal femur and proximal humerus with pathological fracture and their management.

Siddiqui, Yasir Salam; Khan, Abdul Qayyum; Sherwani, MKA

2013-01-01

253

Function and malfunction of hematopoietic stem cells in primary bone marrow failure syndromes.  

PubMed

Hematopoietic stem cells (HSCs) are responsible for the production of mature blood cells in bone marrow; peripheral pancytopenia is a common clinical presentation resulting from several different conditions, including hematological or extra-hematological diseases (mostly cancers) affecting the marrow function, as well as primary failure of hematopoiesis. Primary bone marrow failure syndromes are a heterogeneous group of diseases with specific pathogenic mechanisms, which share a profound impairment of the hematopoietic stem cell pool resulting in global or selective marrow aplasia. Constitutional marrow failure syndromes are conditions caused by intrinsic defects of HSCs; they are due to inherited germline mutations accounting for specific phenotypes, and often involve also organs and systems other than hematopoiesis. By contrast, in acquired marrow failure syndromes hematopoietic stem cells are thought to be intrinsically normal, but subjected to an extrinsic damage affecting their hematopoietic function. Direct toxicity by chemicals or radiation, as well as association with viruses and other infectious agents, can be sometimes demonstrated. In idiopathic Aplastic Anemia (AA) immunological mechanisms play a pivotal role in damaging the hematopoietic compartment, resulting in a depletion of the hematopoietic stem cell pool. Clinical and experimental evidences support the presence of a T cell-mediated immune attack, as confirmed by clonally expanded lymphocytes, even if the target antigens are still undefined. However, this simple model has to be integrated with recent data showing that, even in presence of an extrinsic damage, preexisting mutations or polymorphisms of genes may constitute a genetic propensity to develop marrow failure. Other recent data suggest that similar antigen-driven immune mechanisms may be involved in marrow failure associated with lymphoproliferative or autoimmune disorders characterized by clonal expansion of T lymphocytes, such as Large Granular Lymphocyte leukemia. In this wide spectrum, a unique and intriguing condition is Paroxysmal Nocturnal Hemoglobinuria (PNH); even in presence of a somatic mutation of the PIG-A gene carried by one or more HSCs and their progeny, the typical marrow failure in PNH is likely due to pathogenic mechanisms similar to those involved in AA, and not to the intrinsic abnormality conferred to the clonal population by the PIG-A mutation. The study of hematopoietic stem cell function in marrow failure syndromes provides hints for specific molecular pathways disturbed in many diseases of hematopoietic and non-hematopoietic stem cells. Beyond the specific interest of investigators involved in the field of these rare diseases, marrow failure syndromes represent a model that provides intriguing insight into quantity and function of normal hematopoietic stem cells, improving our knowledge on stem cell biology. PMID:18220891

Risitano, Antonio M; Maciejewski, Jaroslaw P; Selleri, Carmine; Rotoli, Bruno

2007-01-01

254

Cell lines and primary cell cultures in the study of bone cell biology  

Microsoft Academic Search

Bone is a metabolically active and highly organized tissue consisting of a mineral phase of hydroxyapatite and amorphous calcium phosphate crystals deposited in an organic matrix. Bone has two main functions. It forms a rigid skeleton and has a central role in calcium and phosphate homeostasis. The major cell types of bone are osteoblasts, osteoclasts and chondrocytes. In the laboratory,

Vicky Kartsogiannis; Kong Wah Ng

2004-01-01

255

Differential Expression of Surface Markers in Mouse Bone Marrow Mesenchymal Stromal Cell Subpopulations with Distinct Lineage Commitment  

PubMed Central

Bone marrow mesenchymal stromal cells (BM MSCs) represent a heterogeneous population of progenitors with potential for generation of skeletal tissues. However the identity of BM MSC subpopulations is poorly defined mainly due to the absence of specific markers allowing in situ localization of those cells and isolation of pure cell types. Here, we aimed at characterization of surface markers in mouse BM MSCs and in their subsets with distinct differentiation potential. Using conditionally immortalized BM MSCs we performed a screening with 176 antibodies and high-throughput flow cytometry, and found 33 markers expressed in MSCs, and among them 3 were novel for MSCs and 13 have not been reported for MSCs from mice. Furthermore, we obtained clonally derived MSC subpopulations and identified bipotential progenitors capable for osteo- and adipogenic differentiation, as well as monopotential osteogenic and adipogenic clones, and thus confirmed heterogeneity of MSCs. We found that expression of CD200 was characteristic for the clones with osteogenic potential, whereas SSEA4 marked adipogenic progenitors lacking osteogenic capacity, and CD140a was expressed in adipogenic cells independently of their efficiency for osteogenesis. We confirmed our observations in cell sorting experiments and further investigated the expression of those markers during the course of differentiation. Thus, our findings provide to our knowledge the most comprehensive characterization of surface antigens expression in mouse BM MSCs to date, and suggest CD200, SSEA4 and CD140a as markers differentially expressed in distinct types of MSC progenitors.

Rostovskaya, Maria; Anastassiadis, Konstantinos

2012-01-01

256

Comparison of technetium 99m-phytate and technetium 99m-sulphur colloid in primary bone tumours.  

PubMed

Eleven patients with proven primary bone tumour (five Ewing sarcomas, six osteosarcomas) and two cases of metastatic bone involvement (primary other than bone) were investigated with 99mTc-phytate and 99mTc-sulphur colloid to compare the behaviour of the two radiopharmaceuticals at the tumour site. After intravenous administration of the respective radiopharmaceutical, imaging of the tumour site and its contralateral part was carried out at 15 min and 1 h intervals. The data were stored in our computer. Bone scanning was also carried out in all patients. 99mTc-phytate uptake was observed at the tumour site in ten cases. The 99mTc-sulphur colloid study revealed sparse or no significant uptake in eight cases. In two patients, with osteosarcoma 99mTc-sulphur colloid investigation showed uptake at the primary tumour site. However, the distribution pattern is different from that of 99mTc-phytate. No significant uptake of either 99mTc-phytate or 99mTc-sulphur colloid was observed in the two patients with metastatic skeletal disease. It may be concluded that the unusual accumulation of 99mTc-phytate at the tumour site is not due to any generalized reticuloendothelial phenomenon and that the radiopharmaceutical itself is responsible for this. PMID:2209654

Rao, P N; Murthy, S N; Muddukrishna, S N; Devaru, S; Radhakrishnan, E R; Bhargava, M K

1990-01-01

257

INFLUENCE OF LEPTIN ON THE EARLY PHASES OF BONE FORMATION DURING MOUSE SKELETON ORGANOGENESIS  

Microsoft Academic Search

Leptin is a 16-kDa hormone, primarily secreted by adipose tissue, which controls body weight through its effects on food intake and energy expenditure by negative feedback at the hypothalamic nuclei. Leptin is now known to have actions in the immune system, reproduction, development, haemopoiesis, angiogenesis and, most recently, in bone metabolism (1). Concerning the skeleton, it seems to be involved

Rivasi Marianna; Benelli Augusta; Ferretti Marzia; Benincasa Marta; Bertoni Laura; Cavani Francesco; Palumbo Carla

2007-01-01

258

Onset of apoprotein E secretion during differentiation of mouse bone marrow-derived mononuclear phagocytes  

SciTech Connect

A number of macrophage functions were sequentially expressed when the bone marrow precursors of mononuclear phagocytes differentiated in culture in the presence of a specific growth factor, colony-stimulating factor-1. The authors defined the expression of apoprotein E (ApoE), a major secreted protein of resident peritoneal macrophages, during maturation of adherent bone marrow-derived mononuclear phagocytes into macrophages. By 5 d the bone marrow macrophages were active secretory cells, but few cells contained intracellular immunoreactive ApoE, and little, if any, ApoE was secreted. ApoE secretion was initiated at 9 d, and this correlated with an increase in the percentage of macrophages containing intracellular ApoE. The onset of ApoE secretion was selective, and little change occurred in the other major secreted proteins detected by (/sup 35/S)methionine incorporation. In parallel, the high rate of plasminogen activator secretion, which peaked at 7 d, decreased markedly. ApoE secretion was not associated with altered expression of the macrophage surface antigen, la, or with secretion of fibronectin. Virtually all cells in independent colonies of bone marrow-derived macrophages eventually expressed ApoE. The proliferating monocyte/macrophage-like cell lines P388D1, J774.2, WHEI-3, RAW 264.1, and MGI.D/sup +/ secreted little or no ApoE. These data establish that ApoE secretion is developmentally regulated.

Werb, Z.; Chin, J.R.

1983-10-01

259

USP6 and CDH11 Oncogenes Identify the Neoplastic Cell in Primary Aneurysmal Bone Cysts and Are Absent in So-Called Secondary Aneurysmal Bone Cysts  

PubMed Central

Aneurysmal bone cyst (ABC) is a locally recurrent bone lesion that has been regarded as a reactive process. Recently, a neoplastic basis in primary ABC was evidenced by demonstration of clonal chromosome band 17p13 translocations that place the USP6 (TRE2 or TRE17) oncogene under the regulatory influence of the highly active CDH11 promoter. Herein, we report CDH11 and/or USP6 rearrangements in 36 of 52 primary ABCs (69%), of which 10 had CDH11-USP6 fusion, 23 had variant USP6 rearrangements without CDH11 rearrangement, and three had variant CDH11 rearrangements without USP6 rearrangement. USP6 and CDH11 rearrangements were restricted to spindle cells in the ABC and were not found in multinucleated giant cells, inflammatory cells, endothelial cells, or osteoblasts. CDH11 and USP6 rearrangements did not correlate with recurrence-free survival, or with other clinicopathological features. CDH11 and USP6 rearrangements were not found in any of 17 secondary ABC associated with giant cell tumor, chondroblastoma, osteoblastoma, and fibrous dysplasia. These findings demonstrate that primary ABC are mesenchymal neoplasms exhibiting USP6 and/or CDH11 oncogenic rearrangements. By contrast, secondary ABC lack CDH11 and USP6 rearrangements, and although morphological mimics of primary ABC, appear to represent a non-specific morphological pattern of a diverse group of non-ABC neoplasms.

Oliveira, Andre M.; Perez-Atayde, Antonio R.; Inwards, Carrie Y.; Medeiros, Fabiola; Derr, Victoria; Hsi, Bae-Li; Gebhardt, Mark C.; Rosenberg, Andrew E.; Fletcher, Jonathan A.

2004-01-01

260

Exogenous administration of 15d-PGJ2-loaded nanocapsules inhibits bone resorption in a mouse periodontitis model.  

PubMed

The 15-deoxy-(?12,14)-PG J(2) (15d-PGJ(2)) has demonstrated excellent anti-inflammatory results in different experimental models. It can be used with a polymeric nanostructure system for modified drug release, which can change the therapeutic properties of the active principle, leading to increased stability and slower/prolonged release. The aim of the current study was to test a nanotechnological formulation as a carrier for 15d-PGJ(2), and to investigate the immunomodulatory effects of this formulation in a mouse periodontitis model. Poly (D,L-lactide-coglycolide) nanocapsules (NC) were used to encapsulate 15d-PGJ(2). BALB/c mice were infected on days 0, 2, and 4 with Aggregatibacter actinomycetemcomitans and divided into groups (n = 5) that were treated daily during 15 d with 1, 3, or 10 ?g/kg 15d-PGJ(2)-NC. The animals were sacrificed, the submandibular lymph nodes were removed for FACS analysis, and the jaws were analyzed for bone resorption by morphometry. Immunoinflammatory markers in the gingival tissue were analyzed by reverse transcriptase-quantitative PCR, Western blotting, or ELISA. Infected animals treated with the 15d-PGJ(2)-NC presented lower bone resorption than infected animals without treatment (p < 0.05). Furthermore, infected animals treated with 10 ?g/kg 15d-PGJ(2)-NC had a reduction of CD4(+)CD25(+)FOXP3(+) cells and CD4/CD8 ratio in the submandibular lymph node (p < 0.05). Moreover, CD55 was upregulated, whereas RANKL was downregulated in the gingival tissue of the 10 ?g/kg treated group (p < 0.05). Several proinflammatory cytokines were decreased in the group treated with 10 ?g/kg 15d-PGJ(2)-NC, and high amounts of 15d-PGJ(2) were observed in the gingiva. In conclusion, the 15d-PGJ(2)-NC formulation presented immunomodulatory effects, decreasing bone resorption and inflammatory responses in a periodontitis mouse model. PMID:22706081

Napimoga, Marcelo H; da Silva, Carlos A T; Carregaro, Vanessa; Farnesi-de-Assunção, Thais S; Duarte, Poliana M; de Melo, Nathalie F S; Fraceto, Leonardo F

2012-06-15

261

Effect of L-carnitine on erythroid colony formation in mouse bone marrow cells  

Microsoft Academic Search

Background. L-Carnitine can alleviate uraemic anaemia in haemodialysis patients by improving erythrocyte membrane functions or erythropoiesis, which are depressed under uraemic conditions. L-Carnitine and palmitoyl-L-carnitine were reported to increase the formation of colony-forming unit-erythroid (CFU-E) colonies in cultures of fetal mouse liver cells, an effect that depended on the concentration of palmitoyl-L- carnitine but not of L-carnitine. In this study,

Yukika Kitamura; Kazunori Satoh; Takashi Satoh; Manabu Takita; Akihiro Matsuura

262

Conservation and divergence in Toll-like receptor 4-regulated gene expression in primary human versus mouse macrophages.  

PubMed

Evolutionary change in gene expression is generally considered to be a major driver of phenotypic differences between species. We investigated innate immune diversification by analyzing interspecies differences in the transcriptional responses of primary human and mouse macrophages to the Toll-like receptor (TLR)-4 agonist lipopolysaccharide (LPS). By using a custom platform permitting cross-species interrogation coupled with deep sequencing of mRNA 5' ends, we identified extensive divergence in LPS-regulated orthologous gene expression between humans and mice (24% of orthologues were identified as "divergently regulated"). We further demonstrate concordant regulation of human-specific LPS target genes in primary pig macrophages. Divergently regulated orthologues were enriched for genes encoding cellular "inputs" such as cell surface receptors (e.g., TLR6, IL-7R?) and functional "outputs" such as inflammatory cytokines/chemokines (e.g., CCL20, CXCL13). Conversely, intracellular signaling components linking inputs to outputs were typically concordantly regulated. Functional consequences of divergent gene regulation were confirmed by showing LPS pretreatment boosts subsequent TLR6 responses in mouse but not human macrophages, in keeping with mouse-specific TLR6 induction. Divergently regulated genes were associated with a large dynamic range of gene expression, and specific promoter architectural features (TATA box enrichment, CpG island depletion). Surprisingly, regulatory divergence was also associated with enhanced interspecies promoter conservation. Thus, the genes controlled by complex, highly conserved promoters that facilitate dynamic regulation are also the most susceptible to evolutionary change. PMID:22451944

Schroder, Kate; Irvine, Katharine M; Taylor, Martin S; Bokil, Nilesh J; Le Cao, Kim-Anh; Masterman, Kelly-Anne; Labzin, Larisa I; Semple, Colin A; Kapetanovic, Ronan; Fairbairn, Lynsey; Akalin, Altuna; Faulkner, Geoffrey J; Baillie, John Kenneth; Gongora, Milena; Daub, Carsten O; Kawaji, Hideya; McLachlan, Geoffrey J; Goldman, Nick; Grimmond, Sean M; Carninci, Piero; Suzuki, Harukazu; Hayashizaki, Yoshihide; Lenhard, Boris; Hume, David A; Sweet, Matthew J

2012-03-26

263

Expression of Bone Morphogenetic Protein Receptors in the Developing Mouse Metanephros  

Microsoft Academic Search

While bone morphogenetic proteins (BMPs) 2, 4 and 7 have recently been implicated in aspects of metanephric development, and expression patterns of these ligands have been described in the developing metanephros, the distribution of BMP receptors in developing metanephroi remains unknown. In the present study, in situ hybridisation histochemistry was used to localise mRNAs for BMP type-I receptors (BMPR-IA and

Gemma Martinez; Kate L. Loveland; Amander T. Clark; Marie Dziadek; John F. Bertram

2001-01-01

264

Clastogenic potential of Ruta graveolens extract and a homeopathic preparation in mouse bone marrow cells.  

PubMed

Ruta graveolens belonging to family Rutaceae has long been traditionally used as a medicinal plant as well as a flavoring agent in food. However, very little data are available on the toxicity of the plant. This report presents evidence on the genotoxic and clastogenic potential of an extract of Ruta graveolens and Ruta 200C, a homeopathic preparation. Various types of chromosomal aberrations were noted in bone marrow cells after treatment. The percentage of aberrated cells in the 400mg/kgb.wt extract administered group was found to be 21% and with 1,000 mg/kg.b.wt it was 31%. The value for the Ruta 200C treated group was also elevated to 23% as compared to the 3%for untreated animals. In addition, bone marrow cells had higher incidence of micronuclei induction when treated with the extract (400 mg and 1,000 mg/kg body weight) and Ruta 200C for 30 days. Administration of the extract (1,000 mg/kg.b.wt) over a period of 30 days also resulted in damage to cellular DNA as evidenced by comet formation where the comet parameters such as percentage DNA in tail, tail length, tail moment of the bone marrow cells were increased several fold over control values. The comet tail moment of the bone marrow cells increased from 4.5 to 50.2 after the extract treatment. Administration of Ruta 200C for 5 consecutive days increased the tail moment to 11.7. These results indicate that Ruta graveolens and Ruta 200C may induce genotoxicity in animals. PMID:19256773

Preethi, Korengath C; Nair, Cherappally K K; Kuttan, Ramadasan

265

Genotoxic effect of Anovlar 21, an oral contraceptive, on mouse bone marrow.  

PubMed

Anovlar 21, a combination drug containing the oestrogen ethinyloestradiol and the progestin norethisterone acetate, was studied for its in vivo genotoxic effect on the bone marrow cells of Swiss albino mice. The chromosomal aberration assay and the micronucleus test were employed for the study. 0.08, 0.4, 0.8, 1.6, 3.2, 4.8, 6.4 and 8.0 mg/kg/day of the drug was orally administered for 15 consecutive days to mice. Bone marrow preparations were made 24 h after the final feeding. The lowest dose, 0.08 mg/kg, represents the human therapeutic range. Marrow preparations of mice fed 0.8 mg/kg/day for 15 days were made at 6, 12, 24, 48 and 96 h, and 1, 2 and 3 weeks and a time-yield analysis was carried out. Statistically significant increases in chromosomal aberrations were observed in animal groups fed doses of greater than or equal to 0.4 mg/kg/day. In the time-response study, the maximum frequency of aberrations was noted at 24 h, thereafter decreasing gradually with increasing time. But the drug did not induce a significant increase in the number of micronuclei in bone marrow erythrocytes at any of the doses or time intervals studied. PMID:2027340

Shyama, S K; Abdul Rahiman, M; Vijayalaxmi, K K

1991-05-01

266

Differential effects of simvastatin on IL-13-induced cytokine gene expression in primary mouse tracheal epithelial cells  

PubMed Central

Background Asthma causes significant morbidity worldwide in adults and children alike, and incurs large healthcare costs. The statin drugs, which treat hyperlipidemia and cardiovascular diseases, have pleiotropic effects beyond lowering cholesterol, including immunomodulatory, anti-inflammatory, and anti-fibrotic properties which may benefit lung health. Using an allergic mouse model of asthma, we previously demonstrated a benefit of statins in reducing peribronchiolar eosinophilic inflammation, airway hyperreactivity, goblet cell hyperplasia, and lung IL-4 and IL-13 production. Objectives In this study, we evaluated whether simvastatin inhibits IL-13-induced pro-inflammatory gene expression of asthma-related cytokines in well-differentiated primary mouse tracheal epithelial (MTE) cell cultures. We hypothesized that simvastatin reduces the expression of IL-13-inducible genes in MTE cells. Methods We harvested tracheal epithelial cells from naïve BALB/c mice, grew them under air-liquid interface (ALI) cell culture conditions, then assessed IL-13-induced gene expression in MTE cells using a quantitative real-time PCR mouse gene array kit. Results We found that simvastatin had differential effects on IL-13-mediated gene expression (inhibited eotaxin-1; MCP-1,-2,-3; and osteopontin (SPP1), while it induced caspase-1 and CCL20 (MIP-3?)) in MTE cells. For other asthma-relevant genes such as TNF, IL-4, IL-10, CCL12 (MCP-5), CCL5 (RANTES), and CCR3, there were no significant IL-13-inducible or statin effects on gene expression. Conclusions Simvastatin modulates the gene expression of selected IL-13-inducible pro-inflammatory cytokines and chemokines in primary mouse tracheal epithelial cells. The airway epithelium may be a viable target tissue for the statin drugs. Further research is needed to assess the mechanisms of how statins modulate epithelial gene expression.

2012-01-01

267

Discordant hormone receptor and human epidermal growth factor receptor 2 status in bone metastases compared to primary breast cancer.  

PubMed

Abstract Background. In patients with metastatic breast cancer, the evaluation of the biological characteristics of metastatic bone deposits may be a valuable adjunct in clinical practice. We assessed the discordance in expression levels for estrogen receptor (ER), progesterone receptor (PgR) and human epidermal growth factor receptor 2 (HER2) between primary tumor and bone metastases and its clinical impact on patient management. Material and methods. We retrospectively reviewed 363 CT-guided bone biopsies performed from January 1997 to December 2009. The proportions of ER, PgR and HER2 positive tumors at primary diagnosis and bone metastases, determined by IHC and/or FISH, were compared using McNemar's test. The impact of the biopsy reassessment on treatment choice was evaluated with Fisher's exact test. Results. We selected 109 metastatic breast cancer patients with histologically confirmed bone metastases. Among 107 assessable patients the overall discordance rate was detected in 22 (20.5%) and in 47 (43.9%) patients for ER and PgR, respectively, and in six of 86 assessable patients (6.9%) for HER2 status. The indication to change endocrine therapy occurred in 62% and 30% of patients with ER discordance and ER concordance, respectively (p = 0.01). The indication to change targeted therapy occurred in 67% and 8% of patients with HER2 discordance and HER2 concordance, respectively (p = 0.002). Conclusions. We confirm that biopsy of metastases, including bone metastases, for reassessment of biology should be considered, since it is likely to impact on treatment choice. PMID:23327413

Aurilio, Gaetano; Monfardini, Lorenzo; Rizzo, Stefania; Sciandivasci, Angela; Preda, Lorenzo; Bagnardi, Vincenzo; Disalvatore, Davide; Pruneri, Giancarlo; Munzone, Elisabetta; Della Vigna, Paolo; Renne, Giuseppe; Bellomi, Massimo; Curigliano, Giuseppe; Goldhirsch, Aron; Nolè, Franco

2013-01-17

268

Improved Peripheral Cortical Bone Geometry after Surgical Treatment of Primary Hyperparathyroidism in Postmenopausal Women  

Microsoft Academic Search

Objective: The present study was performed to examine the longitudinal effects of treating en- dogenous PTH excess on cortical bone geometry in postmenopausal patients with pHPT by using peripheral quantitative computed tomography. Patients:TwentypostmenopausalpHPTpatientsand30postmenopausalcontrolsubjectsmatched for age participated in this study. Main Outcome Measures: Volumetric bone mineral density (vBMD), cortical bone geometric pa- rameters, polar strength strain index, and polar cross-sectional moment

Hiroshi Kaji; Mika Yamauchi; Rikako Nomura; Toshitsugu Sugimoto

2010-01-01

269

The influence of stem size and extent of porous coating on femoral bone resorption after primary cementless hip arthroplasty.  

PubMed

The influence of stem size and extent of porous coating on femoral bone resorption was examined in 411 cases of primary cementless hip arthroplasty. Moore design, cobalt alloy femoral implants with powder-made sintered porous coating on either one-third, two-thirds, or the full implant length were compared radiographically two years after surgery. A semiquantitative method was adopted for assessing resorption that involved dividing the anteroposterior (AP) and lateral roentgenograms into a total of 16 discrete sites. The 16 sites were qualitatively examined for evidence of resorption by either thinning or darkening of bone relative to the time immediately following surgery. Based on the number of sites that demonstrated resorption, the bone loss was classified as either minor and not likely to cause problems (0 to 4 sites) or pronounced and of potentially harmful clinical consequence (5 or more sites). Pronounced resorption occurred in 18% of the 411 cases. The use of larger stems resulted in increased occurrence of marked bone resorption: stems greater than or equal to 13.5 mm in diameter showed five times the incidence of pronounced resorption compared with stems less than or equal to 12.0 mm in diameter. Stems with two-thirds and full porous coating resulted in a twofold to fourfold increase in the incidence of pronounced bone resorption. The theoretic degree of stress shielding of the femoral shaft in bending was calculated for cases with complete canal filling and a radiographic appearance of bone ingrowth. There was a strong correlation between this theoretic factor and the observed bone resorption. PMID:3370887

Engh, C A; Bobyn, J D

1988-06-01

270

Morphogenesis and proliferation in mono- and organotypic co-cultures of primary human periodontal ligament fibroblasts and alveolar bone cells  

Microsoft Academic Search

Cells of the periodontal ligament and the alveolar bone lie in close vicinity in the periodontium. The goal of this study was to create an in vitro model to facilitate the study of the morphogenesis and proliferation of these two cell types under more in-vivo-like conditions. This was accomplished by the generation of organotypic co-cultures of primary human periodontal ligament

T. Reuther; A. Kohl; G. Komposch; P. Tomakidi

2003-01-01

271

Proliferation effect of He-Ne laser intermittent irradiation on mouse bone-marrow cells  

NASA Astrophysics Data System (ADS)

The effect of He-Ne laser intermittent irradiation on the bone marrow cell suspension of donor mice is presented in this paper. The recipient mice, irradiated with 8.5 GY CO60-(gamma) ray, were then infused, and they were killed different days (1, 3, 5, 7, 9, and 11 days post injection). Spleens were removed and fixed in Bouin's solution for 24 hours, and then the numbers of protruding splenic nodules visible on the surface of the spleens were counted. According to statistics, the number of the splenic nodules increased with laser exposure over the control group.

Chen, Ji; Zhang, Jianjun

1993-03-01

272

Effect of bone marrow depletion on prostaglandin E-producing suppressor macrophages in mouse spleen  

SciTech Connect

The i.p. injection of Corynebacterium parvum (CP) into CBA/J mice effected increases in macrophage colony-forming cells (M-CFC) when spleen cells were cultured with L cell culture filtrate as a source of colony-stimulating factor. Significant increases in phagocytic macrophages (M phi) with Fc receptors for IgG2a and IgG2b immune complexes were additionally noted among the spleen cells in these mice. These M phi effectively inhibited Con A-induced lymphocyte proliferation, probably reflecting a 10-fold increase above normal controls in prostaglandin E to 47 ng/3 X 10(6) spleen cells/ml. To determine whether the suppressor M phi are immediate derivatives of splenic M-CFC, we tried to induce suppressor M phi by the injection of CP into mice depleted of bone marrow M-CFC by the earlier administration of the bone-seeking isotope, 89Sr. This procedure reduced M-CFC in the bone marrow to less than 1% of normal for more than 30 days. Monocytes in the blood fell to 5% of normal by day 10 and were 30% on day 30. Levels of resident peritoneal M phi showed relatively little change in this period. By contrast, splenic M-CFC increased to 20-fold higher than the cold 88Sr controls. CP-induced suppressor M phi activity, however, was sharply reduced in 89Sr marrow-depleted mice on day 10, despite the striking increase in M-CFC. There was a threefold increase in the number of phagocytic M phi binding IgG2a immune complexes, with no significant increase in IgG2b binding M phi. The kinetics of recovery of suppressor M phi activity showed that on days 20, 30, and 50 after 89Sr injection the activities reached 20%, 30%, and 70% of the cold control, respectively, and correlated with the recovery of significant levels of M-CFC in the bone marrow. Taken together, these observations suggest that splenic M-CFC are not an immediate source of PGE-suppressor M phi in vivo.

Shibata, Y.; Volkman, A.

1985-12-01

273

Protection of mouse bone marrow from etoposide-induced genomic damage by dexrazoxane  

Microsoft Academic Search

Purpose  The objective of the current investigation is to determine whether non-toxic doses of the catalytic topoisomerase-II inhibitor,\\u000a dexrazoxane, have influence on the genomic damage induced by the anticancer topoisomerase-II poison, etoposide, on mice bone\\u000a marrow cells.\\u000a \\u000a \\u000a \\u000a Method  The scoring of micronuclei, chromosomal aberrations, and mitotic activity were undertaken as markers of cyto- and genotoxicity.\\u000a Oxidative damage markers such as reduced glutathione

Sabry M. Attia; Alaa A. Al-Anteet; Nouf M. Al-Rasheed; Abdulqader A. Alhaider; Mohammed M. Al-harbi

2009-01-01

274

Water permeability of primary mouse keratinocyte cultures grown at the air-liquid interface  

SciTech Connect

In order to study the development of the epidermal permeability barrier in vitro, tritiated water (HTO) flux was measured across murine keratinocytes cultured at the air-liquid interface. Using a micro-diffusion technique, it was shown that air-liquid cultures form areas where the water diffusion is comparable to that of intact neonatal mouse skin. When water permeability is measured over a large area of the culture surface, however, significantly higher flux is obtained. These results show that under the culture conditions used, areas of water barrier comparable to intact neonatal mouse skin coexist with regions of less complete barrier formation.

Cumpstone, M.B.; Kennedy, A.H.; Harmon, C.S.; Potts, R.O.

1989-04-01

275

Comparison of proliferation and differentiation potential between mouse primary hepatocytes and embryonic hepatic progenitor cells in vitro.  

PubMed

Cell therapy may be a novel and effective treatment strategy for liver diseases, replacing liver transplantation. The potential of two alternative cell types (hepatic progenitor/stem cells and mature hepatocytes) has not yet been fully assessed; the issues of low amplification efficiency and recovery function remain to be resolved. In this study, we investigated the proliferation, differentiation and function of primary mouse mature hepatocytes and embryonic hepatic progenitor cells. Primary cells were obtained from the livers of mouse embryos at 14.5 days post coitus [hepatic progenitor 14.5d (HP14.5d) cells], as well as from the livers of 3-month-old mice [liver cells 3m (LC3m)]. Using trypan blue staining and crystal violet staining to detect cell viability, we found that compared with the limited growth capability of primary LC3m cells, primary HP14.5d cells exhibited an active cell proliferation; however, proliferative ability of passaged HP14.5d cells significantly decreased. After the HP14.5d cells were treated in hepatic induction medium, the expression of progenitor cell markers decreased and that of mature hepatic markers increased, to levels similar to those of LC3m cells. On day 12 of induction, the HP14.5d cells showed comparable indocyanine green (ICG) uptake and glycogen storage to that of the LC3m cells. Therefore, our study demonstrates that primary hepatic progenitor cells have a stronger proliferation capacity and differentiation potential, supporting their clinical application in liver cell transplantation. PMID:23756629

He, Yun; Zhou, Jian-Wu; Xu, Lei; Gong, Meng-Jia; He, Tong-Chuan; Bi, Yang

2013-06-11

276

Thymic repopulation following intrathymic injection of mouse bone marrow cells in MHC matched and mismatched recipients  

SciTech Connect

T cell precursors (pre-T cells) have traditionally been detected by their ability to repopulate the thymus of heavily irradiated mice following intravenous injection. Recently, Goldschneider et. al. developed an assay system which involves the direct injection of pre-T cells into the thymus. The authors used this technique to evaluate the ability of bone marrow cells to repopulate thymuses in various donor-host strain combinations. Sub-lethally irradiated (600R) mice were injected intrathymically with 2 x 10/sup 6/ bone marrow cells which differed from the recipient with respect to their Thy 1 allotype. The percentage of thymus cells expressing either the donor or recipient type Thy 1 marker was determined 14 to 21 days after injection. These experiments showed that in MHC matched donor-host combinations (AKR/cum ..-->.. AKR/J and CBA/J ..-->.. AKR/J), cells derived from the donor inoculum accounted for 40% to 75% of the total thymus population. MHC mismatched donor-host combinations (C57BL/6J ..-->.. AKR/J and Balb/c ..-->.. AKR/J) resulted in significantly less donor-type repopulation of the thymus. In these cases, donor repopulation typically ranged from 0% to 4%. The ability of the pre-T cells detected by intrathymic injection to proliferate in the thymic environment, therefore, appears to be influenced by the MHC. This may reflect commitment of pre-T cells to MHC haplotype recognition prior to their migration to the thymus.

Chervenak, R.

1986-03-01

277

The Use of Structural Allograft in Primary and Revision Knee Arthroplasty with Bone Loss  

PubMed Central

Bone loss around the knee in the setting of total knee arthroplasty remains a difficult and challenging problem for orthopaedic surgeons. There are a number of options for dealing with smaller and contained bone loss; however, massive segmental bone loss has fewer options. Small, contained defects can be treated with cement, morselized autograft/allograft or metal augments. Segmental bone loss cannot be dealt with through simple addition of cement, morselized autograft/allograft, or metal augments. For younger or higher demand patients, the use of allograft is a good option as it provides a durable construct with high rates of union while restoring bone stock for future revisions. Older patients, or those who are low demand, may be better candidates for a tumour prosthesis, which provides immediate ability to weight bear and mobilize.

Kuchinad, Raul A.; Garbedian, Shawn; Rogers, Benedict A.; Backstein, David; Safir, Oleg; Gross, Allan E.

2011-01-01

278

Implication of connexins 40 and 43 in functional coupling between mouse cardiac fibroblasts in primary culture  

Microsoft Academic Search

Cardiac fibroblasts contribute to the structure and function of the myocardium. However their involvement in electrophysiological processes remains unclear; particularly in pathological situations when they proliferate and develop fibrosis. We have identified the connexins involved in gap junction channels between fibroblasts from adult mouse heart and characterized their functional coupling. RT-PCR and Western blotting results show that mRNA and proteins

Claire Louault; Najate Benamer; Jean-François Faivre; Daniel Potreau; Jocelyn Bescond

2008-01-01

279

Bone tissue ultrastructural defects in a mouse model for osteogenesis imperfecta: a Raman spectroscopy study  

NASA Astrophysics Data System (ADS)

Osteogenesis imperfecta (OI) is genetic defect in which the genes that code for the ?1(I) or ?2(I) chains of type I collagen are defective. The defects often result in substitution of a bulky amino acid for glycine, causing formation of collagen that can not form the normal triple helix. Depending on the details of the defects, the outcomes range from controllable to lethal. This study focuses on OI type IV, a more common and moderately severe form of the disease. People with the disease have a substantial increase in the risk and rate of fracture. We examine the spectroscopic consequences of these defects, using a mouse model (BRTL) that mimics OI type IV. We compare Raman images from tibial cortical tissue of wild-type mice and BRTL mice with single copy of mutation and show that both mineral to matrix ratios and collagen inter-fibril cross-links are different in wild-type and mutant mice.

Chen, Tsoching; Kozloff, Kenneth M.; Goldstein, Steven A.; Morris, Michael D.

2004-07-01

280

Effect of parathyroidectomy versus risedronate on volumetric bone mineral density and bone geometry at the tibia in postmenopausal women with primary hyperparathyroidism.  

PubMed

The objective of the study was to evaluate the effect of parathyroidectomy (PTX) versus 35 mg once-weekly (ow) risedronate administration on volumetric bone mineral density (vBMD) and bone geometry at the tibia in postmenopausal women with primary hyperparathyroidism (PHPT). Our open-label prospective observational study included 32 postmenopausal women with PHPT as the study group: 16 underwent PTX and 16 were treated with 35 mg ow risedronate for 2 years. We assessed areal BMD (aBMD) by DXA, and vBMD and bone mineral content (BMC) (cortical and trabecular area) by peripheral quantitative computed tomography (pQCT) at the tibia at baseline and at 2 years. Risedronate did not result in any significant change on vBMD and structural pQCT indices. PTX resulted in significant increase in trabecular (trab) BMC (6.44 %) and vBMD (4.64 %), with percent increase being significantly higher than risedronate (p < 0.05). At cortical sites, there was no significant change following PTX. However, the percent change in cortical (cort) vBMD was higher following PTX versus risedronate (0.39 % vs. -0.26 %, p < 0.05). In conclusion, in postmenopausal women with PHPT, PTX is superior to ow risedronate, in terms of improvement of trabecular mineralization and vBMD at the tibia, whereas the effect at cortical sites is less pronounced. PMID:23700284

Tournis, Symeon; Fakidari, Eleni; Dontas, Ismene; Liakou, Chrysoula; Antoniou, Julia; Galanos, Antonis; Marketou, Helen; Makris, Konstantinos; Katsalira, Katerina; Trovas, George; Lyritis, George P; Papaioannou, Nikolaos

2013-05-23

281

Early stage transplantation of bone marrow cells markedly ameliorates copper metabolism and restores liver function in a mouse model of Wilson disease  

Microsoft Academic Search

Background  Recent studies have demonstrated that normal bone marrow (BM) cells transplantation can correct liver injury in a mouse model\\u000a of Wilson disease (WD). However, it still remains unknown when BM cells transplantation should be administered. The aim of\\u000a this study was to investigate the potential impact of normal BM cells transplantation at different stages of WD to correct\\u000a liver injury

Xi Chen; Shihui Xing; Yanqing Feng; Songlin Chen; Zhong Pei; Chuhuai Wang; Xiuling Liang

2011-01-01

282

Comparative in-vivo genotoxicity of antiviral nucleoside analogues; penciclovir, acyclovir, ganciclovir and the xanthine analogue, caffeine, in the mouse bone marrow micronucleus assay  

Microsoft Academic Search

Three purine nucleoside analogues, penciclovir (PCV), acyclovir (ACV) and ganciclovir (GCV), were assessed for in-vivo genotoxicity in the mouse bone marrow micronucleus assay, together with the xanthine (purine) analogue, caffeine (CAF). All these compounds exhibit anti-viral properties and the first three are marketed anti-viral drugs. All have been shown to be genotoxic in separate in-vitro and in-vivo studies. Because of

P. Haynes; T. R. Lambert; I. de G. Mitchell

1996-01-01

283

[En bloc excision and reconstruction of the defect with pasteurized autograft for femoral primary malignant bone tumor in childen].  

PubMed

Objective: To evaluate the efficacy of femoral primary malignant bone tumors treated by limb salvage, with wide en bloc excision and reconstruction of the defect with recycled pasteurized autograft. Methods: From January 2008 to January 2011, 11 patients (7 males, 4 females), aged (11.0±2.5) years suffering from femoral primary malignant bone tumors were treated with en bloc excision and reconstruction of the defect with recycled pasteurized autograft. Ten patients were histopathologically diagnosed with high-grade osteosarcoma, and 1 with Ewing's sarcoma.According to the Enneking staging system, all patients were in Stage IIb. Results: All patients were followed-up for 24-65(42±20) months and all showed bony union at the last follow-up. The length of tumor bone resection was (17.5±3.2) cm, the operation time was (180±35) min, the intraoperative blood loss was (1200±250) mL, and drainage volume was (650±125) mL. Local skin necrosis occurred in 1 patient (9.1%), which was resolved by debridement. Nonunion occurred in 3 patients (27.3%), who were treated by secondary iliac crest cancellous bone grafting.Pulmonary metastasis occurred in 1 patient (9.1%) who died 35 months post-operatively. According to the function assessment by the Enneking system, 5 patients had excellent results, 4 had good results, 1 fair and 1 poor results, with a satisfaction rate of 81.2%. Conclusion: A pasteurized autograft can be an easily accessible and economical alternative for children's malignant bone tumor of femurs. PMID:23981993

Zhang, Qing; Liu, Tang; Guo, Xiaoning; Lin, Ling

2013-08-01

284

Primary bone carcinosarcoma of the fibula with chondrosarcoma and squamous cell carcinoma components.  

PubMed

Carcinosarcoma is defined as a malignant neoplasm that is composed of both carcinomatous and sarcomatous components. The occurrence of carcinosarcoma in the bone is extremely rare. In this report, we describe the third documented de novo case of carcinosarcoma of the bone. A 59-year-old Japanese female presented with a painful tumor in her right lower leg. Plane radiography revealed an osteolytic destructive lesion with periosteal reaction and mineralization in the right fibula. Resection of the fibula tumor was performed under a clinical diagnosis of chondrosarcoma. Histopathological study revealed that the tumor was comprised of three components. The main component was proliferation of small round to short spindle cells (approximately 50%), and the remaining components were chondrosarcoma (30%) and squamous cell carcinoma (20%). Immunohistochemically, SOX9 was expressed in the small round to spindle cells and chondrosarcoma component, and p63 and p40 were expressed in all three components. Accordingly, an ultimate diagnosis of carcinosarcoma of the bone was made. The clinicopathological analysis of carcinosarcoma of the bone revealed that this type of tumor affects the middle-aged to elderly persons and occurs in the long bone. All three de novo cases had chondrosarcoma and squamous cell carcinoma components. One of the 3 patients died of the disease. The histogenesis of carcinosarcoma of the bone remains a matter of controversy, although a multpotential stem cell theory has been proposed. Additional studies are required to clarify the clinical behavior and histogenesis of carcinosarcoma of the bone. PMID:24133601

Ishida, Mitsuaki; Kodama, Narihito; Takemura, Yoshinori; Iwai, Muneo; Yoshida, Keiko; Kagotani, Akiko; Matsusue, Yoshitaka; Okabe, Hidetoshi

2013-09-15

285

Primary bone carcinosarcoma of the fibula with chondrosarcoma and squamous cell carcinoma components  

PubMed Central

Carcinosarcoma is defined as a malignant neoplasm that is composed of both carcinomatous and sarcomatous components. The occurrence of carcinosarcoma in the bone is extremely rare. In this report, we describe the third documented de novo case of carcinosarcoma of the bone. A 59-year-old Japanese female presented with a painful tumor in her right lower leg. Plane radiography revealed an osteolytic destructive lesion with periosteal reaction and mineralization in the right fibula. Resection of the fibula tumor was performed under a clinical diagnosis of chondrosarcoma. Histopathological study revealed that the tumor was comprised of three components. The main component was proliferation of small round to short spindle cells (approximately 50%), and the remaining components were chondrosarcoma (30%) and squamous cell carcinoma (20%). Immunohistochemically, SOX9 was expressed in the small round to spindle cells and chondrosarcoma component, and p63 and p40 were expressed in all three components. Accordingly, an ultimate diagnosis of carcinosarcoma of the bone was made. The clinicopathological analysis of carcinosarcoma of the bone revealed that this type of tumor affects the middle-aged to elderly persons and occurs in the long bone. All three de novo cases had chondrosarcoma and squamous cell carcinoma components. One of the 3 patients died of the disease. The histogenesis of carcinosarcoma of the bone remains a matter of controversy, although a multpotential stem cell theory has been proposed. Additional studies are required to clarify the clinical behavior and histogenesis of carcinosarcoma of the bone.

Ishida, Mitsuaki; Kodama, Narihito; Takemura, Yoshinori; Iwai, Muneo; Yoshida, Keiko; Kagotani, Akiko; Matsusue, Yoshitaka; Okabe, Hidetoshi

2013-01-01

286

In vitro assays using primary embryonic mouse lymphatic endothelial cells uncover key roles for FGFR1 signalling in lymphangiogenesis.  

PubMed

Despite the importance of blood vessels and lymphatic vessels during development and disease, the signalling pathways underpinning vessel construction remain poorly characterised. Primary mouse endothelial cells have traditionally proven difficult to culture and as a consequence, few assays have been developed to dissect gene function and signal transduction pathways in these cells ex vivo. Having established methodology for the purification, short-term culture and transfection of primary blood (BEC) and lymphatic (LEC) vascular endothelial cells isolated from embryonic mouse skin, we sought to optimise robust assays able to measure embryonic LEC proliferation, migration and three-dimensional tube forming ability in vitro. In the course of developing these assays using the pro-lymphangiogenic growth factors FGF2 and VEGF-C, we identified previously unrecognised roles for FGFR1 signalling in lymphangiogenesis. The small molecule FGF receptor tyrosine kinase inhibitor SU5402, but not inhibitors of VEGFR-2 (SU5416) or VEGFR-3 (MAZ51), inhibited FGF2 mediated LEC proliferation, demonstrating that FGF2 promotes proliferation directly via FGF receptors and independently of VEGF receptors in primary embryonic LEC. Further investigation revealed that FGFR1 was by far the predominant FGF receptor expressed by primary embryonic LEC and correspondingly, siRNA-mediated FGFR1 knockdown abrogated FGF2 mediated LEC proliferation. While FGF2 potently promoted LEC proliferation and migration, three dimensional tube formation assays revealed that VEGF-C primarily promoted LEC sprouting and elongation, illustrating that FGF2 and VEGF-C play distinct, cooperative roles in lymphatic vascular morphogenesis. These assays therefore provide useful tools able to dissect gene function in cellular events important for lymphangiogenesis and implicate FGFR1 as a key player in developmental lymphangiogenesis in vivo. PMID:22792354

Kazenwadel, Jan; Secker, Genevieve A; Betterman, Kelly L; Harvey, Natasha L

2012-07-06

287

In Vitro Assays Using Primary Embryonic Mouse Lymphatic Endothelial Cells Uncover Key Roles for FGFR1 Signalling in Lymphangiogenesis  

PubMed Central

Despite the importance of blood vessels and lymphatic vessels during development and disease, the signalling pathways underpinning vessel construction remain poorly characterised. Primary mouse endothelial cells have traditionally proven difficult to culture and as a consequence, few assays have been developed to dissect gene function and signal transduction pathways in these cells ex vivo. Having established methodology for the purification, short-term culture and transfection of primary blood (BEC) and lymphatic (LEC) vascular endothelial cells isolated from embryonic mouse skin, we sought to optimise robust assays able to measure embryonic LEC proliferation, migration and three-dimensional tube forming ability in vitro. In the course of developing these assays using the pro-lymphangiogenic growth factors FGF2 and VEGF-C, we identified previously unrecognised roles for FGFR1 signalling in lymphangiogenesis. The small molecule FGF receptor tyrosine kinase inhibitor SU5402, but not inhibitors of VEGFR-2 (SU5416) or VEGFR-3 (MAZ51), inhibited FGF2 mediated LEC proliferation, demonstrating that FGF2 promotes proliferation directly via FGF receptors and independently of VEGF receptors in primary embryonic LEC. Further investigation revealed that FGFR1 was by far the predominant FGF receptor expressed by primary embryonic LEC and correspondingly, siRNA-mediated FGFR1 knockdown abrogated FGF2 mediated LEC proliferation. While FGF2 potently promoted LEC proliferation and migration, three dimensional tube formation assays revealed that VEGF-C primarily promoted LEC sprouting and elongation, illustrating that FGF2 and VEGF-C play distinct, cooperative roles in lymphatic vascular morphogenesis. These assays therefore provide useful tools able to dissect gene function in cellular events important for lymphangiogenesis and implicate FGFR1 as a key player in developmental lymphangiogenesis in vivo.

Betterman, Kelly L.; Harvey, Natasha L.

2012-01-01

288

Genetic toxicity evaluation of iodotrifluoromethane (Cf{sub 3}I). Volume 2. Results of in vivo mouse bone marrow erythrocyte micronucleus testing. Final report, March-December 1994  

SciTech Connect

Under subcontract to ManTech Environmental Technology, Incorporated, Genesys Research, Incorporated, examined the potential of odotrifluoromethane (CF3I) to induce structural chromosomes aberrations in erythropoietic cells of the bone marrow. Genesys used the mouse micronucleus test which measures the clastogenic (chromosomes breaking) action of chemicals by the induction of micronuclei in bone marrow cells, as observed in erythrocytes from the peripheral blood of male and female mice obtained approximately 24 hours after steady-state dosing. Based on preliminary toxicity information obtained by ManTech, a mouse bone marrow micronucleus test of CF3I was conducted using 2.6, 5.0, and 7.5% CF3I administered to male and female Swiss Webster mice by inhalation for six hours on each of three consecutive days. Bone marrow cells were obtained from the mice sacrificed 24 hours after the third exposure. Erythrocytes from mice exposed to the test material, and to the negative and positive controls, were evaluated for toxicity and the presence of micronuclei. The positive control, 0.4 mg triethylenemelamine (TEM)/kg (administered intraperitonealy) significantly (pmous` bone marrow micronucleus test and clastogenic in vivo.

Mitchell, A.D.

1995-01-01

289

Bone conducted vibration selectively activates irregular primary otolithic vestibular neurons in the guinea pig.  

PubMed

The main objective of this study was to determine whether bone-conducted vibration (BCV) is equally effective in activating both semicircular canal and otolith afferents in the guinea pig or whether there is preferential activation of one of these classes of vestibular afferents. To answer this question a large number (346) of single primary vestibular neurons were recorded extracellularly in anesthetized guinea pigs and were identified by their location in the vestibular nerve and classed as regular or irregular on the basis of the variability of their spontaneous discharge. If a neuron responded to angular acceleration it was classed as a semicircular canal neuron, if it responded to maintained roll or pitch tilts it was classified as an otolith neuron. Each neuron was then tested by BCV stimuli-either clicks, continuous pure tones (200-1,500 Hz) or short tone bursts (500 Hz lasting 7 ms)-delivered by a B-71 clinical bone-conduction oscillator cemented to the guinea pig's skull. All stimulus intensities were referred to that animal's own auditory brainstem response (ABR) threshold to BCV clicks, and the maximum intensity used was within the animal's physiological range and was usually around 70 dB above BCV threshold. In addition two sensitive single axis linear accelerometers cemented to the skull gave absolute values of the stimulus acceleration in the rostro-caudal direction. The criterion for a neuron being classed as activated was an audible, stimulus-locked increase in firing rate (a 10% change was easily detectable) in response to the BCV stimulus. At the stimulus levels used in this study, semicircular canal neurons, both regular and irregular, were insensitive to BCV stimuli and very few responded: only nine of 189 semicircular canal neurons tested (4.7%) showed a detectable increase in firing in response to BCV stimuli up to the maximum 2 V peak-to-peak level we delivered to the B-71 oscillator (which produced a peak-to-peak skull acceleration of around 6-8 g and was usually around 60-70 dB above the animal's own ABR threshold for BCV clicks). Regular otolithic afferents likewise had a poor response; only 14 of 99 tested (14.1%) showed any increase in firing rate up to the maximum BCV stimulus level. However, most irregular otolithic afferents (82.8%) showed a clear increase in firing rate in response to BCV stimuli: of the 58 irregular otolith neurons tested, 48 were activated, with some being activated at very low intensities (only about 10 dB above the animal's ABR threshold to BCV clicks). Most of the activated otolith afferents were in the superior division of the vestibular nerve and were probably utricular afferents. That was confirmed by evidence using juxtacellular injection of neurobiotin near BCV activated neurons to trace their site of origin to the utricular macula. We conclude there is a very clear preference for irregular otolith afferents to be activated selectively by BCV stimuli at low stimulus levels and that BCV stimuli activate some utricular irregular afferent neurons. The BCV generates compressional and shear waves, which travel through the skull and constitute head accelerations, which are sufficient to stimulate the most sensitive otolithic receptor cells. PMID:16761136

Curthoys, Ian S; Kim, Juno; McPhedran, Samara K; Camp, Aaron J

2006-06-08

290

Bone Marrow-Derived Mesenchymal Stem Cells Improve the Functioning of Neurotrophic Factors in a Mouse Model of Diabetic Neuropathy  

PubMed Central

Diabetic neuropathy is one of the most frequent and troublesome complications of diabetes. Although there has been a continuous increase in the incidence of diabetic neuropathy, treatments have yet to be found that effectively treat diabetic neuropathy. Neurotrophic factors are proteins that promote the survival of specific neuronal populations. They also play key roles in the regeneration of peripheral nervous system. Recent evidence from diabetic animal models and human diabetic subjects suggest that reduced availability of neurotrophic factors may contribute to the pathogenesis of diabetic neuropathy. One way to reverse this effect is to take advantage of the finding that bone marrow derived mesenchymal stem cells (BM-MSCs) promote peripheral nerve repair and the functioning of neurotrophic factors. Therefore, we speculated that treatment with BM-MSCs could be a viable therapeutic strategy for diabetic neuropathy. The present study was designed to examine the possible beneficial effect of BM-MSCs on functions of neurotrophic factors in diabetic neuropathy. To assess this possibility, we used an in vivo streptozotocin-induced diabetic neuropathy mouse model. Quantitative real-time polymerase-chain reacion showed that BM-MSCs significantly increase expression levels of neurotrophic factors. Also, BM-MSCs ameliorated nerve conduction velocity in streptozotocin-treated mice. These results may help to elucidate the mechanism by which BM-MSCs function as a cell therapy agent in diabetic neuropathy.

Kim, Bae Jin

2011-01-01

291

Cell fusion reprogramming leads to a specific hepatic expression pattern during mouse bone marrow derived hepatocyte formation in vivo.  

PubMed

The fusion of bone marrow (BM) hematopoietic cells with hepatocytes to generate BM derived hepatocytes (BMDH) is a natural process, which is enhanced in damaged tissues. However, the reprogramming needed to generate BMDH and the identity of the resultant cells is essentially unknown. In a mouse model of chronic liver damage, here we identify a modification in the chromatin structure of the hematopoietic nucleus during BMDH formation, accompanied by the loss of the key hematopoietic transcription factor PU.1/Sfpi1 (SFFV proviral integration 1) and gain of the key hepatic transcriptional regulator HNF-1A homeobox A (HNF-1A/Hnf1a). Through genome-wide expression analysis of laser captured BMDH, a differential gene expression pattern was detected and the chromatin changes observed were confirmed at the level of chromatin regulator genes. Similarly, Tranforming Growth Factor-?1 (TGF-?(1)) and neurotransmitter (e.g. Prostaglandin E Receptor 4 [Ptger4]) pathway genes were over-expressed. In summary, in vivo BMDH generation is a process in which the hematopoietic cell nucleus changes its identity and acquires hepatic features. These BMDHs have their own cell identity characterized by an expression pattern different from hematopoietic cells or hepatocytes. The role of these BMDHs in the liver requires further investigation. PMID:22457803

Quintana-Bustamante, Oscar; Grueso, Esther; Garcia-Escudero, Ramon; Arza, Elvira; Alvarez-Barrientos, Alberto; Fabregat, Isabel; Garcia-Bravo, Maria; Meza, Nestor W; Segovia, Jose C

2012-03-23

292

Leukemic spleen cells are more potent than bone marrow-derived cells in a transgenic mouse model of CML.  

PubMed

Spleen size ranks among the most important risk factors in chronic myeloid leukemia (CML), but the pathogenic mechanisms of splenic hematopoiesis in CML remain poorly defined. Here, we studied the biology of Bcr-Abl positive leukemia-initiating cells in the spleen, using an inducible transgenic mouse model of CML. Disease kinetics showed greater increases of immature leukemic cells in spleen vs bone marrow (BM). To assess how Bcr-Abl alters the behavior of spleen-derived CML cells, we transplanted these cells either before ('pre-uninduced') or 44 days after ('pre-induced') expression of the oncogene. Mice transplanted with pre-induced spleen cells showed significantly increased neutrophilia and splenomegaly compared with mice receiving pre-uninduced spleen cells, suggesting that Bcr-Abl expression in the donors had increased splenic tumor burden. However, pre-induction also altered the biology of these cells, as shown by a striking increase in erythropoietic potential. These results differ from those of BM-derived CML stem cells where pre-induction of Bcr-Abl had previously been shown to decrease disease transplantability. Moreover, splenic cells were less sensitive to imatinib than BM cells. In conclusion, Bcr-Abl alters the biology of splenic leukemic stem cells by a cell-autonomous mechanism, but the disease phenotype is also influenced by the microenvironment of these cells. PMID:22193968

Schemionek, M; Spieker, T; Kerstiens, L; Elling, C; Essers, M; Trumpp, A; Berdel, W E; Müller-Tidow, C; Koschmieder, S

2011-12-23

293

Triptolide attenuates idiopathic pneumonia syndrome in a mouse bone marrow transplantation model by down-regulation of IL-17.  

PubMed

Idiopathic pneumonia syndrome (IPS) accounts for significant morbidity and mortality in patients following bone marrow transplantation (BMT). However, no effective therapy has been identified to reliably treat IPS. Previous studies using mouse BMT models suggest that the pathology of IPS in IFN-? deficient host involves increased IL-17 levels along with recruitment of donor T cells into lung. Triptolide is a potent immunosuppressive compound isolated from an anti-inflammatory Chinese herbal medicine. Triptolide can significantly inhibit generation of IL-17 by T cells and mediate immunosuppressive effect on autoimmune disease. In the present study, we used a specific murine BMT model (IFN-? deficient B6 to wildtype B6D2F1) to assess the protective effect of Triptolide on the development of IPS. We observed that IL-17 levels were significantly decreased in the lung after triptolide treatment compared with vehicle group. Furthermore, decreased number of Th17 cells in lung was found to be associated with amelioration of lung histological injury and pulmonary dysfunction. Additionally, neutralization of IL-17 also significantly reduced IPS pathology. Our study implied that triptolide could significantly inhibit donor T cell recruitment into lung, and thus prevent lung dysfunction after BMT usefully and effectively. Our study may shed some light on searching for proper strategies to prevent IL-17 mediated IPS and other Th17 cell-mediated immune pathologies. PMID:23102663

Xu, Xiaoli; Xiong, Minjian; Xu, Yafei; Su, Yuan; Zou, Ping; Zhou, Hao

2012-10-24

294

Cell Fusion Reprogramming Leads to a Specific Hepatic Expression Pattern during Mouse Bone Marrow Derived Hepatocyte Formation In Vivo  

PubMed Central

The fusion of bone marrow (BM) hematopoietic cells with hepatocytes to generate BM derived hepatocytes (BMDH) is a natural process, which is enhanced in damaged tissues. However, the reprogramming needed to generate BMDH and the identity of the resultant cells is essentially unknown. In a mouse model of chronic liver damage, here we identify a modification in the chromatin structure of the hematopoietic nucleus during BMDH formation, accompanied by the loss of the key hematopoietic transcription factor PU.1/Sfpi1 (SFFV proviral integration 1) and gain of the key hepatic transcriptional regulator HNF-1A homeobox A (HNF-1A/Hnf1a). Through genome-wide expression analysis of laser captured BMDH, a differential gene expression pattern was detected and the chromatin changes observed were confirmed at the level of chromatin regulator genes. Similarly, Tranforming Growth Factor-?1 (TGF-?1) and neurotransmitter (e.g. Prostaglandin E Receptor 4 [Ptger4]) pathway genes were over-expressed. In summary, in vivo BMDH generation is a process in which the hematopoietic cell nucleus changes its identity and acquires hepatic features. These BMDHs have their own cell identity characterized by an expression pattern different from hematopoietic cells or hepatocytes. The role of these BMDHs in the liver requires further investigation.

Arza, Elvira; Alvarez-Barrientos, Alberto; Fabregat, Isabel; Garcia-Bravo, Maria; Meza, Nestor W.; Segovia, Jose C.

2012-01-01

295

In vivo radiosensitivity and recovery pattern of the hematopoietic precursor cells and stem cells in mouse bone marrow  

SciTech Connect

Survival curves were determined for colony-forming hematopoietic stem cells (CFU-Mix and CFU-S10) as well as precursor cells (CFU-E, CFU-C, CFU-F, and BFU-E) in bone marrow of the mouse at various times up to 4 weeks after whole-body irradiation (1.5 or 3.0 Gy) in order to elucidate in vivo radiosensitivity and recovery patterns. These measurements represented the first attempts to examine CFU-Mix simultaneously with the other cell populations. CFU-E (D0 = 53 rad) and BFU-E (D0 = 68 rad) were the most radiosensitive. CFU-S10 (D0 = 81 rad) had intermediate radiosensitivity. CFU-Mix (D0 = 144 rad) and CFU-C (D0 = 157 rad) were relatively radioresistant. CFU-F (D0 = 257 rad) was the most radioresistant. The precursor and stem cells could be classified into three groups based on the recovery pattern. The first group, consisting of CFU-Mix, BFU-E, and CFU-S10, showed very slow recovery and did not reach normal levels even after day 28. CFU-E, the second group, showed the most severe depletion immediately after irradiation, and recovered most quickly with an overshoot at day 5. CFU-C and CFU-F cells, forming the third group, decreased more gradually and slightly, and recovered to the normal level after a transient rise by day 10-14.

Imai, Y.; Nakao, I.

1987-09-01

296

Convergence of bone morphogenetic protein and laminin-1 signaling pathways promotes proliferation and colony formation by fetal mouse pancreatic cells  

SciTech Connect

We previously reported that bone morphogenetic proteins (BMPs), members of the transforming growth factor superfamily, together with the basement membrane glycoprotein laminin-1 (Ln-1), promote proliferation of fetal pancreatic cells and formation of colonies containing peripheral insulin-positive cells. Here, we further investigate the cross-talk between BMP and Ln-1 signals. By RT-PCR, receptors for BMP (BMPR) (excepting BMPR-1B) and Ln-1 were expressed in the fetal pancreas between E13.5 and E17.5. Specific blocking antibodies to BMP-4 and -6 and selective BMP antagonists partially inhibited colony formation by fetal pancreas cells. Colony formation induced by BMP-6 and Ln-1 was completely abolished in a dose-dependent manner by blocking Ln-1 binding to its {alpha}{sub 6} integrin and {alpha}-dystroglycan receptors or by blocking the Ln-1 signaling molecules, phosphatidyl-inositol-3-kinase (P13K) and MAP kinase kinase-1. These results demonstrate a convergence of BMP and Ln-1 signaling through P13K and MAP kinase pathways to induce proliferation and colony formation in E15.5 fetal mouse pancreatic cells.

Jiang Fangxu [Autoimmunity and Transplantation Division, Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville 3050 (Australia)]. E-mail: jiang@wehi.edu.au; Harrison, Leonard C. [Autoimmunity and Transplantation Division, Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville 3050 (Australia)

2005-08-01

297

Gene Expression Profiles in Human and Mouse Primary Cells Provide New Insights into the Differential Actions of Vitamin D3 Metabolites  

PubMed Central

1?,25-Dihydroxyvitamin D3 (1?,25(OH)2D3) had earlier been regarded as the only active hormone. The newly identified actions of 25-hydroxyvitamin D3 (25(OH)D3) and 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3) broadened the vitamin D3 endocrine system, however, the current data are fragmented and a systematic understanding is lacking. Here we performed the first systematic study of global gene expression to clarify their similarities and differences. Three metabolites at physiologically comparable levels were utilized to treat human and mouse fibroblasts prior to DNA microarray analyses. Human primary prostate stromal P29SN cells (hP29SN), which convert 25(OH)D3 into 1?,25(OH)2D3 by 1?-hydroxylase (encoded by the gene CYP27B1), displayed regulation of 164, 171, and 175 genes by treatment with 1?,25(OH)2D3, 25(OH)D3, and 24R,25(OH)2D3, respectively. Mouse primary Cyp27b1 knockout fibroblasts (mCyp27b1?/?), which lack 1?-hydroxylation, displayed regulation of 619, 469, and 66 genes using the same respective treatments. The number of shared genes regulated by two metabolites is much lower in hP29SN than in mCyp27b1?/?. By using DAVID Functional Annotation Bioinformatics Microarray Analysis tools and Ingenuity Pathways Analysis, we identified the agonistic regulation of calcium homeostasis and bone remodeling between 1?,25(OH)2D3 and 25(OH)D3 and unique non-classical actions of each metabolite in physiological and pathological processes, including cell cycle, keratinocyte differentiation, amyotrophic lateral sclerosis signaling, gene transcription, immunomodulation, epigenetics, cell differentiation, and membrane protein expression. In conclusion, there are three distinct vitamin D3 hormones with clearly different biological activities. This study presents a new conceptual insight into the vitamin D3 endocrine system, which may guide the strategic use of vitamin D3 in disease prevention and treatment.

Tuohimaa, Pentti; Wang, Jing-Huan; Khan, Sofia; Kuuslahti, Marianne; Qian, Kui; Manninen, Tommi; Auvinen, Petri; Vihinen, Mauno; Lou, Yan-Ru

2013-01-01

298

Gene expression profiles in human and mouse primary cells provide new insights into the differential actions of vitamin d3 metabolites.  

PubMed

1?,25-Dihydroxyvitamin D3 (1?,25(OH)2D3) had earlier been regarded as the only active hormone. The newly identified actions of 25-hydroxyvitamin D3 (25(OH)D3) and 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3) broadened the vitamin D3 endocrine system, however, the current data are fragmented and a systematic understanding is lacking. Here we performed the first systematic study of global gene expression to clarify their similarities and differences. Three metabolites at physiologically comparable levels were utilized to treat human and mouse fibroblasts prior to DNA microarray analyses. Human primary prostate stromal P29SN cells (hP29SN), which convert 25(OH)D3 into 1?,25(OH)2D3 by 1?-hydroxylase (encoded by the gene CYP27B1), displayed regulation of 164, 171, and 175 genes by treatment with 1?,25(OH)2D3, 25(OH)D3, and 24R,25(OH)2D3, respectively. Mouse primary Cyp27b1 knockout fibroblasts (mCyp27b1 (-/-)), which lack 1?-hydroxylation, displayed regulation of 619, 469, and 66 genes using the same respective treatments. The number of shared genes regulated by two metabolites is much lower in hP29SN than in mCyp27b1 (-/-). By using DAVID Functional Annotation Bioinformatics Microarray Analysis tools and Ingenuity Pathways Analysis, we identified the agonistic regulation of calcium homeostasis and bone remodeling between 1?,25(OH)2D3 and 25(OH)D3 and unique non-classical actions of each metabolite in physiological and pathological processes, including cell cycle, keratinocyte differentiation, amyotrophic lateral sclerosis signaling, gene transcription, immunomodulation, epigenetics, cell differentiation, and membrane protein expression. In conclusion, there are three distinct vitamin D3 hormones with clearly different biological activities. This study presents a new conceptual insight into the vitamin D3 endocrine system, which may guide the strategic use of vitamin D3 in disease prevention and treatment. PMID:24116037

Tuohimaa, Pentti; Wang, Jing-Huan; Khan, Sofia; Kuuslahti, Marianne; Qian, Kui; Manninen, Tommi; Auvinen, Petri; Vihinen, Mauno; Lou, Yan-Ru

2013-10-08

299

Isolation of mouse mesenchymal stem cells with normal ploidy from bone marrows by reducing oxidative stress in combination with extracellular matrix  

PubMed Central

Background Isolation of mouse MSCs (mMSCs) with normal ploidy from bone marrow remains challenging. mMSCs isolated under 20% O2 are frequently contaminated by overgrown hematopoietic cells, and could also be especially vulnerable to oxidative damage, resulting in chromosomal instability. Culture under low oxygen or extracellular matrix (ECM) improves proliferation of MSCs in several species. We tested the hypothesis that culture under low oxygen in combination with ECM prepared from mouse embryonic fibroblast (MEF-ECM) could be used to purify proliferative mMSCs, and to reduce oxidative damage and maintain their chromosomal stability. Results Optimization of culture conditions under 20% O2 resulted in immortalization of mMSCs, showing extensive chromosome abnormalities, consistent with previous studies. In contrast, culture under low oxygen (2% O2) improved proliferation of mMSCs and reduced oxidative damage, such that mMSCs were purified simply by plating at low density under 2% O2. MEF-ECM reduced oxidative damage and enhanced proliferation of mMSCs. However, these isolated mMSCs still exhibited high frequency of chromosome abnormalities, suggesting that low oxygen or in combination with MEF-ECM was insufficient to fully protect mMSCs from oxidative damage. Notably, antioxidants (alpha -phenyl-t-butyl nitrone (PBN) and N-acetylcysteine (NAC)) further reduced DNA damage and chromosomal abnormalities, and increased proliferation of mMSCs. mMSCs isolated by the combination method were successfully used to generate induced pluripotent stem (iPS) cells by ectopic expression of Oct4, Sox2, Klf4 and c-Myc. Conclusions We have developed a technique that allows to reduce the number of karyotypic abnormalities for isolation of primary mMSCs and for limited culture period by combination of low oxygen, MEF-ECM, antioxidants and low density plating strategy. The effectiveness of the new combination method is demonstrated by successful generation of iPS cells from the isolated mMSCs. However, a culture system for mMSCs still is needed to prevent all the anomalies, especially after a long-term culture period.

2011-01-01

300

BMEC-1: A Human Bone Marrow Microvascular Endothelial Cell Line with Primary Cell Characteristics  

Microsoft Academic Search

Bone marrow microvascular endothelial cells (BMEC) are a functional component of the bone marrow stroma and have been shown to release hematopoietic regulatory factors as well as to selectively adhere and support the proliferation and differentiation of CD34+hematopoietic progenitors. An early passage of these cells was immortalized by transfection with a vector (pSVT) encoding the large T antigen of SV40.

Francisco J. Candal; Shahin Rafii; Jeffery T. Parker; Edwin W. Ades; Barbara Ferris; Ralph L. Nachman; Kathryn L. Kellar

1996-01-01

301

Long-term culture of leukemic bone marrow primary cells in biomimetic osteoblast niche  

Microsoft Academic Search

We constructed a “biomimetic osteoblast niche” with bio-derived bone as a scaffold, on which we seeded marrow mesenchymal\\u000a stem cells (MSCs) from CML patients, and induced the MSCs to differentiate into osteoblasts. Bone marrow mononuclear cells\\u000a from CML patients were cultured in the biomimetic niche (3D culture system) or a 2D culture system with the induced MSCs\\/osteoblasts\\u000a as a feeder

Li Hou; Ting Liu; Jing Tan; Wentong Meng; Li Deng; Hongtao Yu; Xingli Zou; Yuchun Wang

2009-01-01

302

Fibrin Gel-Immobilized Primary Osteoblasts in Calcium Phosphate Bone Cement: In vivo Evaluation with Regard to Application as Injectable Biological Bone Substitute  

Microsoft Academic Search

Osteogenic injectable bone substitutes may be useful for many applications. We developed a novel injectable bone substitute based on osteoblast-fibrin glue suspension and calcium phosphate bone cement (BC). Human osteoblasts were isolated from trabecular bone samples and cultured under standard conditions. Osteoblasts were suspended in fibrinogen solution (FS). BC was cured with thrombin solution. 8 × 4 mm injectable bone

U. Kneser; A. Voogd; J. Ohnolz; O. Buettner; L. Stangenberg; Y. H. Zhang; G. B. Stark; D. J. Schaefer

2005-01-01

303

Caffeine regulates osteogenic differentiation and mineralization of primary adipose-derived stem cells and a bone marrow stromal cell line.  

PubMed

Caffeine consumption reportedly influences bone mineral density and body weight. However, the effects of caffeine on bone metabolism are still controversial, and whether the dosage of caffeine influences osteogenic differentiation is yet to be clarified. In the present study, we cultured primary adipose-derived stem cells (ADSCs) and a bone marrow stromal cell line (M2-10B4) in osteogenic differentiation media containing varying concentrations of caffeine. Caffeine had biphasic effects: 0.1 mM caffeine significantly enhanced mineralization and alkaline phosphatase (ALP) activity. Consistent with these observations, a caffeine concentration of 0.1 mM upregulated the osteogenic differentiation marker genes ALP and osteocalcin (OCN), and elevated osteoprotegerin (OPG), Runt-related transcription factor 2 (RUNX2) and Sirtuin 1 (SIRT1) levels. However, a concentration of caffeine greater than 0.3 mM suppressed the differentiation of both the cell types. These findings indicate that caffeine has a beneficial effect on ADSCs and bone marrow stromal cells, enhancing differentiation to osteoblasts; this effect, which is mediated via RUNX2 activation at low doses is significantly suppressed at high doses. PMID:23301724

Su, Shu-Jem; Chang, Kee-Lung; Su, Shu-Hui; Yeh, Yao-Tsung; Shyu, Huey-Wen; Chen, Kuan-Ming

2013-01-09

304

Isolation and culture of primary osteocytes from the long bones of skeletally mature and aged mice  

PubMed Central

The purpose of this work was to establish a methodology to enable the isolation and study of osteocytes from skeletally mature young (4-month-old) and old (22-month-old) mice. The location of osteocytes deep within bone is ideal for their function as mechanosensors. However, this location makes the observation and study of osteocytes in vivo technically difficult. Osteocytes were isolated from murine long bones through a process of extended collagenase digestions combined with EDTA-based decalcification. A tissue homogenizer was used to reduce the remaining bone fragments to a suspension of bone particles, which were placed in culture to yield an outgrowth of osteocyte-like cells. All of the cells obtained from this outgrowth that displayed an osteocyte-like morphology stained positive for the osteocyte marker E11/GP38.[Q1] The osteocyte phenotype was further confirmed by a lack of staining for alkaline phosphatase and the absence of collagen1a1 expression. The outgrowth of osteocytes also expressed additional osteocyte-specific genes such as Sost and Mepe. This technique facilitates the isolation of osteocytes from skeletally mature bone. This novel enabling methodology should prove useful in advancing our understanding of the roles mature osteocytes play in bone health and disease.

Stern, Amber Rath; Stern, Matthew M.; Van Dyke, Mark E.; Jahn, Katharina; Prideaux, Matthew; Bonewald, Lynda F.

2013-01-01

305

Application of osteomucoperiosteal flap in large unicystic ameloblastoma to promote early bone healing: An alternative to primary bone grafting  

PubMed Central

Background: The unicystic variety of ameloblastoma is reported to be significantly less prone to recurrence in young patients than its conventional counterpart, and therefore can be treated conservatively. This paper describes a technique of using an osteoperiosteal flap to allow complete enucleation of a unicystic ameloblastoma. The technique maintains the continuity of the mandible and restores full thickness as well as strength of the mandible to promote early healing. It also maintains blood supply and proper facial contour so that esthetics is unimpaired. Materials and Methods: We describe two cases of unicystic ameloblastoma in which we used an osteoperiosteal flap. This flap was then infractured at the lower border to obliterate the dead space. Results: The sequential radiographs demonstrate early incorporation of the graft and complete filling of the defect by 3 months. At 5 years of follow-up in our first case, complete healing of bone was observed. Conclusions: We believe that these procedures can be the treatment of choice in such cases, especially with larger lesions, as these rapidly restore the patient's facial contour to normal as well as reduce the healing time.

Khare, Gagan; Kumar, Sanjeev

2011-01-01

306

Graft versus Host Disease in the Bone Marrow, Liver and Thymus Humanized Mouse Model  

PubMed Central

Mice bearing a “humanized” immune system are valuable tools to experimentally manipulate human cells in vivo and facilitate disease models not normally possible in laboratory animals. Here we describe a form of GVHD that develops in NOD/SCID mice reconstituted with human fetal bone marrow, liver and thymus (NS BLT mice). The skin, lungs, gastrointestinal tract and parotid glands are affected with progressive inflammation and sclerosis. Although all mice showed involvement of at least one organ site, the incidence of overt clinical disease was approximately 35% by 22 weeks after reconstitution. The use of hosts lacking the IL2 common gamma chain (NOD/SCID/?c?/?) delayed the onset of disease, but ultimately did not affect incidence. Genetic analysis revealed that particular donor HLA class I alleles influenced the risk for the development of GVHD. At a cellular level, GVHD is associated with the infiltration of human CD4+ T cells into the skin and a shift towards Th1 cytokine production. GVHD also induced a mixed M1/M2 polarization phenotype in a dermal murine CD11b+, MHC class II+ macrophage population. The presence of xenogenic GVHD in BLT mice both presents a major obstacle in the use of humanized mice and an opportunity to conduct preclinical studies on GVHD in a humanized model.

Greenblatt, Matthew B.; Vbranac, Vladimir; Tivey, Trevor; Tsang, Kelly; Tager, Andrew M.; Aliprantis, Antonios O.

2012-01-01

307

Investigation of transcriptional responses of juvenile mouse bone marrow to power frequency magnetic fields.  

PubMed

To seek alterations in gene transcription in bone marrow cells following in vivo exposure of juvenile mice to power frequency magnetic fields, young (21-24-day old) C57BL/6 mice were exposed to a 100?T 50Hz magnetic field for 2h. Transcription was analysed by three methods, High Coverage Expression Profiling (HiCEP), Illumina microarrays and quantitative real-time polymerase chain reaction (QRT-PCR). A pilot HiCEP experiment with 6 exposed (E) and 6 non-exposed (NE) mice identified four candidate responsive transcripts (two unknown transcripts (AK152075 and F10-NED), phosphatidylinositol binding clathrin assembly protein (Picalm) and exportin 7 (Xpo7)). A larger experiment compared 19 E and 15 NE mice using two independent QRT-PCR assays and repeated microarray assays. No significant field-dependent changes were seen, although Picalm showed a trend to significance in one QRT-PCR assay (E/NE=0.91; P=0.06). However, the study was underpowered to detect an effect of this magnitude (52% power at P=0.05). These data indicate the current experimental constraints in detecting small changes in transcription that may occur in response to magnetic fields. These constraints result from technical limitations in the accuracy of assays and biological variation, which together were sufficient to account statistically for the number of differentially expressed transcripts identified in the pilot experiment. PMID:23523963

Kabacik, Sylwia; Kirschenlohr, Heide; Raffy, Claudine; Whitehill, Kevin; Coster, Margaret; Abe, Masumi; Brindle, Kevin; Badie, Christophe; Sienkiewicz, Zenon; Bouffler, Simon

2013-03-20

308

Effect of lead on proliferation and neural differentiation of mouse bone marrow-mesenchymal stem cells.  

PubMed

Bone marrow-mesenchymal stem cells (MSCs) are considered to be an ideal source of stem cells for assessing the effects of environmental toxins on the proliferation, multipotency and differentiation of adult stem cells. The aim of this study was to investigate the effect of lead on the proliferation and neuronal differentiation of murine MSCs. MTT assay used in this study revealed that while the proliferation of MSCs is sensitive to higher than 10 microM lead, a 50% reduction in the rate of their proliferation can be achieved in the presence of 60 microM lead. The results of immunocytochemistry and RT-PCR showed that beta-mercaptoethanol induced-neuronal differentiation is also reduced after the treatment of MSCs by 60 microM lead. Furthermore, the comet assay analysis of MSCs showed a substantial increase in DNA damage in the lead treated cells compared to the control. In conclusion our results revealed for the first time that lead is not only cytotoxic to the survival and proliferation of MSCs but also inhibits their differentiation to neurons in a dose-dependant manner. Therefore, MSCs appear to be an alternative method for assessing the cytotoxic effects of such environmental hazards. PMID:18381235

Kermani, Shabnam; Karbalaie, Khadijeh; Madani, Seyed Hossein; Jahangirnejad, Ali Akbar; Eslaminejad, Mohamadreza Baghaban; Nasr-Esfahani, Mohammad Hossein; Baharvand, Hossein

2008-02-21

309

Primary mouse renal tubular epithelial cells have variable injury tolerance to ischemic and chemical mediators of oxidative stress.  

PubMed

We have developed and evaluated an in vitro culture method for assessing ischemic injury in primary mouse renal tubular epithelial cells (RTEC) in which to explore the pathobiology underlying acute kidney injury. RTEC were predominately of proximal tubule origin which is most susceptible to ischemic injury as compared to other nephron segments. Oxidative stress was induced by chemically depleting ATP using Antimycin A and 2-Deoxy-D-Glucose and by exposing cells to a 1% oxygen environment. Necrotic injury was assessed by measuring LDH released into culture supernatants. Optimal dose and time of exposure to each injury agent was determined for induction of mild, moderate and severe ischemic injury defined as LDH release of /= 50% above baseline respectively. Antimycin A and 2-Deoxy-D-Glucose produced a progressive increase in LDH release which was time dependent but chemical concentration independent. A 1% oxygen environment also induced cell injury over time but only if glucose was absent from the culture media. Antimycin A was most effective at inducing oxidative stress causing a mean LDH release of 61% at 48 hr compared to 19% and 50% LDH release induced by 2-Deoxy-D-Glucose and by exposure to 1% oxygen respectively at the same 48 hour time point.The cell culture method described provides several advantages including the use of serum free media and the ability to grow primary cells without matrix support. The LDH assay for injury assessment is reproducible, cost effective, objective and minimizes background cell death. A simple method for the culture and injury of primary mouse renal tubular epithelial cells has thereby been established and provides a useful tool for future investigations of ischemic kidney injury. PMID:19794906

Breggia, Anne C; Himmelfarb, Jonathan

310

TNF-? Regulates the Effects of Irradiation in the Mouse Bone Marrow Microenvironment  

PubMed Central

Background Secondary bone marrow (BM) myelodysplastic syndromes (MDS) are increasingly common, as a result of radio or chemotherapy administered to a majority of cancer patients. Patients with secondary MDS have increased BM cell apoptosis, which results in BM dysfunction (cytopenias), and an increased risk of developing fatal acute leukemias. In the present study we asked whether TNF-?, known to regulate cell apoptosis, could modulate the onset of secondary MDS. Principal Findings We show that TNF-? is induced by irradiation and regulates BM cells apoptosis in vitro and in vivo. In contrast to irradiated wild type (WT) mice, TNF-? deficient (TNF-? KO) mice or WT mice treated with a TNF-?-neutralizing antibody were partially protected from the apoptotic effects of irradiation. Next we established a 3-cycle irradiation protocol, in which mice were sub-lethally irradiated once monthly over a 3 month period. In this model, irradiated WT mice presented loss of microsatellite markers on BM cells, low white blood cell (WBC) counts, reduced megakaryocyte (MK) and platelet levels (thrombocytopenia) and macrocytic anemia, phenoypes that suggest the irradiation protocol resulted in BM dysfunction with clinical features of MDS. In contrast, TNF-? KO mice were protected from the irradiation effects: BM cell apoptosis following irradiation was significantly reduced, concomitant with sustained BM MK numbers and absence of other cytopenias. Moreover, irradiated WT mice with long term (?5 months) BM dysfunction had increased BM angiogenesis, MMPs and VEGF and NFkB p65, suggestive of disease progression. Conclusion Taken together, our data shows that TNF-? induction following irradiation modulates BM cell apoptosis and is a crucial event in BM dysfunction, secondary MDS onset and progression.

Cachaco, Ana Sofia; Carvalho, Tania; Santos, Ana Cristina; Igreja, Catia; Fragoso, Rita; Osorio, Catarina; Ferreira, Manuela; Serpa, Jacinta; Correia, Sofia; Pinto-do-O, Perpetua; Dias, Sergio

2010-01-01

311

Differentiating Functional Roles of Gene Expression from Immune and Non-immune Cells in Mouse Colitis by Bone Marrow Transplantation  

PubMed Central

To understand the role of a gene in the development of colitis, we compared the responses of wild-type mice and gene-of-interest deficient knockout mice to colitis. If the gene-of-interest is expressed in both bone marrow derived cells and non-bone marrow derived cells of the host; however, it is possible to differentiate the role of a gene of interest in bone marrow derived cells and non- bone marrow derived cells by bone marrow transplantation technique. To change the bone marrow derived cell genotype of mice, the original bone marrow of recipient mice were destroyed by irradiation and then replaced by new donor bone marrow of different genotype. When wild-type mice donor bone marrow was transplanted to knockout mice, we could generate knockout mice with wild-type gene expression in bone marrow derived cells. Alternatively, when knockout mice donor bone marrow was transplanted to wild-type recipient mice, wild-type mice without gene-of-interest expressing from bone marrow derived cells were produced. However, bone marrow transplantation may not be 100% complete. Therefore, we utilized cluster of differentiation (CD) molecules (CD45.1 and CD45.2) as markers of donor and recipient cells to track the proportion of donor bone marrow derived cells in recipient mice and success of bone marrow transplantation. Wild-type mice with CD45.1 genotype and knockout mice with CD45.2 genotype were used. After irradiation of recipient mice, the donor bone marrow cells of different genotypes were infused into the recipient mice. When the new bone marrow regenerated to take over its immunity, the mice were challenged by chemical agent (dextran sodium sulfate, DSS 5%) to induce colitis. Here we also showed the method to induce colitis in mice and evaluate the role of the gene of interest expressed from bone-marrow derived cells. If the gene-of-interest from the bone derived cells plays an important role in the development of the disease (such as colitis), the phenotype of the recipient mice with bone marrow transplantation can be significantly altered. At the end of colitis experiments, the bone marrow derived cells in blood and bone marrow were labeled with antibodies against CD45.1 and CD45.2 and their quantitative ratio of existence could be used to evaluate the success of bone marrow transplantation by flow cytometry. Successful bone marrow transplantation should show a vast majority of donor genotype (in term of CD molecule marker) over recipient genotype in both the bone marrow and blood of recipient mice.

Koon, Hon Wai; Ho, Samantha; Cheng, Michelle; Ichikawa, Ryan; Pothoulakis, Charalabos

2012-01-01

312

Mass spectrometric quantification of glycogen to assess primary substrate accumulation in the Pompe mouse.  

PubMed

Glycogen storage in the ?-glucosidase knockout((6neo/6neo)) mouse recapitulates the biochemical defect that occurs in the human condition; as such, this mouse serves as a model for the inherited metabolic deficiency of lysosomal acid ?-glucosidase known as Pompe disease. Although this model has been widely used for the assessment of therapies, the time course of glycogen accumulation that occurs as untreated Pompe mice age has not been reported. To address this, we developed a quantitative method involving amyloglucosidase digestion of glycogen and quantification of the resulting free glucose by liquid chromatography/electrospray ionization-tandem mass spectrometry. The method was sensitive enough to measure as little as 0.1 ?g of glycogen in tissue extracts with intra- and interassay coefficients of variation of less than 12%. Quantification of glycogen in tissues from Pompe mice from birth to 26 weeks of age showed that, in addition to the accumulation of glycogen in the heart and skeletal muscle, glycogen also progressively accumulated in the brain, diaphragm, and skin. Glycogen storage was also evident at birth in these tissues. This method may be particularly useful for longitudinal assessment of glycogen reduction in response to experimental therapies being trialed in this model. PMID:22239964

Fuller, Maria; Duplock, Stephen; Turner, Christopher; Davey, Philippa; Brooks, Doug A; Hopwood, John J; Meikle, Peter J

2011-12-20

313

Differential expression of collagen types I and III in consequential and primary fibrosis in irradiated mouse colon  

SciTech Connect

These studies were undertaken to understand further the pathogenesis of consequential and primary fibrosis in mouse colon after irradiation. The distal 2.5 cm of colon of C3Hf/Kam mice was irradiated with either a single dose of 27 Gy or a split dose of 2 x 14.75 Gy separated by 10 days to induce a consequential or primary fibrotic lesion, respectively. The amount of total collagen in the two lesions was quantified by hydroxyproline, and tensile strength, an assay of tissue rigidity, was measured as a function of dose and time after irradiation. The relative distribution of collagen types I, III and IV in the colon was visualized by immunohistochemistry. Collagen types I, III and IV were quantified by immunoblot techniques, and in situ hybridization was used to identify and score the cells producing procollagen mRNA types I and III as a function of time after irradiation. The hydroxyproline and tensile strength measurements demonstrated that both lesions contained significantly increased amounts of collagen compared to controls. However, the ulcerated lesion of consequential fibrosis contained three times as much collagen and required a three- to fourfold increase in the peak force to rupture the colon as did the non-ulcerative lesion of primary fibrosis. The fibrosis accompanying the consequential lesion contained elevated levels of both collagen types I and III, but primary fibrosis contained only elevated levels of type I collagen compared to controls. The in situ hybridization studies showed cells producing increased amounts of procollagen mRNA 8 and 25 weeks before the elevated levels of collagen were detected for consequential and primary fibrosis, respectively. The cells producing the excess collagen mRNA were identified as fibroblasts. No distinction between the two lesions could be made based on the cell types producing the collagen. 48 refs., 7 figs.

Followill, D.S.; Travis, E.L. [Univ. of Texas, Houston, TX (United States)

1995-12-01

314

Human primary ductal carcinoma in situ (DCIS) subtype-specific pathology is preserved in a mouse intraductal (MIND) xenograft model  

PubMed Central

Ductal carcinoma in situ (DCIS) is a non-obligate precursor of invasive breast cancer. The current recognition that DCIS lesions exhibit inter- and intra-lesion diversity suggests that the process of evolution to invasive breast cancer is more complex than previously recognized. Here we demonstrate the reproducible growth of primary DCIS cells derived from patient’s surgical and biopsy samples by the mouse intraductal (MIND) model. MIND involves injection of cells into the NOD-SCID IL2Rgammanull (NSG) mouse mammary ducts. Twelve (8 unique and 4 repeats) DCIS and 2 atypical hyperplasia specimens, heterogeneous with respect to biomarker expression and histology, were injected into 48 mouse mammary glands and analyzed for successful xenotransplantation. Overall, 14/34 and 11/14 of MIND xenotransplanted glands contained human DCIS and atypical hyperplastic cells, respectively, after 8 weeks, which formed single and multi-layered epithelium inside the ducts, and were heterogeneous with respect to expression of human cytokeratins, estrogen receptor ? (ER), and HER2. ER protein expression was recapitulated in MIND xenografts at ratios similar to the corresponding patient biopsies. In both patient biopsies and corresponding MIND xenografts HER2 protein expression and nuclear HER2 gene over-expression was restricted to the DCIS lesions and were not found in the surrounding stroma or normal ducts. The xenografted DCIS lesions recapitulate the pathology and heterogeneity of human disease thus providing a powerful tool for the characterization of the distinct cellular and molecular basis of inter- and intra-tumoral heterogeneity and the processes of DCIS to early invasive breast cancer progression.

Valdez, Kelli Elizabeth; Fang, Fan; Smith, William; Allred, D. Craig; Medina, Daniel; Behbod, Fariba

2012-01-01

315

Daily administration of eldecalcitol (ED-71), an active vitamin D analog, increases bone mineral density by suppressing RANKL expression in mouse trabecular bone.  

PubMed

Eldecalcitol (ED-71) is a new vitamin D? derivative recently approved for the treatment of osteoporosis in Japan. Previous studies have shown that the daily administration of ED-71 increases bone mineral density (BMD) by suppressing bone resorption in various animal models. In this study, we examined how ED-71 suppresses bone resorption in vivo, by analyzing bone histomorphometry and ex vivo osteoclastogenesis assays. Daily administration of ED-71 (50 ng/kg body weight) to 8-week-old male mice for 2 and 4 weeks increased BMD in the femoral metaphysis without causing hypercalcemia. Bone and serum analyses revealed that ED-71 inhibited bone resorption and formation, indicating that the increase in BMD is the result of the suppression of bone resorption. This suppression was associated with a decrease in the number of osteoclasts in trabecular bone. We previously identified cell cycle-arrested receptor activator of NF-?B (RANK)-positive bone marrow cells as quiescent osteoclast precursors (QOPs) in vivo. Daily administration of ED-71 affected neither the number of RANK-positive cells in vivo nor the number of osteoclasts formed from QOPs in ex vivo cultures. In contrast, ED-71 suppressed the expression of RANK ligand (RANKL) mRNA in femurs. Immunohistochemical experiments also showed that the perimeter of the RANKL-positive cell surface around the trabecular bone was significantly reduced in ED-71-treated mice than in the control mice. ED-71 administration also increased BMD in 12-week-old ovariectomized mice, through the suppression of RANKL expression in the trabecular bone. These results suggest that the daily administration of ED-71 increases BMD by suppressing RANKL expression in trabecular bone in vivo. PMID:22052469

Harada, Suguru; Mizoguchi, Toshihide; Kobayashi, Yasuhiro; Nakamichi, Yuko; Takeda, Satoshi; Sakai, Sadaoki; Takahashi, Fumiaki; Saito, Hitoshi; Yasuda, Hisataka; Udagawa, Nobuyuki; Suda, Tatsuo; Takahashi, Naoyuki

2012-02-01

316

Induction of terminal deoxynucleotidyl transferase and Lyt antigens with thymosin: identification of multiple subsets of prothymocytes in mouse bone marrow and spleen.  

PubMed Central

Thymosin (fraction 5 and synthetic alpha 1 peptide) induced prothymocytes in mouse bone marrow and spleen to express terminal deoxynucleotidyl transferase (TdT; DNA nucleotidylexotransferase; nucleosidetriphosphate:DNA deoxynucleotidylexotransferase, EC 2.7.7.31) or Lyt-1+, 2+, 3+ alloantigens (or both) after brief incubation in vitro. Three antigenic phenotypes were generated: (i) TdT+ Lyt+, (ii) TdT- Lyt+, and (iii) TdT+ Lyt-. The TdT+ Lyt+ phenotype was expressed by 80% of prothymocytes in bone marrow and 30% of prothymocytes in spleen from normal mice. The TdT- Lyt+ phenotype was expressed by 81% of prothymocytes in bone marrow from athymic mice. More than 80% of TdT+ bone marrow cells from normal and athymic mice expressed Lyt antigens after thymosin treatment. We interpret these observations as suggesting that (i) most TdT+ hemopoietic cells in normal and athymic mice are thymocyte progenitors; (ii) two independent lineages of prothymocytes exist, one that expresses TdT and another that does not, (iii) commitment of prothymocytes to the TdT+ cell pathway is partially regulated by a thymic feedback mechanism; and (iv) the bone marrow preferentially produces TdT+ prothymocytes, whereas the spleen may serve as a repository for TdT- prothymocytes. A model of T-cell development is presented in which the thymus functions as a compound organ to process TdT+ and TdT- thymocytes progenitors and to generate two lines of T cells. Images

Goldschneider, I; Ahmed, A; Bollum, F J; Goldstein, A L

1981-01-01

317

Critical-Size Calvarial Bone Defects Healing in a Mouse Model with Silk Scaffolds and SATB2- Modified iPSCs  

PubMed Central

Induced pluripotent stem cells (iPSCs) can differentiate into mineralizing cells and thus have a great potential in application in engineered bone substitutes with bioactive scaffolds in regeneration medicine. In the current study we characterized and demonstrated the pluripotency and osteogenic differentiation of mouse iPSCs. To enhance the osteogenic differentiation of iPSCs, we then transduced the iPSCs with the potent transcription factor, nuclear matrix protein SATB2. We observed that in SATB2-overexpressing iPSCs there were increased mineral nodule formation and elevated mRNA levels of key osteogenic genes, osterix (OSX), Runx2, bone sialoprotein (BSP) and osteocalcin (OCN). Moreover, the mRNA levels of HoxA2 was reduced after SATB2 overexpression in iPSCs. The SATB2-overexpressing iPSCs were then combined with silk scaffolds and transplanted into critical-size calvarial bone defects created in nude mice. Five weeks post-surgery, radiological and micro-CT analysis revealed enhanced new bone formation in calvarial defects in SATB2 group. Histological analysis also showed increased new bone formation and mineralization in the SATB2 group. In conclusion, the results demonstrate that SATB2 facilitates the differentiation of iPSCs towards osteoblast-lineage cells by repressing HoxA2 and augmenting the functions of the osteoblast determinants Runx2, BSP and OCN.

Ye, Jin-Hai; Xu, Yuan-Jin; Gao, Jun; Yan, Shi-Guo; Zhao, Jun; Tu, Qisheng; Zhang, Jin; Duan, Xue-Jing; Sommer, Cesar A.; Mostoslavsky, Gustavo; Kaplan, David; Wu, Yu-Nong; Zhang, Chen-Ping; Wang, Lin; Chen, Jake

2011-01-01

318

Chemotaxis of bone marrow derived eosinophils in vivo: a novel method to explore receptor-dependent trafficking in the mouse.  

PubMed

Here, we describe a novel method via which ex vivo cultured mouse bone marrow derived eosinophils (bmEos) can be adoptively transferred into recipient mice in order to study receptor-dependent recruitment to lung tissue in vivo. Intratracheal instillation of recombinant human eotaxin-2 (hCCL24) prior to introduction of bmEos via tail vein injection resulted in an approximately fourfold increase in Siglec F-positive/CD11c-negative eosinophils in the lungs of eosinophil-deficient ?dblGATA recipient mice compared with controls. As anticipated, bmEos generated from CCR3-gene-deleted mice did not migrate to the lung in response to hCCL24 in this model, indicating specific receptor dependence. BmEos generated from GFP-positive BALB/c mice responded similarly to hCCL24 in vitro and were detected in lung tissue of BALB/c WT as well as BALB/c ?dblGATA eosinophil-deficient recipient mice, at approximately fourfold (at 5 h post-injection) and approximately threefold (at 24 h postinjection) over baseline, respectively. Comparable results were obtained with GFP-positive C57BL/6 bmEos responding to intratracheal hCCL24 in C57BL/6 ?dblGATA recipient mice. The use of ex vivo cultured bmEos via one or more of these methods offers the possibility of manipulating bmEos prior to transfer into a WT or gene-deleted recipient host. Thus, this chemotaxis model represents a novel and robust tool for pharmacological studies in vivo. PMID:23670593

Sturm, Eva M; Dyer, Kimberly D; Percopo, Caroline M; Heinemann, Akos; Rosenberg, Helene F

2013-06-14

319

Initial binding and recellularization of decellularized mouse lung scaffolds with bone marrow-derived mesenchymal stromal cells.  

PubMed

Recellularization of whole decellularized lung scaffolds provides a novel approach for generating functional lung tissue ex vivo for subsequent clinical transplantation. To explore the potential utility of stem and progenitor cells in this model, we investigated recellularization of decellularized whole mouse lungs after intratracheal inoculation of bone marrow-derived mesenchymal stromal cells (MSCs). The decellularized lungs maintained structural features of native lungs, including intact vasculature, ability to undergo ventilation, and an extracellular matrix (ECM) scaffold consisting primarily of collagens I and IV, laminin, and fibronectin. However, even in the absence of intact cells or nuclei, a number of cell-associated (non-ECM) proteins were detected using mass spectroscopy, western blots, and immunohistochemistry. MSCs initially homed and engrafted to regions enriched in types I and IV collagen, laminin, and fibronectin, and subsequently proliferated and migrated toward regions enriched in types I and IV collagen and laminin but not provisional matrix (fibronectin). MSCs cultured for up to 1 month in either basal MSC medium or in a small airways growth media (SAGM) localized in both parenchymal and airway regions and demonstrated several different morphologies. However, while MSCs cultured in basal medium increased in number, MSCs cultured in SAGM decreased in number over 1 month. Under both media conditions, the MSCs predominantly expressed genes consistent with mesenchymal and osteoblast phenotype. Despite a transient expression of the lung precursor TTF-1, no other airway or alveolar genes or vascular genes were expressed. These studies highlight the power of whole decellularized lung scaffolds to study functional recellularization with MSCs and other cells. PMID:21756220

Daly, Amanda B; Wallis, John M; Borg, Zachary D; Bonvillain, Ryan W; Deng, Bin; Ballif, Bryan A; Jaworski, Diane M; Allen, Gilman B; Weiss, Daniel J

2011-09-23

320

Activation of antioxidant response element in mouse primary cortical cultures with sesquiterpene lactones isolated from Tanacetum parthenium  

PubMed Central

Tanacetum parthenium (Asteraceae) produces biologically active sesquiterpene lactones (SL). Nuclear factor E2-related factor 2 (Nrf2) is a transcription factor known to activate a series of genes termed the antioxidant response element (ARE). Activation of the Nrf2/ARE may be useful for the treatment of neurodegenerative disease. In this study we isolated 11 sesquiterpene lactones from T. parthenium with centrifugal partition chromatography and semi-preparative HPLC. Compounds were screened in-vitro for their ability to activate the ARE on primary mouse cortical cultures as well as for their toxicity towards the cultures. All sesquiterpene lactones containing the ?-methylene-?-lactone moiety were able to activate the ARE although a number of compounds displayed significant cellular toxicity towards the cultures. The structure activity relationship of the sesquiterpene lactones indicate that the guaianolides isolated were more active and less toxic then the germacranolides.

Fischedick, Justin T; Standiford, Miranda; Johnson, Delinda A.; De Vos, Ric C.H.; Todorovic, Sla?ana; Banjanac, Tijana; Verpoorte, Rob; Johnson, Jeffrey A.

2012-01-01

321

Physical activity in the androgen receptor knockout mouse: evidence for reversal of androgen deficiency on cancellous bone.  

PubMed

Disruption of the androgen receptor (AR) in male mice reduces cortical bone expansion and muscle mass during puberty and results in high bone turnover-related cancellous osteopenia. We hypothesized that voluntary wheel running during growth is able to rescue the effects of AR disruption on bone. To this end, 5-week-old AR knockout (ARKO) mice were randomized to a running group (cage with running wheel) and a sedentary group (cage without wheel) and followed-up until 16 weeks of age. Voluntary wheel running in ARKO mice did not influence body weight, muscle mass or periosteal bone expansion. Interestingly, voluntary running significantly reduced bone turnover in ARKO mice and prevented cancellous bone loss due to a preservation of trabecular number. Thus, voluntary running in ARKO mice was able to reduce cancellous bone resorption, suggesting that sustained exercise may potentially compensate the effects of androgen disruption on cancellous bone. PMID:19013130

Ophoff, Jill; Callewaert, Filip; Venken, Katrien; De Gendt, Karel; Ohlsson, Claes; Gayan-Ramirez, Ghislaine; Decramer, Marc; Boonen, Steven; Bouillon, Roger; Verhoeven, Guido; Vanderschueren, Dirk

2008-11-12

322

Arsenic-induced aberrant gene expression in fetal mouse primary liver-cell cultures.  

PubMed

Exposure of maternal mice to inorganic arsenic through the drinking water induces liver tumors and aberrant gene expression in offspring when they reach adulthood. To help define if these are direct fetal effects of arsenic, fetal liver cells were isolated from untreated mice at gestation day 13.5 by mechanical dissection and centrifugation. Two hours after seeding the cells on collagen1-coated plates in William E media containing 10% fetal bovine serum, 1x ITS (insulin, transferrin, and selenium) and antibiotics, inorganic arsenite (0, 0.1, 0.3, and 1.0 microM) was added to the fresh media for 72 h. Cell morphology and viability were not significantly altered by these arsenic concentrations. At the end of arsenic exposure, cells were harvested into Trizol, and total RNA was extracted, purified, and subjected to real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis. Arsenite exposure produced a concentration-dependent induction of heme oxygenase-1 (up to eight-fold) and metallothionein-1 (up to five-fold), indicative of stress response to adapt to arsenic insult. Expression of genes related to steroid metabolism, such as 17beta-hydroxysteroid dehydrogenase-7 (HSD17beta7) and Cyp2a4, were increased approximately two-fold, together with increases in estrogen receptor-alpha (ER-alpha) and ER-alpha-linked genes, such as anterior gradient-2, keratin 1-19, and trefoil factor-3. Arsenic in vitro induced a three-fold increase in the expression of alpha-fetoprotein (AFP), a biomarker associated with transplacental arsenic-induced mouse liver tumors. Thus, exposure of mouse fetal liver cells to arsenic induces adaptive responses and aberrant gene expression, which could alter genetic programming at a very early life stage, potentially contributing to tumor formation much later in life. PMID:18991936

Liu, Jie; Yu, Limei; Tokar, Erik J; Bortner, Carl; Sifre, Maria I; Sun, Yang; Waalkes, Michael P

2008-10-01

323

Prognostic relevance of urokinase plasminogen activator detection in micrometastatic cells in the bone marrow of patients with primary breast cancer.  

PubMed Central

Patients with an elevated level of urokinase plasminogen activator (uPA) in breast cancer tissue have an adverse prognosis. This study evaluated the prognostic relevance of uPA detection in disseminated tumour cells in bone marrow. Bone marrow was sampled intraoperatively from both iliac crests in 280 patients with primary breast cancer. Interphase cells were enhanced and stained immunocytologically with two antibodies: 2E11, which detects TAG 12--a tumour-associated glycoprotein typically expressed by almost all breast cancer cells--and the anti-uPA antibody HD-UK9. Thirty-five of the 2E11-positive women (n = 132, 47%) developed metastatic disease (median follow-up time 44 months). Of these, most were uPA positive (n = 23, 65%) and only 12 were uPA negative. Patients with uPA-positive cells in bone marrow (n = 98, 35%) had a significantly shorter metastasis-free interval (36 months) than women who were uPA negative (44.5 months). The worst prognosis was seen in patients positive for both markers (29.5 months), followed by those who were uPA negative and 2E11 positive (37 months). The detection of uPA on disseminated tumour cells characterizes a subgroup of patients with an even worse prognosis, who should undergo more aggressive adjuvant systemic therapy. For the first time, it was possible to evaluate an important qualitative parameter involved in the process of breast cancer metastases. Images Figure 1

Solomayer, E. F.; Diel, I. J.; Wallwiener, D.; Bode, S.; Meyberg, G.; Sillem, M.; Gollan, C.; Kramer, M. D.; Krainick, U.; Bastert, G.

1997-01-01

324

Multiple quantitative trait loci for cortical and trabecular bone regulation map to mid-distal mouse chromosome 4 that shares linkage homology to human chromosome 1p36.  

PubMed

The mid-distal region of mouse chromosome 4 (Chr 4) is homologous with human Chr 1p36. Previously, we reported that mouse Chr 4 carries a quantitative trait locus (QTL) with strong regulatory effect on volumetric bone mineral density (vBMD). The intent of this study is to utilize nested congenic strains to decompose the genetic complexity of this gene-rich region. Adult females and males from 18 nested congenic strains carrying discrete C3H sequences were phenotyped for femoral mineral and volume by pQCT and for trabecular bone volume (BV), tissue volume (TV), trabecular number (Trab.no), and trabecular thickness (Trab.thk) by MicroCT 40. Our data show that the mouse Chr 4 region consists of at least 10 regulatory QTL regions that affected either or both pQCT and MicroCT 40 phenotypes. The pQCT phenotypes were typically similar between sexes, whereas the MicroCT 40 phenotypes were divergent. Individual congenic strains contained one to seven QTL regions. These regions conferred large positive or negative effects in some congenic strains, depending on the particular bone phenotype. The QTL regions II to X are syntenic with human 1p36, containing from 1 to 102 known genes. We identified 13 candidate genes that can be linked to bone within these regions. Six of these genes were linked to osteoblasts, three linked to osteoclasts, and two linked to skeletal development. Three of these genes have been identified in Genome Wide Association Studies (GWAS) linked to 1p36. In region III, there is only one gene, Lck, which conferred negative pQCT and MicroCT 40 phenotypes in both sexes. This gene is important to development and functioning of T cells, has been associated with osteoclast activity, and represents a novel bone regulatory gene that merits further experimental evaluation. In summary, congenic strains are powerful tools for identifying regulatory regions that influence bone biology and offer models for testing hypotheses about gene-gene and gene-environment interactions that are not available to experimental work in humans. PMID:22031020

Beamer, Wesley G; Shultz, Kathryn L; Coombs, Harold F; Horton, Lindsay G; Donahue, Leah Rae; Rosen, Clifford J

2012-01-01

325

Combination Immunotherapy of Primary Prostate Cancer in a Transgenic Mouse Model Using CTLA-4 Blockade1  

Microsoft Academic Search

We have previously shown that antibodies to CTLA-4, an inhibitory receptor on T cells, can be effective at inducing regression of transplantable murine tumors. In this study, we demonstrate that an effective immune response against primary prostate tumors in transgenic (TRAMP) mice can be elicited using a strategy that combines CTLA-4 blockade and an irradiated tumor cell vaccine. Treatment of

Arthur A. Hurwitz; Barbara A. Foster; Eugene D. Kwon; Tan Truong; Eugene M. Choi; Norman M. Greenberg; Maurice B. Burg; James P. Allison

2000-01-01

326

Expression of genetically determined diabetes and insulitis in the nonobese diabetic (NOD) mouse at the level of bone marrow-derived cells. Transfer of diabetes and insulitis to nondiabetic (NOD X B10) F1 mice with bone marrow cells from NOD mice  

Microsoft Academic Search

The development of autoimmune diabetes in the nonobese diabetic (NOD) mouse is controlled by at least three recessive loci, including one linked to the MHC. To determine whether any of these genetic loci exert their effects via the immune system, radiation bone marrow chimeras were constructed in which (NOD X B10)F1-irradiated recipients were reconstituted with NOD bone marrow cells. Unmanipulated

L. S. Wicker; B. J. Miller; A. Chai; M. Terada; Y. Mullen

1988-01-01

327

Differences in Renal Tubule Primary Cilia Length in a Mouse Model of Bardet-Biedl Syndrome  

Microsoft Academic Search

Background: Bardet-Biedl syndrome (BBS) is a heterogeneous genetic disorder that comprises numerous features, including renal cystic disease. Twelve BBS genes have been identified (BBS1–12). Although the exact functions of the BBS proteins are unknown, evidence suggests that they are involved in cilia assembly, maintenance and\\/or function. Renal primary cilia dysfunction can lead to cystic kidney disease. To test whether lacking

Elaine M. Mokrzan; Jacqueline S. Lewis; Kirk Mykytyn

2007-01-01

328

Natural and radiation-induced degeneration of primordial and primary follicles in mouse ovary  

Microsoft Academic Search

The present study deals with the morphological changes of the degenerating primordial and primary follicles induced by ?-radiation. Prepubertal female mice of 3 weeks old ICR strain were ?-irradiated with the dose of LD80(30) (8.3 Gy). The ovaries were collected at 3, 6 and 12 h after irradiation. The largest cross-sections were prepared by histological semithin sections for microscopical observations.

Chang Joo Lee; Ho Hyun Park; Byung Rok Do; Yong-Dal Yoon; Jin Kyu Kim

2000-01-01

329

Loss of neuronatin promotes "browning" of primary mouse adipocytes while reducing Glut1-mediated glucose disposal.  

PubMed

Failure of white adipose tissue to appropriately store excess metabolic substrate seems to underpin obesity-associated type 2 diabetes. Encouraging "browning" of white adipose has been suggested as a therapeutic strategy to help dispose of excess stored lipid and ameliorate the resulting insulin resistance. Genetic variation at the DNA locus encoding the novel proteolipid neuronatin has been associated with obesity, and we recently observed that neuronatin expression is reduced in subcutaneous adipose tissue from obese humans. Thus, to explore the function of neuronatin further, we used RNAi to silence its expression in murine primary adipocyte cultures and examined the effects on adipocyte phenotype. We found that primary adipocytes express only the longer isoform of neuronatin. Loss of neuronatin led to increased mitochondrial biogenesis, indicated by greater intensity of MitoTracker Green staining. This was accompanied by increased expression of UCP1 and the key genes in mitochondrial oxidative phosphorylation, PGC-1?, Cox8b, and Cox4 in primary subcutaneous white adipocytes, indicative of a "browning" effect. In addition, phosphorylation of AMPK and ACC was increased, suggestive of increased fatty acid utilization. Similar, but less pronounced, effects of neuronatin silencing were also noted in primary brown adipocytes. In contrast, loss of neuronatin caused a reduction in both basal and insulin-stimulated glucose uptake and glycogen synthesis, likely mediated by a reduction in Glut1 protein upon silencing of neuronatin. In contrast, loss of neuronatin had no effect on insulin signaling. In conclusion, neuronatin appears to be a novel regulator of browning and metabolic substrate disposal in white adipocytes. PMID:23482445

Gburcik, Valentina; Cleasby, Mark E; Timmons, James A

2013-03-12

330

High-Efficiency Transfection of Primary Human and Mouse T Lymphocytes Using RNA Electroporation  

PubMed Central

The use of nonviral gene transfer methods in primary lymphocytes has been hampered by low gene transfer efficiency and high transfection-related toxicity. In this report, high gene transfection efficiency with low transfection-related toxicity was achieved by electroporation using in vitro-transcribed mRNA. Using these methods, >90% transgene expression with >80% viable cells was observed in stimulated primary human and murine T lymphocytes transfected with GFP or mCD62L. Electroporation of unstimulated human PBMCs or murine splenocytes with GFP RNA yielded 95 and 56% GFP+ cells, respectively. Electroporation of mRNA for NY-ESO-1, MART-1, and p53 antigen-specific TCRs into human T lymphocytes redirected these lymphocytes to recognize melanoma cell lines in an MHC-restricted manner. The onset of gene expression was rapid (within 30 min) and durable (up to 7 days postelectroporation) using both GFP and TCR-mediated recognition of target cells. There was no adverse effect observed on the T lymphocytes subjected to RNA electroporation evaluated by cell growth rate, annexin-V staining of apoptotic cells, BrdU incorporation, tumor antigen-specific recognition or antigen-specific TCR affinity. The results of this study indicate that mRNA electroporation provides a powerful tool to introduce genes into both human and murine primary T lymphocytes.

Zhao, Yangbing; Zheng, Zhili; Cohen, Cyrille J.; Gattinoni, Luca; Palmer, Douglas C.; Restifo, Nicholas P.; Rosenberg, Steven A.; Morgan, Richard A.

2006-01-01

331

Expressional Analysis of GFP-Tagged Cells in an In Vivo Mouse Model of Giant Cell Tumor of Bone  

PubMed Central

Giant cell tumor of bone in a neoplastic stromal cell which survives for multiple passages in primary cell culture with a stable phenotype. In the pathological environment of GCT, the neoplastic nature of the mesenchymal stromal component drives local hematopoietic precursors to undergo fusion and form multinucleated osteoclast like giant cells. There is currently very limited knowledge about the pathogenesis of GCT due to the lack of suitable in vivo models for this tumor. Here we report stable gene transfer of Green fluorescence protein (GFP) in GCT stromal cells. In the present study, we have used GCT stromal cells that stably express enhanced green fluorescence protein (GFP) that are used in a new in vivo culture model. Our results show the utility of the GFP tagged cell lines that stably express GFP signals up to 52 weeks of continuous growth. The in vivo model described herein can serve as an excellent system for in vivo therapeutic and mechanistic evaluation of existing and novel targets for GCT.

Singh, S; Singh, M; Mak, I; Ghert, M

2013-01-01

332

Necator americanus in the mouse: Histopathological changes associated with the passage of larvae through the lungs of mice exposed to primary and secondary infection  

Microsoft Academic Search

The mouse\\/Necator americanus model was studied to assess the histopathological changes that occur in the lungs following primary and secondary exposure to infective larvae. Groups of BALB\\/c mice were infected percutaneously and killed on various days post infection. Parasite numbers were counted, the bronchoalveolar leukocyte response was quantified and histological sections of lung material were examined for evidence of host

M. J. Wilkinson; C. Wells; J. M. Behnke

1990-01-01

333

Ex vivo gene therapy-induced endochondral bone formation: comparison of muscle-derived stem cells and different subpopulations of primary muscle-derived cells  

Microsoft Academic Search

Muscle-based gene therapy and tissue engineering hold great promise for improving bone healing. However, the relative advantage of muscle-derived stem cells (MDSCs) or primary muscle-derived cells (MDCs) remains to be defined. We compared the ability of MDSCs and different subpopulations of MDCs (PP1 and PP3) to induce bone formation via ex vivo gene therapy. We were able to efficiently transduce

Hsain-Chung Shen; Hairong Peng; Arvydas Usas; Brian Gearhart; James Cummins; Freddie H Fu; Johnny Huard

2004-01-01

334

SCP-2/SCP-x gene ablation alters lipid raft domains in primary cultured mouse hepatocytes.  

PubMed

Although reverse cholesterol transport from peripheral cell types is mediated through plasma membrane microdomains termed lipid rafts, almost nothing is known regarding the existence, protein/lipid composition, or structure of these putative domains in liver hepatocytes, cells responsible for the net removal of cholesterol from the body. Lipid rafts purified from hepatocyte plasma membranes by a nondetergent affinity chromatography method were: i) present at 33 +/- 3% of total plasma membrane protein; ii) enriched in key proteins of the reverse cholesterol pathway [scavenger receptor class B type I (SR-B1), ABCA1, P-glycoprotein (P-gp), sterol carrier protein-2 (SCP-2)]; iii) devoid of caveolin-1; iv) enriched in cholesterol, sphingomyelin, GM1, and phospholipids low in polyunsaturated fatty acid and double bond index; and v) exhibited an intermediate liquid-ordered lipid phase with significant transbilayer fluidity gradient. Ablation of the gene encoding SCP-2 significantly altered lipid rafts to: i) increase the proportion of lipid rafts present, thereby increasing raft total content of ABCA1, P-gp, and SR-B1; ii) increase total phospholipids while decreasing GM1 in lipid rafts; iii) decrease the fluidity of lipid rafts, consistent with the increased intermediate liquid-ordered phase; and iv) abolish the lipid raft transbilayer fluidity gradient. Thus, despite the absence of caveolin-1 in liver hepatocytes, lipid rafts represented nearly one-third of the mouse hepatocyte plasma membrane proteins and displayed unique protein, lipid, and biophysical properties that were differentially regulated by SCP-2 expression. PMID:17609524

Atshaves, Barbara P; McIntosh, Avery L; Payne, H Ross; Gallegos, Adalberto M; Landrock, Kerstin; Maeda, Nobuyo; Kier, Ann B; Schroeder, Friedhelm

2007-07-03

335

Mouse Bone Marrow-Derived Mesenchymal Stromal Cells Turn Activated Macrophages into a Regulatory-Like Profile  

PubMed Central

In recent years it has become clear that the therapeutic properties of bone marrow-derived mesenchymal stromal cells (MSC) are related not only to their ability to differentiate into different lineages but also to their capacity to suppress the immune response. We here studied the influence of MSC on macrophage function. Using mouse thioglycolate-elicited peritoneal macrophages (M) stimulated with LPS, we found that MSC markedly suppressed the production of the inflammatory cytokines TNF-?, IL-6, IL-12p70 and interferon-? while increased the production of IL-10 and IL-12p40. Similar results were observed using supernatants from MSC suggesting that factor(s) constitutively released by MSC are involved. Supporting a role for PGE2 we observed that acetylsalicylic acid impaired the ability of MSC to inhibit the production of inflammatory cytokines and to stimulate the production of IL-10 by LPS-stimulated M. Moreover, we found that MSC constitutively produce PGE2 at levels able to inhibit the production of TNF-? and IL-6 by activated M. MSC also inhibited the up-regulation of CD86 and MHC class II in LPS-stimulated M impairing their ability to activate antigen-specific T CD4+ cells. On the other hand, they stimulated the uptake of apoptotic thymocytes by M. Of note, MSC turned M into cells highly susceptible to infection with the parasite Trypanosoma cruzi increasing more than 5-fold the rate of M infection. Using a model of inflammation triggered by s.c. implantation of glass cylinders, we found that MSC stimulated the recruitment of macrophages which showed a low expression of CD86 and the MHC class II molecule Iab and a high ability to produce IL-10 and IL-12p40, but not IL-12 p70. In summary, our results suggest that MSC switch M into a regulatory profile characterized by a low ability to produce inflammatory cytokines, a high ability to phagocyte apoptotic cells, and a marked increase in their susceptibility to infection by intracellular pathogens.

Maggini, Julian; Mirkin, Gerardo; Bognanni, Ianina; Holmberg, Josefina; Piazzon, Isabel M.; Nepomnaschy, Irene; Costa, Hector; Canones, Cristian; Raiden, Silvina; Vermeulen, Monica; Geffner, Jorge R.

2010-01-01

336

Differential gene expression of bone anabolic factors and trabecular bone architectural changes in the proximal femoral shaft of primary hip osteoarthritis patients  

Microsoft Academic Search

Previous studies have shown a generalised increase in bone mass in patients with osteoarthritis (OA). Using molecular histomorphometry, this study examined the in vivo expression of mRNA encoding bone anabolic factors and collagen type I genes (COL1A1, COL1A2) in human OA and non-OA bone. Bone samples were obtained from the intertrochanteric (IT) region of the proximal femur, a skeletal site

Le-Hoa Truong; Julia S Kuliwaba; Helen Tsangari; Nicola L Fazzalari

2006-01-01

337

Combined zoledronic acid and meloxicam reduced bone loss and tumour growth in an orthotopic mouse model of bone-invasive oral squamous cell carcinoma.  

PubMed

Oral squamous cell carcinoma (OSCC) is common in cats and humans and invades oral bone. We hypothesized that the cyclooxygenase (COX)-2 inhibitor, meloxicam, with the bisphosphonate, zoledronic acid (ZOL), would inhibit tumour growth, osteolysis and invasion in feline OSCC xenografts in mice. Human and feline OSCC cell lines expressed COX-1 and COX-2 and the SCCF2 cells had increased COX-2 mRNA expression with bone conditioned medium. Luciferase-expressing feline SCCF2Luc cells were injected beneath the perimaxillary gingiva and mice were treated with 0.1?mg?kg(-1) ZOL twice weekly, 0.3?mg?kg(-1) meloxicam daily, combined ZOL and meloxicam, or vehicle. ZOL inhibited osteoclastic bone resorption at the tumour-bone interface. Meloxicam was more effective than ZOL at reducing xenograft growth but did not affect osteoclastic bone resorption. Although a synergistic effect of combined ZOL and meloxicam was not observed, combination therapy was well-tolerated and may be useful in the clinical management of bone-invasive feline OSCC. PMID:23651067

Martin, C K; Dirksen, W P; Carlton, M M; Lanigan, L G; Pillai, S P; Werbeck, J L; Simmons, J K; Hildreth, B E; London, C A; Toribio, R E; Rosol, T J

2013-05-01

338

Effects of metabolic inhibition on the membrane properties of isolated mouse primary sensory neurones.  

PubMed Central

1. The patch-clamp technique has been used to investigate the mechanisms that couple membrane excitability to metabolism in neurones isolated from mouse dorsal root ganglia. 2. Blockade of electron transport by cyanide (CN-), reduction of the mitochondrial membrane potential with carbonyl cyanide p-trifluoromethoxyphenyl hydrazone (FCCP), removal of glucose or inhibition of glycolysis with idoacetic acid (IAA), all increased a K+ conductance (gK), which could be sufficient to shunt action potentials. 3. The K+ conductance was reduced by incubation of cells in Ca2(+)-free solutions or by increasing the Ca2+ buffering power of pipette-filling solutions. The Ca2+ ionophore, ionomycin, also increased a K+ conductance, and current fluctuation analysis showed that the channels carrying the current induced by both ionomycin and by CN- had a similar mean conductance of circa 9 pS. Thus, increased gK was a Ca2(+)-dependent K+ conductance, gK(Ca), reflecting a rise in resting [Ca2+]i. 4. The conductance was not affected by inclusion of ATP or an ATP-regenerating system in the pipette, suggesting that the underlying rise in [Ca2+] is not due directly to loss of ATP, and confirming that the increased gK is not carried through ATP-dependent K+ channels. 5. Voltage-gated K+ currents evoked by membrane depolarization were increased by CN- or glucose removal. The current-voltage relation of the increased gK mirrored the voltage dependence of Ca2+ entry, and thus reflects impaired cellular handling of the Ca2+ load imposed by depolarization. 6. The rise in [Ca2+]i and altered Ca2+ buffering capacity induced by metabolic blockade affected several other conductances: (i) a Ca2(+)-dependent chloride current was increased. (ii) Both the low-threshold transient and high-threshold sustained voltage-gated Ca2+ currents were attenuated and their thresholds were shifted in the hyperpolarizing direction. (iii) The inward current activated by hyperpolarization. IH, seen in large cells, was attenuated by either metabolic blockade or ionomycin. 7. The responses of these neurones to impaired metabolism thus depend largely on the effects of raised [Ca2+]i on the populations of channels expressed by the cells. These changes in membrane properties could account for some of the changes in neuronal behaviour seen during the clinical states of hypoxia or hypoglycaemia, underlying changes in central nervous system function.

Duchen, M R

1990-01-01

339

Intravenous Administration of Adenoviruses Targeting Transforming Growth Factor Beta Signaling Inhibits Established Bone Metastases in 4T1 Mouse Mammary Tumor Model in an Immunocompetent Syngeneic Host  

PubMed Central

We have examined the effect of adenoviruses expressing soluble transforming growth factor receptorII-Fc (sTGF?RIIFc) in a 4T1 mouse mammary tumor bone metastasis model using syngeneic BALB/c mice. Infection of 4T1 cells with a non-replicating adenovirus, Ad(E1-).sT?RFc, or with two oncolytic adenoviruses, Ad.sT?RFc and TAd.sT?RFc, expressing sTGF?RIIFc (the human TERT promoter drives viral replication in TAd.sT?RFc) produced sTGF?RIIFc protein. Oncolytic adenoviruses produced viral replication and induced cytotoxicity in 4T1 cells. 4T1 cells were resistant to the cytotoxic effects of TGF?-1 (up to 10 ng/ml). However, TGF?-1 induced the phosphorylation of SMAD2 and SMAD3, which were inhibited by co-incubation with sTGF?RIIFc protein. TGF?-1 also induced IL-11, a well-known osteolytic factor. Intracardiac injection of 4T1-luc2 cells produced bone metastases by day 4. Intravenous injection of Ad.sT?RFc (on days 5 and 7) followed by bioluminescence imaging (BLI) of mice on days 7, 11 and 14 in tumor bearing mice indicated inhibition of bone metastasis progression (p<0.05). X-ray radiography of mice on day 14 showed a significant reduction of the lesion size by Ad.sT?RFc (p<0.01) and TAd.sT?RFc (p<0.05). Replication-deficient virus Ad(E1-).sT?RFc expressing sTGF?RIIFc showed some inhibition of bone metastasis, while Ad(E1-).Null was not effective in inhibiting bone metastases. Thus, systemic administration of Ad.sT?RFc and TAd.sT?RFc can inhibit bone metastasis in the 4T1 mouse mammary tumor model, and can be developed as potential anti-tumor agents for breast cancer.

Zhang, Zhenwei; Hu, Zebin; Gupta, Janhavi; Krimmel, Jeffrey; Gerseny, Helen; Berg, Arthur; Robbins, John; Du, Hongyan; Prabhakar, Bellur; Seth, Prem

2012-01-01

340

Space Radiation and Bone Loss.  

PubMed

Exposure to ionizing radiation may negatively impact skeletal integrity during extended spaceflight missions to the moon, Mars, or near-Earth asteroids. However, our understanding of the effects of radiation on bone is limited when compared to the effects of weightlessness. In addition to microgravity, astronauts will be exposed to space radiation from solar and cosmic sources. Historically, radiation exposure has been shown to damage both osteoblast precursors and local vasculature within the irradiated volume. The resulting suppression of bone formation and a general state of low bone-turnover is thought to be the primary contributor to bone loss and eventual fracture. Recent investigations using mouse models have identified a rapid, but transient, increase in osteoclast activity immediately after irradiation with both spaceflight and clinically-relevant radiation qualities and doses. Together with a chronic suppression of bone formation after radiation exposure, this acute skeletal damage may contribute to long-term deterioration of bone quality, potentially increasing fracture risk. Direct evidence for the damaging effects of radiation on human bone are primarily demonstrated by the increased incidence of fractures at sites that absorb high doses of radiation during cancer therapy: exposures are considerably higher than what could be expected during spaceflight. However, both the rapidity of bone damage and the chronic nature of the changes appear similar between exposure scenarios. This review will outline our current knowledge of space and clinical exploration exposure to ionizing radiation on skeletal health. PMID:22826632

Willey, Jeffrey S; Lloyd, Shane A J; Nelson, Gregory A; Bateman, Ted A

2011-01-01

341

Cross-species bone marrow transplantation: Evidence for tolerance induction, stem cell engraftment, and maturation of T lymphocytes in a xenogeneic stromal environment (rat----mouse)  

SciTech Connect

Transplantation of untreated F344 rat bone marrow into irradiated B10 mouse recipients (non-TCD F344----B10) to produce fully xenogeneic chimeras resulted in stable xenogeneic lymphoid chimerism, ranging from 82% to 97% rat. Survival of animals was excellent, without evidence for GVH disease. The specificity of tolerance which resulted was highly donor-specific; MHC disparate third party mouse and rat skin grafts were promptly rejected while donor-specific F344 grafts were significantly prolonged (MST greater than 130 days). Multi-lineage rat stem cell-derived progeny including lymphoid cells (T- and B-lymphocytes), myeloid cells, erythrocytes, platelets, and natural killer (NK) cells were present in the fully xenogenic chimeras up to 7 months after bone marrow transplantation. Immature rat T-lymphocytes matured and acquired the {alpha}/{beta} T-cell receptor in the thymus of chimeras in a pattern similar to normal rat controls, suggesting that immature T-lymphocytes of rat origin could interact with the murine xenogeneic thymic stroma to undergo normal maturation and differentiation. This model may be useful to study the mechanisms responsible for the induction and maintenance of donor-specific transplantation tolerance across a species barrier.

Ildstad, S.T.; Wren, S.M.; Boggs, S.S.; Hronakes, M.L.; Vecchini, F.; Van den Brink, M.R. (Department of Surgery, University of Pittsburgh, School of Medicine, PA (USA))

1991-08-01

342

Genotoxicity assessment of NIM-76 and its formulation (pessary) in an in vitro Ames Salmonella/microsome assay and in vivo mouse bone marrow micronucleus test.  

PubMed

The possible genotoxic potential of NIM-76, a volatile fraction obtained from neem oil, having promising contraceptive activity, as well as its formulation product, called pessary (7.5% NIM-76 in polyethylene glycol), were evaluated in the Ames assay and mouse bone marrow micronucleus (MN) assay. Genotoxicity of NIM-76 (0.1-1000 µg/plate) and pessary (0.1-10,000 µg/plate) were studied using the liquid preincubation protocol of the Ames assay both in the presence and absence of S9. Likewise, the ability of NIM-76 [1-1000 mg/kg body weight (b.w.)] and its formulation product (18.75-300 mg/kg b.w.) to induce clastogenic effects were studied in the female mouse bone marrow MN test by using a two-dose intraperitoneal treatment protocol. There was no increase in the number of revertant colonies resulting from NIM-76 or pessary at any of their doses over the respective negative control plates, either in the presence or absence of S9. Similarly, in the MN assay, neither of them showed any clastogenic activity because there was no significant increase in the frequency of micronucleated polychromatic erythrocytes, over the negative control group of animals. The use of this compound in humans is therefore not likely to have mutagenic effects and may be considered as safe with regard to genotoxic potential. PMID:23527474

Vijayan, Vinod; Meshram, Ghansham P

2013-03-26

343

Fusion of the primary and accessory navicular bones: a modification of the kidner procedure  

Microsoft Academic Search

Accessory navicular is a congenital anomaly. Present in about 9% of the population, it is symptomatic in only a small percentage. The pertinent anatomy, clinical presentation, classification system, and conservative treatment are discussed. Surgical options are reviewed. The surgical technique for a modified Kidner procedure in which the primary and accessory naviculars are fused is described in detail.

Michael P Swords; John R Shank; Sigvard T Hansen; Bruce J Sangeorzan

2004-01-01

344

Role of TASK2 Potassium Channels Regarding Volume Regulation in Primary Cultures of Mouse Proximal Tubules  

PubMed Central

Several papers reported the role of TASK2 channels in cell volume regulation and regulatory volume decrease (RVD). To check the possibility that the TASK2 channel modulates the RVD process in kidney, we performed primary cultures of proximal convoluted tubules (PCT) and distal convoluted tubules (DCT) from wild-type and TASK2 knockout (KO) mice. In KO mice, the TASK2 coding sequence was in part replaced by the lac-Z gene. This allows for the precise localization of TASK2 in kidney sections using ?-galactosidase staining. TASK2 was only localized in PCT cells. K+ currents were analyzed by the whole-cell clamp technique with 125 mM K-gluconate in the pipette and 140 mM Na-gluconate in the bath. In PCT cells from wild-type mice, hypotonicity induced swelling-activated K+ currents insensitive to 1 mM tetraethylammonium, 10 nM charybdotoxin, and 10 ?M 293B, but blocked by 500 ?M quinidine and 10 ?M clofilium. These currents were increased in alkaline pH and decreased in acidic pH. In PCT cells from TASK2 KO, swelling-activated K+ currents were completely impaired. In conclusion, the TASK2 channel is expressed in kidney proximal cells and could be the swelling-activated K+ channel responsible for the cell volume regulation process during osmolyte absorptions in the proximal tubules.

Barriere, Herve; Belfodil, Radia; Rubera, Isabelle; Tauc, Michel; Lesage, Florian; Poujeol, Chantal; Guy, Nicolas; Barhanin, Jacques; Poujeol, Philippe

2003-01-01

345

[Mechanisms of neural circuit remodeling in the primary somatosensory cortex in mouse models of chronic pain].  

PubMed

Chronic pain is caused by injury or inflammation of the peripheral tissues and by abnormal activities in the central nervous system including the spinal cord area related to generation and maintenance of chronic pain. In the cerebral cortex, several areas respond to noxious stimulation. Since activities of these cortical areas are enhanced in chronic pain conditions, not only a single cortical area, but also the interaction between multiple cortical areas seems to contribute to chronic pain. The primary somatosensory cortex (S1), which is one of the pain-related cortical areas, plays a role in recognition of strength, location, and duration of pain. Since activities of the S1 are enhanced in chronic pain conditions, this area is likely to have an important role in the generation or maintenance of chronic pain. The S1 consists of 6 layers and has both excitatory and inhibitory neurons. The excitatory neurons integrate pain information from the peripheral nerves and send it to other excitatory neurons in the S1 and other cortical areas. On the other hand, the inhibitory neurons modulate activities of the excitatory neurons and contribute to sensory processing. However, how these neuronal structures and functions alter in chronic pain and contribute to chronic pain behavior is unknown. In this review, we highlight recent two-photon microscopy studies that elucidate the mechanisms of neuronal circuit remodeling in the S1 under chronic pain conditions. PMID:23735524

Eto, Kei; Ishikawa, Takuya; Sun Kwang, Kim; Nabekura, Junichi

2013-06-01

346

Primary implant stability in the atrophic sinus floor of human cadaver maxillae: impact of residual ridge height, bone density, and implant diameter.  

PubMed

OBJECTIVES: Simultaneous implant placement in conjunction with lateral or transcrestal maxillary sinus floor augmentation gives the benefit of reduction in healing times and surgical interventions. Primary implant stability, however, may be significantly reduced in resorbed residual ridges. Aim of the present study was to investigate the impact of residual bone height, bone density, and implant diameter on primary stability of implants in the atrophic sinus floor. MATERIAL AND METHODS: A total of 66 NobelActive(™) implants were inserted in the sinus floor of fresh human cadaver maxillae: 22 narrow (3.5 mm), 22 regular (4.3 mm), and 22 wide (5.0 mm) diameter implants in residual ridges of 2-6 mm height. Presurgical computed tomographic scans were acquired to assess bone height and density. Primary implant stability was evaluated by insertion torque values (ITV), Periotest values (PTV), and Osstell implant stability quotients (ISQ). RESULTS: Correlations within outcomes (ITV, PTV, ISQ) were highly significant (P < 0.001). Radiographic bone density was found to significantly impact all three outcome measures (P < 0.001), while no influence of residual bone height and implant diameter could be revealed by multifactorial analysis. Consistent results were seen in all subgroups (including residual ridges of 5-6 mm height). CONCLUSIONS: Bone density seems to represent the major determinant of primary stability in maxillary sinus augmentation with simultaneous implant placement (as well as 5-6 mm short implants in the maxillary sinus floor). Preoperative bone density assessment may help to avoid stability-related complications in one-stage implant treatment of the atrophic posterior maxilla. PMID:23167282

Pommer, Bernhard; Hof, Markus; Fädler, Andrea; Gahleitner, André; Watzek, Georg; Watzak, Georg

2012-11-21

347

A novel mouse model for the study of the inhibitory effects of chronic ethanol exposure on direct bone formation  

Technology Transfer Automated Retrieval System (TEKTRAN)

Excessive alcohol consumption has been reported to interfere with human bone homeostasis and repair in multiple ways. Previous studies have demonstrated that chronic ethanol exposure in the rat via an intragastric dietary delivery system inhibits direct bone formation during distraction osteogenesis...

348

Alendronate affects long bone length and growth plate morphology in the oim mouse model for Osteogenesis Imperfecta  

Microsoft Academic Search

Alendronate, a bisphosphonate drug, has shown promise in reducing remodeling and bone loss in postmenopausal osteoporosis. Alendronate acts directly on the osteoclast, inhibiting its resorption capability. This inhibition of osteoclast activity has led to the use of bisphosphonates in the treatment of the osteogenesis imperfecta condition. Treatment of osteogenesis imperfecta with bisphosphonates enhances bone strength, but the consequences on linear

K. D Evans; S. T Lau; A. M Oberbauer; R. B Martin

2003-01-01

349

Bone Scan Index: a prognostic imaging biomarker for high-risk prostate cancer patients receiving primary hormonal therapy  

PubMed Central

Background The objective of this study was to explore the prognostic value of the Bone Scan Index (BSI) obtained at the time of diagnosis in a group of high-risk prostate cancer patients receiving primary hormonal therapy. Methods This was a retrospective study based on 130 consecutive prostate cancer patients at high risk, based on clinical stage (T2c/T3/T4), Gleason score (8 to 10) and prostate-specific antigen (PSA) (> 20 ng/mL), who had undergone whole-body bone scans < 3 months after diagnosis and who received primary hormonal therapy. BSI was calculated using an automated method. Cox proportional-hazards regression models were used to investigate the association between clinical stage, Gleason score, PSA, BSI and survival. Discrimination between prognostic models was assessed using the concordance index (C-index). Results In a multivariate analysis, Gleason score (p = 0.01) and BSI (p < 0.001) were associated with survival, but clinical stage (p = 0.29) and PSA (p = 0.57) were not prognostic. The C-index increased from 0.66 to 0.71 when adding BSI to a model including clinical stage, Gleason score and PSA. The 5-year probability of survival was 55% for patients without metastases, 42% for patients with BSI < 1, 31% for patients with BSI = 1 to 5, and 0% for patients with BSI > 5. Conclusions BSI can be used as a complement to PSA to risk-stratify high-risk prostate cancer patients at the time of diagnosis. This imaging biomarker, reflecting the extent of metastatic disease, can be of value both in clinical trials and in patient management when deciding on treatment.

2013-01-01

350

[Bone marrow autotransplantation in patients with acute myeloblastic leukemia in primary remission].  

PubMed

Fifteen bone marrow autotransplants (BMAT) in patients with acute myeloblastic leukemia (AML) were performed after the first remission. The mean age was 37 years (range 12 to 60 years). According to the morphological classification FAB, 8 patients had monocytic leukemia (M4, M5) and 7 myeloid leukemia (M1, M2, M3). The mean interval elapsed between the date of complete remission and the BMAT was 3.9 months (range 1 to 5-9 months). In 8 patients this interval was longer than 6 months and in 7 cases it was shorter than 6 months. After achievement of the complete remission all patients underwent certain cycles of intensification before the BMAT. Eight patients received only a cycle whereas 7 patients received more than one cycle (between 2 and 4). The conditioning protocol consisted of cyclophosphamide (CP) (60 mg/kg x 2) and total body radiotherapy (TBR) (10 Gy) in 9 patients; CP and busulfan in five; and CP, cytarabine at high doses and melphalan in one case. Marrow extraction was performed after completion of chemotherapy of intensification. In 5 cases the bone marrow was depleted of leukemic cells by previous in vitro treatment with ASTA-Z. There are at present 8 alive patients. The survival free of illness was 51.8%. Seven patients died: 3 cases because relapse of the leukemia, 3 due to attachment failure of the transplantation, and one patient suffered a viral myocarditis. The survival free of illness was significantly longer in those patients transplanted after 6 months of the complete remission. PMID:2280616

Richard, C; Iriondo, A; Baro, J; Conde, E; Hermosa, V; Alsar, M J; Gómez Casares, M T; Muruzabal, M J; Pérez Encinas, M; Zubizarreta, A

1990-09-22

351

Stimulation of Osteoclast Formation by RANKL Requires Interferon Regulatory Factor-4 and Is Inhibited by Simvastatin in a Mouse Model of Bone Loss.  

PubMed

Diseases of bone loss are a major public health problem. Here, we report the novel therapeutic action of simvastatin in osteoclastogenesis and osteoprotection, demonstrated by the ability of simvastatin to suppress osteoclast formation in vitro and in vivo. We found that in vitro, IRF4 expression is upregulated during osteoclast differentiation induced by RANKL (receptor activator of nuclear factor-?B ligand), while simvastatin blocks RANKL-induced osteoclastogenesis and decreases expression of NFATc1 (nuclear factor of activated T-cells, cytoplasmic, calcineurin-dependent 1), IRF4 and osteoclast markers. We also show that IRF4 acts in cooperation with NFATc2 and NF-?B on the promoter region of NFATc1 to accelerate its initial transcription during the early stage of osteoclastogenesis. Moreover, our study using IRF4 siRNA knockdown directly demonstrates the requirement for IRF4 in NFATc1 mRNA transcription and its necessity in RANKL-induced osteoclast differentiation. Our results suggest that the reduction in osteoclastogenesis is partly due to the inhibition of IRF4 production in RANKL-induced osteoclast differentiation. To investigate the in vivo effects of simvastatin in RANKL-treated mice, we examined the bone mineral density (BMD) of a mouse model of bone loss, and found that simvastatin significantly reduced bone loss by suppressing osteoclast numbers in vivo, even in the presence of high concentrations of RANKL. These results suggest that the depletion of osteoclasts is not due to the reduction in RANKL produced by osteoblasts in vivo. The results are consistent with the hypothesis that simvastatin blocks RANKL-induced IRF4 expression in osteoclastogenesis. We propose that the expression of IRF4 by osteoclasts could be a promising new therapeutic target in bone-loss diseases. PMID:24039733

Nakashima, Yoshiki; Haneji, Tatsuji

2013-09-11

352

RNA Interference-Mediated Silencing of Atp6i Prevents Both Periapical Bone Erosion and Inflammation in the Mouse Model of Endodontic Disease  

PubMed Central

Dental caries is one of the most prevalent infectious diseases in the United States, affecting approximately 80% of children and the majority of adults. Dental caries may lead to endodontic disease, where the bacterial infection progresses to the root canal system of the tooth, leading to periapical inflammation, bone erosion, severe pain, and tooth loss. Periapical inflammation may also exacerbate inflammation in other parts of the body. Although conventional clinical therapies for this disease are successful in approximately 80% of cases, there is still an urgent need for increased efficacy of treatment. In this study, we applied a novel gene-therapeutic approach using recombinant adeno-associated virus (AAV)-mediated Atp6i RNA interference (RNAi) knockdown of Atp6i/TIRC7 gene expression to simultaneously target periapical bone resorption and periapical inflammation. We found that Atp6i inhibition impaired osteoclast function in vitro and in vivo and decreased the number of T cells in the periapical lesion. Notably, AAV-mediated Atp6i/TIRC7 knockdown gene therapy reduced bacterial infection-stimulated bone resorption by 80% in the mouse model of endodontic disease. Importantly, Atp6i+/? mice with haploinsufficiency of Atp6i exhibited protection similar to that in mice with bacterial infection-stimulated bone erosion and periapical inflammation, which confirms the potential therapeutic effect of AAV-small hairpin RNA (shRNA)-Atp6i/TIRC7. Our results demonstrate that AAV-mediated Atp6i/TIRC7 knockdown in periapical tissues can inhibit endodontic disease development, bone resorption, and inflammation, indicating for the first time that this potential gene therapy may significantly improve the health of those who suffer from endodontic disease.

Ma, Junqing; Zhang, Lijie; Tucker, Byron; Zhu, Guochun; Sasaki, Hajime; Hao, Liang; Wang, Lin; Ci, Hongliang; Jiang, Hongbing; Stashenko, Philip

2013-01-01

353

Developmental and cell type-specific expression of thyroid hormone transporters in the mouse brain and in primary brain cells.  

PubMed

Cellular thyroid hormone uptake and efflux are mediated by transmembrane transport proteins. One of these, monocarboxylate transporter 8 (MCT8) is mutated in Allan-Herndon-Dudley syndrome, a severe mental retardation associated with abnormal thyroid hormone constellations. Since mice deficient in Mct8 exhibit a milder neurological phenotype than patients, we hypothesized that alternative thyroid hormone transporters may compensate in murine brain cells for the lack of Mct8. Using qPCR, Western Blot, and immunocytochemistry, we investigated the expression of three different thyroid hormone transporters, i.e., Mct8 and L-type amino acid transporters Lat1 and Lat2, in mouse brain. All three thyroid hormone transporters are expressed from corticogenesis and peak around birth. Primary cultures of neurons and astrocytes express Mct8, Lat1, and Lat2. Microglia specifically expresses Mct10 and Slco4a1 in addition to high levels of Lat2 mRNA and protein. As in vivo, a brain microvascular endothelial cell line expressed Mct8 and Lat1. 158N, an oligodendroglial cell line expressed Mct8 protein, consistent with delayed myelination in MCT8-deficient patients. Functional T(3)- and T(4)-transport assays into primary astrocytes showed K(M) values of 4.2 and 3.7 ?M for T(3) and T(4). Pharmacological inhibition of L-type amino acid transporters by BCH and genetic inactivation of Lat2 reduced astrocytic T(3) uptake to the same extent. BSP, a broad spectrum inhibitor, including Mct8, reduced T(3) uptake further suggesting the cooperative activity of several T(3) transporters in astrocytes. PMID:21264952

Braun, Doreen; Kinne, Anita; Bräuer, Anja U; Sapin, Remy; Klein, Marc O; Köhrle, Josef; Wirth, Eva K; Schweizer, Ulrich

2010-12-29

354

Casein induces the proliferation of bone marrow mononuclear cells, apoptosis of WEHI-3 leukaemic cells and increased survival in a leukaemia mouse model.  

PubMed

Acute myeloid leukaemia results from the neoplastic transformation of haematopoietic stem cells. Although advances have been made in its treatment, the mortality rate remains high. As a result, therapeutic alternatives continue to be explored. In this study, we present evidence that suggests that casein, the principal protein in milk, possesses significant antileukaemic properties. We investigated whether casein inhibited the in vitro proliferation and induced the apoptosis of the mouse myelomonocytic leukaemia cell line WEHI-3. By contrast, under identical conditions, casein markedly promotes the proliferation of mouse normal mononuclear bone marrow cells. Since the selective elimination of leukaemia cells is an ideal therapeutic strategy, we also evaluated the antileukaemic potential of casein in vivo. The results showed that casein increases the survival of mice bearing WEHI-3-induced tumours, suggesting that this molecule is also capable of inhibiting the proliferation of these cells in vivo. The evidence that casein inhibited cell proliferation and induced apoptosis in leukaemia cells in vitro, but increased survival in vivo in a leukaemia mouse model, indicates that casein may be useful in leukaemia therapy. PMID:22970044

Ledesma-Martínez, E; Pérez-Cordero, C; Córdova-Galaviz, Y; Sánchez-Tellez, G; Huerta-Yepez, S; Aguiñiga-Sánchez, I; Miranda-Peralta, E; Monroy-García, A; Weiss-Steider, B; Santiago-Osorio, E

2012-06-14

355

Lycopene Protects against Hypoxia/Reoxygenation-Induced Apoptosis by Preventing Mitochondrial Dysfunction in Primary Neonatal Mouse Cardiomyocytes  

PubMed Central

Background Hypoxia/reoxygenation(H/R)-induced apoptosis of cardiomyocytes plays an important role in myocardial injury. Lycopene is a potent antioxidant carotenoid that has been shown to have protective properties on cardiovascular system. The aim of the present study is to investigate the potential for lycopene to protect the cardiomyocytes exposed to H/R. Moreover, the effect on mitochondrial function upon lycopene exposure was assessed. Methods and Findings Primary cardiomyocytes were isolated from neonatal mouse and established an in vitro model of H/R which resembles ischemia/reperfusion in vivo. The pretreatment of cardiomyocytes with 5 µM lycopene significantly reduced the extent of apoptosis detected by TUNEL assays. To further study the mechanism underlying the benefits of lycopene, interactions between lycopene and the process of mitochondria-mediated apoptosis were examined. Lycopene pretreatment of cardiomyocytes suppressed the activation of the mitochondrial permeability transition pore (mPTP) by reducing the intracellular reactive oxygen species (ROS) levels and inhibiting the increase of malondialdehyde (MDA) levels caused by H/R. Moreover, the loss of mitochondrial membrane potential, a decline in cellular ATP levels, a reduction in the amount of cytochrome c translocated to the cytoplasm and caspase-3 activation were observed in lycopene-treated cultures. Conclusion The present results suggested that lycopene possesses great pharmacological potential in protecting against H/R-induced apoptosis. Importantly, the protective effects of lycopene may be attributed to its roles in improving mitochondrial function in H/R-treated cardiomyocytes.

Yue, Rongchuan; Hu, Houxiang; Yiu, Kai Hang; Luo, Tao; Zhou, Zhou; Xu, Lei; Zhang, Shuang; Li, Ke; Yu, Zhengping

2012-01-01

356

Skp2 promotes adipocyte differentiation via a p27{sup Kip1}-independent mechanism in primary mouse embryonic fibroblasts  

SciTech Connect

Skp2, the substrate-binding subunit of an SCF ubiquitin ligase complex, is a key regulator of cell cycle progression that targets substrates for degradation by the 26S proteasome. We have now shown that ablation of Skp2 in primary mouse embryonic fibroblasts (MEFs) results both in impairment of adipocyte differentiation and in the accumulation of the cyclin-dependent kinase inhibitor p27{sup Kip1}, a principal target of the SCF{sup Skp2} complex. Genetic ablation of p27{sup Kip1} in MEFs promoted both lipid accumulation and adipocyte-specific gene expression. However, depletion of p27{sup Kip1} by adenovirus-mediated RNA interference failed to correct the impairment of adipocyte differentiation in Skp2{sup -/-} MEFs. In contrast, troglitazone, a high-affinity ligand for peroxisome proliferator-activated receptor {gamma} (PPAR{gamma}), largely restored lipid accumulation and PPAR{gamma} gene expression in Skp2{sup -/-} MEFs. Our data suggest that Skp2 plays an essential role in adipogenesis in MEFs in a manner that is at least in part independent of regulation of p27{sup Kip1} expression.

Okada, Mitsuru; Sakai, Tamon; Nakamura, Takehiro [Division of Diabetes, Metabolism, and Endocrinology, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe 650-0017 (Japan); Tamamori-Adachi, Mimi; Kitajima, Shigetaka [Department of Biochemical Genetics, Medical Research Institute, Tokyo Medical and Dental University, Tokyo 113-8510 (Japan); Matsuki, Yasushi; Watanabe, Eijiro; Hiramatsu, Ryuji [Pharmacology Research Laboratories, Dainippon Sumitomo Pharma Co. Ltd., Takarazuka 665-0051 (Japan); Sakaue, Hiroshi [Division of Diabetes, Metabolism, and Endocrinology, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe 650-0017 (Japan); Department of Pharmacology, Kinki University School of Medicine, Osakasayama 589-8511 (Japan)], E-mail: hsakaue@med.kindai.ac.jp; Kasuga, Masato [Division of Diabetes, Metabolism, and Endocrinology, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe 650-0017 (Japan); Research Institute, International Medical Center of Japan, Tokyo 162-8655 (Japan)

2009-02-06

357

Mitochondrial Distribution of Neuroglobin and Its Response to Oxygen-Glucose Deprivation in Primary Cultured Mouse Cortical Neurons  

PubMed Central

Neuroglobin (Ngb) is a new member of the globin family and a novel endogenous neuroprotective molecule, but its neuroprotective mechanisms remain largely undefined. Previous studies suggest Ngb is both physically and functionally related to mitochondria, however without direct evidence. Our recent discovery has shown that Ngb can physically interact with a number of mitochondrial proteins. In this study we aimed to define the physical interaction between Ngb and mitochondria by determining whether there is a mitochondrial distribution of Ngb under both physiological resting and pathological oxygen-glucose deprivation (OGD) conditions. Western blot for the first time revealed a small portion of Ngb was physically localized in mitochondria, and the relative mitochondrial Ngb level was significantly increased after OGD in primary cultured mouse cortical neurons, indicating a translocation of Ngb into mitochondria. Complementary approaches including confocal imaging and immuno-electron microscopy confirmed Ngb distribution in mitochondria under both basal resting condition and OGD. Inhibitors of mitochondria permeability transition pore (mPTP) and Voltage Dependent Anion Channel (VDAC) blocked OGD-induced increase of mitochondrial Ngb level, demonstrating a possible role of mPTP in Ngb’s mitochondrial translocation. We further found that Ngb overexpression-conferred neuroprotection was correlated with increased mitochondrial Ngb level, suggesting the mitochondria distribution of Ngb is clearly associated with and even might contribute to Ngb’s neuroprotection.

Yu, Zhanyang; Xu, Jianfeng; Liu, Ning; Wang, Yi; Li, Xiaokun; Pallast, Stefanie; van Leyen, Klaus; Wang, Xiaoying

2012-01-01

358

Rapid Structural Remodeling of Thalamocortical Synapses Parallels Experience-Dependent Functional Plasticity in Mouse Primary Visual Cortex  

PubMed Central

Monocular lid closure (MC) causes a profound shift in the ocular dominance (OD) of neurons in primary visual cortex (V1). Anatomical studies in both cat and mouse V1 suggest that large-scale structural rearrangements of eye-specific thalamocortical (TC) axons in response to MC occur much more slowly than the shift in OD. Consequently, there has been considerable debate as to whether the plasticity of TC synapses, which transmit competing visual information from each eye to V1, contributes to the early functional consequences of MC or is simply a feature of long-term deprivation. Here, we used quantitative immuno-electron microscopy to examine the possibility that alterations of TC synapses occur rapidly enough to impact OD after brief MC. The effect of short-term deprivation on TC synaptic structure was examined in male C57BL/6 mice that underwent 3 and 7 d of MC or monocular retinal inactivation (MI) with tetrodotoxin. The data show that 3 d of MC is sufficient to induce substantial remodeling of TC synapses. In contrast, 3 d of MI, which alters TC activity but does not shift OD, does not significantly affect the structure of TC synapses. Our results support the hypothesis that the rapid plasticity of TC synapses is a key step in the sequence of events that shift OD in visual cortex.

Coleman, Jason E.; Nahmani, Marc; Gavornik, Jeffrey P.; Haslinger, Robert; Heynen, Arnold J.; Erisir, Alev; Bear, Mark F.

2011-01-01

359

Biphasic regulation of the NADPH oxidase by HGF/c-Met signaling pathway in primary mouse hepatocytes.  

PubMed

Redox signaling is emerging as an essential mechanism in the regulation of biological activities of the cell. The HGF/c-Met signaling pathway has been implicated as a key regulator of the cellular redox homeostasis and oxidative stress. We previously demonstrated that genetic deletion of c-Met in hepatocytes disrupts redox homeostasis by a mechanism involving NADPH oxidase. Here, we were focused to address the mechanism of NADPH oxidase regulation by HGF/c-Met signaling in primary mouse hepatocytes and its relevance. HGF induced a biphasic mechanism of NADPH oxidase regulation. The first phase employed the rapid increase in production of ROS as signaling effectors to activate the Nrf2-mediated protective response resulting in up-regulation of the antioxidant proteins, such as NAD(P)H quinone oxidoreductase and ?-glutamylcysteine synthetase. The second phase operated under a prolonged HGF exposure, caused a suppression of the NADPH oxidase components, including NOX2, NOX4, p22 and p67, and was able to abrogate the TGF?-induced ROS production and improve cell viability. In conclusion, HGF/c-Met induces a Nrf2-mediated protective response by a double mechanism driven by NADPH oxidase. PMID:23333744

Clavijo-Cornejo, Denise; Enriquez-Cortina, Cristina; López-Reyes, Alberto; Domínguez-Pérez, Mayra; Nuño, Natalia; Domínguez-Meraz, Marcela; Bucio, Leticia; Souza, Verónica; Factor, Valentina M; Thorgeirsson, Snorri S; Gutiérrez-Ruiz, María Concepción; Gómez-Quiroz, Luis E

2013-01-16

360

Three-dimensional culture of mouse bone marrow cells within a porous polymer scaffold: effects of oxygen concentration and stromal layer on expansion of haematopoietic progenitor cells.  

PubMed

To establish an ex vivo expansion method of haematopoietic progenitor cells (HPCs) and erythroid cells, three-dimensional (3D) cultures of mouse bone marrow cells were performed, employing a porous polyvinyl formal (PVF) resin as a scaffold. In these cultures, the effects of oxygen concentration and co-cultures with stromal cells on the expansion of HPCs and erythroid cells were investigated. When bone marrow cells were cultured under 3D conditions, HPCs and erythroid cells expanded without supplementation of exogenous cytokines, irrespective of the presence of stromal cells. On the contrary, slight expansion of HPCs or erythroid cells was observed in monolayer cultures as controls, indicating that the 3D cultures using the PVF scaffold were far better in expanding HPCs and erythroid cells than the monolayer cultures. Under hypoxic conditions, bone marrow stromal cells allowed for a 3D culture of erythroid cells and HPCs at higher cell densities compared to cultures without stromal cells, and the duration of the expansion of HPCs and erythroid cells after initiating the 3D co-cultures was prolonged. The number of these cells increased throughout the culture period up to 3 weeks under hypoxic conditions, although the number decreased after 2 weeks under normoxic conditions. In conclusion, the 3D co-culture method of haematopoietic cells with stromal cells under hypoxic conditions was confirmed to be effective in expanding HPCs and erythroid cells, and this method seemed to be useful for developing an ex vivo expansion method for haematopoietic cells. PMID:20653040

Miyoshi, Hirotoshi; Murao, Mariko; Ohshima, Norio; Tun, Thein

2011-02-01

361

Quality of Life in Survivors of a Primary Bone Tumour: A Systematic Review  

PubMed Central

Purpose. We conducted a systematic search of published literature, to assess (i) quality of life (QoL) for survivors of a bone tumour compared with the normal population; (ii) QoL implications following amputation, successful or failed limb salvage; (iii) adaptation of young children to amputation compared with older children or adolescents. Methods. Electronic databases were searched including Medline, PsycLIT and Cinahl covering the years 1982– 1998. Results. We identified 11 studies. Regardless of treatment, physical functioning was poor compared with population norms or healthy siblings.There was less consistent evidence regarding emotional functioning. Seven studies compared functioning in amputees and limb salvage patients.Two reported advantages in physical function for the limb salvage group, one for the amputees and the rest no differences. Evidence about social functioning or marriage is inconclusive, but there are suggestions that amputees report more job discrimination. Discussion. The literature is inconclusive, largely because of methodological problems. These include small and non-representative samples, and lack of sensitive and appropriate measures. Specific gaps in the literature include very little work concerned with psychological outcomes for children, or for those experiencing failed limb salvage. More attention needs to be given to gender differences in emotional response to traumatic surgery.The implications of the results for helping families balance the merits of different treatments are discussed.

Grimer, Robert J.

1999-01-01

362

Thrombopoietin receptor agonist therapy in primary immune thrombocytopenia is associated with bone marrow hypercellularity and mild reticulin fibrosis but not other stromal abnormalities.  

PubMed

Primary immune thrombocytopenia is an acquired autoimmune disorder characterized by platelet count of <100 × 10(9)/l in the absence of other causes of thrombocytopenia. Primary immune thrombocytopenia is defined as 'chronic' when it has been present for more than 12 months without spontaneous remission or maintenance of complete response to therapy. Recently, thrombopoietin receptor agonists became available for treatment of chronic primary immune thrombocytopenia. Anecdotal reports have raised concerns about a possible association between therapy with thrombopoietin receptor agonists and an increase in bone marrow fibrosis. To investigate this association we studied eight patients with primary immune thrombocytopenia in detail comparing fibrosis and other morphological features in pre-therapy and on-therapy bone marrow biopsies, with the longest follow-up reported to date. A slight but significant increase to MF-1 in reticulin fibrosis was observed during therapy, but collagen was never present. On-therapy bone marrows were hypercellular due to panmyelosis with increased trilineage hematopoiesis. Megakaryocytes were increased in number, with acquisition of evident pleomorphism, nuclear hyperlobulation and tendency in some cases to form clusters. The overall picture of the on-therapy marrows was characterized by myeloproliferative neoplasm-like features, resembling essential thrombocythemia or occasionally early primary myelofibrosis. As thrombopoietin receptor agonists are becoming a mainstream treatment for primary immune thrombocytopenia, general pathologists and especially hematopathologists need to be aware of the characteristic morphological changes associated with use of these therapeutic agents, in order to avoid misdiagnosis of a myeloid neoplasm. PMID:21841770

Boiocchi, Leonardo; Orazi, Attilio; Ghanima, Waleed; Arabadjief, Melissa; Bussel, James B; Geyer, Julia Turbiner

2011-08-12

363

Partial laminin alpha2 chain restoration in alpha2 chain-deficient dy/dy mouse by primary muscle cell culture transplantation  

PubMed Central

Laminin-2 is a component of skeletal and cardiac basal lamina expressed in normal mouse and human. Laminin alpha2 chain (LAMA2), however, is absent from muscles of some congenital muscular dystrophy patients and the dystrophia muscularis (dy/dy) mouse model. LAMA2 restoration was investigated following cell transplantation in vivo in dy/dy mouse. Allogeneic primary muscle cell cultures expressing the beta- galactosidase transgene under control of a muscular promoter, or histocompatible primary muscle cell cultures, were transplanted into dy/dy mouse muscles. FK506 immunosuppression was used in noncompatible models. All transplanted animals expressed LAMA2 in these immunologically-controlled models, and the degrees of LAMA2 restoration were shown to depend on the age of the animal at transplantation, on muscle pretreatment, and on duration time after transplantation in some cases. LAMA2 did not always colocalize with new or hybrid muscle fibers formed by the fusion of donor myoblasts. LAMA2 deposition around muscle fibers was often segmental and seemed to radiate from the center to the periphery of the injection site. Allogeneic conditionally immortalized pure myogenic cells expressing the beta-galactosidase transgene were characterized in vitro and in vivo. When injected into FK506- immunosuppressed dy/dy mice, these cells formed new or hybrid muscle fibers but essentially did not express LAMA2 in vivo. These data show that partial LAMA2 restoration is achieved in LAMA2-deficient dy/dy mouse by primary muscle cell culture transplantation. However, not all myoblasts, or myoblasts alone, or the muscle fibers they form are capable of LAMA2 secretion and deposition in vivo.

1996-01-01

364

Partial laminin alpha2 chain restoration in alpha2 chain-deficient dy/dy mouse by primary muscle cell culture transplantation.  

PubMed

Laminin-2 is a component of skeletal and cardiac basal lamina expressed in normal mouse and human. Laminin alpha2 chain (LAMA2), however, is absent from muscles of some congenital muscular dystrophy patients and the dystrophia muscularis (dy/dy) mouse model. LAMA2 restoration was investigated following cell transplantation in vivo in dy/dy mouse. Allogeneic primary muscle cell cultures expressing the beta-galactosidase transgene under control of a muscular promoter, or histocompatible primary muscle cell cultures, were transplanted into dy/dy mouse muscles. FK506 immunosuppression was used in noncompatible models. All transplanted animals expressed LAMA2 in these immunologically-controlled models, and the degrees of LAMA2 restoration were shown to depend on the age of the animal at transplantation, on muscle pretreatment, and on duration time after transplantation in some cases. LAMA2 did not always colocalize with new or hybrid muscle fibers formed by the fusion of donor myoblasts. LAMA2 deposition around muscle fibers was often segmental and seemed to radiate from the center to the periphery of the injection site. Allogeneic conditionally immortalized pure myogenic cells expressing the beta-galactosidase transgene were characterized in vitro and in vivo. When injected into FK506-immunosuppressed dy/dy mice, these cells formed new or hybrid muscle fibers but essentially did not express LAMA2 in vivo. These data show that partial LAMA2 restoration is achieved in LAMA2-deficient dy/dy mouse by primary muscle cell culture transplantation. However, not all myoblasts, or myoblasts alone, or the muscle fibers they form are capable of LAMA2 secretion and deposition in vivo. PMID:8601607

Vilquin, J T; Kinoshita, I; Roy, B; Goulet, M; Engvall, E; Tomé, F; Fardeau, M; Tremblay, J P

1996-04-01

365

Identification of a bone marrow-derived epithelial-like population capable of repopulating injured mouse airway epithelium  

PubMed Central

The bone marrow compartment is enriched in stem and progenitor cells, and an unidentified subpopulation of these cells can contribute to lung epithelial repair. Here we identify this subpopulation and quantitate its relative contribution to injured airway epithelium. A subpopulation of adherent human and murine bone marrow cells that expresses Clara cell secretory protein (CCSP) was identified using flow cytometry. When cultured at the air-liquid interface in ex vivo cultures, Ccsp+ cells expressed type I and type II alveolar markers as well as basal cell markers and active epithelial sodium channels. Ccsp+ cells preferentially homed to naphthalene-damaged airways when delivered transtracheally or intravenously, with the former being more efficient than the latter. Interestingly, naphthalene-induced lung damage transiently increased Ccsp expression in bone marrow and peripheral circulation. Furthermore, lethally irradiated Ccsp-null mice that received tagged wild-type bone marrow contained donor-derived epithelium in both normal and naphthalene-damaged airways. This study therefore identifies what we believe to be a newly discovered cell in the bone marrow that might have airway reconstitution potential in the context of cell-based therapies for lung disease. Additionally, these data could reconcile previous controversies regarding the contribution of bone marrow to lung regeneration.

Wong, Amy P.; Keating, Armand; Lu, Wei-Yang; Duchesneau, Pascal; Wang, Xinghua; Sacher, Adrian; Hu, Jim; Waddell, Thomas K.

2009-01-01

366

Cellular mechanism of decreased bone in Brtl mouse model of OI: imbalance of decreased osteoblast function and increased osteoclasts and their precursors.  

PubMed

The Brtl mouse, a knock-in model for moderately severe osteogenesis imperfecta (OI), has a G349C substitution in half of type I collagen alpha1(I) chains. We studied the cellular contribution to Brtl bone properties. Brtl cortical and trabecular bone are reduced before and after puberty, with BV/TV decreased 40-45%. Brtl ObS/BS is comparable to wildtype, and Brtl and wildtype marrow generate equivalent number of colony-forming units (CFUs) at both ages. However, OcS/BS is increased in Brtl at both ages (36-45%), as are TRACP(+) cell numbers (57-47%). After puberty, Brtl ObS/BS decreases comparably to wildtype mice, but osteoblast matrix production (MAR) decreases to one half of wildtype values. In contrast, Brtl OcS falls only moderately (approximately 16%), and Brtl TRACP staining remains significantly elevated compared with wildtype. Consequently, Brtl BFR decreases from normal at 2 mo to one half of wildtype values at 6 mo. Immunohistochemistry and real-time RT-PCR show increased RANK, RANKL, and osteoprotegerin (OPG) levels in Brtl, although a normal RANKL/OPG ratio is maintained. TRACP(+) precursors are markedly elevated in Brtl marrow cultures and form more osteoclasts, suggesting that osteoclast increases arise from more RANK-expressing precursors. We conclude that osteoblasts and osteoclasts are unsynchronized in Brtl bone. This cellular imbalance results in declining BFR as Brtl ages, consistent with reduced femoral geometry. The disparity in cellular number and function results from poorly functioning osteoblasts in addition to increased RANK-expressing precursors that respond to normal RANKL/OPG ratios to generate more bone-resorbing osteoclasts. Interruption of the stimulus that increases osteoclast precursors may lead to novel OI therapies. PMID:18684089

Uveges, Thomas E; Collin-Osdoby, Patricia; Cabral, Wayne A; Ledgard, Felicia; Goldberg, Leah; Bergwitz, Clemens; Forlino, Antonella; Osdoby, Philip; Gronowicz, Gloria A; Marini, Joan C

2008-12-01

367

Alteration of proteoglycan sulfation affects bone growth and remodeling.  

PubMed

Diastrophic dysplasia (DTD) is a chondrodysplasia caused by mutations in the SLC26A2 gene, leading to reduced intracellular sulfate pool in chondrocytes, osteoblasts and fibroblasts. Hence, proteoglycans are undersulfated in the cartilage and bone of DTD patients. To characterize the bone phenotype of this skeletal dysplasia we used the Slc26a2 knock-in mouse (dtd mouse), that was previously validated as an animal model of DTD in humans. X-rays, bone densitometry, static and dynamic histomorphometry, and in vitro studies revealed a primary bone defect in the dtd mouse model. We showed in vivo that this primary bone defect in dtd mice is due to decreased bone accrual associated with a decreased trabecular and periosteal appositional rate at the cell level in one month-old mice. Although the osteoclast number evaluated by histomorphometry was not different in dtd compared to wild-type mice, urine analysis of deoxypyridinoline cross-links and serum levels of type I collagen C-terminal telopeptides showed a higher resorption rate in dtd mice compared to wild-type littermates. Electron microscopy studies showed that collagen fibrils in bone were thinner and less organized in dtd compared to wild-type mice. These data suggest that the low bone mass observed in mutant mice could possibly be linked to the different bone matrix compositions/organizations in dtd mice triggering changes in osteoblast and osteoclast activities. Overall, these results suggest that proteoglycan undersulfation not only affects the properties of hyaline cartilage, but can also lead to unbalanced bone modeling and remodeling activities, demonstrating the importance of proteoglycan sulfation in bone homeostasis. PMID:23369989

Gualeni, Benedetta; de Vernejoul, Marie-Christine; Marty-Morieux, Caroline; De Leonardis, Fabio; Franchi, Marco; Monti, Luca; Forlino, Antonella; Houillier, Pascal; Rossi, Antonio; Geoffroy, Valerie

2013-01-28

368

Characterisation of the p53-Mediated Cellular Responses Evoked in Primary Mouse Cells Following Exposure to Ultraviolet Radiation  

PubMed Central

Exposure to ultraviolet (UV) light can cause significant damage to mammalian cells and, although the spectrum of damage produced varies with the wavelength of UV, all parts of the UV spectrum are recognised as being detrimental to human health. Characterising the cellular response to different wavelengths of UV therefore remains an important aim so that risks and their moderation can be evaluated, in particular in relation to the initiation of skin cancer. The p53 tumour suppressor protein is central to the cellular response that protects the genome from damage by external agents such as UV, thus reducing the risk of tumorigenesis. In response to a variety of DNA damaging agents including UV light, wild-type p53 plays a role in mediating cell-cycle arrest, facilitating apoptosis and stimulating repair processes, all of which prevent the propagation of potentially mutagenic defects. In this study we examined the induction of p53 protein and its influence on the survival of primary mouse fibroblasts exposed to different wavelengths of UV light. UVC was found to elevate p53 protein and its sequence specific DNA binding capacity. Unexpectedly, UVA treatment failed to induce p53 protein accumulation or sequence specific DNA binding. Despite this, UVA exposure of wild-type cells induced a p53 dependent G1 cell cycle arrest followed by a wave of p53 dependent apoptosis, peaking 12 hours post-insult. Thus, it is demonstrated that the elements of the p53 cellular response evoked by exposure to UV radiation are wavelength dependent. Furthermore, the interrelationship between various endpoints is complex and not easily predictable. This has important implications not only for understanding the mode of action of p53 but also for the use of molecular endpoints in quantifying exposure to different wavelengths of UV in the context of human health protection.

McFeat, Gillian D.; Allinson, Sarah L.; McMillan, Trevor J.

2013-01-01

369

Toxic effects of cobalt in primary cultures of mouse astrocytes. Similarities with hypoxia and role of HIF-1alpha.  

PubMed

Cobalt is suspected to cause memory deficit in humans and was reported to induce neurotoxicity in animal models. We have studied the effects of cobalt in primary cultures of mouse astrocytes. CoCl(2) (0.2-0.8mM) caused dose-dependent ATP depletion, apoptosis (cell shrinkage, phosphatidylserine externalization and chromatin rearrangements) and secondary necrosis. The mitochondria appeared to be a main target of cobalt toxicity, as shown by the loss of mitochondrial membrane potential (DeltaPsi(m)) and release from the mitochondria of apoptogenic factors, e.g. apoptosis inducing factor (AIF). Pre-treatment with bongkrekic acid reduced ATP depletion, implicating the involvement of the mitochondrial permeability transition (MPT) pore. Cobalt increased the generation of oxygen radicals, but antioxidants did not prevent toxicity. There was also an impaired response to ATP stimulation, evaluated as a lower raise in intracellular calcium. Similarly to hypoxia and dymethyloxallyl glycine (DMOG), cobalt triggered stabilization of the alpha-subunit of hypoxia-inducible factor HIF-1 (HIF-1alpha). This early event was followed by an increased expression of HIF-1 regulated genes, e.g. stress protein HO-1, pro-apoptotic factor Nip3 and iNOS. Although all of the three stimuli activated the HIF-1alpha pathway and decreased ATP levels, the downstream effects were different. DMOG only inhibited cell proliferation, whereas the other two conditions caused cell death by apoptosis and necrosis. This points to cobalt and hypoxia not only inducing HIF-1alpha regulated genes but also affecting similarly other cellular functions, including metabolism. PMID:17169330

Karovic, Olga; Tonazzini, Ilaria; Rebola, Nelson; Edström, Erik; Lövdahl, Cecilia; Fredholm, Bertil B; Daré, Elisabetta

2006-11-17

370

Characterisation of the p53-Mediated Cellular Responses Evoked in Primary Mouse Cells Following Exposure to Ultraviolet Radiation.  

PubMed

Exposure to ultraviolet (UV) light can cause significant damage to mammalian cells and, although the spectrum of damage produced varies with the wavelength of UV, all parts of the UV spectrum are recognised as being detrimental to human health. Characterising the cellular response to different wavelengths of UV therefore remains an important aim so that risks and their moderation can be evaluated, in particular in relation to the initiation of skin cancer. The p53 tumour suppressor protein is central to the cellular response that protects the genome from damage by external agents such as UV, thus reducing the risk of tumorigenesis. In response to a variety of DNA damaging agents including UV light, wild-type p53 plays a role in mediating cell-cycle arrest, facilitating apoptosis and stimulating repair processes, all of which prevent the propagation of potentially mutagenic defects. In this study we examined the induction of p53 protein and its influence on the survival of primary mouse fibroblasts exposed to different wavelengths of UV light. UVC was found to elevate p53 protein and its sequence specific DNA binding capacity. Unexpectedly, UVA treatment failed to induce p53 protein accumulation or sequence specific DNA binding. Despite this, UVA exposure of wild-type cells induced a p53 dependent G1 cell cycle arrest followed by a wave of p53 dependent apoptosis, peaking 12 hours post-insult. Thus, it is demonstrated that the elements of the p53 cellular response evoked by exposure to UV radiation are wavelength dependent. Furthermore, the interrelationship between various endpoints is complex and not easily predictable. This has important implications not only for understanding the mode of action of p53 but also for the use of molecular endpoints in quantifying exposure to different wavelengths of UV in the context of human health protection. PMID:24098727

McFeat, Gillian D; Allinson, Sarah L; McMillan, Trevor J

2013-09-30

371

Coactivator PGC-1{alpha} regulates the fasting inducible xenobiotic-metabolizing enzyme CYP2A5 in mouse primary hepatocytes  

SciTech Connect

The nutritional state of organisms and energy balance related diseases such as diabetes regulate the metabolism of xenobiotics such as drugs, toxins and carcinogens. However, the mechanisms behind this regulation are mostly unknown. The xenobiotic-metabolizing cytochrome P450 (CYP) 2A5 enzyme has been shown to be induced by fasting and by glucagon and cyclic AMP (cAMP), which mediate numerous fasting responses. Peroxisome proliferator-activated receptor {gamma} coactivator (PGC)-1{alpha} triggers many of the important hepatic fasting effects in response to elevated cAMP levels. In the present study, we were able to show that cAMP causes a coordinated induction of PGC-1{alpha} and CYP2A5 mRNAs in murine primary hepatocytes. Furthermore, the elevation of the PGC-1{alpha} expression level by adenovirus mediated gene transfer increased CYP2A5 transcription. Co-transfection of Cyp2a5 5' promoter constructs with the PGC-1{alpha} expression vector demonstrated that PGC-1{alpha} is able to activate Cyp2a5 transcription through the hepatocyte nuclear factor (HNF)-4{alpha} response element in the proximal promoter of the Cyp2a5 gene. Chromatin immunoprecipitation assays showed that PGC-1{alpha} binds, together with HNF-4{alpha}, to the same region at the Cyp2a5 proximal promoter. In conclusion, PGC-1{alpha} mediates the expression of CYP2A5 induced by cAMP in mouse hepatocytes through coactivation of transcription factor HNF-4{alpha}. This strongly suggests that PGC-1{alpha} is the major factor mediating the fasting response of CYP2A5.

Arpiainen, Satu; Jaervenpaeae, Sanna-Mari [Department of Pharmacology and Toxicology, University of Oulu, Oulu (Finland); Manninen, Aki [Biocenter Oulu, Department of Medical Biochemistry and Molecular Biology, University of Oulu, Oulu (Finland); Viitala, Pirkko [Department of Pharmacology and Toxicology, University of Oulu, Oulu (Finland); Lang, Matti A. [Division of Pharmaceutical Biochemistry, Uppsala Biomedical Center, Uppsala University, Uppsala (Sweden); Pelkonen, Olavi [Department of Pharmacology and Toxicology, University of Oulu, Oulu (Finland); Hakkola, Jukka [Department of Pharmacology and Toxicology, University of Oulu, Oulu (Finland)], E-mail: jukka.hakkola@oulu.fi

2008-10-01

372

Electrical stimulation enhances neurogenin2 expression through ?-catenin signaling pathway of mouse bone marrow stromal cells and intensifies the effect of cell transplantation on brain injury.  

PubMed

Bone marrow stromal cells (BMSCs) have received significant attention for its use in neural regeneration. However, neural replacement by transplanted BMSCs was not very effective. Recently, the gene transfection method has improved the capability of cell transplantation; however, this method results in canceration and immune rejection. We induced the differentiation of mouse BMSCs into neural cells using electrical stimulation and transplanted the cells into traumatic brain injury (TBI) model mice. We found that the electrically stimulated cells have good potential to differentiate into neural cells and contribute to recovery from TBI without differentiating into astrocytes. In addition, we found that electrical stimulation enhanced neurogenin2 (Ngn2) expression. Ngn2 is involved in neural differentiation and inhibits astrocytic differentiation during cell growth. Furthermore, we found that this enhancement of Ngn2 expression occurred through ?-catenin signaling pathway. This study may contribute to the use of BMSCs for neural replacement in central nervous system diseases. PMID:23142721

Matsumoto, Masaya; Imura, Takeshi; Fukazawa, Takahiro; Sun, Yanan; Takeda, Masaaki; Kajiume, Teruyuki; Kawahara, Yumi; Yuge, Louis

2012-11-06

373

Heavy metal content in the femora of yellow-necked mouse (Apodemus flavicollis) and wood mouse (Apodemus sylvaticus) from different types of polluted environment in Slovakia.  

PubMed

Heavy metal content in the femora of yellow-necked mouse (Apodemus flavicollis) and wood mouse (Apodemus sylvaticus) caught in different polluted biotopes of a low hill level in Slovakia (Nováky and Kolínany) was investigated in the present study. Length, weight and histological structure of mouse bones have also been analysed. According to our results, higher concentrations of Cd, Ni, Fe, Cu and Zn were detected in the femora of A. flavicollis from Kolínany area. Similarly, we observed higher concentrations of Ni, Fe, Cu and Zn in the bones of A. sylvaticus trapped at the same biotope. Significant differences were observed for concentrations of Ni and Zn in both species (P<0.05). The measured values for bone length and bone weight were higher in yellow-necked mice and wood mice from Nováky locality (P<0.01). Histological observation of thin sections from femora of A. flavicollis and A. sylvaticus revealed an outer and inner non-vascular lamellar layer around a poorly developed reticular layer. We did not identify demonstrable changes in qualitative histological characteristics of the femora between the mice (A. flavicollis and A. sylvaticus separately) from different types of polluted environment. Also, no statistically significant differences for all the measured variables of primary osteons' vascular canals were observed. Correlation analysis in yellow-necked mouse showed high positive relation between bone weight and bone length (r=0.66), area and perimeter (r=0.87) and perimeter and maximum diameter (r=0.87). In wood mouse, high positive correlation between bone weight and bone length (r=0.80), area and perimeter (r=0.72), area and maximum diameter (r=0.66) and perimeter and maximum diameter (r=0.74) was found. Our results demonstrate slightly elevated accumulation of some heavy metals in the femora of yellow-necked mouse and wood mouse from Kolínany biotope and thus give an evidence of a contamination of the environment. PMID:20135219

Martiniaková, Monika; Omelka, Radoslav; Jancová, Alena; Stawarz, Robert; Formicki, Grzegorz

2010-02-05

374

Resistance to infection with Eimeria vermiformis in mouse radiation chimeras is determined by donor bone-marrow cells  

SciTech Connect

The course of infection with Eimeria vermiformis was determined in BALB/b, BALB/c, and C57BL/10ScSn (B10) mice and in radiation chimeras prepared from the H-2-compatible BALB/b and