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PubMed2006

Down-regulation of androgen receptor by 3,3'-diindolylmethane contributes to inhibition of cell proliferation and induction of apoptosis in both hormone-sensitive LNCaP and insensitive C4-2B prostate cancer cells.

Despite the initial efficacy of androgen deprivation therapy, most patients with advanced prostate cancer eventually progress to hormone-refractory prostate cancer, for which there is no curative therapy. Previous studies from our laboratory and others have shown the antiproliferative and proapoptotic effects of 3,3'-diindolylmethane (DIM) in prostate cancer cells. However, the molecular mechanism of action of DIM has not been investigated in androgen receptor (AR)-positive hormone-responsive and -nonresponsive prostate cancer cells. Therefore, we investigated the effects of B-DIM, a formulated DIM with greater bioavailability, on AR, Akt, and nuclear factor kappaB (NF-kappaB) signaling in hormone-sensitive LNCaP (AR+) and hormone-insensitive C4-2B (AR+) prostate cancer cells. We found that B-DIM significantly inhibited cell proliferation and induced apoptosis in both cell lines. By Akt gene transfection, reverse transcription-PCR, Western blot analysis, and electrophoretic mobility shift assay, we found a potential crosstalk between Akt, NF-kappaB, and AR. Importantly, B-DIM significantly inhibited Akt activation, NF-kappaB DNA binding activity, AR phosphorylation, and the expressions of AR and prostate-specific antigen, suggesting that B-DIM could interrupt the crosstalk. Confocal studies revealed that B-DIM inhibited AR nuclear translocation, leading to the down-regulation of AR target genes. Moreover, B-DIM significantly inhibited C4-2B cell growth in a severe combined immunodeficiency-human model of experimental prostate cancer bone metastasis. These results suggest that B-DIM-induced cell proliferation inhibition and apoptosis induction are partly mediated through the down-regulation of AR, Akt, and NF-kappaB signaling. These observations provide a rationale for devising novel therapeutic approaches for the treatment of hormone-sensitive, but more importantly, hormone-refractory prostate cancer by using B-DIM alone or in combination with other therapeutics.

Androgen Receptor Antagonists↗
PubMed1993

Leukocytes synthesize angiotensinogen.

To determine whether leukocytes express the angiotensinogen gene, we subjected circulating rat leukocytes and murine bone marrow cells to Northern blot analysis and hybridization with homologous angiotensinogen complementary DNA. Angiotensinogen messenger RNA sequences were detected in circulating adult rat leukocytes, in murine-irradiated and nonirradiated bone marrow stromal cells, and in an adherent stromal cell line (preadipocyte). Western blot analysis of rat leukocyte homogenate showed that rat leukocytes contain two main angiotensinogen isoforms with approximate molecular weights of 46.5 and 53.9 kd. Synthesis and release of angiotensinogen protein by rat leukocytes was confirmed by immunoprecipitation of radiolabeled angiotensinogen from cell lysate and media of rat leukocytes that were metabolically labeled with 35S-L-methionine. In addition, the angiotensinogen protein present in media of rat leukocytes was enzymatically cleaved by hog renin, resulting in generation of angiotensin I (305 +/- 47 pg angiotensin I per milliliter of media per hour). We conclude that circulating rat leukocytes express the angiotensinogen gene and synthesize and release angiotensinogen with the capability to generate angiotensin. Expression of angiotensinogen by leukocytes may provide a mobile angiotensin-generating system of potential importance in the regulation of local inflammatory responses, tissue injury (i.e., myocardial infarction), and arterial hypertension.

Angiotensin I↗
PubMedDate not supplied

Management of infants with prenatal ultrasound diagnosis of airway obstruction by teratoma.

We report a case of a cervical teratoma that was diagnosed prenatally. In anticipation of potential upper airway obstruction, resources were mobilized to the operating room at the time of planned cesarean section. The neonate developed respiratory distress, and her airway was secured. There was no evidence of cerebral anoxia initially, and she is currently thriving with no central or peripheral neurologic defects. As prenatal diagnosis by ultrasound becomes more refined, the otolaryngologist will play an increasing role in perinatal decision-making and anticipated emergencies at the time of delivery. Airway obstruction of various causes will be the most urgent problem.

Airway Obstruction↗
PubMed2000

The MABAL shoe, an alternative method in contact casting for the treatment of neuropathic diabetic foot ulcers.

INTRODUCTION: Total Contact Casting (TCC) is considered the gold standard in the treatment of neuropathic diabetic foot ulcers. To overcome some disadvantages of TCC we developed a removable fiberglass combicast shoe--the MABAL shoe. PATIENTS AND METHODS: The MABAL shoe was used to treat 23 plantar ulcers. RESULTS: Before treatment the mean surface area was 2.0 cm2 (0.5-7.1 cm2). Twenty-one of 23 ulcers healed, with a mean healing time of 34 days (7-75 days). CONCLUSION: The MABAL shoe provides healing of neuropathic diabetic foot ulcers comparable to existing methods of treatment, while offering some potential advantages. The main advantages are mobilization of the ankle, removability of the cast and a less time consuming form of treatment.

Adhesives↗
PubMed2006

Wikis, blogs and podcasts: a new generation of Web-based tools for virtual collaborative clinical practice and education.

BACKGROUND: We have witnessed a rapid increase in the use of Web-based 'collaborationware' in recent years. These Web 2.0 applications, particularly wikis, blogs and podcasts, have been increasingly adopted by many online health-related professional and educational services. Because of their ease of use and rapidity of deployment, they offer the opportunity for powerful information sharing and ease of collaboration. Wikis are Web sites that can be edited by anyone who has access to them. The word 'blog' is a contraction of 'Web Log' - an online Web journal that can offer a resource rich multimedia environment. Podcasts are repositories of audio and video materials that can be "pushed" to subscribers, even without user intervention. These audio and video files can be downloaded to portable media players that can be taken anywhere, providing the potential for "anytime, anywhere" learning experiences (mobile learning). DISCUSSION: Wikis, blogs and podcasts are all relatively easy to use, which partly accounts for their proliferation. The fact that there are many free and Open Source versions of these tools may also be responsible for their explosive growth. Thus it would be relatively easy to implement any or all within a Health Professions' Educational Environment. Paradoxically, some of their disadvantages also relate to their openness and ease of use. With virtually anybody able to alter, edit or otherwise contribute to the collaborative Web pages, it can be problematic to gauge the reliability and accuracy of such resources. While arguably, the very process of collaboration leads to a Darwinian type 'survival of the fittest' content within a Web page, the veracity of these resources can be assured through careful monitoring, moderation, and operation of the collaborationware in a closed and secure digital environment. Empirical research is still needed to build our pedagogic evidence base about the different aspects of these tools in the context of medical/health education. SUMMARY AND CONCLUSION: If effectively deployed, wikis, blogs and podcasts could offer a way to enhance students', clinicians' and patients' learning experiences, and deepen levels of learners' engagement and collaboration within digital learning environments. Therefore, research should be conducted to determine the best ways to integrate these tools into existing e-Learning programmes for students, health professionals and patients, taking into account the different, but also overlapping, needs of these three audience classes and the opportunities of virtual collaboration between them. Of particular importance is research into novel integrative applications, to serve as the "glue" to bind the different forms of Web-based collaborationware synergistically in order to provide a coherent wholesome learning experience.

Audiovisual Aids↗
PubMed2003

External Ca2+ is predominantly used for cytoplasmic and nuclear Ca2+ increases in fertilized oocytes of the marine bivalve Mactra chinensis.

Oocytes of the marine bivalve Mactra chinensis are spawned and arrested at the germinal vesicle stage (first meiotic prophase) until fertilization, without undergoing a process called oocyte maturation. As is the case of other animals, a fertilized oocyte of the bivalve displays increases in intracellular free Ca(2+). We have clarified here the spatiotemporal patterns and sources of the intracellular Ca(2+) changes at fertilization. Shortly after insemination, increased Ca(2+) simultaneously appeared at the whole cortical region of the oocyte and spread inwardly to the center, attaining the maximal Ca(2+) levels throughout the oocyte, including the cytoplasm and nucleus. The initial maximal Ca(2+) peak was followed by a submaximal plateau phase of cytoplasmic and nuclear Ca(2+) elevations, which persisted for several minutes. The nuclear envelope began to break down shortly before the termination of the plateau phase. These sperm-induced Ca(2+) changes were inhibited by suppression of the influx of external Ca(2+) from seawater but not by disturbance of the release of internal Ca(2+) from inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)]-sensitive stores, suggesting that the increased Ca(2+) is from an external source. In contrast to the situation observed at fertilization, an oocyte artificially stimulated with serotonin (5-hydroxytryptamine, 5-HT) displayed repetitive Ca(2+) transients, each of which started from one cortical region and propagated across the oocyte as a Ca(2+) wave. The 5-HT-induced Ca(2+) transients persisted even in the absence of external Ca(2+). Experiments with caged Ins(1,4,5)P(3) revealed that Ca(2+) release from Ins(1,4,5)P(3)-sensitive stores is another pathway that is sufficient to trigger meiosis reinitiation from the first prophase. These results demonstrate that Mactra oocytes can potentially use two different Ca(2+)-mobilizing pathways: Ca(2+) influx producing a centripetal Ca(2+) wave from the whole cortex and Ca(2+) release from Ins(1,4,5)P(3)-sensitive stores producing a point-source propagating Ca(2+) wave. However, it seems likely that the Ca(2+) influx pathway is predominantly activated at fertilization.

Animals↗
PubMed2006

Interplay of PIWI/Argonaute protein MIWI and kinesin KIF17b in chromatoid bodies of male germ cells.

Chromatoid bodies are thought to act as male-germ-cell-specific platforms for the storing and processing of haploid transcripts. The molecular mechanisms governing the formation and function of these germ-cell-specific structures have remained elusive. In this study, we show that the kinesin motor protein KIF17b, which is involved in the nucleocytoplasmic transport of RNA and of a transcriptional coactivator, localizes in chromatoid bodies. The chromatoid body moves actively and non-randomly in the cytoplasm of round spermatids, making frequent contacts with the nuclear envelope. The localization of KIF17b thereby offers a potential mechanism for microtubule-dependent mobility of chromatoid bodies, as well as for the transport of the specific components in and out of the chromatoid body. Interestingly, we demonstrate that KIF17b physically interacts with a testis-specific member of the PIWI/Argonaute family, MIWI, a component of chromatoid bodies implicated in RNA metabolism. A functional interplay between KIF17b and MIWI might be needed for the loading of haploid RNAs in the chromatoid body. Importantly, chromatoid bodies from round spermatids of miwi-null mice are not fully compacted and remain as a diffuse chromatoid material, revealing the essential role played by MIWI in the formation of chromatoid bodies. These results shed new light on the function of chromatoid bodies in the post-transcriptional regulation of gene expression in haploid germ cells.

Animals↗
PubMed2006

User charges in public health facilities in Tanzania: effect on revenues, quality of services and people's health-seeking behaviour for malaria illnesses in Korogwe district.

User charges in public health facilities are aimed at improving efficiency and quality of health services. In Africa, evidence about their effect on patient attendance and community health-seeking behaviour are mixed. This paper reports a study of the effect of user charges on revenue collection, quality of services and people's health-seeking behaviour in relation to malaria in Korogwe district, Tanzania. Data were collected through focus-group discussions with community members, interviewing community leaders and health workers, field observations and review of patient registers. Generally, there was no distinct difference in the trends of patient attendances before and after user fee introduction. Public awareness about cost-sharing policy was high, but had low appreciation in the administration of exemptions and waivers. Shortage of drugs, laboratory facilities, and inhospitality of nurses lowered their confidence in the user-fee system. Autonomy to collect and prioritize expenditure of user-fee revenue at the health-facility level was appreciated by community leaders and health workers who, however, had reservations with funds being held at the district level and delays by the DMO's office in approving budgets submitted in request for expenditure of such revenues. Thus, despite the potential of user charges for revenue mobilization, problems with their administration lowers public confidence in the user-fee system improving quality and accessibility of services to the poor.

Cross-Sectional Studies↗
PubMed1988

Stimulation of prostaglandin release by Ca2+-mobilizing agents from the perfused rat liver. A comparative study on the action of ATP, UTP, phenylephrine, vasopressin and nerve stimulation.

Several Ca2+-mobilizing agents were tested for their potential to elicit the net release of prostaglandins from the isolated perfused rat liver. Among these ATP and UTP only led to an efficient stimulation of PGD2 and PGE2 synthesis. 20 microM ATP or 20 microM UTP increased the release of PGD2 8-fold and that of PGE2 2 to 3-fold. In total, at least 40 times more PGD2 than PGE2 left the liver after stimulation. The time course of prostaglandin release was similar for both nucleotides. Vasopressin had almost no effect on the release of both prostaglandins and on portal vein pressure. But phenylephrine and nerve stimulation while raising the PGD2 efflux only slightly caused an elevation of PGE2 outflow and portal pressure.

Adenosine Triphosphate↗
PubMedDate not supplied

Bioremediation of residual fertilizer nitrate: II. Soil redox potential and soluble iron as indicators of soil health during treatment.

The prospect of using wastewater containing high loads of soluble organic matter (OM) for removing residual agricultural chemicals (fertilizer, pesticide, or herbicide) in farm soil, although promising, could have adverse effects on soil agricultural quality as a result of development of redoximorphic features in the soil profile. In this study, the effect of organic carbon supplement for bioremediation of residual fertilizer nitrate on soil properties, redox potential (Eh), pH, and metal ion mobilization was studied using sandy soils packed in columns. The study was included in a general project, described elsewhere (Ugwuegbu et al., 2000), undertaken to evaluate use of controlled water table management (WTM) systems to supply organic carbon for creating a reduced environment conducive to denitrification of residual fertilizer nitrate leaching from the farm to subsurface water. The columns were subjected to subirrigation with water containing soluble organic carbon in the form of glucose. The work was carried out in two experimental setups and the long-term effect of a range of glucose concentrations on the Eh, pH, and soluble levels of Fe and Mn was investigated. From the results obtained, it could be concluded that excessive organic carbon supplement to soil can have adverse effects on soil quality and that Eh and soluble Fe are the two most practical parameters for monitoring soil health during treatment of farm chemicals.

Agriculture↗
PubMedDate not supplied

Retention and runoff losses of atrazine and metribuzin in soil.

Minimizing herbicide runoff and mobility in the soil and thus potential contamination of water resources is a national concern. Metribuzin [4-amino-6-(1,1-dimethylethyl)-3-(methylthio)-1,2,4-triazin-5(4H)-one] and atrazine [2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine] dynamics in surface soils and in runoff waters were studied on six 0.2-ha sugarcane (Saccharum spp.) plots of a Commerce silt loam (fine-silty, mixed, superactive, nonacid, thermic Fluvaquentic Endoaquept) during three growing seasons under different best management practices. Metribuzin was applied in the spring as a postemergence herbicide and atrazine was applied following winter harvest. Both herbicides were applied on top of the sugarcane rows as 0.6- or 0.9-m band width application, or broadcast application, where the entire area was treated. Maximum effluent concentrations were measured from the broadcast treatment and ranged from 600 to 1100 microg L(-1) for atrazine and 250 to 450 microg L(-1) for metribuzin. Atrazine runoff losses were highest for the broadcast treatment (2.8-11% of that applied) and lowest for the 0.6-m band treatment (1.9-7.6%), with a similar trend for metribuzin losses. Measured extractable herbicides from the surface soil exhibited a sharp decrease with time and were well described with a simple first-order decay model. For atrazine, estimates for the decay rate (lambda) were higher than for metribuzin. Results based on laboratory adsorption-desorption (kinetic-batch) measurements were consistent with field observations. The distribution coefficients (Kd) for atrazine exhibited stronger retention over time in comparison with metribuzin on the Commerce soil. Moreover, discrepancies between adsorption isotherm and desorption indicated slower release and that hysteresis was more pronounced for atrazine compared with metribuzin.

Atrazine↗
PubMed2005

Trace elements analysis in forage samples from a US Navy bombing range (Vieques, Puerto Rico).

Plants are good environmental sensors of the soil conditions in which they are growing. They also respond directly to the state of air. The tops of plants are collectors of air pollutants, and their chemical composition may be a good indicator for contaminated-areas when it is assessed against background values obtained for unpolluted vegetation. Both, aquatic and terrestrial plants are known to bioaccumulate heavy metals and therefore represent a potential source of these contaminants to the human food chain. An evaluation of heavy metals was conducted from vegetation samples collected at the Atlantic Fleet Weapons Training Facilities (AFWTF) in Vieques, Puerto Rico. In order to understand the potential risks associated to heavy metal mobilization through biological systems, it is first necessary to establish background values obtained from reference locations. This information allows a better interpretation of the significance of anthropogenic factors in changing trace elements status in soil and plants. Since Guánica State Forest is located at a similar geoclimatic zone as the AFWTF, samples at this site were used as a standard reference material and as experimental controls. Both sampling and analysis were conducted as previously described in standardized protocols using acid digestion of dry ashes. Then, levels of heavy metals were obtained by air-acetylene flame detection in an atomic absorption spectrophotometer. Our results from the samples taken at the AFWTF indicate mobilization of undesirable trace elements through the marine and terrestrial food web. Since plants naturally remove heavy metals from soils, they could be employed for the restoration of this and similarly contaminated sites.

Environmental Monitoring↗
PubMed2001

N-linked glycosylations at Asn(26) and Asn(114) of human MD-2 are required for toll-like receptor 4-mediated activation of NF-kappaB by lipopolysaccharide.

MD-2 is physically associated with Toll-like receptor 4 (TLR4) and is required for TLR4-mediated LPS signaling. Western blotting analysis revealed the presence of three forms of human (h)MD-2 with different electrophoretic mobilities. After N-glycosidase treatment of the cellular extract prepared from cells expressing hMD-2, only a single form with the fastest mobility was detected. Mutation of either one of two potential glycosylation sites (Asn(26) and Asn(114)) of MD-2 resulted in the disappearance of the slowest mobility form, and only the fastest form was detected in hMD-2 carrying mutations at both Asn(26) and Asn(114). Although these mutants were expressed on the cell surface and maintained its ability to associate with human TLR4, these mutations or tunicamycin treatment substantially impaired the ability of MD-2 to complement TLR4-mediated activation of NF-kappaB by LPS. LPS binding to cells expressing CD14, TLR4, and MD-2 was unaffected by these mutations. These observations demonstrate that hMD-2 undergoes N-linked glycosylation at Asn(26) and Asn(114), and that these glycosylations are crucial for TLR4-mediated signal transduction of LPS.

Acute-Phase Proteins↗
PubMed2004

Chemotaxis and calcium responses of phagocytes to formyl peptide receptor ligands is differentially regulated by cyclic ADP ribose.

Cyclic ADP ribose (cADPR) is a calcium-mobilizing metabolite that regulates intracellular calcium release and extracellular calcium influx. Although the role of cADPR in modulating calcium mobilization has been extensively examined, its potential role in regulating immunologic responses is less well understood. We previously reported that cADPR, produced by the ADP-ribosyl cyclase, CD38, controls calcium influx and chemotaxis of murine neutrophils responding to fMLF, a peptide agonist for two chemoattractant receptor subtypes, formyl peptide receptor and formyl peptide receptor-like 1. In this study, we examine whether cADPR is required for chemotaxis of human monocytes and neutrophils to a diverse array of chemoattractants. We found that a cADPR antagonist and a CD38 substrate analogue inhibited the chemotaxis of human phagocytic cells to a number of formyl peptide receptor-like 1-specific ligands but had no effect on the chemotactic response of these cells to ligands selective for formyl peptide receptor. In addition, we show that the cADPR antagonist blocks the chemotaxis of human monocytes to CXCR4, CCR1, and CCR5 ligands. In all cases, we found that cADPR modulates intracellular free calcium levels in cells activated by chemokines that induce extracellular calcium influx in the apparent absence of significant intracellular calcium release. Thus, cADPR regulates calcium signaling of a discrete subset of chemoattractant receptors expressed by human leukocytes. Since many of the chemoattractant receptors regulated by cADPR bind to ligands that are associated with clinical pathology, cADPR and CD38 represent novel drug targets with potential application in chronic inflammatory and neurodegenerative disease.

ADP-ribosyl Cyclase↗
PubMedDate not supplied

Mobile bearings in primary knee arthroplasty.

Mobile-bearing knee arthroplasty (MBKA) has potential advantages compared with conventional fixed-bearing total knee arthroplasty (TKA). By allowing unconstrained axial rotation, MBKA can offer greater articular conformity without an increased probability of loosening due to increased axial torque. Increased articular conformity minimizes polyethylene contact stresses, thereby reducing linear wear and subsurface fatigue failure. Axial rotation of the platform also enables self-correction of tibial component malrotation. Despite these advantages, the long-term clinical results obtained with current MBKA devices are similar to those obtained with well-designed fixed-bearing TKA prostheses, with no data suggesting their superiority. The disadvantages of MBKA include bearing dislocation and breakage, soft-tissue impingement, a steep technique learning curve, and concerns about volumetric wear. Hypothetically, longer-term follow-up of MBKA results may reveal a significant difference from fixed-bearing TKA results as the fatigue failure threshold of incongruent polyethylene is exceeded.

Biomechanical Phenomena↗
PubMed1986

Community disaster planning and training.

In a major disaster there is no easy way to deal with the multitude of problems that occur unless there are plans prepared and regular exercises to provide practical training. Disaster planning is an ongoing process. Personnel from all response agencies should be trained in their roles and be familiar with the plan. Realistic planning takes into account the need to adjust plans to the people, not vice versa. It should not require people or agencies to deviate sharply from their normal, everyday behavior. Disasters require a prompt response, coordinated activities, and cooperative efforts by all concerned agencies in order to identify and evaluate actual and potential risks, control and eliminate hazards, mobilize resources, and provide effective action to save life and property.

Disaster Planning↗
PubMed2000

Identification of fractalkine, a CX3C-type chemokine, as a direct target of p53.

Fractalkine is a CX3C-type chemokine that induces chemotaxis of monocytes and cytotoxic T cells. Using the differential display method for examining gene expression in p53-defective cells transfected by adenovirus containing wild-type p53, we observed that fractalkine was induced by ectopic expression of p53. An electrophoretic mobility shift assay showed that a potential p53-binding site present in the promoter of the fractalkine gene could bind to p53 protein. Moreover, a heterogeneous reporter assay indicated that this promoter sequence possessed p53-dependent transcriptional activity. The strong induction of fractalkine when p53 protein was expressed by a wild-type transgene in p53-defective cells brought to light a novel role for p53; that is, potential elimination of damaged cells by the host immune response system through transcriptional regulation of fractalkine. Our results imply a pivotal role of p53 in immunosurveillance to prevent cells from undergoing malignant transformation.

Adenoviridae↗
PubMed2001

Serotonin 5-HT1 receptors potentiate histamine and thrombin stimulated prostaglandin synthesis in endothelial cells.

The ability of serotonin 5-HT1 receptors to increase vascular tone was previously found to be activated by vasoconstrictiors such as histamine. In this study, treatment of cultured human aortic endothelial cells (HAEC) with the 5-HT1-selective agonist 5-carboxamidotryptamine (5-CT) alone had no effect on the levels of prostaglandin F2alpha (PGF2alpha) or 6-keto-prostaglandin F1alpha (6-keto PGF1alpha). However, 5-CT potentiated the histamine and thrombin stimulated increases in prostaglandins released by HAEC. In the presence of histamine, increasing doses of 5-CT caused a steep rise in PGF2alpha levels resulting in an increase in the ratio of PGF2alpha over 6-keto PGF1alpha. The ability of 5-CT to potentiate prostaglandin production was correlated with its ability to potentiate the histamine and thrombin mediated mobilization of arachidonic acid. These results demonstrate that the ability of 5-HT1 receptors to stimulate prostaglandin production in endothelial cells is activated by histamine and thrombin.

6-Ketoprostaglandin F1 alpha↗