Targeting elevated heme levels to treat a mouse model for Diamond-Blackfan Anemia.

Diamond-Blackfan anemia (DBA) is a rare genetic disorder in which patients present a scarcity of erythroid precursors in an otherwise normocellular bone marrow. Most, but not all, patients carry mutations in ribosomal proteins such as RPS19, suggesting that compromised mRNA translation and ribosomal stress are pathogenic mechanisms causing depletion of erythroid precursors. To gain further insight to disease mechanisms in DBA, we performed a custom short hairpin RNA (shRNA) based screen against 750 genes hypothesized to affect DBA pathophysiology.

Glucocorticoids improve erythroid progenitor maintenance and dampen Trp53 response in a mouse model of Diamond-Blackfan anaemia.

Diamond-Blackfan anaemia (DBA) is a rare congenital disease causing severe anaemia and progressive bone marrow failure. The majority of patients carry mutations in ribosomal proteins, which leads to depletion of erythroid progenitors in the bone marrow. As many as 40% of all DBA patients receive glucocorticoids to alleviate their anaemia.

Effective density and mixing state of aerosol particles in a near-traffic urban environment.

In urban environments, airborne particles are continuously emitted, followed by atmospheric aging. Also, particles emitted elsewhere, transported by winds, contribute to the urban aerosol. We studied the effective density (mass-mobility relationship) and mixing state with respect to the density of particles in central Copenhagen, in wintertime.

Patient Attachment to Therapist Rating Scale: development and psychometric properties.

Objective: To report on the development and initial psychometric properties of a new rating scale for patent-therapist attachment.

Method: Seventy interviews from the Young Adult Psychotherapy Project (YAPP) were rated.

Results: Excellent internal consistency (Cronbach's α > .

Progress towards mechanism-based treatment for Diamond-Blackfan anemia.

Diamond-Blackfan anemia (DBA) is a congenital erythroid hypoplastic anemia, characterized by macrocytic anemia, reticulocytopenia, and severely reduced numbers of erythroid precursors in the bone marrow. For more than fifty years, glucocorticoids have remained the main option for pharmacological treatment of DBA. While continuous glucocorticoid administration increases hemoglobin levels in a majority of DBA patients, it also causes severe side effects.

NMR-derived models of amidopyrine and its metabolites in complexes with rabbit cytochrome P450 2B4 reveal a structural mechanism of sequential N-dealkylation.

To understand the molecular basis of sequential N-dealkylation by cytochrome P450 2B enzymes, we studied the binding of amidopyrine (AP) as well as the metabolites of this reaction, desmethylamidopyrine (DMAP) and aminoantipyrine (AAP), using the X-ray crystal structure of rabbit P450 2B4 and two nuclear magnetic resonance (NMR) techniques: saturation transfer difference (STD) spectroscopy and longitudinal (T(1)) relaxation NMR. Results of STD NMR of AP and its metabolites bound to P450 2B4 were similar, suggesting that they occupy similar niches within the enzyme's active site. The model-dependent relaxation rates (R(M)) determined from T(1) relaxation NMR of AP and DMAP suggest that the N-linked methyl is closest to the heme.

The influence of small aerosol particles on the properties of water and ice clouds.

In this paper, results are presented of the influence of small organic- and soot-containing particles on the formation of water and ice clouds. There is strong evidence that these particles have grown from nano particle seeds produced by the combustion of oil products. Two series of field experiments are selected to represent the observations made.

The orphan receptor GPR55 is a novel cannabinoid receptor.

Background: The endocannabinoid system functions through two well characterized receptor systems, the CB1 and CB2 receptors. Work by a number of groups in recent years has provided evidence that the system is more complicated and additional receptor types should exist to explain ligand activity in a number of physiological processes.

Experimental Approach: Cells transfected with the human cDNA for GPR55 were tested for their ability to bind and to mediate GTPgammaS binding by cannabinoid ligands.

Secondary organic aerosols from anthropogenic and biogenic precursors.

Secondary organic aerosol (SOA) formation from the photooxidation of an anthropogenic (1,3,5-trimethylbenzene) and a biogenic (alpha-pinene) precursor was investigated at the new PSI smog chamber. The chemistry of the gas phase was followed by proton transfer reaction mass spectrometry, while the aerosol chemistry was investigated with aerosol mass spectrometry, ion chromatography, laser desorption ionization mass spectrometry, and infrared spectroscopy, along with volatility and hygroscopicity studies. Evidence for oligomer formation for SOA from both precursors was given by an increasing abundance of compounds with a high molecular weight (up to 1000 Da) and by an increasing thermal stability with increasing aging time.

Identification and characterisation of a novel splice variant of the human CB1 receptor.

Cannabinoid ligands are implicated in many physiological processes and to date two receptors have been identified. However, a growing body of evidence exists that suggests the presence of additional receptors. Whilst cloning the previously described hCB1a, we have identified a novel variant that we call hCB1b.

Gigantic cannibals driving a whole-lake trophic cascade.

Trophic cascades have been a central paradigm in explaining the structure of ecological communities but have been demonstrated mainly through comparative studies or experimental manipulations. In contrast, evidence for shifts in trophic cascades caused by intrinsically driven population dynamics is meager. By using empirical data of a cannibalistic fish population covering a 10-year period and a size-structured population model, we show the occurrence of a dynamic trophic cascade in a lake ecosystem, in which the community over time alternates between two different configurations.

Prognostic and predictive value of c-erbB-2 overexpression in primary breast cancer, alone and in combination with other prognostic markers.

Purpose: To investigate the prognostic and predictive value of c-erbB-2 overexpression in breast cancer in relation to other prognostic markers.

Patients And Methods: Paraffin-embedded tumors from 315 consecutive primary breast cancer patients were screened for c-erbB-2 protein (p185) overexpression by immunohistochemistry using the monoclonal antibody CB11.

Results: c-erbB-2 protein overexpression was detected in 19% of tumors and was associated with shorter 5-year overall survival (OAS) rate compared with c-erbB-2-negative cases in the total patient material (58% and 77%, respectively; P = .

The p53 gene in breast cancer: prognostic value of complementary DNA sequencing versus immunohistochemistry.

Background: Mutations in the p53 tumor suppressor gene (also known as TP53) have been detected in a wide variety of human cancers. In breast cancer, the presence of p53 gene alterations has been associated with worse prognosis.

Purpose: We compared a complementary DNA (cDNA)-based sequencing method and an immunohistochemical (IHC) method for their abilities to detect p53 mutations in breast cancer specimens.

Overview on human breast cancer with focus on prognostic and predictive factors with special attention on the tumour suppressor gene p53.

A long list of potential prognostic markers has been analysed for breast cancer, some of them will be reviewed in this article. The lymph node status is still the best prognostic marker. The lymph node status combined with information on tumour size, receptor- and proliferation status of the tumour should be analysed as standard for all breast cancer patients.

p53 Status predicts survival in breast cancer patients treated with or without postoperative radiotherapy: a novel hypothesis based on clinical findings.

Purpose And Methods: Primary breast cancer tumors without axillary metastases from 206 consecutive patients in a population-based cohort were investigated with regard to the presence of an intact p53 gene using a cDNA-based sequencing method. Clinical follow-up data and outcome of node-negative patients without any adjuvant systemic therapy (n = 168) were related to locoregional radiotherapy and p53 status.

Results: Mutations in p53 occurred in 31 node-negative breast cancer patients who did not receive any systemic adjuvant treatment, but were treated with postoperative locoregional radiotherapy or nothing.

Complete sequencing of the p53 gene provides prognostic information in breast cancer patients, particularly in relation to adjuvant systemic therapy and radiotherapy.

The complete coding region of the p53 gene was sequenced from 316 consecutively presented breast cancers, of which 97 were lymph node positive and 206 were node negative. The p53 status was related to prognosis and effect of adjuvant therapy. In all, 69 individual mutations, 29 in node-positive tumours, were demonstrated throughout the whole coding sequence.

Changes induced in growing rat bone by immobilization and remobilization.

We studied changes in bone mass and histology in growing rats after different relatively short periods of immobilization and during subsequent remobilization. Immobilization-induced loss of bone weight is mainly due to mineral losses as indicated by changes in wet weight, ash weight, and calcium content. 45Ca2+ incorporation was found to be decreased in immobilized bones and showed strong dependence upon the age of the rats.

Regeneration of cranial suture and bone plate lesions in rabbits. Implications for positioning of osteotomies.

The positioning of osteotomies in intramembranous cranial bone was studied by exploring the pattern of bone regeneration in growth areas (the sutural region) as compared to that of the bone plate proper. Trephine defects in the left coronal suture area and the right parietal bone were produced in fifty-nine young rabbits. A pilot study to refine operative and analytical methods comprised 22 animals.

Calcitonin gene-related peptide in rat salivary glands: neuronal localization, depletion upon nerve stimulation, and effects on salivation in relation to substance P.

Calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibres occurred predominantly around blood vessels and large ducts and, to a minor extent, around acini and small ducts in the parotid, sublingual and submaxillary glands of the rat. Double immunostaining showed most of the CGRP-containing nerve fibres to contain substance P. However, the vast majority of substance P-immunoreactive periacinar nerve fibres in the parotid and submandibular glands lacked CGRP.

Characterization of alkaline phosphatase from buccal mucosa in young rats.

Alkaline phosphatase (APase) is a plasma membrane-integrated protein with unknown function and is found in many different tissues of the body. It is normally not present in squamous epithelium. Nevertheless, it has been demonstrated in developing rat oral epithelium, where it exists together with several different phosphatases.

Effects of bone wax on rabbit cranial bone lesions.

The present study was designed to elucidate the reactions of cranial membranous bone to bone wax. In ten young rabbits, twenty parietal bone defects were created by drilling, the edges of which were partly extended using rongeur forceps to enable investigation of eventual thermal effects. Half of the marginal bone surrounding the lesions was covered by bone wax, the remainder serving as control.

Zinc and alkaline phosphatase in developing rat oral mucosa.

Alkaline phosphatases (AlkPase) of many different tissues and species have been shown to be zinc metalloenzymes. Specific regions of rat oral mucosa have a high activity of AlkPase. Combined autoradiography and enzyme histochemistry showed that they also retained injected radioactive zinc (65Zn).

Lactate dehydrogenase isozymes in developing rat oral mucosa. A comparative study of LDH biochemistry and histochemistry.

Histochemical lactic acid dehydrogenase (LDH) staining methods seem unable to demonstrate the total LDH activity in tissue sections. An analysis was made of LDH tissue staining methods applied on LDH zymograms. The menadione-mediated LDH staining of tissue sections can not possibly reflect true LDH activity.

Acid, neutral, and alkaline phosphatases in developing rat oral mucosa.

Developing rat oral mucosa contains high activities of acid (AcidPase), neutral (adenosine monophosphatase AMPase), and alkaline (AlkPase) phosphatases. This study is concerned with a detailed analysis of the distribution of these enzymes in freeze-dried sections of oral mucosa. The sections were incubated for AcidPase.