Suresh G. Shelat

NuRD mediates activating and repressive functions of GATA-1 and FOG-1 during blood development

GATA transcription factors interact with FOG proteins to regulate tissue development by activating and repressing transcription. FOG‐1 (ZFPM1), a co‐factor for the haematopoietic factor GATA‐1, binds to the NuRD co‐repressor complex through a conserved N‐terminal motif. Surprisingly, we detected NuRD components at both repressed and active GATA‐1/FOG‐1 target genes in vivo. In addition, while NuRD is required for transcriptional repression in certain contexts, we show a direct requirement of NuRD also for FOG‐1‐dependent transcriptional activation. Mice in which the FOG‐1/NuRD interaction is disrupted display defects similar to germline mutations in the Gata1 and Fog1 genes, including anaemia and macrothrombocytopaenia. Gene expression analysis in primary mutant erythroid cells and megakaryocytes (MKs) revealed an essential function for NuRD during both the repression and activation of select GATA‐1/FOG‐1 target genes. These results show that NuRD is a critical co‐factor for FOG‐1 and underscore the versatile use of NuRD by lineage‐specific transcription factors to activate and repress gene transcription in the appropriate cellular and genetic context.

Erythroid dysplasia, megaloblastic anemia, and impaired lymphopoiesis arising from mitochondrial dysfunction.

Recent reports describe hematopoietic abnormalities in mice with targeted instability of the mitochondrial genome. However, these abnormalities have not been fully described. We demonstrate that mutant animals develop an age-dependent, macrocytic anemia with abnormal erythroid maturation and megaloblastic changes, as well as profound defects in lymphopoiesis. Mice die of severe fatal anemia at 15 months of age. Bone-marrow transplantation studies demonstrate that these abnormalities are intrinsic to the hematopoietic compartment and dependent upon the age of donor hematopoietic stem cells. These abnormalities are phenotypically similar to those found in patients with refractory anemia, suggesting that, in some cases, the myelodysplastic syndromes are caused by abnormalities of mitochondrial function.

Mineralocorticoids and Glucocorticoids Cooperatively Increase Salt Intake and Angiotensin II Receptor Binding in Rat Brain

Mineralocorticoids, such as deoxycorticosterone acetate (DOCA), and angiotensin II (AngII) act synergistically in the brain to elicit salt appetite. Glucocorticoids, such as dexamethasone (DEX), also may enhance the behavioral effects of DOCA and AngII. However, the brain regions involved in these behavioral interactions have not been elucidated. This study tested the hypothesis that DEX potentiates the effects of DOCA on AngII binding, especially at the AT1 receptor. We confirmed that DEX potentiated the effects of DOCA on salt appetite. Concomitantly, steroid-specific and region-specific changes in AT1 binding were noted. Specifically, in the hypothalamic paraventricular nucleus, treatment with DEX or DOCA + DEX increased AT1 binding. In the subfornical organ (SFO) and area postrema, there was an increase in AT1 binding when both steroids were combined, but not when given individually. However, there was no change in AT2 binding in any brain region studied and no change in AT1 or AT2 binding to either receptor subtype in the pituitary. The results indicate that DOCA and DEX may increase the sensitivity of the brain to the behavioral and physiological actions of AngII by upregulating AT1 receptors in the SFO and area postrema.

Hematopoietic stem cell function requires 12/15-lipoxygenase–dependent fatty acid metabolism

Fatty acid metabolism governs multiple intracellular signaling pathways in many cell types, but its role in hematopoietic stem cells (HSCs) is largely unknown. Herein, we establish a critical role for 12/15-lipoxygenase (12/15-LOX)-mediated unsaturated fatty acid metabolism in HSC function. HSCs from 12/15-LOX-deficient mice are severely compromised in their capacity to reconstitute the hematopoietic compartment in competitive and serial reconstitution assays. Furthermore, we demonstrate that 12/15-LOX is required for the maintenance of long-term HSC quiescence and number. The defect in HSCs is cell-autonomous and associated with a selective reduction in 12/15-LOX-mediated generation of bioactive lipid mediators and reactive oxygen species and with a decrease in canonical Wnt signaling as measured by nuclear beta-catenin staining. These results have implications for development, aging, and transformation of the hematopoietic compartment.

An Iron Responsive Element-like Stem-Loop Regulates α-Hemoglobin-stabilizing Protein mRNA

Hemoglobin production during erythropoiesis is mechanistically coupled to the acquisition and metabolism of iron. We discovered that iron regulates the expression of α-hemoglobin-stabilizing protein (AHSP), a molecular chaperone that binds and stabilizes free α-globin during hemoglobin synthesis. In primates, the 3′-untranslated region (UTR) of AHSP mRNA contains a nucleotide sequence resembling iron responsive elements (IREs), stem-loop structures that regulate gene expression post-transcriptionally by binding iron regulatory proteins (IRPs). The AHSP IRE-like stem-loop deviates from classical consensus sequences and binds IRPs poorly in electrophoretic mobility shift assays. However, in cytoplasmic extracts, AHSP mRNA co-immunoprecipitates with IRPs in a fashion that is dependent on the stem-loop structure and inhibited by iron. Moreover, this interaction enhances AHSP mRNA stability in erythroid and heterologous cells. Our findings demonstrate that IRPs can regulate mRNA expression through non-canonical IREs and extend the repertoire of known iron-regulated genes. In addition, we illustrate a new mechanism through which hemoglobin may be modulated according to iron status.

Fetal alcohol exposure alters serotonin transporter sites in rat brain.

This study examined the effects of fetal alcohol exposure (FAE) on serotonin transporter (5-HTT) binding sites in the brains of developing male and female rat offspring using the technique of quantitative autoradiography. Time-pregnant dams were fed liquid ethanol diet, isocaloric diet without ethanol or normal rat chow. Male and female offspring were sacrificed at 21, 40 and 60 days of age, brains removed and sectioned for analysis of 5-HTT sites. FAE led to distinct effects on 5-HTT sites depending on the age and gender of the offspring. FAE increased 5-HTT binding sites in cortical layers 5, 6, hippocampal layers CA(2,3), lateral nucleus of the amygdala and in the dorsal raphe nucleus. FAE decreased 5-HTT binding sites in the medial nucleus of amygdala, dorsomedial and ventromedial nuclei of the hypothalamus. FAE decreased 5-HTT binding sites temporarily in the ventromedial nucleus of the hypothalamus in the 21-day-old female; this effect was found to disappear by day 40. In contrast, FAE increased 5-HTT sites in the lateral nucleus of the amygdala in the adult animal, suggesting that ethanol exposure in utero may alter serotonin neurotransmission in discrete brain regions permanently.

Adrenal steroid regulation of central angiotensin II receptor subtypes and oxytocin receptors in rat brain

The neuropeptides angiotensin II (AngII) and oxytocin (OT) play important but opposing roles in the regulation of sodium appetite in the rat, AngII as a stimulatory peptide and OT as an inhibitory peptide. Adrenal steroids increase the density of AngII receptors in brain following in vivo administration, although the neuroanatomical and subtype specificity have not been thoroughly examined. Furthermore, previous studies demonstrate that adrenalectomy (ADX) leads to a reduction in OT receptors, although regions associated with sodium appetite remain to be examined. In the present study, quantitative receptor autoradiography was used to locate regions where perturbations in circulating adrenal steroids affect the density of oxytocin receptors and the angiotensin receptor subtypes AT1 and AT2. The results show that ADX results in a small, but significant decrease in AT1 expression in the paraventricular nucleus of the hypothalamus, subfornical organ, and the area postrema. That this effect is reversed by either aldosterone or low-dose corticosterone replacement suggests that occupancy of the mineralocorticoid receptor is responsible for the steroid effect. No changes were observed in AT2 or OT receptors in nuclei associated with sodium appetite, indicating that perturbations in adrenal steroids did not affect these receptors in brain regions implicated in the control of salt appetite.

Practical Considerations for Planning a Therapeutic Apheresis Procedure

The general medicine and critical care services often care for patients that require therapeutic apheresis. Apheresis procedures are performed for various hematologic, neurological, renal, autoimmune, metabolic, and other indications. To facilitate a prompt start to the procedure, the clinical team must coordinate efforts with several services, including those that perform the apheresis procedure, establish venous access, and provide blood or replacement products, in addition to the pharmacy and clinical laboratory. Some of these tasks are performed typically by the clinical teams, while others are performed typically by the apheresis team. Presented and discussed are the indications for therapeutic apheresis, calculations for the ordering of blood products, and several important and practical details to consider, thus preventing delays in starting the apheresis procedure.

Considerations on the use of adjunct red blood cell exchange transfusion in the treatment of severe Plasmodium falciparum malaria

BACKGROUND: Travelers returning to the United States from malaria‐endemic areas are at increased risk of a potentially fatal infection from Plasmodium falciparum, which requires prompt and aggressive treatment.

Aggressive Epstein-Barr virus-associated, CD8+, CD30+, CD56+, surface CD3-, natural killer (NK)-like cytotoxic T-cell lymphoma.

We report an unusual case of aggressive natural killer (NK)-like cytotoxic T-cell lymphoma in a previously healthy immunocompetent West African male. He presented with a fever of unknown origin, subsequently developed erythematous skin nodules, generalized lymphadenopathy, and hepatosplenomegaly, and then died of multiple organ failure. A skin nodule and lymph node biopsy showed an infiltrate of pleomorphic atypical medium and large lymphoid cells with extensive necrosis and prominent apoptosis. Peripheral blood and ascites also harbored these cells, with cytology revealing irregular nuclear folding and basophilic cytoplasm, and some with azurophilic cytoplasmic granules. Flow cytometry and immunohistochemistry demonstrated the expression of CD2, CD7, CD8, CD30, CD56, and cytoplasmic but not surface CD3. In situ hybridization demonstrated Epstein-Barr virus transcripts. A monoclonal T-cell receptor &ggr; chain gene rearrangement was detected by polymerase chain reaction. This is the first reported case of an NK-like T-cell lymphoma with these unusual features, making precise classification difficult. Some features suggest an NK1.1 or NKT lymphocyte origin. Because the earliest clinical manifestation was splenomegaly and abnormal liver function, the normal cellular counterpart may be a distinct subset of NK1.1 cells normally present in hepatosplenic sinusoids. This tumor disseminated early and pursued a fulminant clinical course, thus emphasizing the importance of early recognition and diagnosis.