Identification and cloning of a novel amplified DNA sequence in human malignant fibrous histiocytoma derived from a region of chromosome 12 frequently rearranged in soft tissue tumors.

Amplification of cellular oncogenes occurs frequently in several human cancers and is an important mechanism of increased gene expression. Identification of amplified genes in tumor cells has proved to be a useful approach for understanding genetic alterations in cancer. Previous procedures for isolating probes from amplified DNA sequences have relied on tissue culture cells, limiting the range of tumors that can be studied and raising questions of in vitro artifact. We have circumvented these problems by combining in gel renaturation of amplified sequences with the polymerase chain reaction. Using this approach, we have identified and partially cloned a DNA amplification unit from biopsies of human malignant fibrous histiocytoma. This amplification unit is derived from chromosome 12q13-14, a site commonly involved in rearrangements in soft tissue tumors, and contains at least one transcribed region (designated SAS, for sarcoma amplified sequence).     

The dopamine D2 receptor: two molecular forms generated by alternative splicing.

Cloned human dopamine D2 receptor cDNA was isolated from a pituitary cDNA library and found to encode an additional 29 amino acid residues in the predicted intracellular domain between transmembrane regions 5 and 6 relative to a previously described rat brain D2 receptor. Results from polymerase chain reactions as well as in situ hybridization revealed that mRNA encoding both receptor forms is present in pituitary and brain of both rat and man. The larger form was predominant in these tissues and, as shown in the rat, expressed by dopaminergic and dopaminoceptive neurons. Analysis of the human gene showed that the additional peptide sequence is encoded by a separate exon. Hence, the two receptor forms are generated by differential splicing possibly to permit coupling to different G proteins. Both receptors expressed in cultured mammalian cells bind [3H]spiperone with high affinity and inhibit adenylyl cyclase, as expected of the D2 receptor subtype.     

Fine needle aspiration cytology in lupus lymphadenopathy. A case report.

This case report describes the aspiration cytologic characteristics of histologically proven acute lupus lymphadenitis. The aspirate contained numerous lymphoid cells and many amorphous, basophilic, hematoxylin-stained bodies dispersed in a granular, necrotic background that lacked polymorphonuclear leukocytes. The lymphadenopathy was diagnosed cytologically as necrotizing lymphadenitis and histologically as acute lupus lymphadenitis.     

Conserved sequences in enzymes of the UDP-GlcNAc/MurNAc family are essential in hamster UDP-GlcNAc:dolichol-P GlcNAc-1-P transferase

The UDP-GlcNAc/MurNAc family of eukaryotic and prokaryotic enzymes use UDP-GlcNAc or UDP-MurNAc-pentapeptide as donors, dolichol-P or polyprenol-P as acceptors, and generate sugar-P-P-polyisoprenols. A series of six conserved sequences, designated A through F and ranging from 5 to 13 amino acid residues, has been identified in this family. To determine whether these conserved sequences are required for enzyme function, various mutations were examined in hamster UDP- GlcNAc:dolichol-P GlcNAc-1-P transferase (GPT). Scramble mutations of sequences B-F, generated by scrambling the residues within each sequence, demonstrated that each is important in GPT. While E and F scrambles appeared to prevent stable expression of GPT, scrambling of B- D resulted in GPT mutants that could be stably expressed and bound tunicamycin, but lacked enzymatic activity. Further, the C and D scramble mutants had an unexpected sorting defect. Replacement of sequences B-F with prokaryotic counterparts from either the B.subtilis mraY or E.coli rfe genes also affected GPT by preventing expression of the mutant protein (B, F) or inhibiting its enzymatic activity (C-E). For the C-E replacements, no acquisition of acceptor activity for polyprenol-P, the fully unsaturated natural bacterial acceptor, was detected. These studies show that the conserved sequences of the UDP- GlcNAc/MurNAc family are important, and that the eukaryotic and prokaryotic counterparts are not freely interchangeable. Since several mutants were efficiently expressed and bound tunicamycin, yet lacked enzymatic activity, the data are consistent with these sequences having a direct role in product formation.      

Trends in incidence of AIDS associated with transfusion of blood and blood products in Europe and the United States, 1985-93.

OBJECTIVE--To quantify patterns and trends in incidence of AIDS associated with transfusion of blood and its products in 14 European countries and the United States. DESIGN--Data were derived from the World Health Organisation''s European non-aggregate AIDS dataset and, for the United States, from the Centers for Disease Control AIDS public information dataset. Rates were standardised by using the world standard populations and adjusted for reporting delays in each country. SUBJECTS--Cases of AIDS in patients with haemophilia and recipients of transfusions. RESULTS--Overall, between 1985 and 1993 almost 6000 cases of AIDS associated with transfusions were registered in the 14 European countries considered and over 8000 in the United States between 1985 and 1992. Most European countries had annual age adjusted rates lower than 0.5 per million children aged 12 or less and between 1 and 2 per million adults. The United States had rates around 1 per million children and 5 per million adults in the most recent period. For children, the highest rates were generally reached in 1985-7, whereas in adults the highest rates were in the late 1980s. France had the highest overall incidence of AIDS related to transfusion in Europe (3.3 per million). Romania had a major epidemic in children (over 30 cases per million children in 1988-90). Incidence rates of AIDS associated with transfusion were still increasing in some southern European countries in the early 1990s. CONCLUSIONS--Apart from in France and Romania it is clear that rates of bloodborne AIDS in European countries are lower than those registered in the United States.     

Optimization of Large-Scale Vogel Spiral Arrays of Plasmonic Nanoparticles

Plasmonics - In this paper, we combine coupled dipole approximation (CDA) theory with optimization codes based on cyclic coordinate descent minimization to obtain the best configurations of...     

BKV QPCR detection and infection monitoring in renal transplant recipients

BKV associated nephropathy (BKVAN) is a cause of renal dysfunction and loss of the graft in transplants. Viral primary infection is usually inapparent and then BKV establishes latency in kidneys. Reactivation occurs in immunocompromised conditions in renal transplant recipients who can develop a subclinical nephritis and eventually a BKV-associated interstitial nephritis or a BKVAN. In this study, we searched for BKV copies in urine and plasma of renal transplants by quantitative assay (QPCR). Results showed that in several patients clearance of viremia is associated with persistent viruria, suggesting that both specimens are necessary to correctly monitor a BKVAN.     

Synthesis of pyrrolo[2,3-d]pyridazinones as potent, subtype selective PDE4 inhibitors

A series of pyrrolo [2,3-d]pyridazinones was synthesized and tested for their inhibitory activity on PDE4 subtypes A, B and D and selectivity toward Rolipram high affinity binding site (HARBS). New agents with interesting profile were reported; in particular compound 9e showed a good PDE4 subtype selectivity, being 8 times more potent (IC50 = 0.32 microM) for PDE4B (anti-inflammatory) than for PDE4D (IC50 = 2.5 microM), generally considered the subtype responsible for emesis. Moreover the ratio HARBS/PDE4B was particularly favourable for 9e (147), suggesting that the best arranged groups around the pyrrolopyridazinone core are an isopropyl at position-1, an ethoxycarbonyl at position-2, together with an ethyl group at position-6. For compounds 8 and 15a the ability to inhibit TNFalpha production in PBMC was evaluated and the results are consistent with their PDE4 inhibitory activity.     

Structure and composition of the assemblage of parasitoids associated to Phyllocnistis citrella Pupae Stainton (Lepidoptera: Gracillariidae) in citrus orchards in Southern Brazil

The structure and composition of the assemblage of pupal parasitoids of Phyllocnistis citrella Stainton, the citrus leafminer, were studied in two citrus orchards (Citrus deliciosa Tenore cv. Montenegrina and Citrus sinensis (L.) Osbeck x Citrus reticulata Blanco hybrid Murcott), in Montenegro County (29 degrees 68'S and 51 degrees 46'W), southern Brazil. At fortnightly samplings, from July 2001 to June 2003, all the new shoots from 24 randomly selected trees were inspected. The species richness reached five native species in the Murcott orchard, and six in Montenegrina. In Murcott, the presence of Ageniaspis citricola (Hymenoptera: Encyrtidae), an exotic species, was detected in the first year of sampling, probably migrating from the nearby areas where it had been released for the miner control. In Montenegrina, its presence was only registered in the second year. A. citricola in both areas was dominant and changed the community structure of parasitoid complex of P. citrella in both orchards.