Histone deacetylase inhibitors induce remission in transgenic models of therapy-resistant acute promyelocytic leukemia.

Acute promyelocytic leukemia (APL) is associated with chromosomal translocations, invariably involving the retinoic acid receptor alpha (RAR alpha) gene fused to one of several distinct loci, including the PML or PLZF genes, involved in t(15;17) or t(11;17), respectively. Patients with t(15;17) APL respond well to retinoic acid (RA) and other treatments, whereas those with t(11;17) APL do not. The PML-RAR alpha and PLZF-RAR alpha fusion oncoproteins function as aberrant transcriptional repressors, in part by recruiting nuclear receptor-transcriptional corepressors and histone deacetylases (HDACs). Transgenic mice harboring the RAR alpha fusion genes develop forms of leukemia that faithfully recapitulate both the clinical features and the response to RA observed in humans with the corresponding translocations. Here, we investigated the effects of HDAC inhibitors (HDACIs) in vitro and in these animal models. In cells from PLZF-RAR alpha/RAR alpha-PLZF transgenic mice and cells harboring t(15;17), HDACIs induced apoptosis and dramatic growth inhibition, effects that could be potentiated by RA. HDACIs also increased RA-induced differentiation. HDACIs, but not RA, induced accumulation of acetylated histones. Using microarray analysis, we identified genes induced by RA, HDACIs, or both together. In combination with RA, all HDACIs tested overcame the transcriptional repression exerted by the RAR alpha fusion oncoproteins. In vivo, HDACIs induced accumulation of acetylated histones in target organs. Strikingly, this combination of agents induced leukemia remission and prolonged survival, without apparent toxic side effects.

Inhibition of transformed cell growth and induction of cellular differentiation by pyroxamide, an inhibitor of histone deacetylase.

PURPOSE: We have synthesized a series of hybrid polar compounds that induce differentiation and/or apoptosis of various transformed cells. These agents are also potent inhibitors of histone deacetylases (HDACs). Pyroxamide (suberoyl-3-aminopyridineamide hydroxamic acid) is a new member of this class of compounds that is currently under development as an anticancer agent. We investigated the activity of pyroxamide as an inducer of differentiation and/or apoptosis in transformed cells. EXPERIMENTAL DESIGN AND RESULTS: Pyroxamide, at micromolar concentrations, induced terminal differentiation in murine erythroleukemia (MEL) cells and caused growth inhibition by cell cycle arrest and/or apoptosis in MEL, prostate carcinoma, bladder carcinoma, and neuroblastoma cells. Administration of pyroxamide (100 or 200 mg/kg/day) to nude mice at doses that caused little evident toxicity significantly suppressed the growth of s.c. CWR22 prostate cancer xenografts. Despite the potent growth-inhibitory effects of pyroxamide in this tumor model, serum prostate-specific antigen levels in control versus pyroxamide-treated mice were not significantly different. Pyroxamide is a potent inhibitor of affinity-purified HDAC1 (ID(50) = 100 nM) and causes the accumulation of acetylated core histones in MEL cells cultured with the agent. Human CWR22 prostate tumor xenografts from mice treated with pyroxamide (100 or 200 mg/kg/day) showed increased levels of histone acetylation and increased expression of the cell cycle regulator p21/WAF1, compared with tumors from vehicle-treated control animals. CONCLUSIONS: The findings suggest that pyroxamide may be a useful agent for the treatment of malignancy and that induction of p21/WAF1 in transformed cells by pyroxamide may contribute to the antitumor effects of this agent.

Leukotriene D4 excitation of rabbit distal colon arises in the region of the muscularis mucosae.

We previously have demonstrated in vivo that intra-arterial administration of leukotriene D4 (LTD4) causes increased myoelectric and mechanical activity in the rabbit distal colon. The aim of this study was to use both in vivo and in vitro techniques to try to elucidate the mechanism underlying this effect. In vivo the excitatory response of the rabbit distal colon to LTD4 was abolished by pretreatment with atropine (0.1 mg/kg iv) or hexamethonium (5 mg/kg iv) or the LTD4 receptor antagonist SK&F 102922 (0.8 micrograms/kg ia). In vitro neither the longitudinal nor the circular muscle layer responded to LTD4 (10(-10) to 10(-7) M) with a contractile response. Over the same concentration range, LTD4 caused contractions of the muscularis mucosae that were attenuated by either SK&F 102922 (10(-9) to 10(-7) M) or indomethacin (10(-6) M) but were unaffected by atropine (10(-6) M), pyrilamine (10(-6) M), or tetrodotoxin (10(-6) M). Full thickness segments of longitudinal muscle, circular muscle, and muscularis mucosae did not contract to LTD4. These data imply that LTD4-induced excitation of the rabbit distal colon in vivo arises as a result of the excitation of LTD4 receptors in the region of the muscularis mucosae and that this leads ultimately to the release of acetylcholine onto the muscularis propria. It is proposed that one possible mechanism leading to the latter effect is an increased excitability of intrinsic nerves resulting from a prostaglandin-induced depression of norepinephrine release from nerves impinging on the submucosal plexus.

Aztreonam therapy for serious gram-negative infections in children.

Fifty-nine children were enrolled in an open trial of aztreonam, a monocyclic beta-lactam, therapy for serious gram-negative infections. Thirty-six infections were microbiologically evaluable and received five or more days of therapy. Patients' ages ranged from 3 days to 12 years, and diagnoses included pyelonephritis or cystitis (20), deep soft tissue or joint infection (seven), septicemia (four), pneumonia (three), peritonitis, and epiglottitis. Causative bacteria included Escherichia coli and other Enterobacteriaceae, Pseudomonas aeruginosa, and Haemophilus influenzae. The standard regimen was 30 mg/kg every six or eight hours intravenously. All isolates were aztreonam-susceptible and were eradicated during therapy. Two patients had microbiologic relapses: a patient with Salmonella choleraesuis meningitis who was initially treated for only ten days and a patient with E coli pyelonephritis. Clinical cure was achieved in 31 of 36 children. Pharmacokinetic studies performed in six children demonstrated no difference in serum concentrations or pharmacokinetic variables between day 1 and day 7 of therapy. Although several patients had transient eosinophilia (eight), elevated levels of aminotransferase (seven), or thrombocytosis (ten), no clinically significant adverse effects were noted. In this initial, uncontrolled study, aztreonam was effective and safe in the treatment of a variety of serious gram-negative infections in children.

Transcutaneous bilirubinometry II. Dermal bilirubin kinetics during phototherapy.

We examined the effect of phototherapy on cutaneous bilirubin using the transcutaneous bilirubinometer (TcB) in 14 white infants at a mean postnatal age of 30 h. Six infants were treated with blue light, eight with white light. An opaque patch, 2.5 cm in diameter, covered the skin and served as a source for TcB control values. Simultaneous TcB measurements were obtained from exposed and patched areas every 15 min during 4 h of phototherapy. After the start of phototherapy, TcB index obtained from patched skin did not change during the course of treatment. Data from exposed skin showed that the initial rate of TcB index fall during the first hour was significantly faster than the successive values during the next 3 h in both groups studied. During the first 2 h of therapy the TcB index decreased faster among infants treated with blue light. Correlation studies indicate that TcB measurements from exposed skin areas may not be clinically useful in predicting serum bilirubin response to phototherapy but data obtained from unexposed sites may fill this role.

Consumo, desempenho e parâmetros econômicos de novilhos Nelore e F1 Brangus x Nelore terminados em pastagens, suplementados com mistura mineral e sal nitrogenado com uréia ou amiréia / Feed intake, performance and profitability of Nellore and crossbred (Brangus x Nellore) steers finished in pastures, supplemented with mineralized salt and nitrogenous salt with urea or starea

Avaliou-se o efeito da suplementação protéica (40 por cento PB) com amiréia ou uréia sobre o consumo de suplemento, desempenho e características econômicas de novilhos terminados em pastagens. Foram utilizados 120 novilhos com 19 meses de idade e 358kg, sendo 60 Nelore e 60 F1 Brangus x Nelore, divididos em três tratamentos com 20 animais, alojados em piquetes de Brachiaria brizantha cv. Marandu de 10 hectares cada, totalizando 120 hectares, sendo dois piquetes por grupo genético e tratamento, pastejados alternadamente a cada pesagem (42 dias). Os tratamentos consistiram em mistura mineral com amiréia-150S (AM), mistura mineral com uréia+milho+enxofre (UR) e mistura mineral (MM). As médias de consumo de suplemento dos animais F1 foram de 206,1; 145,9 e 73,1g/dia, e as dos animais Nelore, 236,0; 205,1 e 94,3g/dia para os tratamentos AM, UR e MM, respectivamente. Para os novilhos Nelore, houve efeito (P<0,05) do suplemento sobre o peso de abate (PA), sendo a média do tratamento UR, 518,85kg, mais alta que a dos demais, 491,89 e 485,20kg, respectivamente, para AM e MM. Para os novilhos F1, foi significativo o efeito da suplementação protéica (P<0,05), com médias de 515,90 e 520,15kg, respectivamente, para os tratamentos UR e AM. A suplementação protéica proporcionou bom desempenho em animais F1 durante períodos de abundância de forragem. O uso de uréia apresentou melhor viabilidade econômica.

The effects of protein supplementation of finishing grazing steers by feeding nitrogenous salts (40 percent CP, urea or starea) or mineralized salt only on supplement intake, growing performance and profitability were evaluated. One hundred and twenty steers (60 Nellore and 60 Brangus x Nellore, 19-month old, 358kg BW) were divided in 12 equal groups which were allotted to one of 12 Brachiaria brizantha pastures (10-ha each) performing two pastures for each breeding group and nutritional treatment. Groups were allowed to graze each pasture for 42 days when they were randomly moved into a new one. Nutritional treatments were as follow: MS - mineralized salt only; ST -mineralized salt plus starea - 150S; and UR - mineralized salt plus urea, corn and sulphur. UR supplement was prepared mixing the same ingredient contents of ST. Crossbred steers consumed 206.1; 145.9 and 73.1g/day whereas Nellore steers consumed 236.0; 205.11 and 94.29g/day of ST, UR and MS; respectively. For Nellore steers, UR increased slaughter weight (518.8kg) compared to ST and MS (491.9 and 485.2kg, respectively). Protein supplementation also increased slaughter weight for crossbred steers (P<0.05) in comparison to animals fed mineralized salt as sole supplementation (515.9 and 520.2kg for the UR and ST, respectively). Protein supplementation enhances growing performance of crossbred steers during periods of forage abundance. Urea supplementation yields a higher profitability in grazing systems for cattle finishing.