Multiple cis elements within the Igf2/H19 insulator domain organize a distance-dependent silencer. A cautionary note.

The 5'-flank of the H19 gene harbors a differentially methylated imprinting control region that represses the maternally derived Igf2 and paternally derived H19 alleles. Here we show that the H19 imprinting control region (ICR) is a potent silencer when positioned in a promoter-proximal position. The silencing effect is not alleviated by trichostatin A treatment, suggesting that it does not involve histone deacetylase functions. When the H19 ICR is separated from the promoter by more than 1.2 +/- 0.3 kb, however, trichostatin A stimulates promoter activity 10-fold. Deletion analyses revealed that the silencing feature extended throughout the ICR segment. Finally, chromatin immunopurification analyses revealed that the H19 ICR prevented trichostatin A-dependent reacetylation of histones in the promoter region in a proximal but not in a distal position. We argue that these features are likely to be side effects of the H19 ICR, rather than explaining the mechanism of silencing of the paternal H19 allele. We issue a cautionary note, therefore, that the interpretation of insulator/silencer data could be erroneous should the distance issue not be taken into consideration.     

Systemic mastocytosis associated with t(8;21)(q22;q22) acute myeloid leukemia

Although KIT mutations are present in 20–25% of cases of t(8;21)(q22;q22) acute myeloid leukemia (AML), concurrent development of systemic mastocytosis (SM) is exceedingly rare. We examined the clinicopathologic features of SM associated with t(8;21)(q22;q22) AML in ten patients (six from our institutions and four from published literature) with t(8;21) AML and SM. In the majority of these cases, a definitive diagnosis of SM was made after chemotherapy, when the mast cell infiltrates were prominent. Deletion 9q was an additional cytogenetic abnormality in four cases. Four of the ten patients failed to achieve remission after standard chemotherapy and seven of the ten patients have died of AML. In the two patients who achieved durable remission after allogeneic hematopoietic stem cell transplant, recipient-derived neoplastic bone marrow mast cells persisted despite leukemic remission. SM associated with t(8;21) AML carries a dismal prognosis; therefore, detection of concurrent SM at diagnosis of t(8;21) AML has important prognostic implications.     

Development and validation of molecular markers linked to an Aegilops umbellulata-derived leaf-rust-resistance gene, Lr9, for marker-assisted selection in bread wheat.

An Aegilops umbellulata-derived leaf-rust-resistance gene, Lr9, was tagged with 3 random amplified polymorphic DNA (RAPD) markers, which mapped within 1.8 cM of gene Lr9 located on chromosome 6BL of wheat. The markers were identified in an F2 population segregating for leaf-rust resistance, which was generated from a cross between 2 near-isogenic lines that differed in the alien gene Lr9 in a widely adopted agronomic background of cultivar 'HD 2329'. Disease phenotyping was done in controlled environmental conditions by inoculating the population with the most virulent pathotype, 121 R63-1 of Puccinia triticina. One RAPD marker, S5550, located at a distance of 0.8+/-0.008 cM from the Lr9 locus, was converted to sequence-characterized amplified region (SCAR) marker SCS5550. The SCAR marker was validated for its specificity to gene Lr9 against 44 of the 50 known Lr genes and 10 wheat cultivars possessing the gene Lr9. Marker SCS5550 was used with another SCAR marker, SCS73719, previously identified as being linked to gene Lr24 on a segregating F2 population to select for genes Lr9 and Lr24, respectively, demonstrating the utility of the 2 markers in marker-assisted gene pyramiding for leaf-rust resistance in wheat.     

Leaching and transformation of urea in dry and wet soils as affected by irrigation water

Summary The leaching losses of urea in dry and wet soils as affected by the irrigation water, were studied in 90 cm long and 5 cm diameter plexiglass columns. In dry sandy loam soil urea leached with irrigation water and peaks of urea were observed with water front but in sandy soil the wetting front moved faster leaving the urea peak behind. In initially wet soils the urea peak did not coincide the wetting front, following infiltration and after redistribution for 24 hours of infiltration. More urea was recovered in sandy soil than in sandy loam soil following infiltration and redistribution for 24 hours. Urea leached only upto 30 cm with 5 and 7.5 cm irrigation water whereas with 10 and 15 cm irrigation water profiles urea penetrated to deeper layers. The sub-surface application of urea did not change the leaching behaviour of urea. Urea behaves as a non-reacting ion like Cl with respect to its movement with soil-water before it is hydrolysed.     

The effect of Streptomyces species on the survival of Salmonella in soil

Abstract A modified Chelex 100 ion-exchange extraction method was used to monitor streptomycete spores, streptomycete mycelia and Salmonella in soil. Salmonella dusseldorf in soil was inhibited by the bactericidal effect of streptomycin and by the growth of Streptomyces bikiniensis . The soil used in the experiments exerted an antimicrobial effect on S. dusseldorf .
Competition between S. dusseldorf, Streptomyces lividans TK24 and Stm. bikiniensis ATCC 11062 was monitored in soil. In sterile amended soil Stm. lividans increased the survival of S. dusseldorf , whereas survival was reduced in the presence of the known streptomycin producer, Stm. bikiniensis . In the presence of S. dusseldorf the production of spores and mycelia by Stm. bikiniensis was reduced, and Stm. lividans sporulation was reduced but mycelia production increased. Evidence was seen for a beneficial effect between S. dusseldorf and Stm. lividans mycelia.
In non-sterile unamended soil the survival of S. dusseldorf was increased in the presence of Stm. lividans , whereas Stm. bikiniensis had no effect. Stm. lividans and Stm. bikiniensis reduced the survival of S. dusseldorf in non-sterile amended soil, with the most dramatic reduction caused by Stm. bikiniensis . No such changes in the survival of S. dusseldorf were observed with non-sterile amended soil that had been treated with sludge. The presence of sludge in unamended soil increased the rate of Salmonella die-off. In unamended soil containing sludge the presence of Stm. lividans increased the survival of S. dusseldorf , whereas survival was reduced in the presence of Stm. bikiniensis . The data provided evidence of antibiosis in soil, relating to the possible production of streptomycin by Stm. bikiniensis .     

Intravenous rehydration for diarrheal dehydration of eutrophic children: survey of protocols provided at Colombian medical schools

Introduction. In all cases of severe dehydration from diarrhea, WHO recommends rapid rehydration. If oral rehydration in children is contraindicated, intravenous rehydration is recommended for immediate administration. However, methods of intravenous rehydration appear to be inadequately addressed in the medical schools of Colombia. Objective. Current approaches to oral rehydration were summarized, and instructors were informed concerning current WHO recommendations. Materials and methods. A survey was designed for pediatric instructors in Colombian medical schools. Direct questions about rehydration methods were included as well as presentation of theoretical clinical situations with dehydrated children. The survey also asked for the conditions necessary for intravenous rehydration and method of administration (volume, solution, concentration and speed of infusion). Results. Forty-one surveys were included (82% of medical schools in Colombia). Inadequate contraindications for oral rehydration therapy were made in 41%. Rapid and slow intravenous rehydration was recommended in 71% and 29%, respectively; 57% recommended fluid bolus to rehydrate. Adequate volumes were recommended by less than half of the respondents and adequate sodium concentration was recommended by 85%. In 56% of medical schools, glucose was not included in solutions and 66% use Ringer lactate. Normal saline solution, dextrose solution with electrolytes and polyelectrolytes solutions are also used. Conclusions. Misconceptions are common concerning the contraindications to oral rehydration therapy. One-third of medical schools promote a slow therapy despite the superiority of the rapid therapy. Uniformity for rapid therapy schemes is lacking. Bolus rehydration is commonly advocated despite the fact that this method is unsupported by the literature. Concepts about rehydration must be updated in medical schools and a national guide for intravenous rehydration is recommended.     

Complex I syndrome in myocardial stunning and the effect of adenosine

Isolated rabbit hearts were exposed to ischemia (I; 15 min) and reperfusion (R; 5-30 min) in a model of stunned myocardium. I/R decreased left-ventricle O₂ consumption (46%) and malate-glutamate-supported mitochondrial state 3 respiration (32%). Activity of complex I was 28% lower after I/R. The pattern observed for the decline in complex I activity was also observed for the reduction in mitochondrial nitric oxide synthase (mtNOS) biochemical (28%) and functional (50%) activities, in accordance with the reported physical and functional interactions between complex I and mtNOS. Malate-glutamate-supported state 4 H₂O₂ production was increased by 78% after I/R. Rabbit heart Mn-SOD concentration in the mitochondrial matrix (7.4 ± 0.7 μM) was not modified by I/R. Mitochondrial phospholipid oxidation products were increased by 42%, whereas protein oxidation was only slightly increased. I/R produced a marked (70%) enhancement in tyrosine nitration of the mitochondrial proteins. Adenosine attenuated postischemic ventricular dysfunction and protected the heart from the declines in O₂ consumption and in complex I and mtNOS activities and from the enhancement of mitochondrial phospholipid oxidation. Rabbit myocardial stunning is associated with a condition of dysfunctional mitochondria named “complex I syndrome.” The beneficial effect of adenosine could be attributed to a better regulation of intracellular cardiomyocyte Ca²⁺ concentration.     

Design and synthesis of 4-arylpiperidinyl amide and N-arylpiperdin-3-yl-cyclopropane carboxamide derivatives as novel melatonin receptor ligands

Two series of 4-arylpiperidinyl amide and N-arylpiperdin-3-yl-cyclopropane carboxamide derivatives exhibiting diverse functionality at rat MT₁ and MT₂ receptors are reported. Compounds 11f and 18b (MT₁/MT₂ agonist) have human microsomal intrinsic clearance comparable to ramelteon.