Effects on crops of irrigation with treated municipal wastewaters.

The fertilizing potential of treated municipal wastewater (oxidation ditch) and crop sanitary acceptability for direct human consumption were evaluated in Mendoza, Argentina. Two experiments were performed on a pilot plot planted with garlic (1998) and onions (1999) using furrow irrigation with three types of water in 10 random blocks: treated effluent (2.5 x 10(3) MPN Escherichia coli/100 ml, 3 helminth eggs/l, and Salmonella (positive); and well water (free of microorganisms), with and without fertilizer. Two responses were evaluated: (1) crop yield, and (2) crop microbiological quality for human consumption at different times after harvest. Crop yields were compared using Variance analysis. Crops' sanitary acceptability was assessed using a two-class sampling program for Salmonella (n=10; c=0), and a three-class program for E. coli (n=5, c=2, M=10(3) and m=10 MPN/g) as proposed by the International Commission on Microbiological Specifications for Foods (ICMSF) for fresh vegetables. Wastewater irrigation acted as well water with fertilizer, increasing garlic and onion yields by 10% and 15%, respectively, compared to irrigation with well water with no fertilizer. Wastewater-irrigated garlic reached sanitary acceptability 90 days after harvest, once attached roots and soil were removed. Onions, which were cleaned immediately after harvest, met this qualification earlier than garlic (55 days). Neither the wastewater-irrigated crops nor the control crops were microbiologically acceptable for consumption raw at harvest.

The catastrophe-promoting activity of ectopic Op18/stathmin is required for disruption of mitotic spindles but not interphase microtubules.

Oncoprotein18/stathmin (Op18) is a microtubule (MT) destabilizing protein that is inactivated during mitosis by phosphorylation at four Ser-residues. Op18 has at least two functions; the N-terminal region is required for catastrophe-promotion (i.e., transition from elongation to shortening), while the C-terminal region is required to inhibit MT-polymerization rate in vitro. We show here that a "pseudophosphorylation" derivative of Op18 (i.e., four Ser- to Glu-substitutions at phosphorylation sites) exhibits a selective loss of catastrophe-promoting activity. This is contrasted to authentic phosphorylation, which efficiently attenuates all activities except tubulin binding. In intact cells, overexpression of pseudophosphorylated Op18, which is not phosphorylated by endogenous kinases, is shown to destabilize interphase MTs but to leave spindle formation untouched. To test if the mitotic spindle is sensitive only to the catastrophe-promoting activity of Op18 and resistant to C-terminally associated activities, N- and C-terminal truncations with defined activity-profiles were employed. The cell-cycle phenotypes of nonphosphorylatable mutants (i.e., four Ser- to Ala-substitutions) of these truncation derivatives demonstrated that catastrophe promotion is required for interference with the mitotic spindle, while the C-terminally associated activities are sufficient to destabilize interphase MTs. These results demonstrate that specific Op18 derivatives with defined activity-profiles can be used as probes to distinguish interphase and mitotic MTs.

The expression of Jagged1 in the developing mammalian heart correlates with cardiovascular disease in Alagille syndrome.

The establishment of the cardiovascular system represents an early, critical event essential for normal embryonic development, and defects in cardiovascular development are a frequent cause of both in utero and neonatal demise. Congenital cardio-vascular malformations, the most frequent birth defect, can occur as isolated events, but are frequently presented clinically within the context of a constellation of defects that involve multiple organs and that define a specific syndrome. In addition, defects can be a primary effect of gene mutations or result from secondary effects of altered cardiac physiology. Alagille syndrome (AGS) is an autosomal dominant disorder characterized by developmental abnormalities of the heart, liver, eye, skeleton and kidney. Congenital heart defects, the majority of which affect the right-sided or pulmonary circulation, contribute significantly to mortality in AGS patients. Recently, mutations in Jagged1 ( JAG1 ), a conserved gene of the Notch intercellular signaling pathway, have been found to cause AGS. In order to begin to delineate the role of JAG1 in normal heart development we have studied the expression pattern of JAG1 in both the murine and human embryonic heart and vascular system. Here, we demonstrate that JAG1 is expressed in the developing heart and multiple associated vascular structures in a pattern that correlates with the congenital cardiovascular defects observed in AGS. These data are consistent with an important role for JAG1 and Notch signaling in early mammalian cardiac development.