Immunocytochemical localization of gamma-glutamyltranspeptidase during fetal development of mouse kidney

In the fully developed kidney, gamma-glutamyltranspeptidase is localized predominantly to the apical plasma membrane of the proximal tubules. The appearance of this activity during murine fetal nephrogenesis was quantitated using a sensitive fluorometric assay, and development of membrane polarity was assessed by immunocytochemistry. Specific activity of the transpeptidase in 13-day fetal kidney was approximately 1 mU/mg protein. Between 13-21 days of gestation, total transpeptidase activity increased 7500-fold, whereas specific activity increased 50-fold. At 13 days of gestation, gamma-glutamyltranspeptidase immunoreactivity is localized to the apical surfaces of developing renal vesicles and the proximal segment of the S-shaped tubules. The organized cell structures have tight tubular junctions but lack a well-defined brush-border membrane. By 15 days of gestation, immunostaining of the apical surface of developing proximal segments is more prominent, and slight reactivity of the basolateral membrane is evident. By 17 days of gestation, the kidney is organized into discrete zones. The large increase in gamma-glutamyltranspeptidase activity correlates with the appearance of increased immunostaining of the developing brush-border membranes of the proximal tubules contained in the inner cortex. A very similar although somewhat delayed pattern of appearance of transpeptidase activity and immunostaining was observed in metanephric organ culture. Induction of proximal tubular cyst formation had no effect on the increase in transpeptidase activity that occurred during organotypic nephrogenesis.     

A graphical method for detecting recombination in phylogenetic data sets

Current phylogenetic tree reconstruction methods assume that there is a single underlying tree topology for all sites along the sequence. The presence of mosaic sequences due to recombination violates this assumption and will cause phylogenetic methods to give misleading results due to the imposition of a single tree topology on all sites. The detection of mosaic sequences caused by recombination is therefore an important first step in phylogenetic analysis. A graphical method for the detection of recombination, based on the least squares method of phylogenetic estimation, is presented here. This method locates putative recombination breakpoints by moving a window along the sequence. The performance of the method is assessed by simulation and by its application to a real data set.      

bilbo, a non-LTR retrotransposon of Drosophila subobscura: a clue to the evolution of LINE-like elements in Drosophila

We used the repetitive character of transposable elements to isolate a non-LTR retrotransposon in Drosophila subobscura. bilbo, as we have called it, has homology to TRIM and LOA elements. Sequence analysis showed a 5' untranslated region (UTR), an open reading frame (ORF) with no RNA-binding domains, a downstream ORF that had structural homology to that of the I factor, and, finally, a 3' UTR which ended in several 5-nt repeats. The results of our phylogenetic and structural analyses shed light on the evolution of Drosophila non-LTR retrotransposons and support the hypothesis that an ancestor of these elements was structurally complex.      

Biofumigant effect of new defatted seed meals against the southern root‐knot nematode,Meloidogyne incognita

In recent years, Brassica carinata defatted seed meals (DSMs) have been successfully applied as an amendment in the control of pests and soil pathogens in several countries with clear advantages for the environment and soil fertility. The effectiveness of this product is clearly linked to the release of bioactive compounds by the well‐known glucosinolate (GL)–myrosinase (MYR) system. Until now, this commercial know‐how has been limited to allyl‐isothiocyanate, the breakdown product of hydrolysis catalysed by MYR of the GL sinigrin. Brassicaceae germplasm is an extremely wide‐ranging family, considering that more than 3500 species have been classified in this family in nature, which contains around 200 different GLs. Therefore, there is a great potential for the availability of new still unexplored bioactive compounds. This study evaluated, in controlled glasshouse conditions, the effect of biofumigation on the nematode Meloidogyne incognita and that of biostimulation on tomato plants of 13 DSMs obtained from different Brassicaceae species at different levels of soil inoculation. Among the tested DSMs, the best results for all inoculations were achieved by Eruca sativa (rocket), Barbarea verna (land cress) and Brassica nigra (black mustard), whereas the other species gave either alternate results or results not different from untreated or sunflower DSM controls. All the DSMs, including sunflower, determined a clear positive effect on plant vigour. These first results open new perspectives for the application of biofumigation in plant protection and management.     

The AP‐2 complex is required for proper temporal and spatial dynamics of endocytic patches in fission yeast

In metazoans the AP‐2 complex has a well‐defined role in clathrin‐mediated endocytosis. By contrast, its direct role in endocytosis in unicellular eukaryotes has been questioned. Here, we report co‐ immunoprecipitation between the fission yeast AP‐2 component Apl3p and clathrin, as well as the genetic interactions between apl3Δ and clc1 and sla2Δ/end4Δ mutants. Furthermore, a double clc1 apl3Δ mutant was found to be defective in FM4‐64 uptake. In an otherwise wild‐type strain, apl3Δ cells exhibit altered dynamics of the endocytic sites, with a heterogeneous and extended lifetime of early and late markers at the patches. Additionally, around 50% of the endocytic patches exhibit abnormal spatial dynamics, with immobile patches and patches that bounce backwards to the cell surface, showing a pervasive effect of the absence of AP‐2. These alterations in the endocytic machinery result in abnormal cell wall synthesis and morphogenesis. Our results complement those found in budding yeast and confirm that a direct role of AP‐2 in endocytosis has been conserved throughout evolution.     

Variation of physico-chemical parameters along a river transect through the Okavango Delta,Botswana

The Okavango Delta depends on water quantity and quality to sustain its ecosystem services. Whereas many studies have been carried out on its hydrology, few have been done on water quality in the delta. Water pH, electrical conductivity (EC), dissolved oxygen (DO), turbidity, total suspended solids (TSS) and dissolved organic carbon (DOC) were monitored at 10 sites along the Okavango–Boro–Thamalakane–Lake Ngami system almost fortnightly from June 2008 to June 2010. Water quality in the delta was generally good, despite high evapotranspiration rates which would normally produce very saline waters. Electrical conductivity and water temperature increased with distance from Mohembo to Lake Ngami, the former most likely due to evapoconcentration. In contrast, pH, DO, turbidity and TSS decreased with distance from Mohembo to Boro at the lower end of the seasonal floodplain, before increasing again to Lake Ngami. Dissolved oxygen and TSS most likely declined due to biological uptake and particle sedimentation, respectively. Strong and significant relationships were observed between TSS and turbidity and between DOC and EC, indicating that turbidity and EC could be useful proxies for routine estimations of TSS and DOC, respectively, in the delta.     

Macroinvertebrates as bioindicators of water quality in the Mkondoa River,Tanzania, in an agricultural area

The suitability of using macroinvertebrates as bioindicators of stream water quality was tested in the Mkondoa River in an agricultural area at Kilosa, using the rapid bioassessment protocol. The family biotic index (FBI) showed marked variation in water quality along the stream from values ranging from 4.1 to 5.0 in the upstream reaches, indicating good water quality, 5.3 to 5.5 in the mid-reaches and 6.0 to 6.5 in the lower reaches. The water quality index (WQI) indicated that water quality was fair (77 ± 0.98) in the upstream reach of the Mkondoa, marginal (55 ± 0.86) in the midstream reach and poor (33 ± 0.45) in the downstream reach. There were significant relationships between biological oxygen demand and dissolved oxygen and the occurrence of specific taxa, mainly Chironomus and Caenis. Significant changes in macroinvertebrate abundance were mostly related to changes in water quality. As in other parts of the world, macroinvertebrate communities proved to be good biological indicators of water quality and they should be used as bioindicators in long-term monitoring of this river.     

Effect of insulin on brain pyruvate dehydrogenase in the rat. (Preliminary note)

The activity of both active and total pyruvate dehydrogenase (E.C.1.2.4.1) is substantially reduced in the rat brain 24h after alloxan administration. Effects are partially removed by insulin administration. Ca++ and Mg++ produce: a) a considerable conversion of the inactive form of pyruvate dehydrogenase into its active form in a preparation from the brain of normal rats and of rats treated with insulin; b) no conversion in a preparation from the brain of rats treated with alloxan; c) some conversion in a preparation from the brain of rats treated with both alloxan and insulin. Active and total pyruvate dehydrogenase from the brain of rats treated with alloxan are activated by a preparation obtained from a mixture of entire plasma membranes-mitochondria from normal and from alloxan-treated rats, or from insulin-treated and alloxan treated rats. The oxygen uptake, the respiratory control index and the ADP/O ratio in mitochondrial preparations obtained from the brain of rats treated with alloxan show no modification at all.     

Activation of pyruvate dehydrogenase by addition of insulin to a mixture of cell membranes and mitochondria from the brain of normal and alloxan treated rats. (Preliminary note)

The effect of insulin and ATP on pyruvate dehydrogenase activity has been studied in a mixture of plasma membranes/mitochondria from normal and diabetic rat brain. In both enzymatic preparations ATP inhibits almost completely pyruvate dehydrogenase activity; insulin on the contrary induces a conspicuous activation and removes the inhibiting action of ATP.