The assessment of enriched apoplastic extracts using proteomic approaches

In plant tissues the extracellular environment or apoplast, incorporating the cell wall, is a highly dynamic compartment with a role in many important plant processes including defence, development, signalling and assimilate partitioning. Soluble apoplast proteins from Arabidopsis thaliana, Triticum aestivum and Oryza sativa were separated by two‐dimensional electrophoresis. The molecular weights and isoelectric points for the dominant proteins were established prior to excision, sequencing and identification by matrix‐assisted laser‐desorption ionisation time of flight mass spectrometry (MALDI ‐ TOF MS). From the selected spots, 23 proteins from O. sativa and 25 proteins from A. thaliana were sequenced, of which nine identifications were made in O. sativa (39%) and 14 in A. thaliana (56%). This analysis revealed that: (i) patterns of proteins revealed by two‐dimensional electrophoresis were different for each species indicating that speciation could occur at the level of the apoplast, (ii) of the proteins characterised many belonged to diverse families reflecting the multiple functions of the apoplast and (iii), a large number of the apoplast proteins could not be identified indicating that the majority of extracellular proteins are yet to be assigned. The principal proteins identified in the aqueous matrix of the apoplast were involved in defence, i.e. germin‐like proteins or glucanases, and cell expansion, i.e. β‐D‐glucan glucohydrolases. This study has demonstrated that proteomic analysis can be used to resolve the apoplastic protein complement and to identify adaptive changes induced by environmental effectors.     

Yield Constant for Acanthamoeba polyphaga in Batch and Continuous Culture1

ABSTRACT. A two-stage chemostat modified to accommodate the growth of adhesive organisms was used to determine the yield constant, Y, of a representative soil amoeba, Acanthamoeba polyphaga, utilizing as its prey Pseudomonas paucimobilis. The first stage consisted of a glucose-limited bacterial culture in steady state. The second stage consisted of a simplified predator-prey system, nongrowing bacteria serving as the limiting substrate for amoebae. A refined methodology to more accurately determine Y was developed, and Y for Acanthamoeba polyphaga in batch and continuous culture was determined to be 19.1%.     

Pneumocystis Carinii Pneumonia: Relationship Between Cyst Size and Response to Treatment

Pneumocystis carinii pneumonia is a common cause of death in patients with AIDS. Diagnosis is based on cytological examination of smears prepared from induced sputum samples and bronchoalveolar lavage (BAL) specimens which have been stained with methenamine silver. We have examined 46 BAL/induced sputum specimens from patients who had clinical symptoms and signs suggestive of P. carinii pneumonia and measured the diameter of a minimum 100 cysts in each specimen. We found that cyst size correlated with response to treatment with co-trimoxazole. This observation has implications for the therapeutic management of patients with this infection. La pneumopathie áPneumocystis carinii est une cause de décès fréquente chez les patients atteints d'un syndrome d'immunodéficience acquise (SIDA). Le diagnostic est basé sur l'examen cytologique des frottis préparés à partir du produit d'expectoration provoquée ou du liquide de lavage bronchoalvéolaire (LBA), après coloration de Grocott. Le diamètre d'au moins 100 kystes par échantillon a été mesuré dans 46 prélèvements de LBA/expectoration provoquée de patients présentant des signes cliniques suggérant une pneumopathie àPneumocystis carinii. Nous avons constaté que le diamètre des kystes est corrélé avec la réponse au traitment par co-trimoxazole, Cette observation a des implications sur la conduite du traitement des patients porteurs de cette infection. PCP ist eine häufige Todesursache bei HIV-Patienten. Die diagnose beruht auf dem cytologischen Nachweis in induzierten Sputumausstrichen oder bronchio-lo-alveolären Lavagen mit der Grokott-Färbung. In 46 positiven Fällen wurde der Durchmesser von mindestens 100 Cysten in jeder Probe gemessen und eine Korrelation zum Behandlungserfolg mit Co-Trimoxazol nachgewiesen.