Fine structural criteria for identifying rat corticotrophs

Summary The fine structural characteristics of normal rat corticotrophs stained with anti-porcine ACTH^1–39 serum were studied. At the ultrastructure level immunoreactive corticotrophs appear to comprise four distinct cell types: (1) large stellate cells (Siperstein cells) containing granules (170–250 nm in diameter) arranged in a peripheral row and usually embracing an acidophil; (2) elongate spindle-shaped cells (Moriarty cells) in which the secretory granules (170–250 nm in diameter) are distributed in a row or in small clusters in the peripheral cytoplasm; (3) oval or polygonal cells filled only with small secretory granules (130–170 nm in diameter), resembling the acidophil of small granules type (Yoshimura et al. 1974); and (4) polygonal or stellate cells filled with secretory granules of varying diameters (180–300 nm in diameter) and occasionally embracing an acidophil. The first type is the most common, but the others are infrequent. It is concluded that the criteria of Siperstein and Miller (1970) do not necessarily include all categories of rat corticotrophs.     

Components of partial resistance to Septoria nodorum in winter wheat

Components of partial resistance to Septoria nodorum were investigated in 10 cultivars of winter wheat having similar field resistances. The components measured were infection frequency, latent period, size, shape and rate of growth of lesions, spore production and its rate of increase. Latent period was found to be lognormally distributed. Some of the components of resistance were found to be significantly different between cultivars. Cluster analysis also showed that cultivars could be distinguished on the basis of their components of resistance. Principal components analysis indicated that resistance could be broken down into four underlying factors, three of which could be readily interpreted. The measurements of the components of resistance were combined in a model, the r-index, based on Van der Plank's r. The amount of variation between cultivars cast some doubt on the predictive value of the index but all the cultivar values were well within the range bounded by two ‘synthetic’ cultivars made up of combinations of either the most resistant or the most susceptible components. It is considered that the r-index has potential in screening for field resistance. The possibility of incorporating the most resistant-type components into one cultivar is discussed. The use of cultivar mixtures containing cultivars having similar field resistances is also discussed in the light of the variability found in this study.     

Induced colistin resistance as an identifying marker for Aeromonas phenospecies groups

AIMS: To investigate the taxonomic interest of colistin resistance as an identifying marker for Aeromonas phenospecies groups. METHODS AND RESULTS: Colistin resistance was investigated in 387 Aeromonas isolates identified at species level using a 14-test format protocol with miniaturized tests combined with determination of urocanic acid utilization whenever necessary. Colistin resistance, determined by the disc diffusion method, was unreliable when compared with minimum inhibitory concentration (MIC) determination. In some strains, the MIC values and resistance rates of colistin could be increased after overnight induction with a 50- microg colistin disc in 20 ml of Mueller-Hinton broth (2.5 mg l(-1)). Colistin-induced resistance level was raised to 85.8% in the Aeromonas hydrophila complex, 2.1% in the A. caviae complex and 2.5% in the A. veronii complex except for A. jandaei (100% colistin resistant). This new marker allowed the identification of 96.2 and 93.6% of Aeromonas isolates to phenospecies and species level, respectively. CONCLUSIONS: Colistin-induced colistin resistance is a new phenotypic marker for Aeromonas isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: With the present protocol, colistin resistance determination may improve the identification of Aeromonas isolates to phenogroup level, when results obtained by conventional biochemical methods are ambiguous.     

Components of partial resistance of barley to Rhynchosporium secalis: use of seedling tests to predict field resistance

Components of partial resistance [disease incidence (DI), infection frequency (IF), latent period (LP), spores per lesion (SPL)] were assessed on glasshouse-grown barley seedlings following inoculation with spore suspensions of Rhynchosporium secalis at growth stage 12 (Zadoks, Chang & Konzak, 1974). Four experiments were carried out at different times during 1988. Three spring barley cultivars [two from Cyprus (cvs Kantara and Athenais) and one from the UK (cv. Triumph)] were used in the first three experiments. In the fourth experiment eight additional UK cultivars with NIAB resistance ratings ranging from 3 to 9 were used. Two races of R. secalis were used in the first three experiments and three in the fourth. The three cultivars, Kantara, Athenais and Triumph, were examined in all four experiments and significant differences detected for virtually all components of partial resistance in each. Differences, however, were often small and ranking of cultivars varied in different experiments. The greater susceptibility of cv. Kantara compared to cv. Athenais, observed under field conditions in Cyprus, would not be anticipated from the small differences in components of partial resistance observed in these experiments, but, for these cultivars, the possibility of a marked genotype x environment interaction cannot be discounted. Mean values for the components of partial resistance differed in the four experiments. LP was correlated with mean glasshouse temperature from inoculation to the onset of sporulation but differences in IF and SPL were not correlated with temperature. For these components, light quality and/or duration appeared to be more important. Overall, there were no differences between races but significant race X cultivar interactions were observed in two experiments. In the fourth experiment, examining 11 cultivars, there were significant differences between cultivars for all components of partial resistance. IF and LP were correlated but neither of these components was correlated with SPL indicating independent control of this latter component. Both IF and LP were correlated with field performance (NIAB ratings) but there was no correlation with SPL. However, combining IF with mean values of SPL restricted to the 5 days following the end of the LP, produced the best correlation (r= 0.92) with NIAB ratings. Problems of assessing components of partial resistance and possible means of improving assessments are discussed.     

Sources and sinks: revisiting the criteria for identifying reservoirs for American cutaneous leishmaniasis

Molecular-based approaches might be misleading for assessing the 'potential importance' of possible reservoirs for parasites unless used in conjunction with population dynamics and ecological studies that provide an understanding of the linkages between the enzootic transmission cycle (in wild animals) and that in domestic animals. Better understanding of ecological constraints on Leishmania transmission is needed to assess fully the passage of the parasite between its sandfly vector and different mammalian host species. We use a straightforward mathematical framework to illustrate that misuse of association patterns, as guidance for implementation of control measures, can in fact increase the endemism of leishmaniasis.     

Proteomics for identifying mechanisms and biomarkers of drug resistance in cancer

A major problem in chemotherapy of cancer patients is drug resistance as well as unpredictable response to treatment. During chemotherapy, multiple alterations of genetics and epigenetics that contribute to chemoresistance take place, eventually impacting on disease outcome. A more complex picture of the mechanisms of drug resistance is now emerging through application of high-throughput proteomics technology. We have entered an exciting time where proteomics are being applied to characterize the mechanisms of drug resistance, and to identify biomarkers for predicting response to chemotherapy, thereby leading to personalized therapeutic strategies of cancer patients. Comparative proteomics have identified a large number of differentially expressed proteins associated with chemoresistance. Although roles and mechanisms of such proteins in chemoresistance need to be further proved, at least some of them may be potential biomarkers for predicting chemotherapeutic response. Herein, we review the recent advancements on proteomic investigation of chemoresistance in human cancer, and emphasize putative biomarkers for predicting chemotherapeutic response and possible mechanisms of chemoresistance identified through proteomic approaches. Suggested avenues for future work are discussed.     

Spontaneous bacteriocin resistance in Listeria monocytogenes as a susceptibility screen for identifying different mechanisms of resistance and modes of action by bacteriocins of lactic acid bacteria

A practical system was devised for grouping bacteriocins of lactic acid bacteria (LAB) based on mode of action as determined by changes in inhibitory activity to spontaneously-acquired bacteriocin resistance (Bac^R). Wild type Listeria monocytogenes 39-2 was sensitive to five bacteriocins produced by 3 genera of LAB: pediocin PA-1 and pediocin Bac3 (Pediococcus), lacticin FS97 and lacticin FS56 (Lactococcus), and curvaticin FS47 (Lactobacillus). A spontaneous Bac^R derivative of L. monocytogenes 39-2 obtained by selective recovery against lacticin FS56 provided complete resistance to the bacteriocin made by Lactococcus lactis FS56. The lacticin FS56-resistant strain of L. monocyotgenes 39-2 was also cross-resistant to curvaticin FS47 and pediocin PA-1, but not to lacticin FS97 or pediocin Bac3. The same pattern of cross-resistance was also observed with Bac^R isolates obtained with L. monocytogenes Scott A-2. A spontaneous mutation that renders a strain cross-resistant to different bacteriocins indicates that they share a common mechanism of resistance due to similar modes of action of the bacteriocins. Spontaneous resistance was acquired to other bacteriocins (in aggregate) by following the same procedure against which the Bac^R strain was still sensitive. In subsequent challenge assays, mixtures of bacteriocins of different modes of action provided greater inhibition than mixtures of bacteriocins of the same mode of action (as determined by our screening method). This study identifies a methodical approach to classify bacteriocins into functional groups based on mechanism of resistance (i.e., mode of action) that could be used for identifying the best mixture of bacteriocins for use as biopreservatives.     

A partial ranking method for identifying repeated inclusion of individuals in anonymized HIV infection reports

Diagnoses of HIV infection are reported to the Public Health Laboratory Service (PHLS) AIDS Centre under a voluntary surveillance scheme. Names are not held in the data set, but the date of birth of the individual concerned is usually available. This paper describes a statistical method for identifying whether there are likely to be individuals repeatedly represented in the resulting data set, which is considered by birth year. A partial ordering method is used that is especially useful for years where the number of birth years in the sample is too small for chi2 tests to be used. At the 5% level, one of the five birth years tested in the data supplied to us by the PHLS shows evidence of more replication than would be expected from independent random sampling from the population. The results are compared with an alternative maximum-likelihood-based test that reaches the same conclusions. Maximum likelihood methods are further used to estimate the percentage of overcounting of individuals in the sample at 2.7%.     

Peptide factors as pharmaceuticals: criteria for application

The increasing interest in the pharmaceutical use of peptide factors in human medicine presents formidable challenges for peptide chemistry. Fully reproducible multistage syntheses with a definite assessment of the degree of purity represent the crucial premise for the introduction of peptide factors as pharmaceuticals. Extensive studies of the stability of peptidic material on storage allows identification of the most suitable form of administration. Nevertheless, the high clearance rate of peptides as pharmaceuticals presents new challenges for improvement by structural modification of resistance to enzyme degradation without creating new problems related to metabolites.     

Antiviral susceptibility testing of cytomegalovirus: criteria for detecting resistance to antivirals

Testing cytomegaloviruses for antiviral susceptibility is increasing especially since the reports of 'resistance' to ganciclovir and foscarnet (Erice et al., 1989; Knox et al., 1991). There is however no standardized method for susceptibility testing nor are there criteria for designating an isolate as sensitive or resistant. In a previous paper we utilized a plaque reduction assay and suggested that a resistant strain be defined as one requiring > 12 microM ganciclovir for inhibition of 50% of viral plaques. (Drew et al., 1991) This concentration was chosen because it was at least four-fold greater than the mean concentration required to inhibit pretherapy isolates. In this paper we present the results of testing a large number of isolates prior to and during therapy with either ganciclovir or foscarnet. By analyzing the results of these assays we propose revised criteria for susceptibility of cytomegalovirus 相似文献   

An assessment of a method based on intrinsic antibiotic resistance for identifying Rhizobium strains

A method based on intrinsic antibiotic resistance (IAR) for identifying large numbers of Rhizobium strains was assessed and found to be unsatisfactory for R. phaseoli and isolates from Cicer arietinum (Rhizobium spp.). Our data showed that the number of different IAR patterns always exceeded the number of strains tested. With 90 nodule isolates from plants inoculated with a mixture of three strains of R. Phaseoli, the technique gave 18 different resistance patterns. When 24 strains of Rhizobium spp., each replicated three times, were examined 68 different resistance patterns were obtained. Single colony isolates from one strain also gave several different IAR patterns. All strains tested with fluorescent“ antibody were readily identified. Attempts to obtain correct strain identification with IAR by simplifying the scoring systems or allowing up to two differences in the resistance patterns were unsuccessful. We were unable to define the source of this variation although incubation time and inoculum concentration were shown to affect the IAR patterns     

Isolate-specific QTLs for partial resistance to Puccinia hordei in barley

By using a high-density AFLP marker linkage map, six QTLs for partial resistance to barley leaf rust (Puccinia hordei) isolate 1.2.1. have been identified in the RIL offspring of a cross between the partially resistant cultivar ’Vada’ and the susceptible line L94. Three QTLs were effective at the seedling stage, and five QTLs were effective at the adult plant stage. To study possible isolate specificity of the resistance, seedlings and adult plants of the 103 RILs from the cross L94×’Vada’ were also inoculated with another leaf rust isolate, isolate 24. In addition to the two QTLs that were effective against isolate 1.2.1. at the seedling stage, an additional QTL for seedling resistance to isolate 24 was identified on the long arm of chromosome 7. Of the eight detected QTLs effective at the adult plant stage, three were effective in both isolates and five were effective in only one of the two isolates. Only one QTL had a substantial effect at both the seedling and the adult plant stages. The expression of the other QTLs was developmental-stage specific. The isolate specificity of the QTLs supports the hypothesis of Parlevliet and Zadoks (1977) that partial resistance may be based on a minor-gene-for-minor-gene interaction. Received: 16 February 1999 / Accepted: 20 February 1999     

In situ extension as an approach for identifying novel alpha-amylase inhibitors

A new approach for the discovery and subsequent structural elucidation of oligosaccharide-based inhibitors of alpha-amylases based upon autoglucosylation of known alpha-glucosidase inhibitors is presented. This concept, highly analogous to what is hypothesized to occur with acarbose, is demonstrated with the known alpha-glucosidase inhibitor, d-gluconohydroximino-1,5-lactam. This was transformed from an inhibitor of human pancreatic alpha-amylase with a K(i) value of 18 mm to a trisaccharide analogue with a K(i) value of 25 mum. The three-dimensional structure of this complex was determined by x-ray crystallography and represents the first such structure determined with this class of inhibitors in any alpha-glycosidase. This approach to the discovery and structural analysis of amylase inhibitors should be generally applicable to other endoglucosidases and readily adaptable to a high throughput format.     

Application of toxico-kinetic criteria as a basis for toxicometric parameters

In an experiment with single exposure to benzene and styrene at a level corresponding, to the threshold of harmful effect established by functional indices the intensity of metabolism was found to decrease. Toxico-kinetic characteristics of the substance in a single experiment permit us to prognosticate the risk of developing chronic intoxication. In comparison with indices characterizing general state of the organism, toxicokinetic indices enhance the reliability of toxicometric parameters used in determining Mac's for the air of the working area.     

Comparing solid tumors with cell lines: implications for identifying drug resistance genes in cancer

Gene expression arrays allow researchers to profile the differences between cell lines or tissues and they may identify genetic markers of development, organ maturation, or tumor progression. Although a primary tumor that grows in a host and a tumor-cell-line derived from that primary tumor and grown in vitro share similar gene expression profiles, there are, not unexpectedly, some important differences. In fact, Stein and colleagues have found that genes that are differentially expressed in primary tumors as compared to the specific genes expressed in their cell-line derivatives are more reliably predictive of tumor tractability. Thus, sensitivity in vitro might not reflect sensitivity in vivo. Because anti-tumor compounds are largely evaluated in cell culture assays, these compounds' therapeutic utility must be judged in light of genes described by Stein et al. that better predict tractability.     

Unusual phyletic distribution of peptidases as a tool for identifying potential drug targets

Profound changes in the phosphorylation state of many proteins occur during mitosis. It is well established that many of these mitotic phosphorylations are carried out by archetypal mitotic kinases that are activated only during mitosis, shifting the equilibrium of kinases and phosphatases towards phosphorylation. However, many studies have also detailed the phosphorylation of proteins at mitosis by kinases that are constitutively active throughout the cell cycle. In most cases, it is uncertain how kinases and phosphatases that appear to be constitutively active can induce phosphorylations specifically at mitosis. In this issue of the Biochemical Journal, Escargueil and Larsen provide evidence of an interesting alternative mechanism to attain specific mitotic phosphorylation. A mitosis-specific phosphorylation site in DNA topoisomerase IIalpha, which is recognized by the MPM-2 antibody, is phosphorylated by protein kinase CK2. The authors found that phosphorylation of this site is suppressed during interphase due to competing dephosphorylation by protein phosphatase 2A. Interestingly, protein phosphatase 2A is excluded from the nucleus during early mitosis, allowing CK2 to phosphorylate topoisomerase IIalpha. It is possible that similar mechanisms are used to regulate the phosphorylation of other proteins.