ANALYSIS OF SPATIAL DISTRIBUTIONS OF CELLULAR MOLECULES DURING MECHANICAL STRESSING OF CELL SURFACE RECEPTORS USING CONFOCAL MICROSCOPY

Confocal laser scanning microscopy represents a suitable technique to study the localization of cellular components in three dimension. The authors used this technique to analyse cellular events related to mechanical stimulation of integrin receptors on the cell surface. By performing optical sections the distribution of integrin receptors on the apical surface of an osteoblastic cell was determined. Concerning intracellular compartimentalization of signal transduction events, it was demonstrated that mechanical stimulation of integrins induced their linkage to the cytoskeleton. Cytoskeletally associated proteins like vinculin and talin accumulated in the vicinity of the site where the mechanical stress was applied to integrins on the cell surface. Optical sections revealed that clustering of these proteins proceeded to the base of the cell with gradually decreasing extent. In summary, it was demonstrated that the local distribution of cellular components is an important factor in mechanically induced signal transduction.     

Extracellular enzymatic activity of aquatic and aero-aquatic conidial fungi

Nineteen species of aquatic and areo-aquatic conidial fungi were tested for their ability to produce extracellular enzymes which degrade cellulose, starch, lipids, proteins and tannic acid. The cellulolytic activity was determined by using both solid and liquid media. The activity of other enzymes was examined using solid media. Two-thirds of the species were able to hydrolyze soluble cellulose (CMC) incorporated in solid and liquid media with varying degrees of activity. Extracellular culture filtrates ofAegerita candida, Helicodendron giganteum andH. tubulosum contained a Cl-Cx enzyme complex that could degrade both soluble cellulose (CMC) and crystalline cellulose (filter paper). Lipase activity was demonstrated by 11 species. Fourteen of the species showed activity for amylase and protease, but only 11 of the 16 were capable of degrading tannic acid.     

Comparative study of desert truffles from kuwait and other countries in the middle east for radionuclide concentration

Concentrations of some radionuclides, including¹³⁷Cs, in desert truffles in Kuwait were studied and compared with similar samples from other countries in the Middle East, namely Iran, Egypt, and Tunisia. In addition, sand samples from Kuwait were assayed to calculate the transfer factor of the radionuclides under consideration. The measured concentrations of⁴⁰K,²²⁶Ra, and¹³⁷Cs show that¹³⁷Cs is much higher in Egyptian samples, whereas⁴⁰K is much lower in samples from Tunisia. The average effective dose equivalent calculated for the Kuwaiti population according to their diet habits was found to be in the range 0.14-0.23 ΜS_v/a. The results are compared with values from other countries.     

D-(+)-lactate dehydrogenase from Lactobacillus murinus

D-(+)-Lactate dehydrogenase from Lactobacillus murinus was purified 670-fold. The Mr was 140,000 as determined by gel filtration. Maximum enzymatic activity was observed at 25 degrees C and pH 6.0 in 200 mM Na2KPO4 buffer. When the temperature was increased from 60 to 65 degrees C, the enzyme was completely inactive in 5 min. The apparent Km for pyruvate and NADH were 4.7 x 10(-4) and 1 x 10(-5) M, respectively. Pyruvate analogs such as oxalate, oxamate, 2-oxobutyrate, and malonate acted as a competitive inhibitors. L-Lactate and L-malate were noncompetitive inhibitors.     

Repellency of pheromones released by females of Heliothis armigera and H. zea to females of both species

An olfactometer was used to determine the effect of pheromones released by females of the bollworms Heliothis armigera (Hübner) and H. zea (Boddie) on females of the same species. Four combinations of virgin and mated females were tested for repellency of one to the other. Evidence is presented that females of the two bollworms were repelled by females of the same species. In addition, extracts of virgin female abdomens of each species repelled virgin females of the other species.

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Résumé L'examen en olfactomètre a porté sur les réactions face à d'autres femelles de la même espèce, de femelles vierges ou ayant copulé d'Heliothis armigera Hübner et H. zea Boddie. Le lot comprenait 8 femelles, vierges ou ayant copulé en présence d'une femelle vierge ou ayant copulé. Les 4 combinaisons possibles de femelles vierges et de femelles ayant copulé ont été examinées avec 12 répétitions pour chaque espèce. Un extrait de l'extrémité de l'abdomen de femelles vierges d'une espèce a été présenté aux femelles de l'autre espèce pour examiner les possibilités de réactions interspécifiques aux phéromones.Pour chaque espèce, les réactions interspécifiques de répulsion entre femelles ont été hautement significatives par rapport aux témoins, à l'exception toutefois des réactions de femelle ayant copulé face à des femelles ayant elles aussi copulé. Les répulsions moyennes chez H. armigera et H. zea pour les 8 femelles de chaque expérience ont été: a) vierges en présence d'une vierge: 7,33 et 7,66; b) vierges en présence d'une femelles ayant copulé: 5,76 et 5,58; c) femelles ayant copulé en présence d'une vierge: 4,67 et 4,83. Les différences sont hautement significatives entre chaque paire de moyennes et entre chaque paire et le lot témoin; 3,17; 3,17; 3,42; 4,00 pour H. armigera; 3,17; 3,50; 2,83 et 3,75 pour H. zea.Les femelles vierges des deux espèces, H. armigera et H. zea ont présenté une réaction de répulsion en présence d'un extrait de l'abdomen de l'autre espèce; les répulsions moyennes étant respectivement 5,53 et 5,33 contre 3,83 et 3,58 pour le lot trémoin.On peut en conclure que ces répulsions doivent entraîner une tendance à la répartition uniforme.

     

Phenol β-glucosyltransferase and β-glucosidase activities in the tobacco hornworm larva Manduca sexta (L.): Properties and tissue localization

Phenol β-glucosyltransferase (PGT; EC 2.4.1.35) was studied in feeding fifth stadium larvae of the tobacco hornworm, Manduca sexta, using reversed-phase HPLC with absorbance detection to separate and quantify both the model substrate, p-nitrophenol (PNP), and the product, p-nitrophenyl β-D-glucopyranoside (PNP-Glc). About 90% of total PGT activity in tissue homogenates was associated with the particulate fraction (15,000g), with the remainder in the microsomal fraction. PGT activity was observed in all tissues, with highest activities in the labial gland and fat body. Appreciable activity occurred in midgut and hindgut tissue but none was found in hemolymph. PGT activity was also observed in eggs and larval fat body at different times during development. Activity was optimal at pH 7.5–9 and was highest with UDP-Glc as a glucose donor. However, appreciable PGT activity was observed with dTDP-Glc or GDP-Glc in place of UDP-Glc. The divalent cations Ca²⁺, Co²⁺, Mg²⁺, and Mn²⁺ stimulated activity, whereas Zn²⁺ and Hg²⁺, as well as pretreatment with the detergent Triton X-100, were inhibitory. Endogenous β-glucosidase in PGT-enriched fractions, especially from the midgut, antagonized the β-glucossylation process, and interference was minimized with higher pH and the addition of D-gluconic acid lactone to the incubation mixtures. The possible role of transglucosylation in the detoxication of phenolic xenobiotics and biotransformation of endogenous phenolic compounds in insects is discussed. Comparisons of PGT with some glycosyltransferases involved in endogenous and xenobiotic conjugation in insects and other organisms are reviewed. © 1992 Wiley-Liss, Inc.     

Predation ofCyrtorhinus lividipennis Reuter on eggs of the green leafhopper and brown planthopper in rice

The functional response of Cyrtorhinus lividipennis feeding on brown planthopper (BPH) and green leafhopper (GLH) eggs was found to be Holling's Type II. The Random Predator Equation fitted the data satisfactorily. Using Manly's preference index, α, both the male and female C. lividipennis were found to prefer BPH eggs. There was, however, no evidence of switching and the ratio of the respective searching efficiencies could account for the preference.     

A combination of urinary biomarker panel and PancRISK score for earlier detection of pancreatic cancer: A case–control study

BackgroundPancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers, with around 9% of patients surviving >5 years. Asymptomatic in its initial stages, PDAC is mostly diagnosed late, when already a locally advanced or metastatic disease, as there are no useful biomarkers for detection in its early stages, when surgery can be curative. We have previously described a promising biomarker panel (LYVE1, REG1A, and TFF1) for earlier detection of PDAC in urine. Here, we aimed to establish the accuracy of an improved panel, including REG1B instead of REG1A, and an algorithm for data interpretation, the PancRISK score, in additional retrospectively collected urine specimens. We also assessed the complementarity of this panel with CA19-9 and explored the daily variation and stability of the biomarkers and their performance in common urinary tract cancers.Methods and findingsClinical specimens were obtained from multiple centres: Barts Pancreas Tissue Bank, University College London, University of Liverpool, Spanish National Cancer Research Center, Cambridge University Hospital, and University of Belgrade. The biomarker panel was assayed on 590 urine specimens: 183 control samples, 208 benign hepatobiliary disease samples (of which 119 were chronic pancreatitis), and 199 PDAC samples (102 stage I–II and 97 stage III–IV); 50.7% were from female individuals. PDAC samples were collected from patients before treatment. The samples were assayed using commercially available ELISAs. Statistical analyses were performed using non-parametric Kruskal–Wallis tests adjusted for multiple comparisons, and multiple logistic regression. Training and validation datasets for controls and PDAC samples were obtained after random division of the whole available dataset in a 1:1 ratio. The substitution of REG1A with REG1B enhanced the performance of the panel to detect resectable PDAC. In a comparison of controls and PDAC stage I–II samples, the areas under the receiver operating characteristic curve (AUCs) increased from 0.900 (95% CI 0.843–0.957) and 0.926 (95% CI 0.843–1.000) in the training (50% of the dataset) and validation sets, respectively, to 0.936 in both the training (95% CI 0.903–0.969) and the validation (95% CI 0.888–0.984) datasets for the new panel including REG1B. This improved panel showed both sensitivity (SN) and specificity (SP) to be >85%. Plasma CA19-9 enhanced the performance of this panel in discriminating PDAC I–II patients from controls, with AUC = 0.992 (95% CI 0.983–1.000), SN = 0.963 (95% CI 0.913–1.000), and SP = 0.967 (95% CI 0.924–1.000). We demonstrate that the biomarkers do not show significant daily variation, and that they are stable for up to 5 days at room temperature. The main limitation of our study is the low number of stage I–IIA PDAC samples (n = 27) and lack of samples from individuals with hereditary predisposition to PDAC, for which specimens collected from control individuals were used as a proxy.ConclusionsWe have successfully validated our urinary biomarker panel, which was improved by substituting REG1A with REG1B. At a pre-selected cutoff of >80% SN and SP for the affiliated PancRISK score, we demonstrate a clinically applicable risk stratification tool with a binary output for risk of developing PDAC (‘elevated’ or ‘normal’). PancRISK provides a step towards precision surveillance for PDAC patients, which we will test in a prospective clinical study, UroPanc.

Silvana Debernardi and colleagues establish a clinical risk score and a biomarker panel for early detection of pancreatic cancer.