Solid-Supported Oligonucleotide Systems for Special Biomedical Applications

Abstract

The use of composite beads consisting of a 6 μm polystyrene core with 30 nm surface-bound silica particles to routine automatic oligodeoxynucleotide (ODN) synthesis is described.     

Inhibitory effect of oregano and thyme essential oils on moulds and foodborne bacteria

The essential oil of oregano ('origanum oil'; thymol type oil from Origanum vulgare) inhibited completely the mycelial growth of Aspergillus niger and A. flaous at 400 μg/ml, while A. ochraceus was inhibited at 600 μg/ml. At 700 μg/ml, thyme oil inhibited the mycelial growth of A. flavus and A. niger but not that of A. ochraceus . Fungal spore germination was inhibited by 600 μg/ml of origanum oil and (with the exception of A. ochraceus) by 700 μg/ml of thyme oil. Under aerobic conditions, the essential oils of oregano (250 μg/ml) and thyme (350 μg/ml) inhibited to some extent the growth of Staphylococcus aureus and Salmonella typhimurium. Pseudomonas aeruginosa was not affected by either oregano or thyme oil at concentrations up to 500 μg/ml. The origanum oil was very effective against Campylohacter jejuni and Clostridiurn sporogenes and thyme oil was very effective against C. jejuni. The antagonistic effect of the two oils on Staph. aureus and Salm. typhimuriutn was greatly enhanced when those organisms were incubated in atmospheres of low oxygen tensions     

The mechanism of photodynamic inactivation ofStaphylococcus aureus by deuteroporphyrin

The mechanism ofStaphylococcus aureus inactivation by deuteroporphyrin (DT) and light was studied with singlet oxygen quenchers or hydroxyl radical scavengers. The light-activated DT (10 /ml) reduced the viability of the culture to less than 1%, whereas methionine, tryptophan, and 1,4-diazabicyclo-2,2,2-octane (DBCO) used as singlet oxygen quenchers provided almost 60% protection. Propylgallate, which is a hydroxyl free radical scavenger, also provided 60% protection. The presence of a singlet oxygen quencher and propylgallate provided almost complete protection from inactivation (96%). Photoinactivation in the absence of culture media (in saline) increased the killing rate and decreased the ability of the singlet oxygen quenchers to protect. In the same conditions damage from hydroxl free radicals was well protected by propyl gallate. The present results indicate thatS. aureus photoinactivation by DT and light is mediated by both singlet oxygen and hydroxyl free radicals.     

Temperature dependence of beta receptor, adenosine receptor, and sodium fluoride stimulated adenylate cyclase from turkey erythrocytes

The individual temperature dependencies of the process which control the activity of turkey erythrocyte adenylate cyclase have been determined. The temperature dependence of the fraction of activable cyclase units experiences a thermal transition at 24 degrees C for all three modes of enzyme activation: l-epinephrine, adenosine, and NaF. This thermal transition probably reflects the phase transition in the inner monolayer of the membrane which influences the behavior of the GTP regulatory unit which is involved in all three modes of enzyme activation. The "rate constant" of enzyme activation by adenosine reflects two thermal transitions, at 24 and at 35 degrees C; the apparent rate constant of cyclase activation by NaF activation experiences a transition only at 24 degrees C whereas the rate constant of the beta-receptor-bound agonist decreases monotonously with no "breaks" on the Arrhenium plot. Following the temperature dependence of the fluorescence intensity of dansylphosphatidylethanolamine embedded in both sides of the membrane and exclusively in the outer monolayer, one can assign the thermal transition of 24 degrees C to the inner monolayer and the other two transitions to the outer monolayer (10 and 35 degrees C). We interpret these results as follows. (a) The monomolecular rate constant characterizing the activation of cyclase by the precoupled adenosine receptor experiences both the transition at 24 and 35 degrees C, indicating that the latter may span the bilayer. (b) The bata receptor activates the cyclase units only in fluid areas since it can diffuse exclusively in the fluid areas of the membrane and is unable to interact with cyclase units in "frozen" areas. the linear dependence of the logarithm of the rate constant on 1/T for the bata receptor reflects the change of membrane fluidity as a function of temperature.     

Hereditary heat-labile hexosaminidase B: its implication for recognizing Tay-Sachs genotypes

Two pairs of alleles, at the two loci of hexosaminidase (HEX), were found to segregate in an Arab inbred family: the normal and the mutant Tay-Sachs (TSD) alleles of HEX A, and the normal and a mutant allele of HEX B. Since the mutant HEX B is heat labile, no reliable identification of TSD genotypes can be obtained in its presence, as long as the proportions of HEX A and B are estimated by the routinely used heat-inactivation method. The genotypes may be correctly identified in such cases by separation of the two isoenzymes on ion-exchange chromatography, estimating their individual activities, and calculating the ratio between them. Of the nine genotype combinations possible with these two pairs of alleles, five have been identified in the reported family by this procedure.     

Predicting the Maximum Earthquake Magnitude from Seismic Data in Israel and Its Neighboring Countries

This paper explores several data mining and time series analysis methods for predicting the magnitude of the largest seismic event in the next year based on the previously recorded seismic events in the same region. The methods are evaluated on a catalog of 9,042 earthquake events, which took place between 01/01/1983 and 31/12/2010 in the area of Israel and its neighboring countries. The data was obtained from the Geophysical Institute of Israel. Each earthquake record in the catalog is associated with one of 33 seismic regions. The data was cleaned by removing foreshocks and aftershocks. In our study, we have focused on ten most active regions, which account for more than 80% of the total number of earthquakes in the area. The goal is to predict whether the maximum earthquake magnitude in the following year will exceed the median of maximum yearly magnitudes in the same region. Since the analyzed catalog includes only 28 years of complete data, the last five annual records of each region (referring to the years 2006–2010) are kept for testing while using the previous annual records for training. The predictive features are based on the Gutenberg-Richter Ratio as well as on some new seismic indicators based on the moving averages of the number of earthquakes in each area. The new predictive features prove to be much more useful than the indicators traditionally used in the earthquake prediction literature. The most accurate result (AUC = 0.698) is reached by the Multi-Objective Info-Fuzzy Network (M-IFN) algorithm, which takes into account the association between two target variables: the number of earthquakes and the maximum earthquake magnitude during the same year.     

Quality of life following parotidectomy for malignant and benign disease

Parotidectomy is performed for benign or malignant tumors and for selected benign inflammatory and autoimmune conditions. Possible associated complications include facial nerve paralysis, pain, loss of sensation, gustatory sweating, and facial scarring. Global quality of life in patients undergoing parotidectomy has not been reported. The implications of facial surgery with the catastrophic potential of facial nerve paralysis may severely affect quality of life. A quality-of-life study was conducted in patients undergoing parotidectomy for benign and malignant diseases to define the significance of associated morbidity and its impact on quality of life. A quality-of-life instrument was specifically created, based on the principles of the University of Washington Quality of Life questionnaire, and mailed to the patients. Questions addressed recognized complications of parotidectomy. Patient group results were compared for age above and below 45 years, sex, benign versus malignant disease, presence or absence of Frey syndrome, and presence or absence of benign pleomorphic adenoma. Forty-six percent of 125 patients meeting the study criteria fully replied to the questionnaire. The global health score was 3.5, corresponding with "good" to "very good." Except for local sensation, which had a score of 50, all other domains scored above 76. Change in appearance, gustatory sweating, and pain were reported by 70 percent, 57 percent, and 30 percent, respectively. Importance attributed to all domains except facial function was low. Pain was encountered significantly less in patients younger than 45 years of age, and scores for appearance were also highly significant in this age group. Postoperative sequelae were noted in the majority of patients. The dominant sequelae were altered sensation, change in appearance, Frey syndrome, and pain. A degree of permanent postoperative facial nerve impairment was reported by 10 patients. Nevertheless, overall, parotidectomy does not seem to severely affect quality of life.     

Proximity of cytoplasmic and periplasmic loops in NhaA Na+/H+ antiporter of Escherichia coli as determined by site-directed thiol cross-linking

The unique trypsin cleavable site of NhaA, the Na(+)/H(+) antiporter of Escherichia coli, was exploited to detect a change in mobility of cross-linked products of NhaA by polyacrylamide gel electrophoresis. Double-Cys replacements were introduced into loops, one on each side of the trypsin cleavage site (Lys 249). The proximity of paired Cys residues was assessed by disulfide cross-linking of the two tryptic fragments, using three homobifunctional cross-linking agents: 1,6-bis(maleimido)hexane (BMH), N,N'-o-phenylenedimaleimide (o-PDM), and N,N'-p-phenylenedimaleimide (p-PDM). The interloop cross-linking was found to be very specific, indicating that the loops are not merely random coils that interact randomly. In the periplasmic side of NhaA, two patterns of cross-linking are observed: (a) all three cross-linking reagents cross-link very efficiently between the double-Cys replacements A118C/S286C, N177C/S352C, and H225C/S352C; (b) only BMH cross-links the double-Cys replacements A118C/S352C, N177C/S286C, and H225C/S286C. In the cytoplasmic side of NhaA, three patterns of cross-linking are observed: (a) all three cross-linking reagents cross-link very efficiently the pairs of Cys replacements L4C/E252C, S146C/L316C, S146C/R383C, and E241C/E252C; (b) BMH and p-PDM cross-link efficiently the pairs of Cys replacements S87C/E252C, S87C/L316C, and S146C/E252C; (c) none of the reagents cross-links the double-Cys replacements L4C/L316C, L4C/R383C, S87C/R383C, A202C/E252C, A202C/L316C, A202C/R383C, E241C/L316C, and E241C/R383C. The data reveal that the N-terminus and loop VIII-IX that have previously been shown to change conformation with pH are in close proximity within the NhaA protein. The data also suggest close proximity between N-terminal and C-terminal helices at both the cytoplasmic and the periplasmic face of NhaA.