Level Of Residual Activity Research Articles

Establishment of the cat flea (Ctenocephalides felis felis) on the ferret (Mustela putorius furo) and its control with imidacloprid

As the ferret, Mustela putorius furo L. (Carnivora: Mustelidae), is becoming increasingly popular as a pet animal and as it is susceptible to the cat-flea, Ctenocephalides felis felis Bouché (Siphonaptera: Pulicidae), an experimental model was established for evaluating insecticidal treatments on this host. A high establishment rate (76.7-91.8%) was recorded when 60 unfed adult C. felis were placed on ferrets. This provided an adequate infestation for chemotherapeutic evaluation without causing undue discomfort to the host. Twelve ferrets were allocated to two groups matched for sex and individual ability to sustain a flea population. One group was treated topically with an imidacloprid spot-on formulation at a dose rate of 10 mg/kg body-weight on Day 0. All ferrets were infested with C. felis on Days -1, 7, 14, 21 and 28, and flea counts were performed 8 and 24 h post-treatment and one day after each subsequent infestation. Fleas were removed at all but the 8 h count (when they were returned to their host). Flea burdens were reduced by 95.3% (P < 0.001) within 8 h of treatment and 100% efficacy was recorded at 24 h. At 1, 2, 3 and 4 weeks post-treatment, protection against re-infestation was 92.9% (P < 0.001), 55.7% (P < 0.02), 18.3% (NS) and 7.4% (NS), respectively. Thus, at this dose rate, imidacloprid gave excellent efficacy against a resident C. felis population and provided a high level of residual activity for at least one week after treatment.

Constitutive expression of a transcription termination factor by a repressed prophage: promoters for transcribing the phage HK022 nun gene.

Lysogens of phage HK022 are resistant to infection by phage lambda. Lambda resistance is caused by the action of the HK022 Nun protein, which prematurely terminates early lambda transcripts. We report here that transcription of the nun gene initiates at a constitutive prophage promoter, P(Nun), located just upstream of the protein coding sequence. The 5' end of the transcript was determined by primer extension analysis of RNA isolated from HK022 lysogens or RNA made in vitro by transcribing a template containing the promoter with purified Escherichia coli RNA polymerase. Inactivation of P(Nun) by mutation greatly reduced Nun activity and Nun antigen in an HK022 lysogen. However, a low level of residual activity was detected, suggesting that a secondary promoter also contributes to nun expression. We found one possible secondary promoter, P(Nun)', just upstream of P(Nun). Neither promoter is likely to increase the expression of other phage genes in a lysogen because their transcripts should be terminated downstream of nun. We estimate that HK022 lysogens in stationary phase contain several hundred molecules of Nun per cell and that cells in exponential phase probably contain fewer.

Ependymal development, proliferation, and functions: a review.

A survey of the literature shows that proliferation of ependyma occurs largely during the embryonic and early postnatal periods of development in most species. Differentiation of these cells proceeds along particular regional and temporal gradients as does the expression of various cytoskeletal (vimentin, cytokeratins, glial fibrillary acidic protein) and secretory proteins (S-100). Turnover declines significantly postnatally, and only low levels of residual activity persist into adulthood under normal conditions. Although the reported response of ependyma to injury is somewhat equivocal, only limited regenerative capacity appears to exist and to varying degrees in different regions of the neuraxis. Proliferation has been most often observed in response to spinal cord injury. Indeed, the ependyma plays a significant role in the initiation and maintenance of the regenerative processes in the spinal cord of inframammalian vertebrates. In the human, however, ependyma appears never to regenerate at any age nor re-express cytoskeletal proteins characteristic of immature cells. The functions of ependyma including tanycytes, a specialized form of ependymal cell that persists into adulthood within circumscribed regions of the nervous system, are still largely speculative. Fetal unlike mature ependyma is believed to be secretory and is believed to play a role in neurogenesis, neuronal differentiation/axonal guidance, transport, and support. In the adult brain, mature ependyma is not merely an inert lining but may regulate the transport of ions, small molecules, and water between the cerebrospinal fluid and neuropil and serve an important barrier function that protects neural tissue from potentially harmful substances by mechanisms that are still incompletely understood.

Immortalization and characterization of a cell line exhibiting a severe multiple sulphatase deficiency phenotype.

Multiple sulphatase deficiency (MSD) is a rare genetic defect that causes a simultaneous deficiency of all known sulphatases. All available evidence suggests that the deficient gene product is normally responsible for the post-translational modification of a conserved cysteine residue to 2-amino-3-oxopropionic acid and that this modification is necessary for sulphatase activity. MSD often has an enzymically mild phenotype, with significant levels of residual sulphatase activity being detectable. Here we identify an MSD cell line in which the residual activity of the sulphatases assayed was generally very low. To characterize the phenotype of this cell line further, immortalized lines were established after transformation with simian virus 40 (SV40) T antigen. Immortalized cell lines representing normal and MSD phenotypes were then transduced with a retroviral vector carrying the gene encoding human N-acetylgalactosamine-4-sulphatase. Analysis of N-acetylgalactosamine-4-sulphatase protein synthesis and enzyme activity showed that transduced cell lines expressed large amounts of enzyme and that the specific activity of this enzyme was approx. 0.5-1.5% of normal, confirming that this cell line defines a severe phenotype for MSD. N-Acetylgalactosamine-4-sulphatase purified from a transduced MSD cell line seemed normal on denaturing PAGE. Kinetic analysis of the purified enzyme suggests that the residual activity is due to small amounts of normal enzyme rather than unmodified enzyme with low levels of residual activity. These cell lines and the availability of large amounts of inactive N-acetylgalactosamine-4-sulphatase from MSD cells should facilitate the further study of this disorder.

MODELLING THE DEACTIVATION KINETICS OF CYCLOHEXANE DEHYDROGENATION ON PLATINUM/ALUMINA CATALYST

Two reaction deactivation kinetic models of the deactivation of cyclohexane on platinum/alumina catalyst have been proposed based on the coking mechanism of Corella and Asua and the reaction mechanism of Susu el al. The difference in the models was in the number of active sites participating in the coking reaction represented by h with h = 1 in model 1 and h ≠ 1 in model 2. The models were dynamic models and parameter estimates were carried out by a method that did not require decoupling of the main from the coking reaction. The model formulation has made possible the estimation of the level of residual activity by the incorporation of a factor, ƒ, that represents incomplete deactivation. For the deactivation kinetic data studied, a high value of ƒ; was obtained indicating a low residual activity. The models were found to give a good fit with the data. The standard deviation of the models from experimental were 10.21 % for model 1 (h ≠ 1) and 12.63% for model 2 (h ≠ 1), None of the models is superior to t...

Prenatal diagnosis by enzyme analysis in 15 pregnancies at risk for the Lesch-Nyhan syndrome.

Fifteen pregnancies at risk for Lesch-Nyhan syndrome were investigated between 8 and 17 weeks' gestation by measurement of hypoxanthine-guanine phosphoribosyl transferase (HGPRT) and adenine phosphoribosyl transferase (APRT) enzyme activities in chorionic villus samples (cultured and uncultured) or in cultured amniotic fluid cells. Ten pregnancies had normal enzyme levels and a normal outcome while a further two predicted to be normal miscarried later in the pregnancy. Three pregnancies had low levels of residual HGPRT activity in chorionic villi. Comparable levels of residual activity in the index case in two pregnancies and in cells from the abortus in the third case confirmed that the pregnancies were affected.

Clinical and molecular heterogeneity of cytochromec oxidase deficiency in the newborn

We report 8 cases of severe cytochrome c oxidase deficiency with onset in the neonatal period. Clinical symptoms were heterogeneous: antenatal cerebral malformations, neurological distress with ketoacidosis, severe myopathy, or isolated respiratory control failure. Lactic acid was elevated in blood and/or CSF in 7 cases. Muscle biopsy (7 patients), liver biopsy (4 patients), and cultured skin fibroblasts (7 patients) were used to assess the cytochrome c oxidase deficiency. Among the patients, the enzymatic defect differed in the level of residual activity, expression in different tissues and subunit composition in muscle (as analysed by immunohistochemistry). Southern blot analysis of the mitochondrial DNA was normal in 7 patients. The heterogeneity of cytochrome c oxidase deficiency was therefore demonstrated by these clinical presentations and by the biochemical assessment of the enzyme defect. This reflects, most probably, the diverse nature of the causal mutations.

Post injection transmission scanning in a volume imaging PET camera

Quantitative, low noise, measured attenuation correction has been established for the PENN PET 240H Volume Imaging Camera. This is achieved, without septa, using a narrow energy (450-570 keV) and sinogram position (2 cm wide mask) gate to minimize scatter contamination. Twelve minute transmission acquisitions with a 0.5 mCi rod source are adequate for this purpose. Post injection transmission scans (with emission activity in the FOV) suffer from emission contamination simulating transmitted gamma ray flux. This emission contamination may be measured by performing a transmission acquisition with normal transmission energy and position gating but without a transmission source. This contamination is then subtracted from the measured post injection transmission scan. Emission activity within the FOV adds to detector deadtime, whereas only a small fraction of events are accepted into the transmission position gate, resulting in a net loss of scan statistics when compared to pre-injection transmission scanning. Removal of emission contamination and compensation for this excess deadtime results in corrected attenuation coefficients close to pre-injection values. For residual activity levels typical of FDG whole-body cancer and cardiac studies the post injection measurements are within 4% of pre-injection values.< <ETX xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">&gt;</ETX>

Truncation of the COOH-terminal domain of the psbE gene product in Synechocystis sp. PCC 6803: requirements for photosystem II assembly and function.

The COOH-terminal domain of the 80-residue cytochrome b559 alpha-subunit (psbE gene product) in Synechocystis sp. PCC 6803 was sequentially truncated in order to determine the minimum polypeptide length needed for function and assembly. A stop codon was introduced into the Arg-50, Arg-59, or Tyr-69 codons of the psbE gene, generating mutants truncated by 31, 22, and 12 residues, respectively. Removal of 12 residues caused a decrease of 20% in PSII function. Truncation of 22 or 31 residues caused a large decrease (60-85%) in the photoautotrophic growth rate, the rate of O2 evolution, and the amplitude of the 77 K 696-nm fluorescence, and a concomitant increase in the constant yield fraction (F0/Fmax) of the chlorophyll fluorescence. The level of residual activity in the Arg50-stop mutant was 10-20% of the wild type, which was reflected in a similar low level of immunochemically detected D2 polypeptide. Quantitation of the PSII reaction center stoichiometry of the Arg50-stop mutant by analysis of [14C]DCMU binding also showed a 5-fold decrease (1:910 Chl in wild type and 1:5480 Chl in R50) in the PSII reaction center concentration. However, the KD value for DCMU in the residual 15% of the complexes to which it bound was approximately equal to that (25 nM) of the wild type. Northern blot analysis showed no decrease in the b559 psbE mRNA level. Chemical difference spectral analysis of heme content indicated that the level of native cytochrome b559 heme in the Arg50-stop mutant (1:640 Chl) was 80% that of wild type (1:510 Chl).(ABSTRACT TRUNCATED AT 250 WORDS)

In Vitro Release of Doxycycline From Bioabsorbable Materials and Acrylic Strips

Treatment of marginal periodontitis may include use of local antibiotics. In the present in vitro study the bioabsorbable materials Surgicel, Tissell, and CollaCote and acrylic strips were examined for release of doxycycline into liquids and residual antibacterial activity of the materials. Pieces of material were incubated in 1 ml of water or human serum which was exchanged every day for 9 days. The concentration of doxycycline in the liquids was measured spectrophotometrically while the residual antibacterial activity of the materials was determined microbiologically. Surgicel constantly showed very high concentrations and level of residual activity. The acrylic strip and CollaCote decreased to low levels of both concentration and residual activity in a few days. Tissell was intermediate with a continuous decrease in concentration but rather high level of residual activity throughout the study. In serum the acrylic strip was partly dissolved and Surgicel was totally dissolved after 2 days. Left undisturbed in serum Surgicel was not dissolved for 2 weeks. These results indicate that Surgicel and Tissel may be capable of prolonged release of doxycycline in vivo.

Radionuclide transit in patients with colon interposition

To assess radionuclide transit in interposed segments of the colon, we examined 25 patients with colon interposition for benign esophageal disease. No such assessment has been reported previously. The most common indications for operation were esophageal strictures that developed after lye ingestion and reflux strictures not responding to other treatment. The operations were performed without thoracotomy by blunt esophageal dissection in 80% of the patients. There were 18 antiperistaltic and seven isoperistaltic colon grafts. A large-field gamma camera and computer system were used. Data were collected at time intervals of 0.5 second during the first 30 seconds and at intervals of 30 seconds up to 20 minutes. The 5% and 90% stomach filling times, times to 50% and 25% activity levels, and residual activity levels as a percentage of the maxima were calculated in the upper, middle, and lower thirds of the colon grafts and of the normal esophagus of 10 healthy control subjects. The examinations were performed with the subject in a sitting position. All parameters showed that emptying of the colon graft was markedly slower than that of the normal esophagus. The intra-abdominal third of the graft had a residual activity of 50.5% +/- 15.7% after 20 minutes' observation. No differences between antiperistaltic and isoperistaltic grafts were observed. Reconstruction with proximal cologastric anastomosis and a short intra-abdominal colon graft segments is suggested.

Xanthine oxidase activity in human intestines. Histochemical and radiochemical study.

Xanthine oxidase (xanthine oxygen-oxidoreductase EC 1.2.3.2.) catalyses oxidation of hypoxanthine and xanthine to uric acid. In man, small intestines and liver are the only tissues that normally show abundant xanthine oxidase (XO) activity. But localization, level, form of catalytic activity and physiological function in the different tissues are not clearly known. In xanthinuria, characterized by a gross deficiency of XO in the tissues it is not known whether the variations in the level of residual activity of XO represent methodologic limitations or heterogeneity of the genetic and molecular defect.

Galactose-1-phosphate uridyltransferase activities in erythrocytes from a patient with galactosemia: Discrepancy between two methods

When measuring with the spectrophotometric UDP-Glu consumption test, the galactose-1-phosphate uridyltransferase (Gal-PUT) activity in erythrocyte lysates from a 22-month-old infant with a late onset form of galactosemia was found to be approximately 25% of normal. With a radiochemical assay only a very low residual activity could be detected (± 1% of normal). Preincubation of the patient's lysate with purified NADase caused a marked decrease of residual Gal-PUT activity as judged from the data obtained with the consumption test. The radiochemical assay was not influenced by a similar pre-treatment. The high level of residual activity found with the consumption test in this patient was attributed to the consumption of UDP-Glu by other reactions than Gal-PUT. Because it is a direct, simple and generally applicable assay, the radiochemical procedure is suggested to be the best method for the more detailed enzymological characterisation of the Gal-PUT deficient state in galactosemics.

Adenosine Triphosphate-dependent Deoxyribonuclease in Bacillus subtilis: I. PURIFICATION AND SOME PROPERTIES

Abstract ATP-dependent deoxyribonuclease was first purified up to 580-fold from a crude extract of Bacillus subtilis W168 by ammonium sulfate fractionation, agarose and DEAE-cellulose column fractionation. Enzyme instability was reduced by use of Tris-HCl media and an in-line dialysis apparatus directly connected to a chromatographic column. During the course of the enzyme purification, an inhibitor to this enzyme was detected in the crude extract prepared by sonication and was partially purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. Based on various criteria, it was concluded that the inhibitor is fragmented DNA or DNA in complex with some other compounds. The ATP-dependent DNase of B. subtilis has relatively narrow limits of ATP concentration for its optimal activity, and there is a marked inhibition above the optimal ATP concentration (20 µm). However, a rather constant level of residual activity is found above 100 µm, up to at least 1 mm ATP. The enzyme also requires Mg2+ for its activity; Km for this is 1.02 mm. The ATP-dependent DNase of B. subtilis has unusal substrate specificities for native and denatured DNA. The enzyme (Fraction VI) requires ATP to hydrolyze native DNA, but is not at all necessary for the hydrolysis of denatured DNA. The activity on the native DNA with optimal concentration of ATP (20 µm) was 3-fold greater than on the denatured DNA with or without ATP, while at a higher ATP concentration (500 µm), the factor was 0.5-fold. Among chemical inhibitors tested, β,γ-methyleneadenosine triphosphate inhibited the enzyme activity with native DNA as a substrate, while with denatured DNA the activity remained unaltered. The possibility of a contaminating enzyme which would hydrolyze single strand DNA preferentially appears unlikely because of the coincidence of the two activities on both agarose and DEAE-cellulose column chromatographies, together with the salt inactivation behavior. A glycerol gradient centrifugation of the above partially purified enzyme raised the purity to approximately 4800-fold. This procedure, however, made a drastic change in the property of the enzyme. Thus, the ratio of activities on native DNA to denatured DNA at 20 µm ATP increased from 3 in Fraction VI to more than 30. A possibility of removing a contaminating enzyme specific to denatured DNA as the substrate was made unlikely. DNA-dependent adenosine triphosphatase activity was found inseparable from the purified enzyme.

RESIDUAL ACTIVITY OF THERMALLY DENATURED TRANSFORMING DEOXYRIBONUCLEIC ACID FROM HAEMOPHILUS INFLUENZAE.

Barnhart, Benjamin J. (Johns Hopkins University School of Hygiene and Public Health, Baltimore, Md.). Residual activity of thermally denatured transforming deoxyribonucleic acid from Haemophilus influenzae. J. Bacteriol. 89:1271-1279. 1965.-The level of residual transforming activity of heated deoxyribonucleic acid (DNA) (i.e., 1 to a few per cent of native DNA-transforming activity) was found to be independent of the heating and quenching temperatures and less susceptible than native or renatured DNA to heat inactivation upon prolonged heating above or below the critical melting temperature. Similar dose-response curves were obtained for inactivation by formamide of native and renatured DNA, but the residual-active material was much more resistant. Heating DNA above the T(m) in the presence of 1% formaldehyde resulted in a level of residual activity 4 logs lower than that obtained without formaldehyde. Residual-active material was not inactivated by Escherichia coli phosphodiesterase, but it was susceptible to snake venom phosphodiesterase. A new genetic marker was induced in heated-quenched DNA but not in purified residual-active material following nitrous acid treatment. Residual activity was found to be less susceptible to ultraviolet inactivation and to band at a higher density region in CsCl than native DNA. In conclusion, it is suggested that the residual-active material is a structure formed by intrastrand hydrogen bonding of the separated units of heated-quenched DNA. Such a configuration would result in at least a partially double-stranded structure, which is probably the essential characteristic of the residual-active material endowing it with biological activity.