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At least 271 records · Page 15Linked to original sources
PubMed2002

Healing of alveolar bone in resorbable and non-resorbable membrane-protected defects. A histologic pilot study in dogs.

Two main types of membrane barriers are used for bone regeneration, non-resorbable and resorbable. Polytetrafluorethilene non-resorbable membranes have been extensively studied but they require a second surgical step for removal. Although polylactic acid (PLA) resorbable membranes avoid this problem, they have not been sufficiently evaluated on bone defects. The purpose of this pilot study was to compare the healing events of bone regeneration after placement of non-resorbable or resorbable membranes and to evaluate the amount of newly formed bone 2 and 4 months after membrane placement. Mandibular second, third and fourth premolars of four adult mongrel dogs were extracted bilaterally. Two rectangular bone defects (8 mm corono-apical and 12 mm mesial-distal) were created bilaterally 3 months after tooth extractions. Each dog received two resorbable membranes and one non-resorbable membrane; one defect was left untreated. Two dogs were killed at 2 months and the remaining two at 4 months following surgery. Undecalcified sections were obtained and stained with toluidine blue and pyronin G. Histomorphometric analysis was performed using the NIH Image software. Newly formed bone was observed under both resorbable and non-resorbable membranes. The amount of regenerated bone was similar between both treatments at 2 and 4 months after surgery. At 2 months, the newly formed bone was still immature whereas at 4 months some areas of woven bone were observed. The bone formation observed in the untreated defects was significantly lower than that observed in both resorbable and non-resorbable membrane-protected defects. In summary, the present study suggests that PLA membranes can yield good results when used on bone defects while avoid a second surgical procedure.

Animals↗
PubMed2001

Differences in cell cycle kinetics of candidate engrafting cells in human bone marrow and mobilized peripheral blood.

Patients undergoing hematopoietic stem cell transplantation (HSCT) with mobilized peripheral blood (MPB) engraft quicker than those receiving bone marrow (BM). Our objective was to determine whether candidate engrafting cells--primitive hematopoietic progenitors (PHPs)--from MPB and BM exhibit different responses to cytokines that could explain this observation. We compared the cell cycle kinetics and ex vivo expansion of PHP-enriched cells obtained from MPB (n = 12) and BM (n = 10) by fluorescence-activated sorting of CD90+, AC133+ or CD38(dull) subsets of pre-selected CD34(+) cells. Cell cycle status, before and after 40 hours of serum-free culture with a cytokine cocktail, was assessed by multiparameter flow cytometry following incubation with Hoechst 33342 and pyronin Y. We found that 0.2% +/- 0.3% of MPB CD34(+)CD90(+) cells were in S/G(2)/M phases at hour 0, compared with 5% +/- 2.5% of those from BM (p = 0.0001), and 86.3% +/- 9.7% were in G(0), compared with 65.3% +/- 10% of those in BM (p = 0.0001). After 40 hours of culture, CD34(+)CD90(+) cells from MPB were more mitotically active than those from BM, with 29% +/- 4.9% in S/G(2)/M and 20% +/- 11.4% in G(0), compared to 19% +/- 6.5% (p = 0.001) and 39.2% +/- 22% (p = 0.027) of cells from BM. There was greater expansion of both total CD34(+) cells and the CD90(+) subset from MPB samples (p = 0.001 and 0.0001, respectively). Results from PHPs defined on the basis of AC133 expression correlated well with results obtained in CD90(+) subsets (r(2) = 0.81; p = 0.014).MPB PHPs appear to be primed for a greater acceleration in mitotic activity upon cytokine exposure. This qualitative difference may contribute to the earlier engraftment seen after HSCT using MPB grafts.

AC133 Antigen↗
PubMed1997

Use of an in vitro infectivity assay in comparison with histological techniques in the study of Theileria parva sporozoite maturation.

Adult male and female Rhipicephalus appendiculatus ticks infected with Theileria parva (Muguga 3087) were fed on rabbits and the development of infection was monitored daily using light microscopy and an in vitro titration technique able to quantify the infectivity of sporozoite suspensions. The salivary glands stained with methyl green pyronine showed presence of infection in some unfed ticks. The intensity of staining was shown to increase with the number of days the ticks had fed. The in vitro technique, on the other hand, could detect infection only in ticks which had fed for 3-5 days. Feeding of ticks on rabbits for 4 days produced significantly more sporozoites than any other lengths of feeding (P = 0.001). The in vitro assay was also able to demonstrate differences between male and female R. appendiculatus in production of infective sporozoites. Female ticks produced significantly more sporozoites than male ticks (P = 0.002).

Analysis of Variance↗
PubMed1990

Laryngeal epithelial changes on effects of smoking and drinking.

The human larynx is essential not only for speech, but also for swallowing and respiration. Its tissues are affected by the presence of alcohol and cigarette smoke. In this report we present our findings on the effects of smoking and drinking on histological changes in laryngeal tissue. The larynges studied were those of autopsy patients: cases with damaged tissue were excluded. We studied the larynges of 84 men (non-smoker and non-drinker, 22 cases; smoker and drinker, 62 cases) and 48 women (non-smoker and non-drinker, 40 cases; smoker and drinker, 8 cases) between the ages of 12 and 88. The squamous metaplasia of laryngeal epithelium was examined using the gross staining method (Pyronin Y) of STELL et al. (J. Laryngeal. Otol. 86: 589-594, 1972). The squamous epithelium is unstained, whereas the respiratory epithelium is a brilliant red. The area of metaplasia in the supraglottic region increased with aging, tobacco and alcohol consumption. Microscopic changes of the laryngeal epithelium were investigated using the hematoxylin-eosin staining method. The thickness of epithelium of the supraglottic region and vocal cord increased with age. There is a significant difference in the thickness of the epithelium of the supraglottic region and vocal cords of smoking and drinking patients, when compared with the same tissues of non-smokers and non-drinkers. Furthermore, there is a significant difference in those tissues of the supraglottic region of heavy smokers, when compared with light smokers, but no significant difference when compared with drinkers. However, there is no significant difference in the thickness of vocal cord tissues when comparing light smokers and drinkers on the one hand, and heavy smokers and drinkers on the other.

Adolescent↗
PubMed1998

Tyrosine hydroxylase- and/or aromatic L-amino acid decarboxylase-containing cells in the suprachiasmatic nucleus of the Syrian hamster (Mesocricetus auratus).

Catecholamines, including dopamine (DA), affect the activity of cells in the suprachiasmatic nucleus (SCN) of the hypothalamus, the principal circadian clock in mammals. This study examined the distribution of dopaminergic cells in the SCN of the male Syrian hamster, using both single- and double-label immunocytochemistry for tyrosine hydroxylase (TH), the rate-limiting enzyme in DA synthesis and for aromatic L-amino acid decarboxylase (AADC), the second enzyme needed to produce DA. Some neurons immunopositive for TH (TH + ) were found in the SCN, but most of the TH + cells of the region were located just outside the borders of the nucleus, as defined by pyronin Y staining. In the SCN, 91% of these cells were also immunopositive for AADC and thus, likely to be dopaminergic. Cells positive for AADC, many of which were not TH +, were found throughout the SCN, with the highest concentration seen in the ventral aspects of the nucleus. Cells containing AADC, but lacking TH may synthesize products other than DA, such as trace amines. These anatomical observations suggest that local neurons that produce DA and perhaps trace amines, may play a role in SCN function and in the neural control of circadian rhythms.

Animals↗
PubMed2003

Phenylpiperidine selective serotonin reuptake inhibitors interfere with multidrug efflux pump activity in Staphylococcus aureus.

Structural variants of phenylpiperidine selective serotonin reuptake inhibitors (P-SSRIs) inhibited the function of two unique Staphylococcus aureus multidrug efflux pumps. The most active compound was the paroxetine isomer NNC 20-7052, which had an IC(50) for ethidium, acriflavine, and pyronin Y efflux of 9, 53, and 18% of its MIC, respectively, against the NorA pump. The unbalanced effect of NNC 20-7052 on the efflux of different substrates suggests the possibility that P-SSRIs function by a physical interaction with NorA. Under the conditions employed pump inhibition partially extended to the resistance-nodulation-division (RND) pump AcrAB-TolC, but not to the Pseudomonas aeruginosa RND pumps MexAB-OprM or MexCD-OprJ.

Biological Transport, Active↗
PubMed1981

Monitoring Theileria parva infection in adult Rhipicephalus appendiculatus ticks.

A rapid method is described for preparing and staining salivary glands of Rhipicephalus appendiculatus ticks infected with Theileria parva. The technique, involving the use of a modified methyl green pyronin stained minimizes the risk of losing material and allows examination of stained glands within minutes of preparation. The technique was applied in a series of studies in which ticks were either infected with T. parva under different conditions, or maturation of parasites in adult ticks was stimulated by different means. When nymphal ticks were fed on the ears of cattle the subsequent infection rate of the adult ticks showed no correlation with the parasitaemia of the cattle at the time of nymphal engorgement. There was no difference in infection rates between adult ticks in which parasite maturation had been stimulated either by incubation at 37 degree C or by feeding on rabbits. However, parasite maturation took about 1 day longer in incubated ticks than in rabbit-fed ticks. Female ticks were consistently more highly infected than males, both in terms of the percentage of ticks infected and the mean number of infected acini/tick. Ticks were infected with T. parva by injection of nymphs with parasitaemic bovine blood, but the resultant adult infection was lower than that in ticks which had been infected naturally by feeding on cattle.

Animals↗
PubMed1998

Theileria lestoquardi--maturation and quantification in Hyalomma anatolicum anatolicum ticks.

The maturation and quantification of Theileria lestoquardi (T. hirci) parasites in unfed and partially fed adult Hyalomma anatolicum anatolicum ticks was studied using (1) methyl green pyronin (MGP) staining of salivary glands, (2) in vitro infection of peripheral blood mononuclear cells (PBM) with parasites harvested from infected ticks and (3) a semi-quantitative polymerase chain reaction (PCR). With MGP staining the greatest infection rate was seen in unfed ticks. Feeding resulted in a gradual reduction in the number of infected acini with a concomitant increase in the maturity of the parasites. In vitro infection of sheep PBM with titrated group-up tick supernate (GUTS) demonstrated that infectivity peaked between 2 and 4 days of tick feeding whereas GUTS prepared from unfed ticks was not infective. The polymerase chain reaction (PCR) was both sensitive and specific, detecting T. lestoquardi DNA in unfed and partially fed ticks, with a maximum sensitivity of 0.022 infected acinus/tick in 2-day fed ticks, though it gave no indication of the infectivity of the parasite.

Animals↗
PubMed2002

Environmental remediation by an integrated microwave/ UV-illumination method. 1. Microwave-assisted degradation of rhodamine-B dye in aqueous TiO2 dispersions.

The photocatalytic decomposition of the cationic rhodamine-B (RhB) dye was examined in aqueous TiO2 dispersions using an integrated microwave/UV-illumination (PD/MW) method. This procedure proved to be superior in the degradation of the dye than the TiO2 photocatalytic degradative method alone. With few exceptions, the integrated PD/MW method also proved superior for other chemical systems. The greater efficacy of the PD/MW technique appears to be the result of the following two considerations: (i) there is enhanced formation of reactive oxygen species (.OH radicals), as attested to by DMPO spin-trap ESR methods and their attack on the dye; and although speculative at this time, (ii) the activity of bulk water or the TiO2 particle surface is somehow affected by microwave radiation. The greater efficacy of the PD/MW degradation method was also observed at low concentrations of molecular oxygen and at low radiant excitance of the light source. A brief mechanistic description is given on the basis of results obtained on the two model compounds, (i) benzoic acid and (ii) pyronin-B dye.

Coloring Agents↗
PubMed2006

Transfer of electronic excitation energy between randomly mixed dye molecules in the channels of zeolite L.

Host-guest materials containing strongly fluorescent donor and acceptor molecules have been prepared. Fine-tuning of the donor to acceptor distance in this material allows beautiful visible and quantitative observation of electronic excitation energy transfer phenomena. Oxonine and pyronine have been used as guest molecules and zeolite L as host. The dyes have been inserted by ion exchange. Stationary state and time-resolved experiments have been carried out with zeolite crystals of 300 and 700 nm size in the dye concentration range of 10(-4) mol/L up to 0.042 mol/L. The fluorescence decay of the donor and the pumping of the acceptor via energy transfer, which can be well observed, became faster with increasing loading. The behavior of the system follows requirements expected for Förster energy transfer material.

Journal Article↗
PubMed2002

On the mechanism of substrate specificity by resistance nodulation division (RND)-type multidrug resistance pumps: the large periplasmic loops of MexD from Pseudomonas aeruginosa are involved in substrate recognition.

Tripartite efflux systems of Gram-negative bacteria that contain an inner membrane transporter belonging to the resistance nodulation division (RND) superfamily can extrude a large variety of structurally diverse compounds. To gain an insight into the molecular mechanisms of substrate recognition by these multidrug resistance (MDR) transporters, we isolated spontaneous mutations that altered the substrate specificity of the MexCD-OprJ pump from Pseudomonas aeruginosa. These mutations enabled the pump to extrude the normally non-transported beta-lactam antibiotic carbenicillin. All amino acid substitutions were mapped to the large periplasmic loops (LPLs) of the RND proper, MexD. Q34K, E89K, A292V and P328L were found in the first LPL, located between transmembrane domains (TMD) 1 and 2, whereas F608S and N673K were contained in the second LPL, located between TMD7 and TMD8. These mutations also had a substantial impact on the MexCD-OprJ-mediated transport of numerous other substrates. Subsequent replacement of amino acid residues identified above by cysteines rendered MexCD-OprJ susceptible to inhibition by a thiol-reactive agent, MIANS. Interestingly, MIANS inhibited the transport of some (pyronin, EtBr) but not other (ANS, Leu-Nap) substrates of the pump. Our results suggest that the precise structure of the periplasmic loops of MexD determines the rate of transport of individual substrates. These results are consistent with the hypothesis that, in the case of RND transporters, the LPLs are directly implicated in substrate recognition and contain multiple sites of interaction for various structurally diverse compounds.

Amino Acid Substitution↗
PubMed1999

Bioenergetics of the staphylococcal multidrug export protein QacA. Identification of distinct binding sites for monovalent and divalent cations.

The multidrug efflux pump QacA from Staphylococcus aureus confers resistance to an extensive range of structurally dissimilar compounds. Fluorimetric analyses demonstrated that QacA confers resistance to the divalent cation 4',6-diamidino-2-phenylindole, utilizing a proton motive force-dependent efflux mechanism previously demonstrated for QacA-mediated resistance to the monovalent cation ethidium. Both the ionophores nigericin and valinomycin inhibited QacA-mediated export of ethidium, indicating an electrogenic drug/nH+ (n >/= 2) antiport mechanism. The kinetic parameters, Km and Vmax, were determined for QacA-mediated export of four fluorescent substrates, 4',6-diamidino-2-phenylindole, 3', 3'-dipropyloxacarbocyanine, ethidium, and pyronin Y. Competition studies showed that QacA-mediated ethidium export is competitively inhibited by monovalent cations, e.g. benzalkonium, and non-competitively inhibited by divalent cations, e.g. propamidine, which suggests that monovalent and divalent cations bind at distinct sites on the QacA protein. The quaternary ammonium salt, 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene, was used as a membrane-specific fluorescence probe and demonstrated that the amount of substrate entering the inner leaflet was significantly reduced in QacA-containing strains, supporting the notion that the substrate is extruded directly from the membrane.

Bacterial Proteins↗
PubMed1996

Abnormal cutaneous nerve outgrowth: a secondary effect of methyl triazene teratology in developing mouse limb buds.

The cutaneous branches originating from the superficial distal division of the ulnar nerve exhibit abnormal developmental features in 70.2% of the forelimb buds from embryos submitted 2 or 3 days earlier to methyl triazene administered to their pregnant mother. Similar abnormalities characterize the preaxial ventral cutaneous nerve of the thumb in 17.8% of forelimb primordia. The affected nerves undergo anticipated growth with respect to the normal schedule, follow abnormal pathways through areas of extensive cell death, and finally reach the apical ectoderm where they run in close contact with the basement membrane without forming a plexus. Histological observations gathered in pyronin-methyl green stained serial sections as well as in whole limb buds after cholinesterase method suggest that three factors probably contribute to modify nerve outgrowth: (1) a discrepancy between the rate of nerve progression and that of mesodermal growth in the prospective zeugopod territory which is preferentially affected by the teratogen; (2) facilitated nerve pathfinding into areas strongly hit by triazene-induced mesodermal cell death; and (3) alteration or abolition of some unknown ectodermal influence necessary to stimulate selective guidance of terminal sensory afferents and to maintain them transiently at some distance from the epidermis.

Alkylating Agents↗
PubMed1984

The development of Theileria annulata in the salivary glands of the vector tick Hyalomma anatolicum anatolicum.

The development at 28 degrees C of Theileria annulata (Hissar) in the salivary glands of its tick vector, Hyalomma anatolicum anatolicum, was studied using Giemsa-stained smears, methyl green-pyronin-stained preparations of whole glands, and electron microscopy. Nymphs which had engorged on T. annulata-infected calves showed kinetes in the haemolymph from Day 7 to Day 14 post engorgement, when all ticks had completed moult. Intracellular sporonts were observed within salivary gland acini from Day 7 onwards and these developed by rapid nuclear division and cytoplasmic proliferation to form primary sporoblasts. Further development was stimulated by feeding on a rabbit or by incubation at 36 degrees C. The primary sporoblasts appeared to become organized into membrane-bound subunits. Within 48 hours of attachment to the host, or 72 hours of 36 degrees C incubation, these units dissociated to form secondary sporoblasts. The final phase of development resulted in the progressive formation of discrete, uninuclear sporozoites within these secondary sporoblasts. No morphological differences were observed in parasite maturation between the fed and incubated groups although development was retarded by at least 24 hours in the latter. In the incubated group there was also a marked decrease in the degree of synchrony of development which resulted in fewer sporozoites being present at any one time.

Animals↗
PubMed1985

The presence of Theileria sergenti in Haemaphysalis longicornis overwintering in pasture in Japan.

Unfed Haemaphysalis longicornis ticks were collected from the soil of pasture during the winter and their salivary glands were examined by the methyl green pyronin staining method in order to detect overwintering of Theileria sergenti, the causative agent of bovine theileriosis in Japan. A transmission experiment was carried out using these ticks applied to a splenectomized calf. The protozoa which appeared in the peripheral blood were identified parasitologically and seroimmunologically as T. sergenti. The results suggested that T. sergenti in Japan might overwinter in H. longicornis which moulted after repletion on infected cattle in the autumn and hid in the soil during the winter. Moreover, it seemed possible that T. sergenti might develop to the infective stage in H. longicornis during the overwintering period.

Animals↗
PubMed1989

Correlation between the infection rate of the vector tick, Haemaphysalis longicornis and the parasitaemia of cattle infected with Theileria sergenti.

The infection rate of Theileria sergenti in salivary glands of nymphal Haemaphysalis longicornis which had dropped from cattle showing different piroplasm parasitaemias was examined by the methyl green-pyronin staining method. The results suggested that there was some correlation between parasitaemia and the subsequent infection rate in salivary glands of ticks. It seemed possible that the parasitaemia in cattle, especially the number of parasitized erythrocytes in blood imbibed by ticks, might be an important influence on the number of infected acini of ticks.

Animals↗
PubMedDate not supplied

The mast cells in semen: their effects on sperm motility.

This study was conducted to evaluate any possible association between mast cells and sperm concentration, morphology, and motility. The study comprised 400 patients who had applied for semen analysis. To evaluate mast cells, 6 smear slides were prepared for each subject and stained with 1% toluidin blue-pyronine (pH 4). The slides revealing any mast cells were labeled as mast+. Concentration and motility was evaluated through a Makler chamber. Kruger's strict criteria were used in morphometric analysis. The mean age of 86 mast+ cases (21% of total patients) was 31+/-6.7; progressive sperm motility rate was 33+/-21.2. The mean concentration was 32+/-30.2 x 10(6)/mL, and normal sperm percentage was 11.8+/-6.5. Progressive sperm motility rate in the mast- cases were 53+/-25. The mean age of mast cell+ patients was higher than that of mast cell- patients (t=3.57, p<.001), while they had lower sperm concentration (p>.05) and lower normal morphologic sperm rate (t=2.26, p<.024), compared to mast cell- patients. The relation between mast cell+ and mast cell- cases and sperm progressive motility was statistically significant (t=6.44, p<.001). It was concluded that sperm parameters were negatively affected by mast cells.

Adolescent↗
PubMed1985

Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry.

Quantitative, correlated determinations of DNA, RNA, and protein, as well as RNA to DNA and RNA to protein ratios, were performed on three-color stained cells using a multiwavelength-excitation flow cytometer. DNA-bound Hoechst 33342 (blue), protein-fluorescein isothiocyanate (green), and RNA-bound pyronin Y (red) fluorescence measurements were correlated as each stained cell intersected three spatially separated laser beams. The analytical scheme provided sensitive and accurate fluorescence determinations by minimizing the effects of overlap in the spectral characteristics of the three dyes. Computer analysis was used to generate two-parameter contour density profiles as well as to obtain numerical data for subpopulations delineated on the basis of cellular DNA content. Such determinations allowed for analysis of RNA to DNA and RNA to protein ratios for cells within particular regions of the cell cycle. The technique was used to study the interrelationship of DNA, RNA, and protein contents in exponentially growing Chinese hamster ovary cells as well as in cell populations progressing the cell cycle after release from arrest in G1 phase. The sensitivity of the method for early detection of conditions of unbalanced growth is demonstrated in the comparison of the differential effects of the cycle-perturbing agent, adriamycin, on cells treated either during exponential growth or while reversibly arrested in G1 phase.

Animals↗