Search PubMed⌕ Search

Biomedical subjects

P Talbot

Publications and source records attributed to P Talbot.

At least 37 records · Page 2Linked to original sources
PubMed1998

Identification of cigarette smoke components that alter functioning of hamster (Mesocricetus auratus) oviducts in vitro.

Previous studies have shown that solutions of mainstream and sidestream cigarette smoke decrease both ciliary beat frequency and oocyte cumulus complex pick-up rate of hamster oviducts in vitro. The purpose of this study was to identify the component(s) in smoke that produces these effects. Chemicals reported to be ciliotoxic in other systems were tested on hamster infundibula at various concentrations to determine whether a dose-response inhibition of ciliary beat frequency occurred. In addition, the concentration of each test component was measured in mainstream and sidestream smoke solutions (whole, particulate, and gas phase). All test components (acrolein, formaldehyde, phenol, acetaldehyde, and potassium cyanide [KCN]) inhibited ciliary beat frequency in a dose-dependent manner. Inhibition of ciliary beat frequency was at least partially reversible for all test compounds except acrolein. The concentrations of acrolein, formaldehyde, and phenol that were required to inhibit beat frequency were at least 3-50 times higher than their corresponding concentrations in smoke solutions. In contrast, cyanide was present in all smoke solutions at concentrations sufficient to inhibit ciliary beat frequency. Cilia on the outer surface of the infundibulum function in picking up the oocyte cumulus complex. Oocyte cumulus complex pick-up rate was measured in vitro at KCN concentrations shown to inhibit ciliary beat frequency. Pick-up rate was likewise inhibited by KCN and remained significantly depressed after washout of KCN. These data show that cyanide is a potent inhibitor of both ciliary beat frequency and oocyte cumulus complex pick-up rate and that its concentration in smoke solutions is sufficiently high to explain the previously reported inhibition of these oviductal processes.

Acetaldehyde↗
PubMed1997

In vitro analysis of oocyte cumulus complex pickup rate in the hamster Mesocricetus auratus.

In mammals, the oocyte and its surrounding cumulus cells constitute on oocyte cumulus complex (OCC). During ovulation, OCCs are extruded into the peritoneal or bursal cavity, depending on the species, and are then rapidly picked up by the fimbria on the outer surface of the oviductal infundibulum and transported to the ampulla, where fertilization occurs. We developed a method to measure OCC pickup rates quantitatively in vitro, and we used this method to evaluate the effects of viscosity and temperature on pickup rates. Hamster infundibula are placed in a holding pipette in a chamber modified to study OCC pickup. Ciliary beat frequencies (CBF) can be measured in the same preparation. Pickup rates vary depending on the pathway on which the OCC travels over the surface of the infundibulum; however, rates for a given pathway are very consistent. The average pickup rate at room temperature calculated from three different pathways/infundibulum was 55.2 +/- 10.6 microns/sec. Both rates between infundibula from the same female and rates among infundibula from different females were in most cases similar. Preparations preincubated in vitro for 2.75 hr produced rates similar to nonpreincubated samples, while longer preincubation resulted in decreased rates. Inclusion of Ficoll in culture medium to increase viscosity caused a concentration-dependent decrease in both OCC pickup rate and in CBF. However, a significant decrease in OCC pickup rate was only observed at viscosities higher than those found in bursal fluid. When trials were run at physiological temperature (36.4 degrees C) rather ambient temperature, rates increased to 136.7 +/- 29.9 (SD) microns/sec. Linear regression analysis demonstrated a strong positive correlation (r = 0.94) between OCC pickup rate and temperature. The OCC pickup rate assay can be used experimentally, and should be valuable in evaluating factors that affect rate and in studies dealing with the mechanism of OCC pickup.

Animals↗
PubMed1997

Intensive nutritional counselling in bulimia nervosa: a role for supplementation with fluoxetine?

OBJECTIVE: The aims of the paper are to determine whether nutritional counselling is associated with an improvement in bulimic symptomatology, whether this improvement is maintained during post-treatment follow-up, and whether the addition of fluoxetine 3 x 20 mg/day confers additional benefit. METHOD: Psychological, pharmacological and combined psychopharmacological treatments of bulimia nervosa were reviewed briefly. Sixty-seven patients referred to specialist eating disorder services who fulfilled strict diagnostic criteria were treated with intensive nutritional counselling and randomly assigned to either fluoxetine 3 x 20 mg/day or placebo. After a 1-week 'wash-out', active treatment was given over 8 weeks, followed by post-treatment interviews at 12 and 20 weeks. RESULTS: Both groups of patients improved significantly during treatment. In some respects, the fluoxetine group did slightly better as demonstrated by the items 'restraint', 'weight concern' and 'shape concern' (p < 0.05 vs p < 0.0001) on the Eating Disorder Examination (EDE). Fluoxetine patients decreased their energy intake and lost a modest amount of weight. They went on to regain weight during the follow-up period, returning to levels higher than they were initially. These patients also appeared more likely to have a recurrence of symptoms, as shown by the fall in percentage of binge-free patients and by changes in the EDE. CONCLUSION: Nutritional counselling is an effective means of treating bulimia nervosa, with improvement maintained up to 3 months follow-up. The addition of fluoxetine may confer some benefit during active treatment, but its discontinuation may contribute to a higher rate of recurrence of symptoms post treatment. Of course, this study cannot be extrapolated to the efficacy of fluoxetine when used as the only form of treatment in patients for whom intensive nutritional counselling or other structured psychological programs are not available.

Adolescent↗
PubMed1996

Multiple lobster tubulin isoforms are encoded by a simple gene family.

Microtubule proteins isolated from pleopod tegumental gland (PTG) tissue of the American lobster, Homarus americanus, reveal a complex tubulin (Tub) profile. To determine whether Tub heterogeneity in PTG is due to expression of a large tub gene family or the result of post-translational modification, a PTG cDNA library was constructed. Clones coding for both alpha- and beta-Tub were isolated, sequenced and found to contain open reading frames (ORFs) of 451 amino acids (aa). Alignments reveal phylogenetic clustering with other arthropods and identify unique changes in primary structure which may have functional significance. These clones, when used to probe restriction enzyme-digested lobster genomic DNA in transfer-hybridization experiments, revealed a simple banding pattern indicating a lobster tub gene family of limited complexity. Lobsters appear to make use of a small tub gene family and fulfill the varied functional requirements imposed upon cellular microtubules through post-translational modifications of relatively few gene products.

Amino Acid Sequence↗
PubMed1995

Analysis of ciliary beat frequencies in hamster oviducal explants.

We have developed a simple direct method, requiring minimal manipulation, to measure beat frequencies of the cilia on the external surface of hamster oviducal infundibula in vitro. Two perfusion chambers (closed and open) were used; both can be hand-made in a few minutes and discarded after use. Ciliary beat frequencies were determined by measuring variations in light intensity with time in a single pixel positioned over a video image of the beating cilia. Data files were collected using Image 1 software and later transferred to PSI Plot or Lotus 123 spreadsheets for analysis by counting the number of brightness peaks recorded per second or by subjecting the data to Fourier transformation with or without smoothing. These methods of analysis gave similar results. To verify that Image 1 data files contain accurate representations of CBF, videotapes of beating cilia were made and subjected to frame-by-frame analysis. Image 1 interfaced with a standard video camera was found to collect reliable data over a beat frequency range of 0-15 cycles/sec. In some Fourier transforms, secondary peaks were observed and were shown to represent cilia beating at more than one frequency in a sampled region. Coefficients of variation for repeated measurements taken on the same region varied from 4.1% to 9.0%. Small but significant differences were found between beat frequencies at different regions of the same oviduct. When chambers were perfused discontinuously and measurements of beat frequency were made at least 5 min after each perfusion, no effect of perfusion on frequencies was observed. However, during continual perfusion of the open chamber, a slight but significant increase in beat frequency was observed after perfusion was initiated. Muscle contraction, which sometimes occurs in the open chamber, did not affect beat frequency measurements. Infundibula could be stored at 4 degrees C overnight without any negative effect on beat frequencies. Cold storage also reduced muscle contraction. Placement of a small coverslip on infundibula in the open chambers was also found to reduce muscle contraction and facilitate beat frequency measurements. Coverslipping did not affect beat frequencies. This method of beat frequency analysis will be valuable for analyzing factors that regulate or influence cilia in mammalian oviducts.

Analysis of Variance↗
PubMed1995

Perivitelline space of marsupial oocytes: extracellular matrix of the unfertilized oocyte and formation of a cortical granule envelope following the cortical reaction.

The purpose of this study was to characterize the structure of the vestments surrounding unfertilized and cortical granule-reacted oocytes from a marsupial, the grey short-tailed opossum Monodelphis domestica and to determine if a cortical granule envelope (CGE) forms in the perivitelline space (PVS) following the cortical reaction. Unfertilized oocytes collected from mature ovarian follicles and oviducal oocytes that had undergone a cortical reaction were fixed for electron microscopy in the presence of ruthenium red which stabilizes extracellular matrices (ECM) and facilitates demonstration of a CGE. Unfertilized oocytes were surrounded by a zona pellucida and had a PVS which contained a thick ECM comprised of granules and filaments. This matrix appeared to attach to the oolemma and was structurally similar to matrices reported previously in the PVS of unfertilized oocytes from eutherian mammals and two other marsupials, the Virginia opossum and the fat-tailed dunnart. The cortex of unfertilized oocytes contained cortical granules which were absent in oocytes recovered from the oviducts of mated females. Oviducal oocytes which lacked cortical granules exhibited a new coat within the PVS between the zona pellucida and the tips of the oocyte microvilli. This coat, the CGE, appeared structurally similar to CGEs described previously around fertilized eutherian oocytes. The CGE of the grey short-tailed opossum is approximately 1 microns thick and is made up of numerous small dense granules. The coats of the opossum oocyte are compared to those present around other marsupial and eutherian oocytes.

Animals↗
PubMed1995

Ciliary beat frequency of hamster oviducts is decreased in vitro by exposure to solutions of mainstream and sidestream cigarette smoke.

Epidemiological data support a correlation between smoking and increased incidence of ectopic pregnancy, yet the causal mechanism responsible for this relationship is unknown. The purpose of this study was to examine the effect of solutions containing dissolved mainstream (MS) or sidestream (SS) cigarette smoke on ciliary beat frequencies (CBF) in explants of hamster oviducts. MS smoke is the puff inhaled by an active smoker, while SS smoke leaves the burning end of cigarette. SS smoke is inhaled by both active and passive smokers. Experiments were performed in handmade perfusion chambers using infundibula from hamster oviducts. After a short incubation in Earle's balanced salt solution containing HEPES buffer (EBSS-H), chambers were flushed with one of six types of smoke solution prepared in EBSS-H, and incubation continued 19 min. A second perfusion (washout) was then done using EBSS-H alone to determine whether effects induced by the smoke solutions could be reversed. CBF were determined at three times in both the smoke and washout solutions, and means were compared to values obtained in the initial EBSS-H incubation. All smoke solutions except the SS particulate solution inhibited CBF in a dose-dependent manner. Whole MS and whole SS smoke solution at the highest strength tested caused the greatest inhibition and in some cases completely stopped ciliary beating. Both single-strength and 0.1-strength MS gas phase solutions, which contained concentrations of nicotine in the range found in typical human smokers, produced about 50% inhibition of ciliary beating. Inhibition was generally seen within 2-12 min of adding smoke solutions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Mammalian cortical granules: contents, fate, and function.

Recent studies have extended our knowledge regarding the contents of mammalian cortical granules (CG) and their function in postfertilization events. Cytochemical staining has demonstrated the presence of carbohydrates within mammalian CG, and lectin-binding studies have shown that these carbohydrates include alpha-D-mannose, alpha-D-GalNAc, and galactose residues in the hamster, alpha-D-mannose in the mouse and cat, and beta-D-Gal(1,3)-D-GalNAc in the pig. Following fertilization and artificial activation, mannosylated material is released from CG and can be found on the oolemma and within the perivitelline space (PVS) of hamster oocytes. Fertilized or artificially activated rabbit, mouse, and human oocytes also release mannosylated, fucosylated and sialylated, and fucosylated material, respectively, which localizes to the oolemma. These glycosylated materials are probably of CG origin, although they have not been directly localized to the CG in rabbit, mice, and humans. The function(s) of the glycosylated material released from mammalian oocytes is not known, although it may participate in blocking polyspermy at the level of the plasma membrane, PVS, and/or zona pellucida (ZP), or it may facilitate preimplantation embryonic development. Proteinases, including tissue plasminogen activator, are also released from mammalian oocytes following fertilization and artificial activation, suggesting that they are of CG origin. These proteinases modify the ZP such that it is no longer receptive to sperm, and some proteinases have been suggested to bring about ZP hardening (an increased resistance to denaturing agents) by an unknown mechanism. Mouse ZP may also be hardened by an ovoperoxidase (cross-links tyrosine residues) cytochemically identified in mouse CG and CG exudate. The phenomena of ZP hardening in mammalian zygotes is not well understood but is likely to function in blocking polyspermic penetration of the ZP and/or in protecting embryos during preimplantation development. Recently, a 75 kD protein (p75) has been immunocytochemically localized to mouse CG and to the PVS of fertilized oocytes and two-cell embryos. The identity and function of p75 remains to be determined. Heparin binding placental protein may also be a CG component, since it is released from hamster oocytes following fertilization. It has not, however, been directly demonstrated to be a CG component, and its functions in fertilization and/or early embryonic development have yet to be defined.

Animals↗
PubMed1994

The protease of adenovirus serotype 2 requires cysteine residues for both activation and catalysis.

Sequence analysis and site-directed mutagenesis were used to study the mechanisms of activation and catalysis of the adenovirus type 2 (Ad2) protease. Primary structure alignments of proteases from 12 serotypes and previously elucidated inhibition profiles were used to target residues for mutagenesis. All conserved serine and cysteine residues were mutated separately and following expression in Escherichia coli their activity in a synthetic peptide assay was compared to that of wild-type recombinant protease. Mutants containing altered serine residues were active while mutations to cysteine-104 and cysteine-122 reduced activity by more than 95%. These results taken together with the known inhibition profile of the adenovirus protease confirm that it is a cysteine protease and suggest that one of these residues provides the active site nucleophile while the other is a part of the thiol-disulphide interchange mechanism previously reported to be involved in its activation.

Adenoviruses, Human↗
PubMed1993

Video microscopic analysis of ionophore induced acrosome reactions of lobster (Homarus americanus) sperm.

Sperm from the American lobster (Homarus americanus) are normally nonmotile. However, during fertilization, the sperm undergo a calcium-dependent acrosome reaction that propels them forward about 18 microns. The reaction occurs in two phases, eversion and ejection, which take place too quickly to permit analysis by direct observation. The purposes of this study were to examine the structural changes occurring in sperm during the normal acrosome reaction and to determine the rate of the reaction using video microscopy. The reaction was induced in vitro by ionophore A23187 and recorded using a video system attached to a Nikon Nomarski interference microscope. Videotapes were played back frame by frame (30 frames/sec), and images of reactions from 10 sperm were analyzed. The acrosome reaction, including the eversion of the acrosomal vesicle and ejection of the subacrosomal material and nucleus, can be divided into 4 steps: (1) expansion of the apical cap followed by expansion of the remainder of the acrosomal cylinder; expansion of the cylinder begins at its apical end and proceeds toward its base, (2) eversion of the apical half of the acrosomal vesicle and initial contraction of the apical cap, (3) eversion of the basal half of the acrosomal vesicle, continued contraction of the apical cap, and ejection of the subacrosomal material and nucleus, and (4) final contraction of the apical cap and ejection of the acrosomal filament. During steps 2, 3, and 4, the mean forward movement of sperm is 12.7, 3.9, and 1.1 microns, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrosome↗
PubMed1993

How well are 'cured' anorexia nervosa patients? An investigation of 16 weight-recovered anorexic patients.

Eating behaviour, nutritional intake and psychopathology were examined in 16 weight-recovered anorexia nervosa patients. The Eating Disorders Examination (EDE) and a food diary were the main outcome measures, but body fat content and psychosocial adjustment were also assessed. Although body fat and overall psychosocial adjustment had returned to normal in most patients, 12 still had a restricted eating pattern with nutritional intake below 90% of their energy requirements. These results suggest that weight, menstruation, and psychosocial criteria are not sufficient to determine full recovery from anorexia nervosa, and that attention should be given also to measures of the specific behavioural and attitudinal disturbance.

Adult↗
PubMed1993

Measuring the specific psychopathology of eating disorder patients.

Cooper and Fairburn's Eating Disorder Examination (EDE) is a semi-structured interview designed to assess the specific psychopathology of eating disorder subjects. It was employed in a study of 116 Sydney patients for 2 purposes: first, to determine its usefulness in an Australian context; and second, to compare patients with anorexia nervosa, bulimia and atypical eating disorder. The instrument appears to be quite appropriate for studies in Australia. With respect to the second aim, the results emphasize the essential similarity in psychopathology between the three diagnostic groups. The relevance of this latter finding to the categorisation of eating disorders in the DSM-IV proposals is discussed.

Adult↗
PubMed1992

Perivitelline space of mammalian oocytes: extracellular matrix of unfertilized oocytes and formation of a cortical granule envelope following fertilization.

Extracellular matrices (ECM) present around unfertilized and fertilized mammalian oocytes were studied ultrastructurally in samples prepared in the presence of ruthenium red to facilitate stabilization of extracellular materials. Unfertilized mouse, hamster, and human oocytes have an ECM comprising granules and filaments in their perivitelline spaces (PVS). This matrix is more abundant in the human than in hamsters and mice. The granule/filament matrix appears identical to the matrix seen between cumulus and corona radiata cells following ruthenium red processing and previously shown to comprise protein and hyaluronic acid. By including ruthenium red during fixation, it is possible to demonstrate the existence of cortical granule exudate in the PVS of fertilized oocytes from hamsters, mice, and humans. Much of the cortical granule exudate is trapped in the PVS and forms a new coat around the fertilized oocyte. This material is particulate when stained with ruthenium red and appears to be uniformly dispersed around the entire oocyte surface. We refer to this new coat as the cortical granule envelope. This envelope is observed in the PVS of all developmental stages up to and including blastocysts in all three species. Following hatching of mouse and hamster blastocysts, the cortical granule envelope is no longer present. Possible functions of this envelope are discussed.

Animals↗
PubMed1992

Structure, distribution and composition of the extracellular matrix of human oocytes and cumulus masses.

The structure, distribution and composition of the extracellular matrix present around the human oocyte and in the cumulus was examined following fixation in the presence of ruthenium red. An extracellular matrix comprising granules and filaments is present in the cumulus layer, in the corona radiata, in the outer pores of the zona pellucida and in the perivitelline space surrounding unfertilized oocytes. In replicate samples, the extracellular matrix comprised filaments which were mostly very long, occasionally cross-connected by shorter filaments, and usually decorated with numerous small granules. Enzymatic digestion with affinity-purified trypsin or Streptomyces hyaluronidase removes the granules and filaments, respectively, at all levels of the oocyte-cumulus complex. These results are interpreted to mean that protein and hyaluronic acid are present in all extracellular compartments of the human oocyte-cumulus complex. The significance of this distribution of hyaluronic acid with respect to the role of sperm hyaluronidase in fertilization is discussed.

Extracellular Matrix↗