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C D Ockleford

Publications and source records attributed to C D Ockleford.

At least 19 recordsLinked to original sources
PubMed2004

Healthy and pre-eclamptic placental basal plate lining cells: quantitative comparisons based on confocal laser scanning microscopy.

Immunocytochemical confocal laser scanning microscope images of the monolayer of cells lining the intervillus space at the basal plate of term placentae were analysed using stereology. Immunoreactively-distinct regions of this mosaic layer were measured. In basal plate from healthy pregnancies, trophoblast epithelium occupied 18.91% of the surface area and endothelium 60.81%. In pre-eclampsia the equivalent areas were 15.57% and 67.63%. Acellular fibrinoid covers the remaining area and this component decreases in area in pre-eclampsia. The statistically significant increase in the cellular endothelial compartment may be relevant to the hypertensive pathology of pre-eclampsia as endothelial signalling plays a major role in regulation of blood pressure.

Endothelial Cells↗
PubMed2004

Confocal laser scanning microscope study of cytokeratin immunofluorescence differences between villous and extravillous trophoblast: cytokeratin downregulation in pre-eclampsia.

Pre-eclampsia is a disease characterized by failures in interstitial implantation. One product of the implantation process is the basal plate; a structure whose complexity makes it hard to fully appreciate the pathological changes in significant diseases of pregnancy. This article describes our use of CLSM immunofluorescence to examine the cytokeratin composition of the cells of trophoblastic origin in the term placental basal plate. Large differences in the content of the structural polymeric protein were compared using analysis of digital images. We show that greater pancytokeratin immunofluorescence is observed in extravillous cytotrophoblast cells as compared with villous trophoblast. There is a >30-fold difference in the mean area percent of the most intensely immunofluorescent pixels in the tissue containing these cells. This is a very high, statistically significant difference as defined by the Wilcoxon Signed Ranks Test Asym. Sig. (two-tailed): P < 0.001. The most invasive population of cells of the trophoblast lineage (the extravillous trophoblast) exhibits a significant reduction in cytokeratin immunofluorescence when comparisons of healthy and pre-eclamptic pregnancies are made. This ratio was 2.4:1. It was tested using the Mann-Whitney U-test. From healthy to pre-eclamptic the reduction was from mean rank 83.42((healthy)) to 51.13((pre-eclamptic)). The difference was very highly statistically significant (n = 53 + 75 = 128; U = 984.500; Z = -4.852; P < 0.001). There was also less cytokeratin-related immunofluorescence in villous trophoblast when healthy villi were compared with pre-eclamptic villi. The observed alterations in trophoblastic cytoskeletal components are expected to damage the anchorage and motility of cells. The extravillous trophoblast is known to be necessary for implantation. This leads to a cellular hypothesis of the failure of implantation resulting in reduced depth of uterine invasion and failure to adapt the spiral arterioles for low-pressure perfusion of the intervillus space, two well-known features of pre-eclampsia. The reduction in cytokeratin-related immunofluorescence in the villus trophoblast seen on comparing healthy term placentae with those from pre-eclamptics implies that the trophoblast is a weaker epithelial layer in the hypertensive pregnancy. This could account for the rise in deported trophoblast associated with pre-eclampsia. Deported trophoblast has been invoked as the systemic messenger that leads to generalized maternal hypertension seen in this condition.

Cell Differentiation↗
PubMed2002

The distribution of caveolin-3 immunofluorescence in skeletal muscle fibre membrane defined by dual channel confocal laser scanning microscopy, fast Fourier transform and image modelling.

Membrane domains rich in caveolin-3 overlie sarcomeric actin in skeletal muscle. The membrane exhibits a regular array of caveolin-3 immunofluorescence using confocal laser scanning microscopy (CLSM). Fourier analysis of tissue imaged by CLSM accurately defines a repeating intensity with a long-axis spacing of 1.48 microm confirmed by measurement of direct images. Reverse fast Fourier transform (FFT) and image-modelling allow reconstruction of the pattern. Mathematical modelling has allowed replication of several features of the FFT, including the second order maxima that confirm the relatively high information content of the original images. Measurements of membrane-pattern primary long-axis spacings are consistent with our measurements of the I-band sarcomere repeat in similarly prepared specimens labelled with fluorescent phalloidin or imaged using differential interference contrast microscopy. Dual-channel CLSM analysis of the sarcomeric banding pattern of actin and the repeating pattern of muscle fibre membrane caveolin showed that caveolae overlie the I-band. The anti-caveolin immunofluorescence is deficient over the Z-disc and maximal toward each of the I-band extremities. A mechanism of membrane shape change in which membrane-lipid molecules are interposed between more stable anchored rafts associated with caveolae can be envisaged. Thus, increasing girth and reducing length of the sarcolemma in rapid contraction may be explained.

Animals↗
PubMed2001

Immunocytochemical localization of a caveolin-1 isoform in human term extra-embryonic membranes using confocal laser scanning microscopy: implications for the complexity of the materno-fetal junction.

This immunochemical, immunocytochemical, histological and ultrastructural study demonstrates the presence of caveolin 1 in a number of locations in term human extra-embryonic membranes. Strong expression was observed in fetal blood vessel endothelial cells of chorionic villi (cv) and in cv, amniotic and chorionic plate mesenchymal cells, but weak expression was characteristic of trophoblast. Expression in the amniotic epithelium indicated a stronger association with apical as opposed to baso-lateral membranes. Strong immunoreactivity in the thin lining layer of the maternal blood space of the basal plate was a surprising finding. Previously defined as trophoblast, we argue that this is at least partly endothelium based on this new histological, ultrastructural and immunocytochemical data.

Amnion↗
PubMed2000

Characterization of an in vitro model for the study of the short and prolonged effects of myocardial ischaemia and reperfusion in man.

The mechanisms underlying myocardial ischaemia and reperfusion-induced injury have been investigated, mainly by using animal experimental preparations in vitro and in vivo, but little is known of the process in human myocardium. The present studies characterize an in vitro model using human myocardium for the study of early and delayed effects of ischaemia and reperfusion. The right atrial appendage was manually sliced and incubated in buffer through which was bubbled O(2)/CO(2) (19:1, v/v) for various time periods. Lactate dehydrogenase (LDH) leakage, 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl-2H-tetrazolium bromide (MTT) reduction, oxygen consumption, nucleotide levels and tissue morphology were all investigated as markers of myocardial injury. The specimens remained stable and viable up to 24 h, but had significantly deteriorated by 48 h. The preparation responded to ischaemia in a time-related manner. Tissue viability was reduced by 25% after 30 min ischaemia, declined to 60% after 60 min ischaemia and to 75% after 120 min ischaemia. Interestingly, the tissue was more susceptible when ischaemia was induced after 24 h of aerobic incubation. The effects of the duration of reperfusion were investigated after a fixed 60 min ischaemic insult. The results of LDH leakage suggest that reperfusion injury is mainly sustained within the first 2 h of reperfusion. However, the results of MTT reduction show that there is a progressive decrease in tissue viability over the 24 h reperfusion period, possibly reflecting the occurrence of tissue necrosis and apoptosis at different reperfusion times. In conclusion, the data provide evidence that the incubation of human atrial tissue in vitro is stable, and slices are viable for at least 24 h, which permits the study of early and delayed consequences of ischaemia and reperfusion in the human myocardium.

Buffers↗
PubMed1999

Expression of tissue type and urokinase type plasminogen activators as well as plasminogen activator inhibitor type-1 and type-2 in human and rhesus monkey placenta.

The distribution of mRNAs and antigens of tissue type (t) and urokinase type (u) plasminogen activators (PA) plus their corresponding inhibitors, type-1 (PAI-1) and type-2 (PAI-2) were studied in human and rhesus monkey placentae by in situ hybridisation and immunocytochemistry. Specific monkey cRNA and antibodies against human tPA, uPA, PAI-1 and PAI-2 were used as probes. The following results were obtained. (1) All the molecules tPA, uPA, PAI-1 and PAI-2 and their mRNAs were identified in the majority of the extravillous cytotrophoblast cells of the decidual layer between Rohr's and Nitabuch's striae and in cytotrophoblast cells of the chorionic plate, basal plate, intercotyledonary septae and cytotrophoblast cells of the chorionic villous tree. (2) Expression of uPA and PAI-2 was noted in villous trophoblast whereas tPA and PAI-1 were mainly concentrated where detachment from maternal tissue occurs. (3) No expression of tPA, uPA, PAI-1 and PAI-2 was observed in the basal plate endometrial stromal cells, chorionic plate connective tissue cells, septal endometrial stromal cells or villous core mesenchyme. (4) The distribution of probes observed following in situ hybridisation is generally consistent with the immunofluorescence pattern of the corresponding antigens and no significant interspecies differences were noted. It is possible that both decidual and extravillous trophoblast cells of placentae of human and rhesus monkey are capable of producing tPA, uPA, PAI-1 and PAI-2 to differing extents. Coordinated expression of these genes in the tissue may play an essential role in the maintenance of normal placentation and parturition. The differences in distribution we observed are consistent with the suggestion that coordinated expression of tPA and its inhibitor PAI-1 may play a key role in fibrinolytic activity in the early stages of placentation and separation of placenta from maternal tissue at term. On the other hand, uPA with its inhibitor PAI-2 appears mainly to play a role in degradation of trophoblast cell-associated extracellular matrix, and thus may be of greatest importance during early stages of placentation.

Animals↗
PubMed1996

Leucocyte and endothelial cell adhesion molecule expression in porcine histiocytic leiomyofibrosarcoma.

A histiocytic sarcoma was present at birth in a pig. On the basis of ultra-structure and structural-protein composition (presence of alpha-smooth-muscle actin but not keratin), the sarcoma component was identified as a leiomyofibrosarcoma. Lipid-laden macrophages (histiocytes), which permeated the tumour in an apparently random fashion, were somewhat atypical in that they were negative for some macrophage markers; they gave a reaction, however, for CDw14. Despite its aggressive metastatic capacity, this tumour occurred almost exclusively in the subcutis, dermis and skeletal muscle. The tumour was extensively vascularized with many small capillaries which did not express E-selectin (CD69E), MHC class II or the L-selectin (CD69L) ligand, markers characteristic of inflamed (activated) endothelial cells in pig skin. Significant numbers of the histiocytes were positive for the integrins CD18 and VLA-4 (CD49d), indicating involvement of integrin pathways in the spread or growth, or both, of the leiomyofibrosarcoma. Most of the fibrous sarcoma cells also had extensive reactivity with an antibody to the standard variant form of CD44 (CD44s).

Animals↗
PubMed1996

Behaviour of two IgG subclasses in transport of immunoglobulin across the human placenta.

The human IgG subclasses are a family of highly related yet distinct molecules. Each of these four subclasses performs a discrete function within the human immune system. Previous studies have shown that one of these molecules, hIgG2, may be discriminated against in transport across the human placenta. We have aimed to elucidate the mechanism of this discrimination in order to gain a more comprehensive understanding of the process of transport of immunoglobulin across the human placenta. We have used a combination of immunocyctochemical localisation and biochemical analysis to detail the behaviour of hIgG2. Confocal laser scanning microscopy was used to compare the localisation of hIgG1 (chosen as representative of the efficiently transported subclasses) and hIgG2 in term and first trimester chorionic villi. Complementary evidence was provided from immunoblot analysis of isolated placental coated vesicles. The data presented here suggest that the hIgG2 is transported into the syncytiotrophoblast and appears to accumulate in the stroma of the villi. This leads us to the hypothesis that the fetal capillary endothelium is the cellular impediment to the transport of hIgG2 into the fetal circulation.

Biological Transport, Active↗
PubMed1995

Ultrastructural distribution of endogenous immunoglobulin-G in human term amniochorion.

Maternal immunoglobulin-G (IgG) is known to be transported across the placental syncytiotrophoblast during the period when the human fetus is incapable of manufacturing these defensive molecules. In this study we investigated the possible role of the amniochorion, that surrounds the amniotic cavity in which the fetus lies, in the transfer of immunoglobulin. Endogenous IgG was localised in the amniochorion by confocal immunofluorescence microscopy and by ultrastructural labelling of ultrathin frozen tissue sections using the protein A-gold technique. Immunoreactivity was identified in the extracellular matrix tissues and necrotic amniotic epithelial cells. Healthy amniotic epithelial cells and cytotrophoblast cells of the chorion laeve were devoid of endogenous IgG. These results suggest a possible non-specific paracellular transport pathway between cytotrophoblast cells, which may conceivably contribute to the acquisition of passive immunity by the fetus, and offer a rational explanation for the presence of small quantities of maternal IgG in the amniotic fluid.

Amnion↗
PubMed1995

Cytotrophoblast cells: a barrier to maternofetal transmission of passive immunity.

The human fetus receives passive immunity via the chorioallantoic placenta in the form of maternal immunoglobulin G (IgG) class antibodies. This provides protection against pathogens at a time when the fetus is immunologically naive. We localized endogenous human IgG using confocal laser scanning fluorescence microscopy and immunoelectron microscopy of frozen sections of chorionic villi from early and late gestation. With confocal microscopy we also investigated the distribution of a receptor for IgG (Fc gamma RIII; CD16) that is typically expressed on the surface of human leukocytes. Endogenous IgG was present in the syncytiotrophoblast that surrounds chorionic villi but underlying cytotrophoblast cells were devoid of endogenous antibody. Fc gamma RIII immunoreactivity was confined to the syncytiotrophoblast and was also absent from cytotrophoblast cells. We propose that cytotrophoblast cells present a barrier to the transmission of maternally derived IgG across the human placenta. This accounts for the paradox that there are low levels of transport in the first trimester when the syncytiotrophoblast is known to express receptors for IgG. Cytotrophoblast cells form an almost complete epithelial layer underlying the syncytiotrophoblast at this stage of gestation, but this becomes discontinuous as the placenta matures, thus removing the cellular impediment to IgG transmission.

Cesarean Section↗
PubMed1994

The mode of insertion of umbilical cord and vessels: association with maternal haemoglobin genotype, neonatal factors, and placental component volumes.

We investigated whether variations in mode of attachment of umbilical cords and vessels coincided with differences in maternal haemoglobin genotype and neonatal factors or placental micro-volumetric composition. The incidence of placentae with marginally inserted cords, or those in which umbilical vessels separated prior to insertion, was not statistically different in samples from sickle cell patients having haemoglobin-SS and haemoglobin-SC genotypes, as compared with haemoglobin-AA controls. Results obtained from analysis of variance (ANOVA) suggest that the mode of insertion (status) of umbilical vessels may have clinical significance, because it produced differences in the main effects associated with neonatal gestational age, placental weight, and placental index. Point counting stereology was employed to estimate the microscopic compartment volumes of placentae prior to an assessment of statistical association between the data obtained and the status of umbilical vessels and maternal haemoglobin genotype. Furcate placentae (with separated umbilical vessels) had statistically greater than normal volumes of villi, villous trophoblast, and syncytial knots. We deduce that furcate placentae are prone to early delivery, because they are heavier, having more voluminous villi with more trophoblast and syncytial knots than controls.

Analysis of Variance↗
PubMed1994

Cryofracture of human term amniochorion.

By use of cryofracture and scanning electron microscopy of human amniochorion we have captured images of all the major layers of the tissue. Correlation of confocal and electron-microscope data has allowed greater understanding of how these cellular and acellular layers interconnect in order to maintain their integrity as a multilaminar tissue. This is not straightforward as mutual sliding or area change is required of concentric curved surfaces which expand and contract as does the amnion. In this paper we suggest a mechanism by which the amnion is able to slide with respect to the chorion and still maintain continuity as a structural unit. It is based on the observation of complementary gyri and sulci on surfaces facing the spongy layer which is a shear plane. Cellular detail at higher resolution of the amniotic epithelium and acellular layers provides a more complete description of structural composition than was previously available.

Amnion↗
PubMed1994

Heterogeneity of Fc gamma receptor-bearing cells in human term amniochorion.

Confocal microscopy has been used to study immunoreactivity to monoclonal antibodies directed against the three characterized human leukocyte Fc gamma receptors in term amniochorion. Fc gamma RI (CD64), Fc gamma RII (CDw32) and Fc gamma RIII (CD16) are expressed by cells of the maternal decidual layer associated with the amniochorion. These three subtypes of receptor are also expressed by cells of the fibroblast and reticular layers although not all cells of these layers express receptors which suggest that they are heterogeneous cell populations. We were not able to demonstrate Fc gamma RII immunoreactivity in decidual layer endothelial cells. Human IgG specific immunofluorescence was detected in the extracellular matrix of the tissue and associated with the Fc gamma receptor-bearing cells of mesenchymal origin and the maternal decidua, as demonstrated by dual labelling immunofluorescence studies. Two epithelial layers of cells (amniotic and trophoblast) lack Fc gamma receptors that are present in placental chorionic villi, and which may be necessary for transepithelial transport of IgG. The cells of these layers also contain no demonstrable endogenous IgG. These data support the hypothesis that maternally derived IgG is acquired by the fetus across the vascular system of the placenta only and not via a transamniochorion route. The demonstration of Fc gamma receptors on cells of extraembryonic mesodermal lineage is a novel finding that may suggest a potential immunological role within the amniochorion.

Amnion↗
PubMed1994

Laser scanning confocal examination and comparison of nidogen (entactin) with laminin in term human amniochorion.

Indirect immunofluorescence confocal microscopy was used to compare the distribution of nidogen (entactin) and laminin in term human amniochorion. Rabbit antiserum raised against recombinant mouse nidogen was immunoreactive with components of the amniotic epithelial basement membrane, chorion laeve basement membrane and maternally derived capillary basement membranes. Overall anti-nidogen immunoreactivity co-distributes with laminin except in the extracellular matrix of the maternally derived decidua, the embryonically derived trophoblast and the apical surfaces of the amniotic epithelia. These differences lead us to suggest that nidogen and laminin may be independently expressed and are not always complexed together. Original observations include dramatic parallel nidogen rich striations in the decidual layer, the fact that the trophoblastic basement membrane appears to extend inwards forming a boundary layer to degenerate chorionic villi and the finding that the recently discovered collagen type IV rich microtrabeculae of the fibroblast layer also contain nidogen and laminin.

Amnion↗
PubMed1994

Ontogeny and distribution of Fc gamma receptors in the human placenta. Transport or immune surveillance?

The human fetus acquires maternal IgG via the chorioallantoic placenta. Utilising antibodies against 3 characterised subtypes of IgG receptor (Fc gamma R) expressed by human leucocytes, we show by confocal immunofluorescence microscopy that these molecules are also expressed by cells of the placenta. Fc gamma RI (CD64) is expressed by undifferentiated mesenchymal or fibroblast cells of 1st trimester and term chorionic villi. Punctate immunoreactivity for Fc gamma RII (CDw32) is found on capillary endothelial cells of term and 1st trimester villi. Fc gamma RIII (CD16) expression is observed in the trophoblast surrounding chorionic villi that forms the functional 'barrier' between mother and fetus. In 1st trimester villi this receptor is associated with a population of marginated vesicular inclusions of the syncytiotrophoblast. In term villi the receptor is concentrated in the apex of the syncytiotrophoblast, suggesting a possible role in the maternofetal transmission of passive immunity. All 3 subtypes of receptor are expressed by Hofbauer cells. We have been unable to demonstrate these receptors in cytotrophoblast cells. Results obtained using immunofluorescence and immunoelectron microscopic detection of endogenous IgG are consistent with the hypothesis that IgG is internalised into clathrin-coated pits and vesicles. Endogenous IgG was not demonstrable in cytotrophoblast cells. The significance of our inability to demonstrate IgG or specific receptor molecules for IgG in cytotrophoblast cells, and possible roles of Fc gamma receptor-bearing cells of the placenta are discussed.

Chorionic Villi↗
PubMed1990

A quantitative interference light microscope study of human first trimester chorionic villi.

Using a Jamin-Lebedeff-type interference microscope an analysis of frozen sections of human first trimester chorionic villi reveals regional differences down to subcellular resolution. The evidence indicates compositional differences between villus cell types and shows that the syncytiotrophoblast is differentiated into at least three layers, one of which corresponds positionally to the previously described syncytioskeletal layer. Quantitative measurements have been made of specimen thickness, refractive index and dry mass of regions in the tissue. Local differences in syncytiotrophoblast have been noted with respect to the content of a population of organelles with distinctive optical properties. These may correspond to stored forms of steroid hormone or their precursors.

Chorionic Villi↗
PubMed1990

A confocal and conventional epifluorescence microscope study of the intermediate filaments in chorionic villi.

An immuno-epifluorescence microscopic study of first trimester human chorionic villi has revealed different patterns of distribution for intermediate filament proteins. Keratin staining was restricted to the trophoblastic epithelium. The protein was shown to be concentrated into desmosome-containing apical and basal cytoplasm. Strands of keratin-rich cytoplasm extended across the interior of the syncytium, integrating basal and apical layers. This arrangement integrates the epithelial cytoskeleton in a manner well adapted to resisting shearing forces. Vimentin was found in the cells of the villous core and was most strongly expressed in endothelial cells. Desmin was also restricted in distribution of spindle-shaped cells of the villous core. It was excluded from terminal villi.

Chorionic Villi↗