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Biomedical subjects

W R Jones

Publications and source records attributed to W R Jones.

At least 73 records · Page 4Linked to original sources
PubMed1985

Antibodies to spermatozoa and seminal plasma antigens detected by various enzyme-linked immunosorbent (ELISA) assays.

Various ELISA methods have been applied by different research centers to test the efficiency of this approach for the diagnosis of sperm-immune infertility cases. The antigens used were either whole spermatozoa or solubilized spermatozoal membrane preparations and were immobilized on microtiter plates, except in one case where plastic beads were employed. Polyvalent second antibodies or protein-A labelled with enzymes served as tracers. A high frequency of positive sera was found in all groups including fertile controls with tests using whole spermatozoa as antigen. The methods using solubilized antigen preparations showed fewer positives on the whole and correlated better with the various clinical categories of the WHO sera. Whilst there was some agreement in the results between the various laboratories on a few sera, most of the positive sera found by one laboratory were reported as negative by others. More investigative work is needed to improve reproducibility between different laboratories and to reduce non-specific reactions with normal controls. A more precise definition of the proper cut-off levels for positives and negatives is also needed. Despite these short-comings, the development of an ELISA for the diagnosis of sperm-immune infertility cases seems to be justified in the long term.

Antibodies↗
PubMed1985

Identification of an SP-1-like protein in non-pregnancy serum: isolation using a monoclonal antibody.

Human SP-1, a glycoprotein synthesized by the placenta during pregnancy, was shown to exist as polymers in maternal serum by a rapid passive transfer immunoblotting technique following conventional agarose electrophoresis. Moreover, the SP-1 polymers in serum were shown to dissociate into one main component upon treatment with 8 M urea prior to electrophoresis. However, an unexpected observation was the existence of an SP-1-like immunoreactive species in male serum with the sensitive immunoblotting technique. This SP-1-like protein in male serum had similar properties to its placentally derived counterpart in pregnancy serum, namely a propensity for complex formation and a reduced electrophoretic mobility following neuraminidase treatment. The relationship between the two SP-1 proteins was demonstrated by isolating them from their respective sera using an immobilized monoclonal antibody raised to purified SP-1 from pregnancy serum. Immunoblotting after sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed the existence of two major SP-1 species in pregnancy serum. These two SP-1 species had apparent molecular weights of 68,000 and 64,000. In addition, there were two minor bands at 35,000 and 32,000. These smaller SP-1 species did not appear to be subunits of the larger entities since they were detectable in the absence of reducing conditions. SDS-PAGE and immunoblotting showed that the immunoaffinity-purified SP-1 species from male and pregnancy sera had similar, but not identical, molecular weights.

Animals↗
PubMed1985

An improved indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antisperm antibodies.

An indirect enzyme-linked immunosorbent assay (ELISA) was used for the detection of antisperm antibodies which provides a sensitivity and ease of quantitation that is not available with conventional bioassay systems. A standardized protocol was developed in which washed whole sperm were coated onto polystyrene microelisa plates at a density of 1 X 10(5) sperm per well using a commercially available spray fixative. Urease was employed in the enzyme-antiimmunoglobulin conjugate to minimize nonspecific background reactions. Diluted positive and negative human sera were incubated at 37 degrees C and the results were read on an ELISA auto reader. Using mouse antihuman sperm antisera and sera from selected infertile patients, it was found that the ELISA method was significantly more sensitive than the sperm microimmobilization test and the microtray agglutination test. The results also confirmed that the ELISA detected a different spectrum of sperm antibodies compared with the other two techniques.

Animals↗
PubMed1985

Human milk cell migration and production of monocyte chemotactic factor: lack of activity.

The lymphocytes of human breast milk have been previously shown to be immunologically competent while the monocytes have been described as actively motile in some reports but not in another. Cells were separated from milk samples collected by lactating women 2 to 9 days postpartum. Milk cell migration and production of the lymphokine, monocyte chemotactic factor, by milk cells were assayed using membrane filters. Milk cells were poorly motile although prior culturing improved their migratory ability. Cell-free milk was chemotactic for blood monocytes. Chemotactic factor production was detected in only two of 16 phytohemagglutinin-stimulated milk cell cultures.

Cell Movement↗
PubMed1985

Danazol: objective assessment in the treatment of endometriosis.

The objective assessment of Danocrine (Danazol, Winthrop) in the treatment of endometriosis was hampered in the 1970s by the absence of a universally accepted classification. The American Fertility Society (AFS) classification, proposed in 1979, has now become widely accepted. However, there is a lack of recorded objective data utilizing the AFS score before and after Danazol therapy. In a series of 51 patients with endometriosis, who underwent laparoscopy, then received Danazol, 800 mg daily for 6 months, and had a repeat laparoscopy, there was a statistically significant improvement in the AFS score in Stage I and II disease.

Danazol↗
PubMed1984

Monocyte chemotactic factor production in mixed cultures of maternal and neonatal human lymphocytes.

The immunological interaction between related maternal and neonatal human lymphocytes was investigated by assaying monocyte chemotactic factor production in mixed cell culture. Lymphokine was detected in a significantly lower proportion of maternal-neonatal cell cultures compared to mixed cultures of cells from unrelated adults and/or neonates. It is possible that a reduced inflammatory mediator production at the feto-maternal interface may contribute to the survival of the fetal allograft.

Adult↗
PubMed1984

Temperature dependence of protein transport across lymphatic endothelium in vitro.

The purpose of the work was to develop an in vitro model for the study of lymphatic endothelium and to determine, using this model, whether or not a cytoplasmic process may be involved in transendothelial transport. Segments of canine renal hilar lymphatics were dissected clean, cannulated at both ends, and transferred to a perfusion chamber for measurement of transendothelial protein transport and for ultrastructural tracer studies. The segments were subsequently processed for light and electron microscopy. By both structural and functional criteria the lymphatics were judged to have retained their integrity. At 37 degrees C, 36 lymphatics showed a mean rate of protein transport of 3.51 +/- 0.45 (SEM) micrograms/min per cm2 of lymphatic endothelium. The rate was influenced by the temperature of the system, being significantly reduced by 49% +/- 4.8, 31% +/- 5.3, and 29% +/- 3.9 when the temperature was lowered to 4 degrees, 24 degrees, and 30 degrees C, respectively. When the temperature was raised to 40 degrees C, the rate was significantly increased by 48% +/- 12.2. The vesicular system and the intercellular regions in vessels with increased or reduced rates of transport were analyzed quantitatively to ascertain whether the rate changes could be correlated with ultrastructurally demonstrable changes in either of these postulated pathways. No significant changes in junctional or vesicular parameters were found between the control lymphatics and those perfused at 24 degrees, 30 degrees, and 40 degrees C. At 4 degrees C, the temperature at which the rate of protein transport was maximally reduced, vesicular size decreased, and the number of free cytoplasmic vesicles increased, whereas the number associated with the abluminal and luminal surfaces decreased. We concluded that isolated perfused lymphatic segments transport protein at a relatively constant rate under control conditions, and that this transendothelial transport comprises both temperature-dependent and temperature-independent mechanisms. The findings were considered in terms of the different theories of lymph formation and were interpreted as providing support for the vesicular theory.

Animals↗
PubMed1984

Variations in rabbit oviduct microvascular architecture after ovulation induced by hCG.

Rabbits were induced to ovulate by injection with hCG and vascular corrosion casts of the oviducts were examined by scanning electron microscopy after 24 and 48 h, when the ova would be expected to be at the ampullary-isthmic junction, and traversing the isthmus respectively. At 24 h there was dilatation of the isthmic subserosal venous plexus. It is suggested that venous distension in the isthmic subserosal venous plexus, due to raised venous pressure or to reduced venous wall tone, may occlude the isthmic lumen to ova, and thus explain the known pre-isthmic delay in ovum transport. By 48 h after hCG, distension was no longer evident, consistent with the possibility of ovum transport.

Animals↗
PubMed1984

Microvascular architecture of the pregnant rabbit oviduct.

Methyl-methacrylate vascular corrosion casts of the oviducts were prepared in 7 rabbits which were 2-3 weeks pregnant. Scanning electron microscopy of the acrylic casts revealed little change in tubal microvascular connections when compared with control oviducts. Venous distension in the isthmic subserosal venous plexus, ampullary subserosal vasculature and in the fimbrial core was substantially greater than that observed in controls. These changes are interpreted as indicating a sensitivity of tubal microvasculature to the increased levels of circulating placental hormones in pregnancy. The implications of this interpretation in the role of tubal microvasculature at the time of ovulation are discussed.

Animals↗
PubMed1983

Ultrastructural evidence for a reabsorptive role by intrarenal veins.

Intrarenal veins, although known to have thin walls, are considered to be simple conducting vessels. Using light- and electron microscopy, the distribution and structure of named intrarenal veins was examined qualitatively and quantitatively in rat kidneys fixed by retrograde arterial perfusion. Although the venous system follows the pattern of arterial branching in general, a class of intracortical veins similar in appearance to interlobular veins but without a companion artery was found in the present study. It is suggested that these vessels be designated intralobular veins. Structurally, we have found intrarenal veins to be surprisingly similar to peritubular capillaries both in respect to their spatial relations to renal tubules as well as ultrastructurally. Like peritubular capillaries, the majority of the wall of intrarenal veins is intimately apposed to renal tubules. This fraction decreases centrally, values of 0.83 being obtained in intralobular veins, 0.69 in interlobular veins, and 0.56 in arcuate veins. The walls of intrarenal veins are comprised of little more than an endothelium, which, like peritubular capillaries, is remarkable for its thinness, high density of fenestrae, and lack of extraintimal elements. Endothelial thickness was not significantly greater in either interlobular or arcuate veins than in peritubular capillaries; the fenestrae were, however, about twice as frequent in peritubular capillaries as in interlobular or arcuate veins and 35 times more frequent than in interlobar veins. The size and numerical and volume densities of uncoated endocytotic vesicles did not differ significantly between peritubular capillaries and any of the intrarenal veins. Based on their marked qualitative and quantitative similarities to peritubular capillaries, we conclude that, like the latter, intrarenal veins are capable of sustaining passive transport between plasma and interstitium.

Absorption↗
PubMed1983

The organization of endocytotic vesicles in lymphatic endothelium.

The lymphatic endothelium from renal hilar lymphatics in the rat was subjected to qualitative and quantitative ultrastructural analysis with emphasis on the extent and disposition of its vesicular component. The uncoated endocytotic vesicles had an average maximum diameter of 0.073 micron and occupied 7% of the cytoplasm. There were approximately 21 vesicles in each cubic micrometer of cytoplasm. In standard electron microscopic preparations of glutaraldehyde-fixed endothelium 50% of the vesicles appeared to lie free within the cytoplasm. The remainder were seen to touch or open onto the luminal or abluminal surface of the endothelium. The degree to which intracytoplasmic endocytotic vesicles remained discrete or communicated with the plasma membrane was examined using tannic acid and ruthenium red. These substances specifically bind to charged molecules on the cell surface and identify membranes continuous with it. When this technique was applied to aldehyde-fixed tissue nearly 90% of the vesicles that were apparently free within the cell could be shown to retain a connection with the surface, approximately equal numbers communicating with either the luminal or abluminal surface. At least 15% of these vesicles existed as intercommunicating clusters. These results suggest that many vesicles are not simple discrete units, but rather form parts of chains that reach either luminal or abluminal surface. Thus, apparently discrete vesicles may be parts of vesicular chains cut in cross section. The possible relation between chemical fixation and this plan of vesicular organization is discussed and it is concluded that while chemical fixation may result in an overestimation of the numbers of intercommunicating vesicles, the qualitative aspects of vesicle disposition seem largely unaffected. Although the functional significance of our observations has yet to be determined, should this plan of vesicular organization apply to initial lymphatics as well, the concept of vesicular transport solely by random movement of discrete vesicles across lymphatic endothelium should be modified.

Animals↗
PubMed1983

Morphological characteristics and protein profile of isolated human decidual cells.

Isolated decidual cells were prepared from human decidual tissue obtained during early pregnancy by digesting the tissue fragments with 0.1% collagenase solution. Morphological studies of the cells were carried out using morphometric and flow cytometric analysis while the protein profile was analysed by polyacrylamide gel electrophoresis and gel filtration column chromatography. An average of 90% cell viability was achieved and the results showed that decidual cells constitute up to 70% of cell number and 89% of cell area of the isolated decidual cell suspension. The presence of serum proteins in uterine tissue extracts is due to blood contamination. However, the similarities of the non-serum protein profiles in tissue and cellular extracts validates previous studies performed in uterine fluids and tissue extracts. Finally, at least one unique uterine protein appeared to be a sub-unit of a larger molecule.

Cell Count↗
PubMed1983

Pre-eclampsia in pregnancies from donor inseminations.

The outcome of 584 AID pregnancies was analysed in order to examine the possible role of immune mechanisms in the development of pre-eclampsia (PE). The overall incidence of PE was high (9.3%) compared with the expected incidence of 0.5-5.0%. The incidence was similar in both multigravid and primigravid women. The expected protective effect of a previous pregnancy was not seen, with a 47-fold increase in PE (observed versus expected) in AID pregnancies after a previous full-term pregnancy and a 15-fold increase after a previous pregnancy of short duration. In the context of AID pregnancies these findings tend to support the proposed 'fresh mating' concept of PE although the incidence is lower in multigravidas after a full-term (4.7%) than after a partial pregnancy (15.7%). This suggests that it may only be full-term pregnancy which protects against the development of PE. THe category and duration of infertility and the duration of marriage did not affect the incidence of PE except where the male partner was oligozoospermic. In this case the incidence of PE was higher where infertility was 3 years or longer. Further analysis of the findings failed to provide clear evidence to discriminate the role of HLA and tissue-specific non-HLA antigens in the feto-maternal relationship as determinants of the development of PE. Neither did the results support the operation of immunological enhancement induced by prior coital antigen exposure in the genesis of PE, but raised instead the possibility of the operation of a mechanism of sensitisation of the female partner to seminal components over a long period of time when the husband was oligozoospermic compared with one whose husband was azoospermic.

Female↗
PubMed1983

Fallopian tube microvasculature in the rabbit.

The microvascular architecture of rabbit Fallopian tube isthmic, ampullary and fimbrial regions was examined by the corrosion vascular cast-scanning electron microscope method. The aorta and inferior vena cava of virgin adult female rabbits (2.5-3.5 kg) were cannulated; after blood wash out, methacrylate casting medium was infused via the aorta. After plastic hardening, Fallopian tubes were excised, tissues corroded and casts prepared for scanning electron microscopy. The isthmic myosalpinx is encircled subserosally by an interlocking venous plexus. We suggest that a rise in pelvic venous pressure could lead to a decrease in isthmic luminal diameter and thus act as a sphincter contributing to the known delay in ovum transport at the ampullary isthmic junction. Stereo pairs of photomicrographs showed that isthmic plicae are supplied by arterioles which change little in diameter passing through the tubal wall. Near the apex of a fold, these arterioles terminate in a subepithelial capillary plexus which drains to deep mucosal and plical core venules. In contrast, the ampulla is supplied by smaller and less frequent arterioles. Frequent large venules extending high into the plicae drain the subepithelial capillary plexus. Based on this microvascular architecture, we suggest that the subepithelial capillary plexus of the apical region of the plicae probably contains high pressure blood in the isthmus and lower pressure blood in the ampulla. Therefore, plicae in these regions may be specialised for net luminal fluid secretion and absorption respectively, implying a role for the microvasculature in tubal transport by initiation or modification of fluid flow along the lumen.

Animals↗