Sultan Al-Sedairy

Prediction of successful embryo implantation by measuring interleukin-1-alpha and immunosuppressive factor(s) in preimplantation embryo culture fluid*

To find a better predictor of pregnancy after in vitro fertilization (IVF), supernatant fluids from embryo culture media were analyzed after 24 hours and 48 hours for the presence of interleukin-1-alpha (IL-1), interleukin-2, and the percent of immunosuppression. The measurements were performed on 108 consecutive IVF cycles between June 1989 and October 1989. The IL-1 level +/- SD in the 24-hour aliquots of the supernatant of embryo culture fluid was 66.2 +/- 10.2 pg/mL in all viable pregnancy cycles and 35.4 +/- 9.01 pg/mL in unsuccessful cycles. The percent of immunosuppression after 24 hours was 22.06% +/- 4.5% in viable pregnancy cycles and 7.3 +/- 5.5% in unsuccessful cycles. The percent of immunosuppression 48 hours after ovum pick-up was generally decreased in all embryo culture fluid, showing 17.5% +/- 4.4% in viable pregnancy cycles and 3.8% +/- 3.6% in unsuccessful cycles. Interleukin-1 levels in the 48-hour aliquots were moderately decreased, being 39.0 +/- 6.3 pg/mL in viable pregnancy cycles and 34.3 +/- 4.7 pg/mL in the unsuccessful cycles. In 24 hours, embryo culture aliquots IL-1 level greater than 60 pg/mL was seen in 17 of 21 (80.9%) pregnancy cycles, and the combined data of IL-1 level greater than 60 pg/mL and/or greater than 20 percent of immunosuppression predicted 21 of 21 (100%) pregnancy cycles.

Immunomodulatory effect of Nigella sativa proteins fractionated by ion exchange chromatography.

Whole Nigella sativa (N. sativa) proteins were purified on a DEAE Sephadex A50 ion exchange column. Complete fractionation was achieved in four peaks. Analysis of the purified peaks was carried out by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Whole N. sativa showed a number of protein bands ranging from 94-10 kDa molecular mass. In mixed lymphocyte cultures (MLC), whole N. sativa and its purified proteins were found stimulatory as well as suppressive and this effect varied from one donor to another. Maximum stimulation (mean + S.E. of % relative index was 63.73 + 20.78) was observed with fractionated N. sativa proteins (P1) (10 microg/ml) in MLC. In MLC, also N. sativa peaks (P1 and P2) were stimulatory at all concentrations (10 microg/ml, 1 microg/ml or 0.1 microg/ml) used. However, a uniformly suppressive effect of N. sativa and its all four peaks at a concentration of 10 microg/ml was noticed when lymphocytes were activated with pokeweed mitogen (PWM). The effect of N. sativa proteins was further evaluated on the production of cytokines which were measured by using specific enzyme-linked immunosorbent assay. Large quantities of IL-1beta were secreted by whole N. sativa in culture medium with non-activated peripheral blood mononuclear cells (PBMC) (450 pg/ml) and with allogeneic cells (410 pg/ml). Fractionated N. sativa was less effective when compared with whole N. sativa proteins. No effect on IL-4 secretion was seen either by using non-activated, PWM-activated or allogeneic-cells. Whole N. sativa suppressed as well as stimulated the production of IL-8 in non-activated and PWM-activated PBMC respectively. All N. sativa peaks with protein concentration of 2 microg/ml were stimulatory for the induction of IL-8 by PWM-activated cells. However, no effect on IL-8 was seen either with whole N. sativa or its peaks when allogeneic PBMC were used. Stimulatory effect of whole N. sativa and fractionated proteins was also noticed on the production of TNF-alpha either using non-activated or mitogen activated cells.

Concentrations of soluble tumor necrosis factor and interleukin-6 receptors in heatstroke and heatstress

OBJECTIVE Increased proinflammatory cytokine concentrations have been implicated in the pathogenesis of heatstroke. Soluble cytokine receptors can modulate circulating cytokine activities. We examined the possible role of soluble tumor necrosis factor receptors (sTNFR 60, sTNFR 80) and interleukin-6 receptor (sIL-6R) in heatstroke by determining their concentrations before and after cooling, as well as in heatstressed controls. DESIGN Prospective controlled study. SETTING Heatstroke Center, Makkah, Saudi Arabia (1993 pilgrimage). PATIENTS Twenty-five consecutive heatstroke patients before and after cooling, 14 heatstressed controls (HSC), and 13 normal controls (NC). MEASUREMENTS AND MAIN RESULTS Concentrations of sTNFR 60, sTNFR 80, and sIL-6R, as well as their ligands, were measured using commercially available enzyme-linked immunosorbent assay kits. Mean sTNFR 60 concentration was increased in heatstroke (p <.0001, vs. NC; p < .0001, vs. HSC) and in HSC (p = .004, vs. NC). Mean sTNFR 80 concentration was increased in heatstroke and decreased in HSC (p = .01, heatstroke vs. HSC). Mean sIL-6R concentration was decreased in heatstroke and increased in HSC (p = .04, heatstroke vs. NC; p = .001, heatstroke vs. HSC). IL-6 was undetectable in NC and mean IL-6 concentration was more increased in heatstroke than in HSC (p = .001). Rectal temperature and creatinine concentrations correlated significantly with sTNFR 60, sTNFR 80, sIL-6R, and IL-6 concentrations. After cooling, mean concentrations of sIL-6R and sTNFR 80 increased significantly, whereas the mean sTNFR 60 concentration did not change. Residual neurologic deficits were associated with higher precooling IL-6 (p = .002) and postcooling sTNFRs (p < .0001) concentrations. CONCLUSIONS Significant changes in cytokine receptor concentrations are associated with heatstress. In heatstroke, the changes are more pronounced, and for some cytokine receptors, the changes are in the opposite direction (compared with changes in heatstress). Concentrations of IL-6 and sTNFRs correlate with hyperthermia and outcome. Cooling did not normalize sTNFR concentrations, suggesting failure to control the inflammatory response.

Evidence for endothelial cell activation/injury in heatstroke.

OBJECTIVES We treated the hypothesis that heatstroke is associated with endothelial cell activation/injury and examined the possibility that the markers of endothelial cell activation/injury may be associated with its severity and complications such as disseminated intravascular coagulation, lung injury, and renal dysfunction. DESIGN Prospective analyses. SETTING Heatstroke Center in Makkah, Saudi Arabia. PATIENTS Twenty-two adult patients with heatstroke. INTERVENTIONS The plasma concentration of endothelin, circulating intercellular adhesion molecule-1 (ICAM-1), and von Willebrand factor-antigen values were measured, respectively, by radioimmunoassay, enzyme-linked immunosorbent assay, and rocket electroimmunoassay, in heatstroke patients on admission (precooling) and after complete cooling (postcooling), and in ten normal control patients. MEASUREMENTS AND MAIN RESULTS Precooling heatstroke patients (rectal temperature 40.9 +/- 1.1 [SD] degrees C) had increased circulating concentrations of endothelin, c-ICAM-1, and von Willebrand factor-antigen in 100%, 80%, and 77% of patients to 126.4 +/- 11.2 pmol/L, 523.1 +/- 154.4 ng/mL, and 3.85 +/- 2.3 U/mL, respectively (control values: 13.7 +/- 4.2 pmol/L [p < .001]; 247.4 +/- 68.2 ng/ml [p < .001]; and < 1.5 U/mL, respectively). There was a significant (r2 = .68, p < .01) correlation between circulating ICAM-1 and endothelin concentrations. Plasma endothelin concentration correlated negatively with temperature (r2 = .35, p < .05). Mean endothelin concentration was similar in patients with or without renal dysfunction, and mean von Willebrand factor-antigen concentration was similar in patients with or without lung injury or disseminated intravascular coagulation. There were no significant correlations between circulating ICAM-1, endothelin, or von Willebrand factor-antigen concentration and the Simplified Acute Physiology core. After cooling, mean circulating ICAM-1 and endothelin concentrations decreased significantly to 400 +/- 109 ng/mL and 93 +/- 38.5 pmol/L, respectively, whereas the mean von Willebrand factor-antigen concentration increased to 5.55 +/- 2.18 U/mL (p > .05). CONCLUSIONS Our findings of increased circulating concentrations of circulating ICAM-1, endothelin, and von Willebrand factor-antigen are consistent with the hypothesis that heatstroke is associated with endothelial cell activation/injury. Whether the endothelial cell activation/injury is implicated in the pathophysiology of this disorder merits further studies.

Nigella sativa: effect on human lymphocytes and polymorphonuclear leukocyte phagocytic activity.

The effects of Nigella sativa (N. sativa) seeds and their soluble fractions were studied in vitro on lymphocyte response to different mitogens and on polymorphonuclear leukocyte phagocytic activity. No stimulatory effect of N. sativa was detected on lymphocyte response to phytohemagglutinin, concanavalin-A or pokeweed mitogen. A stimulatory effect of N. sativa was noticed on the lymphocyte response to pooled allogeneic cells. This effect was more pronounced when the low molecular weight (< 10 kDa) fraction was used and varied from one normal individual to another (25% to 825%). N. sativa enhanced the production of interleukin-3 by human lymphocytes when cultured with pooled allogeneic cells or without any added stimulator. N. sativa did not, however, enhance or suppress interleukin-2 secretion by mitogen activated peripheral blood mononuclear cells. Interestingly, N. sativa increased interleukin-1 beta, suggesting therefore, that it has an effect on macrophages. It also suppressed the leukocyte chemiluminescence activity using phorbol myristate acetate and Zymosan as stimulants. No effect of N. sativa or its fractions was, however, noticed on bacterial phagocytosis or killing when Staphylococcus aureus was used, indicating that the decrease in chemiluminescence activity in the presence of N. sativa is not relevant to the bactericidal activity.

Aeroallergens and viable microbes in sandstorm dust: Potential triggers of allergic and nonallergic respiratory ailments

Aeroallergens and antigens in sandstorm dust, extracts of which were skin prick test (SPT) positive in allergic patients, were detected by roeket immunoelectrophoresis and ELISA. Fungi and bacteria isolated by agar settle plates and soil dilution and soil washing methods were enumerated and identified. Cat dander. Acacia, Alternaria. Aspergillus. Chenopodium. Ciadosporium, Bermuda grass, Pitheceilobiiim, Prosopis. Rumex, cultivated rye. and Washingtonia palm allergens were detected by both methods. Viable microbes including 1892±325 eolony‐forming units (cfu) of bacteria, and 869±75 cfu of fungi were isolated per gram of dust by the soil dilution method. Randomly selected microbial colonies on streaking and subculture were found to consist of between two and seven mixed colonies. Fungi including Alternaria, Aspergiiius, Botrytis, Ciadosporium, Mortierelia, Mucor, Mycelia sterilia, Peniciilium, Pythium, Uiodadium, Verticiliium, and some yeasts were isolated. Actinomyces, Bacillus, Pseudomonas. and mostly coagulase‐negative Staphylococcus species were identified, but the bulk of unidentified bacterial isolates were mainly mixed colonies of rods, cocci, coccobacilli. and some filamentous types. Six‐hour agar settle‐plate counts during sandstorms were 1(X) and 40% higher for bacteria and fungi, respectively, than without sandstorms. The most abundant aeroallergens were those of Acacia, Alternaria, Aspergillus, Bermuda grass, Ciadosporium, cultivated rye, Prosopis, and cat dander. Pithecellobium duke, Rumex crispus, and Washingtonia palm allergens were detectable for the first time in Riyadh. IgE reactivities ofthe dust in man were demonstrated by ELISA using sera from atopic, exposed, and normal subjects. These results indicate that sandstorm dust is a prolific source of potential triggers of allergic and nonallergic respiratory ailments, and the methods mentioned here should be routinely used for quick sampling of the environment.

Cladosporium and respiratory allergy: Diagnostic implications in Saudi Arabia

An allergological study to evaluate allergenicity to Cladosporium, Burkard 7-Day Volumetric Spore Trap and Personal Volumetric air sampler (viable mode) were employed to conduct air sampling for 12 months in three regions of Saudi Arabia. The study was extended for a continuous 3rd year at one site. Skin prick testing (SPT) was also conducted on 605 allergic individuals using commercial extracts of C. herbarum. Cladosporium emerged to be the most prevalent genus in the outdoor environment constituting up to 25% of all fungal spores in the dry region and 37.1 and 41.2% in two coastal cities respectively. Amongst the species C. sphaerospermum, C. macrocarpum, C. cladosporioides and C. herbarum were noted. Maximum hourly concentrations up to 14 × 103 m−3 were recorded in coastal region during winter months. Morning concentrations were higher at both city sites compared to afternoon concentration. SPT result revealed an overall 19.67% positive reactions with majority showing mild reactions.

Changes in peripheral blood lymphocyte subsets associated with marathon running.

The percentage of peripheral blood mononuclear leukocytes that reacted with monoclonal antibodies specific for T-lymphocytes (CD3 cells), the helper/inducer subsets (CD4 cells), and cytotoxic/suppressor subsets (CD8 cells) of T-lymphocytes, and cells with NK activity (CD16 cells) were enumerated by fluorescence-activated flow cytometry for samples obtained immediately before and after the marathon running. It was found that long-term physical exercise resulted in a significant (P < 0.04 for relative and P < 0.008 for absolute lymphocytes) reduction in CD3 cells. A significant (P < 0.009) percentage change was also observed in B lymphocytes (CD19 cells) right after the marathon. The number of NK (CD16 cells) lymphocyte subsets was significantly (P < 0.05 for relative and P < 0.03 for absolute lymphocytes) changed. No significant changes were recorded for CD4, CD8, or CD4/CD8 ratios after the marathon run. A marked leukocytosis was noticed after the endurance exercise and the mean white blood cell (WBC number was increased from 7.8 +/- 2.6 to 22.9 +/- 2.8 x 10(9) cells x 1-1) count was changed by a factor of 2.9. The mean serum cortisol was significantly (P < 0.0001) increased. No hematocrit change was recorded in subjects pre- to post-run. The results of this study demonstrated that long-term physical exercise (marathon running) influenced the T-cell subsets remarkably and produced leukocytosis that was stress dependent and correlated with the increased serum cortisol levels and not the hemoconcentration.

Presence of leukemia inhibitory factor and interleukin-12 in human Follicular fluid during Follicular growth

PROBLEM: Cytokines have been shown to be present in human follicular fluid and have regulatory functions on follicular maturation. The presence of leukemia inhibitory factor (LIF) and interleukin (IL)‐12 in human follicular fluid obtained at different stages of maturation was investigated.

Fresh Frozen Plasma Contains Viable Progenitor Cells – Should We Irradiate?

While it is well established [I] that cellular blood products may be the cause of transfusion-associated graft-versus-host disease (TAGVHD), acellular products such as fresh plasma, fresh frozen plasma (FFP), and cryoprecipitate are widely regarded as containing too few immunocompetent cells to pose a serious threat even to the immunocompromised recipient. Only one report has identified passenger leukocytes in fresh plasma as the likely cause of TAGVHD [2]. The freeze-thaw procedure required for FFP and cryoprecipitate makes it even more unlikely that such products may contain sufficient numbers of viable lymphocyted and progenitor cells. The minimal dose of immunocompetent cells needed to induce TAGVHD is not fully established but a figure of lo7 lymphocytes per kilogram has been suggested, based on animal studies [3]. FFP is obtained from whole-blood donations after an initial spin to create platelet-rich plasma, followed by a second spin to create platelet concentrate. FFP is expressed after the second spin, leaving approximately 50 ml plasma with the platelet component and stored below -18°C. We looked at nucleated cell counts, viability, and colony-forming capacity in cells isolated from a series of FFP. FFP was transferred to plastic tubes and spun, and all sedimented cells were collected and resuspended in RPMI-1640 medium. Viability was assessed by trypan blue dye exclusion. Cell concentration was adjusted to lo6 per ml and seeded onto Terry Fox complete methyl cellulose media fortified with interleukin-3, stem cell factor and human placcnta conditioning media. Plates were incubated at 37°C in a humid atmosphere containing 5% C 0 2 . Colonies were counted after 2 weeks of incubation. Results are shown in table 1. Fresh Frozen Plasma Contains Viable Progenitor Cells Should We Irradiate?