The impact of circulating exosomes derived from early and late onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells.

OBJECTIVES: The pathogenesis of pre-eclampsia (PE) is associated with significant maternal and neonatal complications, an increased inflammatory response, placental hypoxia, and endothelial dysfunction, coupled with differential exosomal release profiles with immune modulation effects. Hence, this study evaluated the impact of circulating exosomes derived from early and late-onset pre-eclamptic pregnancies on inflammatory cytokine secretion by BeWo cells. STUDY DESIGN: Exosomes were isolated from plasma obtained from early-onset pre-eclamptic (EOPE; n = 15), late-onset pre-eclamptic (LOPE; n = 15), and gestational age-matched normotensive pregnancies (N ≤ 33 weeks; n = 15 and N ≥ 34 weeks; n = 15). Human BeWo cells were treated with characterized and quantified exosomes (100 µg/mL exosomal protein per pregnant group) for 24 h. The immunoassay method was used to measure the concentration of IL-8, IL-10, leptin, and HIF-α. RESULTS: Exosome administration from women with EOPE and LOPE increased IL-8 and decreased IL-10 expression in BeWo cells. CONCLUSION: Cumulatively, our data demonstrated that circulating exosomes from the placenta and activated immune cells potentially influence inflammatory cytokine production in pre-eclamptic pregnancies.

The Screening of Nails for Selected Essential and Toxic Elements in Normotensive and Pre-Eclamptic Women.

To compare the concentrations of 13 different elements in nail samples from pre-eclamptic and normotensive pregnant women. The study site was a regional hospital in Durban, KwaZulu Natal. Nail samples were collected from normotensive (n = 33) and pre-eclamptic (n = 33) pregnant women. Approximately 0.02 g of nail samples were digested in 70% nitric acid and analyzed using inductively coupled plasma-optical emission spectrometry. Analytes of interest were the following essential elements calcium (Ca), chromium (Cr), cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), nickel (Ni), selenium (Se) and Zinc (Zn) as well as toxic elements, arsenic (As), cadmium (Cd) and lead (Pb). The observed concentrations of bioelements (mean, µg/g), Ca: normotensive (N) 3467 ± 197 vs (PE) 2897 ± 190; Mg: (N) 736 ± 61 vs (PE) 695 ± 59, were lower in pre-eclampsia albeit not statistically significant. Similarly, the observed concentrations of bioelements (mean, µg/g), Cd: (N) 3 ± 0.3 vs (PE) 2 ± 0.4; Co: (N) 3 ± 0.3 (PE) not detected; Mn: (N) 7 ± 1 (PE) 4 ± 0.8, were significantly lower in pre-eclampsia (p = 0.004, 0.0001 and 0.022, respectively). Therefore, this study demonstrated significantly lower levels of Cd, Co and Mn in pre-eclampsia which justifies the need for further research on these elements towards the effective management or prevention of pre-eclampsia which could ultimately also aid in establishing its pathogenesis.

Elemental analysis of serum and hair from pre-eclamptic South African women.

Pre-eclampsia is a hypertensive disorder that is associated with adverse maternal and perinatal outcomes. It has been proposed that specific trace and macro elements associated with antioxidant activities may also play a contributory role in aetiology of pre-eclampsia. The aim of this study was to measure the concentrations of thirteen different elements in hair and serum samples from women with a diagnosis of pre-eclampsia and compare them with normotensive controls. Venous blood and pubic hair samples were collected from forty-three pre-eclamptic and twenty-three normotensive pregnant women. In each sample, the concentration of arsenic (As); calcium (Ca); cadmium (Cd); chromium (Cr); cobalt (Co); magnesium (Mg); manganese (Mn); iron (Fe); copper (Cu); lead (Pb); selenium (Se); nickel (Ni); zinc (Zn) were measured using inductively coupled plasma-optical emission spectrometry. Cobalt concentration in hair was significantly lower in the pre-eclampsia group (1.56±0.74µg/g) compared to the normotensive group (2.89±4.99µg/g) (p=0.02). The concentrations of Zn and Cr were significantly higher in hair samples from the pre-eclamptic group, compared to the normotensive control group (Zn, 395.99±48.60 vs 330.88±29.70µg/g; Cr, 13.31±2.67 vs 11.05±7.62µg/g: p≤0.05). There were no significant differences in the hair levels of other elements between groups. Serum Zn was significantly higher in the pre-eclamptic group (0.16-253.4mg/L) compared to the normotensive group (0.2-48.4mg/L) (p=0.01). Serum Ca, Co, Cu, Mg, Mn and Se levels were found to be significantly lower in the pre-eclamptic group compared to the normotensive group (p<0.05). This study confirms the association between pre-eclampsia and maternal trace as well as macro element levels.

Activation of CD35 and CD55 in HIV associated normal and pre-eclamptic pregnant women.

OBJECTIVE: The delicate balance which exists between complement activation and its regulation is altered in HIV infection and pregnancy disorders such as pre-eclampsia. Therefore, the purpose of this study was to investigate the expression of complement regulatory (Creg) proteins (CD35 and CD55) in HIV associated normal and pre-eclamptic pregnancies. STUDY DESIGN: The total study population (n=100) consisted of normotensive pregnant (n=50) and pre-eclamptic (n=50) women. These groups were equally sub-stratified into HIV infected and uninfected groups (n=25 per group). Standard haematological tests were conducted. Flow cytometric analysis of isolated neutrophils were performed using fluorescein isothiocyanate-conjugated anti-CD35 and phycoerythrin-cyanine 5 conjugated anti-CD55. RESULTS: HELLP syndrome characteristics of increased lactate dehydrogenase enzymes levels, low platelet counts, cell morphological abnormalities (red cell fragmentation) and anaemia were observed in 40% of the HIV infected pre-eclamptic group. Red cell fragmentation inclusive of burr cells and schistocytes were also noted. Activated partial thromboplastin time and fibrinogen differed significantly between the HIV uninfected pre-eclamptic compared to the HIV infected pre-eclamptic groups (p<0.01). Irrespective of HIV status, the mean fluorescence intensity of CD35 and CD55 were significantly higher in the pre-eclamptic compared to the normotensive pregnant (p=0.0001; p=0.0001 respectively) groups. In the pre-eclamptic groups, the expression of both CD35 and CD55 did not significantly differ between HIV infected and uninfected women (p=0.486; p=0.767 respectively). CONCLUSIONS: This study demonstrates an up-regulation of complement regulatory proteins, CD35 and CD55 in HIV associated pre-eclamptic compared to normotensive pregnancy. This elevation of the Creg proteins is an adaptive immune response to the high complement-mediated cell lysis that occurs in HIV infection and further aggravated by the complement activated state of pre-eclampsia.

Morphometrical analysis of placental functional efficiency in normotensive versus preeclamptic South African black women.

OBJECTIVE: To assess the umbilical cord centrality, placental morphometrics, and functional efficiency in preeclampsia. METHODS: Placental morphometry of normotensive (n = 69) and preeclamptic (n = 69) patients was evaluated. RESULTS: There was a significant reduction in mean placental surface area (p = 0.0001), length (p = 0.0001), thickness (p = 0.016), and volume (p = 0.0001) in the preeclamptic than in the normotensive groups. Umbilical cord insertion was predominantly eccentric with marginal in early (29%) and late-onset preeclampsia (16%). Placental and birth weight was lower (p = 0.0001) in preeclampsia than in the normotensive group. Placental efficiency was reduced in early-onset preeclampsia. CONCLUSION: This study demonstrates reduced placental morphometrics with impaired placental efficiency in preeclampsia.

The in vitro photodynamic effect of laser activated gallium, indium and iron phthalocyanine chlorides on human lung adenocarcinoma cells.

Metal-based phthalocyanines currently are utilized as a colorant for industrial applications but their unique properties also make them prospective photosensitizers. Photosensitizers are non-toxic drugs, which are commonly used in photodynamic therapy (PDT), for the treatment of various cancers. PDT is based on the principle that, exposure to light shortly after photosensitizer administration predominately leads to the production of reactive oxygen species for the eradication of cancerous cells and tissue. This in vitro study investigated the photodynamic effect of gallium (GaPcCl), indium (InPcCl) and iron (FePcCl) phthalocyanine chlorides on human lung adenocarcinoma cells (A549). Experimentally, 2 × 10(4)cells/ml were seeded in 24-well tissue culture plates and allowed to attach overnight, after which cells were treated with different concentrations of GaPcCl, InPcCl and FePcCl ranging from 2 µg/ml to 100 µg/ml. After 2h, cells were irradiated with constant light doses of 2.5 J/cm(2), 4.5 J/cm(2) and 8.5 J/cm(2) delivered from a diode laser (λ = 661 nm). Post-irradiated cells were incubated for 24h before cell viability was measured using the MTT Assay. At 24h after PDT, irradiation with a light dose of 2.5 J/cm(2) for each photosensitizing concentration of GaPcCl, InPcCl and FePcCl produced a significant decrease in cell viability, but when the treatment light dose was further increased to 4.5 J/cm(2) and 8.5 J/cm(2) the cell survival was less than 40%. Results also showed that photoactivated FePcCl decreased cell survival of A549 cells to 0% with photosensitizing concentrations of 40 µg/ml and treatment light dose of 2.5 J/cm(2). A 20 µg/ml photosensitizing concentration of FePcCl in combination with an increased treatment light dose of either 4.5 J/cm(2) or 8.5 J/cm(2) also resulted in 0% cell survival. This PDT study concludes that low concentrations on GaPcCl, InPcCl and FePcCl activated with low level light doses can be used for the effective in vitro killing of lung cancer cells.

In vitro photodynamic effect of aluminum tetrasulfophthalocyanines on melanoma skin cancer and healthy normal skin cells.

Photodynamic therapy is a medical treatment that uses an inactive dye/drug and lasers as a light source to activate the dye/drug to produce a toxic form of oxygen that destroys the cancer cells. This study aimed at investigating the cytotoxic effects of different concentrations of aluminum tetrasulfophthalocyanines in its inactive and active state (laser induced) on melanoma skin cancer cells, healthy normal skin fibroblast and keratinocyte cells. Experimentally, 3 × 104 cells/ml were seeded in 24-well plates before treatment with different concentrations of aluminum tetrasulfophthalocyanines. After 2h, cells were irradiated with a light dose of 4.5 J/cm². Post-irradiated cells were incubated for 24h before cell viability was measured using the CellTiter-Blue Viability Assay. Results showed that aluminum tetrasulfophthalocyanines at high concentrations were cytotoxic to melanoma cells in the absence of laser activation. In the presence of laser activation of aluminum tetrasulfophthalocyanines at a concentration of 40 µg/ml decreased cell viability of melanoma cells to 45%, fibroblasts to 78% and keratinocytes to 73%. At this photosensitizing concentration of aluminum tetrasulfophthalocyanines the efficacy of the treatment light dose 4.5 J/cm² and the cell death mechanism induced by photoactivated aluminum tetrasulfophthalocyanines was evaluated. A light dose of 4.5 J/cm² was more efficient in killing a higher number of melanoma cells and a lower number of fibroblast and keratinocyte cells than the other light doses of 2.5 J/cm², 7.5 J/cm² and 10.5 J/cm². Apoptosis features such as blebbing, nucleus condensation, nucleus fragmentation and the formation of apoptotic bodies were seen in the photodynamic therapy treated melanoma skin cancer cells. This in vitro photodynamic therapy study concludes that using aluminum tetrasulfophthalocyanines at a photosensitizing concentration of 40 µg/ml in combination with a laser dose of 4.5 J/cm² was potentially lethal for melanoma skin cancer cells and less harmful for the normal healthy skin cells.

In vitro toxicity testing of zinc tetrasulfophthalocyanines in fibroblast and keratinocyte cells for the treatment of melanoma cancer by photodynamic therapy.

A series of water-soluble tetrasulfonated metallophthalocyanines (MPcs) dyes have been studied to be used as a drug or photosensitizer (PS) in photodynamic therapy (PDT) for the treatment of cancers. During PDT the PS is administrated intravenously or topically to the patient before laser light at an appropriate wavelength is applied to the cancerous area to activate the PS. The activated PS will react with oxygen typically present in the cancerous tissue to generate reactive oxygen species for the destruction of the cancerous tissue. This in vitro study aimed at investigating the cytotoxic effects of different concentrations of zinc tetrasulfophthalocyanines (ZnTSPc) activated with a diode laser (λ = 672 nm) on melanoma, keratinocyte and fibroblast cells. To perform this study 3 × 104 cells/ml were seeded in 24-well plates and allowed to attach overnight, after which cells were treated with different concentrations of ZnTSPc. After 2h, cells were irradiated with a constant light dose of 4.5J/cm². Post-irradiated cells were incubated for 24 h before cell viability was measured using the CellTiter-Blue Viability Assay. Data indicated high concentrations of ZnTSPc (60-100 µg/ml) in its inactive state are cytotoxic to the melanoma cancer cells. Also, results showed that photoactivated ZnTSPc (50 µg/ml) was able to reduce the cell viability of melanoma, fibroblast and keratinocyte cells to 61%, 81% and 83% respectively. At this photosensitizing concentration the efficacy the treatment light dose of 4.5J/cm² against other light doses of 2.5J/cm², 7.5J/cm² and 10J/cm² on the different cell lines were analyzed. ZnTSPc at a concentration of 50 µg/ml activated with a light dose of 4.5J/cm² was the most efficient for the killing of melanoma cancer cells with reduced killing effects on healthy normal skin cells in comparison to the other treatment light doses. Melanoma cancer cells after PDT with a photosensitizing concentration of 50µg/ml and a treatment light dose of 4.5J/cm² showed certain apoptosis characteristics such as chromatin condensation and fragmentation of the nucleus. This concludes that low concentrations of ZnTSPc activated with the appropriate light dose can be used to induce cell death in melanoma cells with the occurrence of minimal damage to surrounding healthy tissue.