The potential of extracellular microvesicles of mesenchymal stromal cells in obstetrics.

BACKGROUND: The rate of cesarean deliveries is steadily growing worldwide as a result of increasing maternal age at first delivery. Ensuring optimal recovery after surgery, specifically the development of a functionally competent uterine scar to facilitate vaginal birth after a cesarean delivery (VBAC), is one of the challenges in modern obstetrics. Extracellular microvesicles (EMVs) are secreted by multiple cell types and act as mediators of intercellular interaction during tissue reparation. The immunomodulatory and regenerative effects of EMVs of mesenchymal stromal cells (MSCs) have been studied shown in pre-clinical studies. AIM OF THE STUDY: To evaluate the safety profile of EMVs of mesenchymal stromal placental cells (MSPCs) injected during the cesarean delivery and the impact of this pilot approach on post-surgery recovery. MATERIALS AND METHODS: This pilot study included 53 women undergoing cesarean delivery with (n = 23) or without (n = 30) an injection of 500 µl of MSC EMVs after closing the uterine incision with a single continuous Vicryl suture. RESULTS: All study participants had uncomplicated post-surgery period. The mean inpatient stay duration in women receiving the EMV injection was 4.26 ± 0.09 days vs. 5.33 ± 0.38 in the control group (p<.05). There were no postpartum inflammatory complications in the study group compared with two cases (6.7%) by postpartum endometritis/myometrial infection and one case (3.3%) of lochiometra in the control group. SUMMARY: Intra-surgery injection of MSC EMVs was well-tolerated and associated with a lower rate of infectious post-partum complications in women undergoing cesarean delivery.

Blood Serum Levels of Proinflammatory Cytokines (IL-1ß, IL-6, TNFα, IL-8, IL-12p70, and IFNγ) in Patients with Uterine Myoma.

We analyzed cytokine profile in blood serum of patients with uterine myoma and revealed significantly reduced level of IFNγ and a tendency towards a decrease in the levels of IL-1ß and TNFα; the levels of IL-6, IL-8, and IL-12p70 did not differ from those in healthy women. The drop in the concentrations of factors responsible for inflammation and angiogenesis in tissues are unfavorable for proliferation and differentiation of the uterine tissues.

Ganglioside biosynthesis in mouse embryos: sialyltransferase IV and the asialo pathway.

The in vitro activity of sialyltransferase IV (SAT-IV), which catalyzes the transfer of sialic acid to the terminal galactose of different gangliotetraosylceramides (GA1, GM1a and GD1b), was examined in membrane-enriched preparations from mouse embryos at embryonic day 12 (E-12). Gangliosides GD1a and GT1b were the only reaction products using GM1a and GD1b as substrates, respectively. The Km values for GM1a and GD1b were 53 microM and 42 microM, respectively. Competitive inhibition experiments showed that the same enzyme (SAT-IV) catalyzed sialic acid transfer to the terminal galactose residues of both GM1a and GD1b. Two labeled ganglioside products were obtained, however, using GA1 as a substrate. One product was identified as ganglioside GM1b and the enzymatic reaction for its formation was maximal at pH 6.0, similar to that for GD1a and GT1b formation. The second product, synthesized by a different sialyltransferase, was identified as GD1 alpha based on results from TLC immunostaining, neuraminidase digestion, and periodate oxidation-borohydride reduction. The pH dependence curve for GD1 alpha formation had a different shape than that for GM1b formation with a maximum at pH 6.3. GD1 alpha is apparently synthesized from GM1b by an endosialyltransferase that catalyzes the transfer of a second sialic acid to the internal N-acetylgalactosamine of GM1b. The formation of both GM1b and GD1 alpha was linear over protein concentration. The ratio of GD1 alpha/GM1b formation varied from 0.25 to 1.20 depending on the GA1 substrate concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

Ganglioside GD3 biosynthesis in normal and mutant mouse embryos.

CMP-sialic acid:GM3 sialyltransferase (GD3 synthase; EC 2.4.99.8) was characterized in a membrane-enriched preparation (P2 pellet) from mouse embryos at embryonic day 12 (E-12). Gangliosides GD3 and GM3 were the major radiolabeled products of the reaction. Optimum GD3 synthase activity was obtained at pH 6.0 using 0.1% detergent Triton CF-54. The Km values for GM3 and CMP-sialic acid were 55 and 80 microM, respectively. The Vmax value was calculated as 622 pmol/mg protein/hr. Ganglioside GD3, as end product, induced a two-step reduction of enzyme activity in the range of concentrations from 0 to 34 microM (40%) and from 150 to 300 microM (65%). The rate of GD3 formation was similar in whole embryos and in embryo head and body regions. GD3 synthase activity in tw1/tw1 mutant mouse embryos, which express defects in neuronal differentiation, was only 40% of that in normal wild-type (+/+) embryos. Enzyme activity in heterozygous (+/twl) embryos was similar to that in +/+ embryos. These findings suggest that the reduced GD3 synthase activity in the mutants might arise as a consequence of failed nervous system development and might reflect a secondary rather than a primary effect of the mutation.

[The immunological cellular phenotype and the prognosis in neuroblastoma in children]. / Immunologicheskii fenotip kletok i prognoz pri neiroblastome u detei.

Ganglioside profile was evaluated in 19 samples of tumor tissue obtained from 13 surgical patients with various morphological patterns of neuroblastoma. In six of those cases, two samples from each proving most different in terms of cell maturity were selected for examination. The relative content of GD2 gangliosides was 27.0-37.6% in sympathoblastoma and as low as 5.1-14.8% in ganglioneuroblastoma. Ganglioneuroblastomas showed fairly high levels of GM1 and GD1a gangliosides which were almost completely absent in sympathoblastomas. Ganglioside profile variations seen within each tumor type were incomparable with differences in profile established between morphological patterns of neuroblastoma studied.

[The membrane sialoglycolipids of tumor cells and the metastasis of bone tumors]. / Sialoglikolipidy membran opukholevykh kletok i metastazirovanie opukholei kostei.

Level and profile of gangliosides were studied in osteogenic and chondrosarcoma cells. Level of lipid-binding sialic acids in bone- and cartilage-producing tumors proved different. Most osteogenic sarcoma samples showed higher level of lipid-binding sialic acids as compared to chondrosarcoma. In the latter tumor, level of lipid-binding sialic acids was related to grade of tumor cell differentiation, peak levels being observed in undifferentiated neoplasms as compared to those showing grade I-II cell anaplasia. Chondro- and osteogenic sarcoma revealed different profiles of sialoglycolipids, particularly, due to markedly reduced set of gangliosides and nearly complete loss of polysialogangliosides in the latter tumor.

[Ganglioside content and profile in the ovarian tumor tissues and in the biological body fluids of patients]. / Soderzhanie i profil' gangliozidov v tkaniakh opukholei iaichnikov i biologicheskikh zhidkostiakh organizma bol'nykh.

Lipid-bound sialic acid (LSA) content and ganglioside composition in resected tumours and body fluids of patients with benign ovarian tumours, borderline ovarian tumours, ovarian cancer and also in unaltered ovaries of patients with uterine cancer were examined. LSA levels in tissues and relation of the main gangliosides GD3/GM3 progressively decreased from unaltered ovaries to ovarian cancer. Distribution of gangliosides GD3 and GM3 in the borderline tumours was not uniform. Absolute content of GD3 increased more than twice in profuse growth from the inner surface and decreased almost three times in the cyst capsule as compared to the intact tumour. Ganglioside GD3 content decreased in malignant ovarian tumours but increased in ascitic fluid of cancer patients as compared to GM3. These results suggest that ganglioside GD3 is shed more intensively in the borderline ovarian tumours.

[ICO-13 monoclonal antibodies to the differentiation antigen of human hemopoietic cells]. / Monoklonal'nye antitela IKO-13 k differentsirovochnomu antigenu gemopoéticheskikh kletok cheloveka.

Monoclonal antibodies (Mab) ICO-13 of IgM isotype reacted in indirect surface immunofluorescence test with 88.5 +/- 3.4% of thymocytes and 7.1 +/- 2.4% of T cells, but failed to react with other peripheral blood cells from healthy donors. The antigen disappeared in cells stored at 4 degrees C overnight or at -196 degrees C in liquid nitrogen. The antigen detected by Mab ICO-13 was expressed on blast cells of acute lymphoblast leukemias and lymphomas. The antigen was absent in chronic lymphoblast leukemia and blast crisis of chronic myeloid leukemia.

[Selective shedding of gangliosides by cells of mouse ascitic sarcoma-37]. / Izbiratel'noe sbrasyvanie gangliozidov kletkami astsitnoi sarkoma-37 myshei.

The profile and content of gangliosides associated with tumour cells of mouse sarcoma-37 and exfoliated from the cell surface were determined. It was shown that 20% of tumour gangliosides shed from the cell surface and only about 7% of all the shed gangliosides were contained in plasma membrane fragments. When incubated in vitro at 37 degrees C for 1 hour, the tumour cells were found to release less than 2% of cellular gangliosides. The main components of cellular gangliosides corresponded in their chromatographic behaviour to GM2 (31-32% of the total ganglioside content), GD3 (17-18%), GD1a (9-11%), GD2 (16-17%) and hematosides (19-21%). The profiles of in vivo and in vitro shed gangliosides were similar but strongly differed from those of cellular gangliosides: the relative content of GM2 was 70-75% and the other gangliosides were found in negligible amounts. The data show that GM2 accumulation in extracellular spaces is rather the result of its selective shedding than the shedding of products of "incomplete" cellular ganglioside synthesis.

[Ganglioside GD3 in the serum of tumor patients]. / Gangliozid GD3 v syvorotke krovi opukholenositelei.

Gangliosides of normal human blood serum and of blood serum of tumour hosts were studied. It has been shown the first time that ganglioside (NeuAc)2LacCer is present in the blood serum of different tumour hosts, but is absent in normal blood serum. The reasons for the appearance of this ganglioside in blood serum are discussed.

[Effect of cyclophosphamide on the ganglioside content and profile in metastasizing and nonmetastasizing mammary carcinoma in mice]. / Vliianie tsiklofosfamida na soderzhanie i profil' gangliozidov v metastaziruiushchei i nemtastaziruiushchei kartsinome molochnoi zhelezy myshei.

The content and profile of gangliosides (Gs) were investigated in metastatic (HMC-L) and nonmetastatic (HMC-O) mouse mammary carcinomas before and after cyclophosphamide (CP) treatment. It was shown that the Gs content did not alter markedly during HMC-O growth, whereas in HMC-L, it increased to maximum on day 15 after tumor inoculation and then dropped. CP treatment led to a steady rise in the Gs content in both tumors. The main Gs in HMC-O corresponded to GDIa, GMI, GDIb and hematosides, while in HMC-L, the main Gs at days 9-15 after tumor inoculation corresponded to GDIa (50-65% of total Gs). The action of CP on HMC-O was not accompanied by any essential alterations in the Gs profile, whereas in HMC-L on days 3-6 after CP treatment the relative content of GDIa was significantly lowered and the GMI and hematoside content markedly increased. These findings indicate that there is an obvious relation between the Gs profile and tumor ability to form metastases.

[Circulating lipid-bound sialic acid levels in patients with malignant tumors]. / Urovni tsirkuliruiushchikh lipidno-sviazannykh sialovykh kislot u bol'nykh so zlokachestvennymi novoobrazovaniiami.

Lipid-bound sialic acids were estimated in blood serum of 118 patients with tumors, of 13 donors and of 38 patients with non-tumoral diseases. Content of the sialic acids in blood serum of the patients with growing tumors constituted 23.1 +/- 1.1 mg/100 ml, which was 2-2.5-fold higher as compared with healthy donors (10.5 +/- 0.4 mg/100 ml) or with the patients with non-tumoral diseases (13.8 +/- 0.5 mg/100 ml). In the oncologic patients during the remission period the content of lipid-bound sialic acids in blood serum approached the normal level - 12.1 +/- 0.5 mg/100 ml. The highest level of the sialic acids was found in patients with lymphogranulomatosis II-IV step, with melanoma during the period of growth, with kidney tumor and with liver tissue primary tumor. Estimation of circulating lipid-bound sialic acids in the oncologic patients may serve as a test for evaluation of the tumor growth as well as of therapeutic treatment efficiency.

[Changes in the content and composition of gangliosides of tumors as affected by chemotherapeutic agents]. / Izmenenie soderzhaniia i sostava gangliozidov opukholei pri deistvii khimioterapevticheskikh preparatov.

The lipid-bound sialic acid (LSA) levels increased in the metastasizing Lewis lung carcinoma (3LL) and significantly decreased in blood plasma of the tumour-bearing mice after cyclophosphamide treatment correlating with the drug therapeutic effect. 5-fluorouracil that was less active in the used therapeutic doses did not cause similar changes in the LSA levels. No correlation was found between the LSA levels in adenocarcinoma 755 and the drug antitumour activity. It was observed that the content of hematoside, GM1 and GD1b sharply increases and GD3 and GD1a levels significantly decrease under the cyclophosphamide treatment of the Lewis tumour.

[Serum lipids in epithelial ovarian tumor patients]. / Lipidy syvorotki krovi u bol'nykh épitelial'nymi opukholiami iaichnikov.

The report deals with the studies of lipid profile of blood serum and alpha-lipoproteins in 59 cases of epithelial ovarian tumors as well as the effects of surgery and chemotherapy on the serum levels of major lipids. Triglyceride and phospholipid levels In the serum of cancer patients appeared to be much higher than in healthy controls. Serum lipid distribution patterns were normal in patients with benign tumors of ovaries. Serum levels of total cholesterol and alpha-lipoprotein phospholipids in cancer patients were much lower than in healthy controls thus pointing to a considerably decreased level of this fraction of lipoproteins. Surgery was shown to be followed by a noticeable rise in triglyceride levels and a decrease in serum phospholipid concentration in cancer patients. Post-operative polychemotherapy was followed by reversed changes in triglyceride and phospholipid concentrations. It was found that polychemotherapy is not an important factor affecting the serum level of alpha-lipoproteins in untreated patients.