A cell line, RTgutGC, was developed from the intestine of Oncorhynchus mykiss. RTgutGC has an epithelial‐like shape, been passaged over 100 times, and cryopreserved successfully. A rainbow trout origin was confirmed by sequencing a 652 bp region of the mitochondrial cytochrome c oxidase I gene. RTgutGC is grown routinely in Leibovitz’s L15 without glutamine supplemented with 10% fetal bovine serum (FBS). Cell viability was evaluated with Alamar blue (AB) for metabolic activity and carboxyfluorescein diacetate acetoxymethyl ester (CFDA AM) for membrane integrity. Viability was unchanged by lipopolysaccharide (LPS) for cultures in FBS. For cultures at low cell densities in L15 without FBS or glutamine, cell viability declined in a LPS dose‐dependent manner, allowing calculation of the concentration causing a 50% decline in viability (EC50). When glutamine was present, the EC50 was increased for both AB and CFDA AM. As the cell density increased, LPS became much less cytotoxic and no EC50 could be calculated for very confluent cultures. Only high‐density cultures had alkaline phosphatase (AP) activity. Thus, glutamine and possibly AP protect against LPS cytotoxicity. RTgutGC should be a useful in vitro tool for studying problems of nutrition and gastrointestinal health in fish.

CD44 proteins are cell surface receptors for hyaluronic acid (HA), a component of the extracellular matrix that has multiple effects on cell behavior. CD44 can be shed from the cell surface by proteolytic cleavage. The resulting soluble form can interfere with the interaction between HA and membrane-bound CD44. Soluble CD44 can abolish the cell proliferation-promoting effect of HA on melanoma cell lines, suggesting that a better understanding of the shedding process might identify ways of blocking tumor cell proliferation. ADAM10, ADAM17, and MMP14 have previously been implicated in the shedding of CD44 from various tumor cells. Using immunohistochemistry we demonstrate that ADAM10 and ADAM17 but not MMP14 are significantly expressed on melanoma cells in histological sections. In human melanoma cell lines expression of these proteases could be blocked by transfection with appropriate siRNAs. However, only blocking of ADAM10 expression led to decreased shedding of CD44. In parallel, cell proliferation was promoted. Confocal microscopy demonstrated that ADAM10 and CD44 colocalize on the cell surface. We conclude that ADAM10 is the predominant protease involved in the constitutive shedding of endogenous CD44 from melanoma cells, and that enhancement of ADAM10 activity could be an approach to decrease the proliferation of melanoma cells.

Background: The classic mid-trimester preterm premature rupture of membranes (PPROM) is defined as a rupture of the fetal membranes prior to 28 weeks of gestation (WG) with oligo/anhydramnion; it complicates approximately 0.4–0.7% of all pregnancies and is associated with very high neonatal mortality and morbidity. Antibiotics have limited success to prevent bacterial growth, chorioamnionitis and fetal inflammation. The repetitive amnioinfusion does not work because fluid is lost immediately after the intervention. The continuous amnioinfusion through the transabdominal port system or catheter in patients with classic PPROM shows promise by flushing out the bacteria and inflammatory components from the amniotic cavity, replacing amniotic fluid and thus prolonging the PPROM-to-delivery interval. Objective: This multicenter trial aims to test the effect of continuous amnioinfusion on the neonatal survival without the typical major morbidities, such as severe bronchopulmonary dysplasia, intraventricular hemorrhage, cystic periventricular leukomalacia and necrotizing enterocolitis one year after the delivery. Study Design: We plan to conduct a randomized multicenter trial with a two-arm parallel design. Randomization will be between 22/0 and 26/0 SSW. The control group: PPROM patients between 20/0 and 26/0 WG who will be treated with antibiotics and corticosteroids (from 22/0 SSW) in accordance with the guidelines of German Society of Obstetrics and Gynecology (standard PPROM therapy). In the interventional group, the standard PPROM therapy will be complemented with the Amnion Flush Method, with the amnioinfusion of Amnion Flush Solution through the intra-amnial catheter (up to 100 mL/h, 2400 mL/day). Subjects: The study will include 68 patients with classic PPROM between 20/0 and 26/0 WG. TRIAL-registration: ClinicalTrials.gov ID: NCT04696003. German Clinical Trials Register: DRKS00024503, January 2021.

No abstract

Background: The classic mid-trimester preterm premature rupture of membranes (PPROM), is defined as rupture of fetal membranes prior to 28 weeks’ gestation (WG) with oligo/ anhydramnion, complicates approximately 0.4-0.7% of all pregnancies and is associated with very high neonatal mortality and morbidity.Antibiotics have limited success to prevent bacterial growth, chorioamnionitis and fetal inflammation. The repetitive amnioinfusion doesn’t work because of immediately fluid lost after the intervention. The continuous amnioinfusion through the transabdomianal port system or catheter in patients with classic PPROM shows promise by flush out of bacteria and inflammatory components from the amniotic cavity, replacing amniotic fluid and thus prolonging PPROM-to-delivery interval.Aim: This multicenter trial tests the effect of continuous amnioinfusion on the neonatal survival without the typical major morbidities, like severe bronchopulmonary dysplasia, intraventricular hemorrhage, cystic periventricular leukomalacia and necrotizing enterocolitis one year after the delivery.Methods/Design: randomized multicenter trial; two-arm parallel design. Control group: PPROM patients between 22/0 (20/0) -26/0 WG treated with antibiotics and corticosteroids in accordance to guidelines of German Society of Obstetrics and Gynecology (standard PPROM therapy). In the interventional group the standard PPROM therapy will be complemented by “Amnion Flush Method” with the amnioinfusion of artificial amniotic fluid (up to 100 ml/h, 2400 ml/day).Subjects: 68 patients with classic PPROM between 22/0 (20/0)-26/0 WG.TRIAL-registration: ClinicalTrials.gov ID: NCT04696003 and German Clinical Trials Register: DRKS00024503, January 2021.The trial is approved by the Ethic committee of the Martin-Luther University Halle-Wittenberg (2020-185, January 25, 2021).