Should the modified natural cycle protocol for frozen embryo transfer be modified? A prospective case series proof of concept study.

OBJECTIVE: Modified natural cycles for frozen embryo transfer utilize an ovulation trigger which assists in embryo transfer scheduling and simplifies cycle monitoring. There have been conflicting results with this protocol and modifications may be sought. We wanted to ascertain whether a modified natural protocol for frozen embryo transfer without triggered ovulation but with luteal progesterone support disconnecting the timing of embryo transfer from the timing of the LH surge can achieve a high pregnancy rate. STUDY DESIGN: Candidates for frozen embryo transfer of 48-h cleavage cell embryos were recruited from May 2016 to April 2018. The patients were monitored for endometrial growth, follicle formation and estradiol, progesterone, and LH hormone levels. After meeting the predetermined criteria, embryo transfer was scheduled. The patients began progesterone treatment 48 h before embryo transfer, regardless of identification of the LH surge if ovulation had not commenced. The predetermined primary outcome was the biochemical pregnancy rate while the secondary outcome included the clinical pregnancy rate and the ongoing pregnancy rate. Patients were monitored to the eighth week of pregnancy, but data was collected from the medical records to provide the live birth rate as well. RESULTS: Fifty-six women were screened. Eleven women declined or did not meet the inclusion criteria. Three had anovulatory cycles and were excluded. Forty-two women were included in the statistical analysis. The implantation rate was 42.9 % [95 %CI 29.3 %-56.4 %). Of the 42 participants, 25 (59.5 %) conceived [95 % CI 44.0 %-75 %]. Two pregnancies ended in first trimester miscarriage leaving 23 (54.7 %) ongoing pregnancies [95 % CI 39.1 %-70.5 %]. One patient experienced a late abortion such that the live birth rate was 22 of 42 patients or 52.4 % [95 % CI 36.4 %-68.0 %]. CONCLUSION: The proposed modified natural protocol which utilizes progesterone luteal support but does not trigger ovulation, maintains a high pregnancy rate while providing flexibility regarding the day of transfer disconnected from the day of the LH surge. This was a prospective, proof of concept study. This protocol may be suitable for smaller or public in-vitro fertility units whose resources are limited and facilities are not available daily. The high pregnancy and live birth rate that we found provides confidence that this protocol can be part of the armament of protocols the clinician may offer to his patients. Larger studies should confirm these findings.

Planned oocyte cryopreservation (Planned OC): systematic review and meta-analysis of cost-efficiency and patients' perspective.

BACKGROUND: Advances in vitrification techniques have enabled planned oocyte cryopreservation ('Planned OC'). OBJECTIVES: To explore the cost-efficiency and utilisation of planned OC, as well as patients' perspectives on the process. SEARCH STRATEGY: A systematic search in PubMed/MEDLINE, Embase, Cochrane Database and PsychINFO, for all relevant studies published between January 2007 and December 2019. SELECTION CRITERIA: The protocol followed PRISMA guidelines in PECO format, and was registered with PROSPERO. DATA COLLECTION AND ANALYSIS: Two independent reviewers evaluated all manuscripts for inclusion eligibility. Authors were contacted for missing data. Included studies were assessed for risk of bias and for heterogeneity. Weighted effects were measured and plotted. MAIN RESULTS: The search yielded 12 545 records, of which 43 were included. Planned OC is cost-efficient at 35, assuming 60% utilisation; and at 37 assuming utilising donor sperm when necessary. At 38 it is cost-efficient to defer planned OC in favour of undergoing 2 IVF cycles. Currently, utilisation of banked-oocytes within 22-58 months, is up to 15%. Nine percent of warmed banked oocytes result in life births. Online resources and treating physicians are equally important sources of information regarding planned OC. Most patients think planned OC is ideal before age 35 and are not fully aware of what the process entails and tend to overestimate the success rates. The main barrier to wider endorsement of planned OC is being wary of potential health implications or of limited success. CONCLUSION: Planned OC is an adequate method for preserving fertility. However, knowledge gaps result in under-utilisation leading to reduced cost-efficiency. TWEETABLE ABSTRACT: Identifying facilitators and barriers for wider adoption of banking oocytes can enhance the cost-efficiency of this modality.

Assessing the impact of dairy waste lagoons on groundwater quality using a spatial analysis of vadose zone and groundwater information in a coastal phreatic aquifer.

Dairy waste lagoons are considered to be point sources of groundwater contamination by chloride (Cl(-)), different nitrogen-species and pathogens/microorganisms. The objective of this work is to introduce a methodology to assess the past and future impacts of such lagoons on regional groundwater quality. The method is based on a spatial statistical analysis of Cl(-) and total nitrogen (TN) concentration distributions in the saturated and the vadose (unsaturated) zones. The method provides quantitative data on the relation between the locations of dairy lagoons and the spatial variability in Cl(-) and TN concentrations in groundwater. The method was applied to the Beer-Tuvia region, Israel, where intensive dairy farming has been practiced for over 50 years above the local phreatic aquifer. Mass balance calculations accounted for the various groundwater recharge and abstraction sources and sinks in the entire region. The mass balances showed that despite the small surface area covered by the dairy lagoons in this region (0.8%), leachates from lagoons have contributed 6.0% and 12.6% of the total mass of Cl(-) and TN (mainly as NO3(-)-N) added to the aquifer. The chemical composition of the aquifer and vadose zone water suggested that irrigated agricultural activity in the region is the main contributor of Cl(-) and TN to the groundwater. A low spatial correlation between the Cl(-) and NO3(-)-N concentrations in the groundwater and the on-land location of the dairy farms strengthened this assumption, despite the dairy waste lagoon being a point source for groundwater contamination by Cl(-) and NO3(-)-N. Mass balance calculations, for the vadose zone of the entire region, indicated that drying of the lagoons would decrease the regional groundwater salinization process (11% of the total Cl(-) load is stored under lagoons). A more considerable reduction in the groundwater contamination by NO3(-)-N is expected (25% of the NO3(-)-N load is stored under lagoons). Results demonstrate that analyzing vadose zone and groundwater data by spatial statistical analysis methods can significantly contribute to the understanding of the relations between groundwater contaminating sources, and to assessing appropriate remediation steps.

Infiltration mechanism controls nitrification and denitrification processes under dairy waste lagoon.

Earthen waste lagoons are commonly used to store liquid wastes from concentrated animal feeding operations. The fate of ammonium (NH) and nitrate (NO) was studied in the vadose zone below earthen-clay dairy farm waste lagoons using three independent vadose zone monitoring systems. The vadose zone was monitored from 0.5 to 30 m below land surface through direct sampling of the sediment porewater and continuous measurement of the sediment profile's water content variations. Four years of monitoring revealed that wastewater infiltration from the lagoon is controlled by two mechanisms: slow (mm d), constant infiltration from the lagoon bed; and rapid (m h) infiltration of wastewater and rainwater via preferential flow in desiccation cracks formed in the unsaturated clay sediment surrounding the lagoon banks. The preferential flow mechanism is active mainly during wastewater-level fluctuations and intensive rain events. The vadose zone below the waste sources remained unsaturated throughout the monitoring period, and all infiltrating NH was oxidized in the upper 0.5 m. The NH oxidation (nitrification) was coupled with NO reduction (denitrification) and depended on the sediment water content, which was controlled by the infiltration mechanism. Coupled nitrification-denitrification (CND) resulted in 90 to 100% reduction in the total nitrogen mass in the vadose zone, with higher removal under high water content (∼0.55 m m). Mass balance of nitrogen and isotopic composition of NO indicated that CND, rather than cation exchange capacity, is the key factor regulating nitrogen's fate in the vadose zone underlying earthen waste lagoons.

The abnormal contralateral atrioventricular valve in mitral and tricuspid atresia in neonates: An echocardiographic study.

Abnormalities of the mitral valve (MV) or the tricuspid valve (TV) morphology and/or function in patients with functional single ventricle may result in early morbidity and death. The purpose of this study was to determine the incidence of contralateral atrioventricular valve (AVV) pathologies in mitral valve atresia (MA) and tricuspid valve atresia (TA). We retrospectively reviewed the echocardiographic data of 50 neonates with MV and 20 with TA. Appearance of the papillary muscles, chordae tendinae, and valve leaflets was assessed. AVV regurgitation was semiquantitated by color-flow Doppler and the AVV annulus diameter was measured and indexed to body surface area. MV abnormalities were found in 9 of 20 (45%) of patients with TA. The MV was myxomatous in 9 patients, the leaflets were redundant in 5 patients, and prolapsing occurred in 4 patients. Mild regurgitation was found in 2 patients. In 18 of 20 (90%) patients MV annulus size was larger than 95% of predicted normal values. TV abnormalities were found in 12 of 50 (24%) patients with MA. The TV was myxomatous in 4 patients, prolapsing in 2, and redundant in 3, and moderate TV regurgitation was found in 3 patients. In 29 of 50 (58%) patients TV annulus size was larger than 95% of predicted normal values. Contralateral AVV abnormalities in tricuspid and mitral valve atresia are common and should be assessed carefully before surgical procedures.

Alterations in membrane lipid dynamics of leukemic cells undergoing growth arrest and differentiation: dependency on the inducing agent.

The effect of various differentiation inducers on membrane cell dynamics was studied using HL-60 and K562 leukemic cell lines. Membrane lipid dynamics was measured by the steady-state fluorescence polarization (P) method utilizing either 1,6-diphenyl-1,3,5-hexatriene (DPH) or the trimethyl ammonium derivative of DPH (TMA-DPH), which ascertains anchorage of the label to the membrane-water-lipid interface. Decrease in membrane microfluidity was observed in HL-60 cells undergoing differentiation into macrophages by 1,25-dihydroxyvitamin D3 and by K562 cells induced to differentiate by DMSO. Sodium butyrate caused an increase in membrane fluidity in K562 cells undergoing differentiation into erythroid-like cells while in HL-60 cells a dual effect was observed. At 0.4 mM concentration, in which the cells were induced to differentiate along the monocyte pathway, a decrease in membrane fluidity was observed, while at 1 mM concentration an increase in membrane fluidity occurred. Interferon-gamma (IFN-gamma) induced an increase in membrane fluidity in both cell lines. Using HL-60 cells fluorescently labeled by TMA-DPH, similar results indicating fluidization of the membrane following IFN-gamma treatment were obtained. Advanced fluorescence lifetime measurements, evaluated either by phase modulation spectrofluorometry or by single photon correlation fluorometry confirmed that the decrease in fluorescence polarization by IFN-gamma resulted from membrane fluidization and not from elongation of the probe's excited state lifetime. It is suggested that the inducer mode of action, and not the differentiation route, determine the outcome of changes in membrane microviscosity.

Effect of sinusoidally varying magnetic fields on cell proliferation and adenosine deaminase specific activity.

The effect of sinusoidally varying magnetic fields (SVMF) on chick embryo fibroblasts (CEF) was examined by two independent methods: 1) measurement of cell proliferation at 0.06-0.7 mT (100, 60 and 50 Hz) using a colorimetric assay (MTT); 2) monitoring of specific activity of adenosine deaminase (ADA) at 0.3 and 0.7 mT, 60 Hz. Both increased cell proliferation and reduced ADA specific activity are associated with cell transformation. The MTT test showed an increase in cell proliferation of up to 64% after a 24 h exposure to SVMF at 100 Hz, 0.7 mT. Cell proliferation at constant frequency (100 Hz) depended on SVMF intensity. Cell proliferation at constant intensity (0.7 mT) increased with increasing field frequency. At 0.7 mT, 60 Hz cell proliferation increased by 31%, 28%, and 26% when measured by hemocytometry, 3H-thymidine incorporation, and the MTT assay, respectively. ADA specific activity in CEF decreased by circa 48% on exposure to SVMF at 60 Hz, 0.3 mT for 24 h; only a statistically insignificant trend was seen at 0.7 mT, 60 Hz. Our findings showed that CEF cell proliferation and ADA specific activity were modified by SVMF. Both methods, independently, qualitatively detect a magnetic field effect.

Interaction of a novel fluorescent analog of interferon-gamma with transformed cells.

A fluorescent analog of human recombinant interferon-gamma (IFN-gamma) was prepared for the first time. The recovered pyrene-labeled IFN-gamma (py-IFN-gamma), with an estimated seven pyrene molecules per IFN-gamma, retained over half of its original biological activity. Binding of py-IFN-gamma to human amnion WISH cells showed appreciable enhancement in fluorescence polarization from 0.055 to 0.215 and in fluorescence lifetime from 56 to 80 ns. The ratio of the vibronic peaks did not change, indicating that the pyrene molecules remained in water environment even after binding. Py-IFN-gamma provides a novel tool for unraveling the mechanism of the initial interaction between this antiproliferative lymphokine and its target, cancer cell membrane receptors. Its fluorescence could provide the means to follow receptor recycling when it occurs.

Painless intussusception. Giving conservative treatment another chance.

Treatment options for intussusception in children range from reduction by barium enema to surgical intervention. The authors describe a case in which a conservative option--a fifth attempt to reduce an ileocolic intussusception by barium enema, this time using general anesthesia--successfully resolved the problem.