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The major rate-limiting step in in vitro fertilization (IVF) success appears to be the implantation of the semi-allogeneic embryo into the maternal endometrium. To determine possible risk factors of recurrent failure of embryos to implant, we investigated immunogenetic determinants as level of human leukocyte antigen (HLA) histocompatibility, frequency of killer-cell immunoglobulin-like receptors (KIR) and HLA-C alleles and HLA-G polymorphism. We DNA typed women with recurrent implantation failure (RIF) and their partners for classical HLA Class I, HLA Class II, HLA-G and KIR alleles and compared these results with couples with successful embryo implantation after their first IVF and normal fertile couples. No association was found between RIF and the degree of histocompatibility between partners or sharing of a specific antigen. Also, no significant difference in KIR haplotype or combination of HLA-C group and KIR was observed. We did find a higher frequency of HLA-C2 and a higher frequency of 14 base pair (bp) insertion in HLA-G in women with RIF. Therefore we conclude that the degree of histocompatibility between partners is not a determining factor for the occurrence of RIF. However, presence of the HLA-C2 allotype and the HLA-G allele with a 14 bp insertion is a significant risk factor. [ABSTRACT FROM AUTHOR]

Background: Systemic hypothermia with bilateral antegrade selective cerebral perfusion (ASCP) is the preferred cerebral protective strategy for type A aortic dissection surgery. The optimal ASCP flow rate remains uncertain and the target flow cannot always be reached due to pressure limitations. The aim of this study was to assess the correlation between ASCP flow and regional cerebral oxygen saturation (rSO2)., Methods: A retrospective analysis was performed on 140 patients with acute type A aortic dissection who underwent surgery with moderate hypothermic circulatory arrest and bilateral ASCP between 2015 and 2021. Pearson correlation analysis was performed between ASCP flow and rSO2., Results: The median circulatory arrest duration was 46.5 (IQR:37.0-61.0) minutes. There was no significant correlation between ASCP flow and rSO2 for both the right (r = -.02, p = .851), and the left hemisphere (r = - .04, p = .618). The rSO2 values for ten patients who received > 10 mL/kg/min flow did not differ significantly from 130 patients who received 10 mL/kg/min or less for both the left hemisphere ( p = .135), and the right hemisphere ( p = .318). The ASCP flow was 5.1 (IQR:5.0- 6.5) mL/kg/min in five patients with, and 7.2 (IQR:5.8-8.3) mL/kg/min in 135 patients without a watershed infarction ( p = .098)., Conclusions: There was no correlation between ASCP flow rate and rSO2 in patients with acute type A aortic dissection. Furthermore, ASCP flow below 10 mL/kg/min was not associated with a reduction in rSO2. Definitive associations between ASCP flow and neurological outcome after type A aortic dissection surgery need further investigation., Competing Interests: Declaration of conflicting interestsThe authors received no financial support for the research, authorship and/or publication of this article.

Introduction: Carbon dioxide production (VCO2i), oxygen consumption and oxygen delivery can be monitored during cardiopulmonary bypass (CPB) as markers for tissue perfusion. This study examines if inline venous pCO2 (PvCO2) monitoring can be used as an alternative to exhaust gas pCO2 (ExCO2) to calculate VCO2i., Methods: PvCO2 and ExCO2 were monitored continuously during 40 elective coronary artery bypass grafting (CABG) procedures. VCO2i was calculated with ExCO2 as well as PvCO2., Results: Mean PvCO2 was 0.27 mmHg higher than mean ExCO2 (p < .001). The 95% limits of agreement of PvCO2 and ExCO2 were [-2.99, 3.53] mmHg which is within the limits proposed by the Clinical Laboratory Improvement Amendments of 2019. VCO2i was calculated using both PvCO2 and ExCO2 (PvVCO2i; ExVCO2i). A strong linear correlation was found for ExVCO2i and PvVCO2i (R2= .94, p < .001)., Conclusion: In conclusion, the differences in VCO2i calculation between the two methods are unlikely to be clinically relevant during normothermic CABG procedures. VCO2i can be calculated with either a capnograph or inline venous pCO2 monitoring.

Do cities accelerate COVID-19 transmission? Increased transmission arising from population density prompts spatial policies for financial support and containment, and poorer prospects for recovery. Using daily case counts from over 3,000 counties in the U.S. from February to September 2020, I estimate a compartmental transmission equation. Rational sheltering behavior plausibly varies by location, so I propose two instruments that exploit unanticipated variation in exposure to potential infection. In the first month of local infections, an additional log point of population density raises the expected transmission parameter estimate by around 3%. After the first month, the relation vanishes: density effects occur only in the outbreaks. Public transport, work-from-home jobs and income explain additional variation in transmission but do not account for the density effects. Consistent with location-varying optimal sheltering behavior, I document stronger mobility declines in denser areas, but only after the first month of infections. These results suggest that differences in transmission between cities and other places do not motivate spatial policies for recovery or containment, or poorer prospects after the pandemic., (© 2022 The Author(s).)

Objective: Despite the positive effects of decision aids (DAs), implementation remains a significant challenge. The aim of the current study was to determine what barriers clinicians experience using a DA for pelvic organ prolapse (POP)., Methods: This study was conducted with a qualitative descriptive design including in-depth semi-structured interviews according to COREQ-criteria. Participants included clinicians and patients. Grounded theory analysis was used to describe the main themes., Results: A total of 9 clinicians and 4 patients participated. Four major themes (1) opinions about shared decision making (SDM), (2) current practice, (3) experience with the DA, (4) suggestions for improvement and one minor theme (5) experience with the study, emerged. Clinicians were predominantly positive about the DA., Conclusion: Despite the positive attitudes of the clinicians in this study, the implementation of a DA is still challenging. The DA is forgotten regularly as improvement of logistics is needed, clinicians assume they already provide good care which might result in a reluctance to change and more engagement of physicians is needed., Practice Implications: Regular contact with clinicians to remind, help and increase engagement and a decrease of the logistic burden is needed to ensure all patients can fully benefit of the DA., Competing Interests: Declaration of Competing Interest None., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A high HLA expression in uveal melanoma (UM) is part of the prognostically unfavorable inflammatory phenotype. We wondered whether the presence of soluble HLA (sHLA) in the aqueous humour is associated with clinical, histopathological or genetic tumour characteristics, and represents tumour HLA expression and intratumoural inflammation. Aqueous humour from 108 UM patients was analysed for the presence of sHLA, using a Luminex assay specific for HLA Class I. Clinical and genetic parameters were compared between sHLA-positive and negative eyes. A qPCR analysis was performed on tumour tissue using a Fluidigm assay. In 19/108 UM-containing eyes, the sHLA level in the aqueous was above the detection limit. Tumours in sHLA-positive eyes were significantly larger, more frequently involved the ciliary body, and more often showed monosomy 3, gain of chromosome 8q and loss of BAP1 staining. Melanoma-related survival was worse in patients with sHLA-positive aqueous humour. sHLA in the aqueous did not represent the tumour's HLA expression and did not relate to immune cell infiltration in the tumour. We conclude that UM-containing eyes may contain sHLA in the aqueous humour, where it is a prognostically-unfavourable sign and may influence local immune responses.

Background: Quantitative (q) PCR by amplification of nucleic acid with a fluorescent dye is widely used. Selection of adequate PCR reagents and devices is relevant to achieve reliable and consistent data. Our main objective was to test the robustness of different commercial SYBR green PCR mixes with respect to specificity and sensitivity of the PCR assay, across various PCR machines (Light Cycler 96, ViiA7) and amplification protocols. Herein, we applied PCR protocols for determining mRNA transcript levels, DNA copy numbers, and DNA genotype., Results: First, we set up 70 primer-based assays that targeted immune-related mRNA transcripts. Of the 70 assays 66 (94.3 %) resulted in a single melting curve peak, indicating specificity of the amplification, with PCR mixes from large vendors (Roche, ABI, Bio-Rad). But this was only seen when the PCR protocol that was indicated in the vendor's guidelines for each particular mix was applied. When deviating from the prescribed protocol, suboptimal melting curves were most often seen when using Roche SYBR green. With respect to PCR yields, the use of ABI mix more often led to lower Cq values. Second, we set up 20 primer-selective PCR assays to target different insertion-deletion and single nucleotide polymorphism regions throughout the genome. The variation in delta Cq between positive and negative DNA samples among the PCR assays was the lowest when using ABI master mix. Finally, the quality of high resolution melting (HRM) assays for DNA genotyping was compared between four commercial HRM PCR mixes (Roche, Bioline, PCR Biosystems, ABI). Only Roche and ABI mixes produced optimal clusters of melting profiles that clearly distinguished genotype variants., Conclusions: The current results show a preference for the use of ABI mix when it comes to obtaining higher sensitivity in cDNA analysis and a higher consistency among assays in distinguishing DNA genotypes among different individuals. For HRM assays, it is advisable to use master mix from a relatively large vendor.

Oocyte donation (OD) enables women with various causes of reproductive failure to conceive, but is accompanied by a high risk of certain pregnancy disorders. Possibly, the allogeneic nature of the fetus in OD pregnancies plays a role in the development of these disorders. In this study, we investigated whether there is a selection for some degree of HLA matching in successful and uncomplicated OD pregnancies. Mothers and children from OD pregnancies that used unrelated donors (n=75) were typed for HLA-A, -B, -C, -DR, and -DQ and the observed number of HLA matches of the child was compared with the expected number of HLA matches. Moreover, we studied the possibility of a preferential selection for maternal KIR and fetal C combinations. We observed a significantly higher level of HLA matching between mother and child than expected by chance. In particular, the incidence of children with 5 or more HLA matches, which is the situation in autologous pregnancy, was higher than expected. A higher level of matching was shown, especially for HLA class I, while no significant differences were observed for the individual HLA loci. With respect to maternal KIR and fetal HLA-C no selection for a favorable combination was found. Larger observational studies including uncomplicated, preeclamptic, and aborted pregnancies are essential to determine to what extent HLA matching affects the outcome of OD pregnancies., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Background: Minor Histocompatibility (H) antigen mismatches significantly influence the outcome of HLA-matched allogeneic stem cell transplantation. The molecular identification of human H antigens is increasing rapidly. In parallel, clinical application of minor H antigen typing has gained interest. So far, relevant and simple tools to analyze the minor H antigens in a quick and reliable way are lacking., Methodology and Findings: We developed a uniform PCR with sequence-specific primers (PCR-SSP) for 10 different autosomal minor H antigens and H-Y. This genomic minor H antigen typing methodology allows easy incorporation in the routine HLA typing procedures. DNA from previously typed EBV-LCL was used to validate the methodology. To facilitate easy interpretation for clinical purposes, a minor H database named dbMinor (http://www.lumc.nl/dbminor) was developed. Input of the minor H antigen typing results subsequently provides all relevant information for a given patient/donor pair and additional information on the putative graft-versus-host, graft-versus-tumor and host-versus-graft reactivities., Significance: A simple, uniform and rapid methodology was developed enabling determination of minor H antigen genotypes of all currently identified minor H antigens. A dbMinor database was developed to interpret the genomic typing for its potential clinical relevance. The combination of the minor H antigen genomic typing methodology with the online dbMinor database and applications facilitates the clinical application of minor H antigens anti-tumor targets after stem cell transplantation.

From a panel of mouse monoclonal antibodies reactive with a repeating epitope of the schistosome circulating anodic antigen, an IgG1 monoclonal antibody was selected. This monoclonal antibody was applied in a sandwich enzyme-linked immunosorbent assay as capture antibody and as alkaline phosphatase labeled conjugate. This assay allowed a sensitive quantitation of circulating anodic antigen in serum samples of infected individuals, detecting less than 1 ng antigen/ml serum. In Schistosoma mansoni infected individuals from Zaire, the level of antigen in serum correlated with fecal egg output. The lower detection level of the immunoassay corresponded to a level of about 10 eggs/gm feces.