Background: The introduction of the vaccination against SARS-CoV-2 infection creates the need for precise tools for the quality control of vaccination procedures, detection of poor humoral response, and estimation of the achieved protection against the disease. Thus, the study aimed to compare the results of the anti-SARS-CoV-2 tests to evaluate the application of the WHO standard unitage (the binding antibody units; BAU/mL) for a measurement of response to the vaccination. Methods: Patients undergoing vaccination against SARS-CoV-2 with Pfizer/BioNTech BNT162b2 (BNT162b2) (n = 79), referred for SARS-CoV-2 antibody measurement prior to vaccination and 21 days after dose 1, and 8, 14, and 30 days after dose 2 were included. The sera were tested with three assays: Elecsys SARS-CoV-2 S (Roche), LIAISON® SARS-CoV-2 TrimericS IgG (DiaSorin), and SARS-CoV-2 IgG II Quant (Abbott). Results: The three assays showed varying correlations at different time points in the study. The overall agreement for all samples was moderate to high (ρ = 0.663–0.902). We observed the most uniform agreement for the day of dose 2 (ρ = 0.775–0.825), while it was least consistent for day 8 (ρ = −0.131–0.693) and 14 (ρ = −0.247–0.603) after dose 2. The dynamics of changes of the SARS-CoV-2 antibody levels in patients without history of prior SARS-CoV-2 infection appears homogenous based on the Roche results, more heterogenous when considering the DiaSorin results, and in between for the Abbott results. Conclusions: The results highlight the need for further work on the international standard of measurement of SARS-CoV-2 Ig, especially in the era of vaccination. The serological assays can be useful to detect IgG/IgM antibodies to assess the response to the vaccination. However, they cannot be used interchangeably. In terms of the evaluation of the immune response to the BNT162b2 vaccine, Roche and Abbott kits appear to be more useful.
A retrospective case control study was undertaken at the molecular biology department of a private center for reproductive medicine in order to determine whether any correlation exists between mitochondrial DNA (mtDNA) content of cleavage-stage preimplantation embryos and their developmental potential. A total of 69 couples underwent IVF treatment (averaged women age: 36.5, SD 4.9) and produced a total of 314 embryos. A single blastomere was biopsied from each embryo at the cleavage stage (day-3 post-fertilization) subjected to low-pass next generation sequencing (NGS), for the purpose of detecting aneuploidy. For each sample, the number of mtDNA reads obtained after analysis using NGS was divided by the number of reads attributable to the nuclear genome. The mtDNA copy number amount was found to be higher in aneuploid embryos than in those that were euploid (mean mtDNA ratio ± SD: 6.3 ± 7.5 versus 7.1 ± 5.8, p < 0.004; U Mann–Whitney test), whereas no statistically significant differences in mtDNA content were seen in relation to embryo morphology (6.6 ± 4.8 vs. 8.5 ± 13.6, p 0.09), sex (6.6 ± 4.1 vs. 6.2 ± 6.8, p 0.16), maternal age (6.9 ± 7.8 vs. 6.7 ± 4.5, p 0.14) or its ability to implant (7.4 ± 6.6 vs. 5.1 ± 4.6, p 0.18). The mtDNA content cannot serve as a useful biomarker at this point in development. However, further studies investigating both quantitative and qualitative aspects of mtDNA are still required to fully evaluate the relationship between mitochondrial DNA and human reproduction.
Mitochondria are well known as ‘the powerhouses of the cell’. Indeed, their major role is cellular energy production driven by both mitochondrial and nuclear DNA. Such a feature makes these organelles essential for successful fertilisation and proper embryo implantation and development. Generally, mitochondrial DNA is exclusively maternally inherited; oocyte’s mitochondrial DNA level is crucial to provide sufficient ATP content for the developing embryo until the blastocyst stage of development. Additionally, human fertility and early embryogenesis may be affected by either point mutations or deletions in mitochondrial DNA. It was suggested that their accumulation may be associated with ovarian ageing. If so, is mitochondrial dysfunction the cause or consequence of ovarian ageing? Moreover, such an obvious relationship of mitochondria and mitochondrial genome with human fertility and early embryo development gives the field of mitochondrial research a great potential to be of use in clinical application. However, even now, the area of assessing and improving DNA quantity and function in reproductive medicine drives many questions and uncertainties. This review summarises the role of mitochondria and mitochondrial DNA in human reproduction and gives an insight into the utility of their clinical use.
BackgroundThe introduction of vaccination against SARS-CoV-2 infection needs precise instruments for quality control of vaccination procedure, detection of poor immunological response and estimation of the achieved protection against the disease but also against infection and being infective.ObjectiveTo compare new automated SARS-CoV 2 Ig assay performance characteristics from the automated Elecsys SARS-CoV-2 S (Roche) with the new LIAISON® SARS-CoV-2 TrimericS IgG (DiaSorin) assay and their compatibility with WHO International Standard for anti-SARS-CoV-2 immunoglobulin. In the context of the mass vaccination programs, we undertook the investigation of clinical utility of the two new automated assays by analyzing results in samples collected at specified time points relative to the vaccination time.DesignProspective assay evaluation.PatientsMedical staff undergoing vaccination with BioNTech/Pfizer Comirnaty vaccine between January and March 2021 (n = 79) and referred for serum antiSARS-CoV 2 Ig testing prior to vaccination, 21 days after the first dose, and 8, 14 and 30 days after the second dose. Main Outcome Measure(s): Serum antibody levels measured with Roche and DiaSorin assays.ResultsIntra-assay imprecision was low with DiaSorin at 3.46%; and Roche at 2.5%. The Passing-Bablok regression equation for all tested samples was y (DiaSorin) = 184.61 + (1.03 x Roche) and the correlation between the assays (r=0.587; p < 0.0001).ConclusionsThe novel automated assays exhibit strong concordance in calibration, with assay-specific interpretation required for routine clinical use. These results highlight the need for further work on the international standard of measurement of SARS-CoV 2 Ig especially in era of vaccination. The serological assays can be useful to detect IgG/IgM antibodies, to assess the degree of immunization, to trace the contacts, and to support the decision to readmit people to work or vaccinate them again. However, the values generated by both assays can be markedly different, and assay-specific and personalized interpretation is required.
A nonsense mutation, p.Q548X, in the BLM gene has recently been associated with an increased risk for breast cancer. In the present work, we investigated the prevalence of this Slavic founder mutation in 2,561 ovarian cancer cases from Russia, Belarus, Poland, Lithuania or Germany and compared its frequency with 6,205 ethnically matched healthy female controls. The p.Q548X allele was present in nine ovarian cancer patients of Slavic ancestry (0.5 %; including one case with concurrent BRCA1 mutation). The mutation was not significantly more frequent in cases than in controls (Mantel-Haenszel OR 1.14, 95 % CI 0.49; 2.67). Ovarian tumours in p.Q548X carriers were mainly of the serous subtype, and there was little evidence for an early age at diagnosis or pronounced family history of cancer. These findings indicate that the BLM p.Q548X mutation is not a strong risk factor for ovarian cancer.
Breast cancer is the most commonly diagnosed cancer worldwide and the fifth leading cause of cancer death. In 2020, there were 2.3 million new cases, and 685,000 women died from it. Breast cancer among young women under 40 years of age accounts for 5% to 10% of all cases of this cancer. The greater availability of multi-gene sequence analysis by next-generation sequencing has improved diagnosis and, consequently, the possibility of using appropriate therapeutic approaches in BRCA1/2 gene mutation carriers. Treatment of young breast cancer patients affects their reproductive potential by reducing ovarian reserve. It can lead to reversible or permanent premature menopause, decreased libido, and other symptoms of sex hormone deficiency. This requires that, in addition to oncological treatment, patients are offered genetic counseling, oncofertility, psychological assistance, and sexological counseling. Given the number of BRCA1/2 gene mutation carriers among young breast cancer patients, but also thanks to growing public awareness, among their healthy family members planning offspring, the possibility of benefiting from preimplantation testing and performing cancer-risk-reduction procedures: RRM (risk-reducing mastectomy) and RRSO (risk-reducing salpingo-oophorectomy) significantly increase the chance of a genetically burdened person living a healthy life and giving birth to a child not burdened by the parent’s germline mutation. The goal of this paper is to show methods and examples of fertility counselling for BRCA1/2 gene mutation carriers, including both patients already affected by cancer and healthy individuals.
Media reports have caused a significant drop in confidence in the AstraZeneca ChAdOx1 nCoV-19 COVID-19 vector vaccine (Vaxzevria, AstraZeneca Södertälje, Sweden). This has caused many people, already vaccinated with the first dose of AstraZeneca, to refuse vaccination with this product. On the other hand, the increased demand for mRNA vaccines has resulted in a greater shortage of mRNA vaccines and cases of people being vaccinated with the AstraZeneca vaccine after the first dose of the Pfizer/BioNTech BNT162b2 COVID-19 vaccine (Comirnaty, Pfizer/BioNTech, Mainz, Germany). Moreover, currently, 60.9% of the global population have received at least one dose of a COVID-19 vaccine, while only 10% of people in low-income countries have received at least one dose. Even less people are fully vaccinated. The present pilot study evaluated the administration of half doses of AstraZeneca and Pfizer vaccines and included the enrollment of 26 subjects who were vaccinated with a different vaccine the first and second time. The reference group included individuals undergoing vaccination with two full doses of the Pfizer vaccine (21-day interval) monitored for their antibody levels as part of a parallel study. The distribution of antibody levels was not significantly different between those who received the Pfizer vaccine alone and those receiving the AstraZeneca vaccine plus Pfizer or Pfizer and AstraZeneca. To prepare for the next pandemic waves, solving the problem of the matching of booster vaccine to the previously received doses would be advisable. The topic is important and emerging as most of the population in low-income countries is still not vaccinated. We strongly believe that vaccine equity is the most important aspect of vaccination strategies.
Intracytoplasmic sperm injection (ICSI) is a widely used and accepted treatment of choice for oocyte fertilization. However, the quality of sperm selection depends on the accurate visualization of the morphology, which can be achieved with a high image resolution. We aim to correct the conviction, shown in a myriad of publications, that an ultra-high magnification in the range of 6000×–10,000× can be achieved with an optical microscope. The goal of observing sperm under the microscope is not to simply get a larger image, but rather to obtain more detail—therefore, we indicate that the optical system’s resolution is what should be primarily considered. We provide specific microscope system setup recommendations sufficient for most clinical cases that are based on our experience showing that the optical resolution of 0.5 μm allows appropriate visualization of sperm defects. Last but not least, we suggest that mixed research results regarding the clinical value of IMSI, comparing to ICSI, can stem from a lack of standardization of microscopy techniques used for both ICSI and IMSI.
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