Technical Aspects and Clinical Limitations of Sperm DNA Fragmentation Testing in Male Infertility: A Global Survey, Current Guidelines, and Expert Recommendations

PURPOSE: Sperm DNA fragmentation (SDF) is a functional sperm abnormality that can impact reproductive potential, for which four assays have been described in the recently published sixth edition of the WHO laboratory manual for the examination and processing of human semen. The purpose of this study was to examine the global practices related to the use of SDF assays and investigate the barriers and limitations that clinicians face in incorporating these tests into their practice. MATERIALS AND METHODS: Clinicians managing male infertility were invited to complete an online survey on practices related to SDF diagnostic and treatment approaches. Their responses related to the technical aspects of SDF testing, current professional society guidelines, and the literature were used to generate expert recommendations via the Delphi method. Finally, challenges related to SDF that the clinicians encounter in their daily practice were captured. RESULTS: The survey was completed by 436 reproductive clinicians. Overall, terminal deoxynucleotidyl transferase deoxyuridine triphosphate Nick-End Labeling (TUNEL) is the most commonly used assay chosen by 28.6%, followed by the sperm chromatin structure assay (24.1%), and the sperm chromatin dispersion (19.1%). The choice of the assay was largely influenced by availability (70% of respondents). A threshold of 30% was the most selected cut-off value for elevated SDF by 33.7% of clinicians. Of respondents, 53.6% recommend SDF testing after 3 to 5 days of abstinence. Although 75.3% believe SDF testing can provide an explanation for many unknown causes of infertility, the main limiting factors selected by respondents are a lack of professional society guideline recommendations (62.7%) and an absence of globally accepted references for SDF interpretation (50.3%). CONCLUSIONS: This study represents the largest global survey on the technical aspects of SDF testing as well as the barriers encountered by clinicians. Unified global recommendations regarding clinician implementation and standard laboratory interpretation of SDF testing are crucial

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection

Artificial intelligence in andrology: From Semen Analysis to Image Diagnostics

Artificial intelligence (AI) in medicine has gained a lot of momentum in the last decades and has been applied to various fields of medicine. Advances in computer science, medical informatics, robotics, and the need for personalized medicine have facilitated the role of AI in modern healthcare. Similarly, as in other fields, AI applications, such as machine learning, artificial neural networks, and deep learning, have shown great potential in andrology and reproductive medicine. AI-based tools are poised to become valuable assets with abilities to support and aid in diagnosing and treating male infertility, and in improving the accuracy of patient care. These automated, AI-based predictions may offer consistency and efficiency in terms of time and cost in infertility research and clinical management. In andrology and reproductive medicine, AI has been used for objective sperm, oocyte, and embryo selection, prediction of surgical outcomes, cost-effective assessment, development of robotic surgery, and clinical decision-making systems. In the future, better integration and implementation of AI into medicine will undoubtedly lead to pioneering evidence-based breakthroughs and the reshaping of andrology and reproductive medicine

Technical aspects and clinical limitations of sperm DNA fragmentation testing in male infertility: A global survey, current guidelines, and expert recommendations

Purpose Sperm DNA fragmentation (SDF) is a functional sperm abnormality that can impact reproductive potential, for which four assays have been described in the recently published sixth edition of the WHO laboratory manual for the examination and processing of human semen. The purpose of this study was to examine the global practices related to the use of SDF assays and investigate the barriers and limitations that clinicians face in incorporating these tests into their practice. Materials and Methods Clinicians managing male infertility were invited to complete an online survey on practices related to SDF diagnostic and treatment approaches. Their responses related to the technical aspects of SDF testing, current professional society guidelines, and the literature were used to generate expert recommendations via the Delphi method. Finally, challenges related to SDF that the clinicians encounter in their daily practice were captured. Results The survey was completed by 436 reproductive clinicians. Overall, terminal deoxynucleotidyl transferase deoxyuridine triphosphate Nick-End Labeling (TUNEL) is the most commonly used assay chosen by 28.6%, followed by the sperm chromatin structure assay (24.1%), and the sperm chromatin dispersion (19.1%). The choice of the assay was largely influenced by availability (70% of respondents). A threshold of 30% was the most selected cut-off value for elevated SDF by 33.7% of clinicians. Of respondents, 53.6% recommend SDF testing after 3 to 5 days of abstinence. Although 75.3% believe SDF testing can provide an explanation for many unknown causes of infertility, the main limiting factors selected by respondents are a lack of professional society guideline recommendations (62.7%) and an absence of globally accepted references for SDF interpretation (50.3%). Conclusions This study represents the largest global survey on the technical aspects of SDF testing as well as the barriers encountered by clinicians. Unified global recommendations regarding clinician implementation and standard laboratory interpretation of SDF testing are crucial

Utility of a modified silver staining technique for detection of Leptospira

Background: Leptospira are spiral thin and highly motile pathogenic bacteria that are best visualized by dark ground microscopy. Although these bacteria are not stained by the Gram’s stain, the Fontana stain, which is a silver impregnation staining method, can be used successfully for light microscopy. It is important to investigate the usefulness of Fontana stain method for direct demonstration of Leptospira in human body fluids.Objectives: To determine the usefulness and sensitivity of a modified Fontana silver staining method for microscopic detection of Leptospira in clinical specimens.Methodology: 6×108 organisms/ml of Leptospira interrogans serovar Icterohaemorrhagiae and Canicola were spiked into PBS (Phosphate Buffered Saline), alkalinized urine and serum in triplicate and serial dilutions were made (6×106 to 6×101 organisms/ml). Smears were prepared using 10 µl of each dilution. In addition, centrifuged sediment of urine were also used to prepare smears. Slides were stained by modified Fontana method as reported by Gangadhar et al.(1998) and examined. Numbers of leptospires per field (×100) were recorded.Results: Leptospira spiked in PBS and urine appeared as thin slender bacteria with characteristic hooked ends after Fontana staining under the light microscope. Serum could not be used for the detection of Leptospira by this method. Leptospires could be detected by staining the spiked PBS and urine at 6x103 – 6x106 organisms/ml.Conclusion: Leptospires could be detected by Fontana staining in spiked PBS, urine (uncentrifuged and sediment). Serum was not suitable for detection of leptospires by Fontana staining. The detection limit of leptospires in PBS and urine by Fontana stain was found to be 6000 organisms/ml.</p

Preliminary study on the effect of Munronia pinnata (Wall) Theob. (Meliaceae) aqueous extract on functional change in endothelial cells infected by dengue virus

Infection of endothelial cells (ECs) and their dysfunction has been implicated in the immunopathogenesis leading to severity in dengue virus (DENV) disease and rationalize the therapeutic targeting of the endothelium. Munronia pinnata is a rare medicinal plant which is commonly used to treat fever in traditional systems of medicine in Sri Lanka. The objective of the present study was to investigate the effect of aqueous plant extract (APE) of M. pinnata on the interaction between DENV-3 and ECs in inducing any changes. The effect of DENV-3 on metabolic activity of ECs was evaluated by using MTT. The effect of APE of M. pinnata on interaction between DENV-3 and ECs was assessed in three different ways to determine the potential of M. pinnata to protect the endothelial cells in a dengue infection; i) pre-treatment of DENV-3 with APE on ECs, ii) sequential treatment of DENV-3 followed by APE on ECs and iii) sequential treatment of APE followed by DENV-3 on ECs. A 49.8% reduction of EC viability was recorded following interaction with DENV-3 as opposed to that in the absence of DENV-3 (p < 0.001). A significant protection of ECs was observed at almost all concentrations of APE in treatment strategies. Interestingly, an increased significant protection was observed during the pre-treatment of DENV-3 with APE prior to the interaction with ECs (lowest p = 0.0003), and highest protection of EC at 15.6 and 31.3 µg/mL of APE (p < 0.0001). The present study suggests that the APE of M. pinnata exhibits an antiviral effect against DENV-3 by protecting the metabolic activity of ECs. Further studies are needed to understand the underlying mechanism of the direct inhibitory activity of M. pinnata against DENVs and to characterize the active compounds in the origin of its efficacy

A preliminary study on efficacy of rupatadine for the treatment of acute dengue infection

Abstract Currently there are no specific treatments available for acute dengue infection. We considered that rupatadine, a platelet-activating factor receptor inhibitor, might modulate dengue-associated vascular leak. The effects of rupatadine were assessed in vitro, and in a dengue model, which showed that rupatadine significantly reduced endothelial permeability by dengue sera in vitro, and significantly inhibited the increased haematocrit in dengue-infected mice with dose-dependency. We conducted a randomised, placebo-controlled trial in 183 adult patients in Sri Lanka with acute dengue, which showed that rupatadine up to 40 mg daily appeared safe and well-tolerated with similar proportions of adverse events with rupatadine and placebo. Although the primary end-point of a significant reduction in fluid leakage (development of pleural effusions or ascites) was not met, post-hoc analyses revealed small but significant differences in several parameters on individual illness days - higher platelet counts and lower aspartate-aminotransferase levels on day 7 in the rupatadine group compared to the placebo group, and smaller effusions on day 8 in the subgroup of patients with pleural effusions. However, due to the small sample size and range of recruitment time, the potential beneficial effects of rupatadine require further evaluation in large studies focused on recruitment during the early febrile phase