Level Of Fragmentation Research Articles

Modeling honeybee flower visitation rates in the fragmented agricultural landscapes based on Lévy-flight behavior

AbstractTypically, honeybees (Apis mellifera L.), rely on waggle dances performed by scout bees to communicate information about fruitful nectar and pollen sources across the landscape. However, when this communication is absent, inaccurate, or when resources become depleted, bees resort to alternative search strategies. Field experiments utilizing harmonic radar have revealed that honeybees follow flight patterns that demonstrate a scale-free (Lévy-flight) behavior, representing an optimal search strategy for relocating the original feeder location. If honeybees adhere to a Lévy flight pattern to discover resources, where would honeybees demonstrate the highest flower visitation rates in agricultural landscapes? We generated simulated landscapes with varying proportions of forest cover scenarios, ranging from 5 to 50% of the total landscape area, along with different levels of fragmentation per se. Subsequently, we constrained the richness of flower farm cells in each landscape. To predict honeybee visitation rates, three different methodologies based on random movement were utilized: (1) moving window, (2) random walk, and (3) Lévy flight. We found that honeybee visitation rates were influenced by the degree of forest fragmentation in each scenario. Across all visitation scenarios, the highest average number of visited flowers per cell was observed in landscapes with maximum fragmentation per se. In landscapes with lower forest cover and higher fragmentation, honeybees were more likely to visit a greater number of flowers due to the increased probability of traversing the landscape and encountering more flower cells. honeybee visitation rates in agricultural landscapes are significantly influenced by the degree of forest fragmentation. The study highlights the importance of considering landscape structure, specifically forest fragmentation, when predicting honeybee visitation rates and underscores the need for further research to better understand the intricate relationship between landscape characteristics and pollinator behavior.

TLR2 and TLR4 Are Expressed in Epiretinal Membranes: Possible Links with Vitreous Levels of Complement Fragments and DAMP-Related Proteins.

Previous studies reported the expression of toll-like receptors (TLRs), merely TLR2 and TLR4, and complement fragments (C3a, C5b9) in vitreoretinal disorders. Other than pathogens, TLRs can recognize endogenous products of tissue remodeling as damage-associated molecular pattern (DAMPs). The aim of this study was to confirm the expression of TLR2 and TLR4 in the fibrocellular membranes and vitreal fluids (soluble TLRs) of patients suffering of epiretinal membranes (ERMs) and assess their association with disease severity, complement fragments and inflammatory profiles. Twenty (n = 20) ERMs and twelve (n = 12) vitreous samples were collected at the time of the vitrectomy. Different severity-staged ERMs were processed for: immunolocalization (IF), transcriptomic (RT-PCR) and proteomics (ELISA, IP/WB, Protein Chip Array) analysis. The investigation of targets included TLR2, TLR4, C3a, C5b9, a few selected inflammatory biomarkers (Eotaxin-2, Rantes, Vascular Endothelial Growth Factor (VEGFA), Vascular Endothelial Growth Factor receptor (VEGFR2), Interferon-γ (IFNγ), Interleukin (IL1β, IL12p40/p70)) and a restricted panel of matrix enzymes (Matrix metalloproteinases (MMPs)/Tissue Inhibitor of Metallo-Proteinases (TIMPs)). A reduced cellularity was observed as function of ERM severity. TLR2, TLR4 and myD88 transcripts/proteins were detected in membranes and decreased upon disease severity. The levels of soluble TLR2 and TLR4, as well as C3a, C5b9, Eotaxin-2, Rantes, VEGFA, VEGFR2, IFNγ, IL1β, IL12p40/p70, MMP7 and TIMP2 levels were changed in vitreal samples. Significant correlations were observed between TLRs and complement fragments and between TLRs and some inflammatory mediators. Our findings pointed at TLR2 and TLR4 over-expression at early stages of ERM formation, suggesting the participation of the local immune response in the severity of disease. These activations at the early-stage of ERM formation suggest a potential persistence of innate immune response in the early phases of fibrocellular membrane formation.

Landscape fragmentation and connectivity as key variables on occurrence of human-wildlife interactions

Human-wildlife interactions (HWI) are one of the most highly studied topics from the fields of wildlife management and are reported to be increasing globally as anthropogenic lands uses expand into wild lands, especially in mountainous areas where forest habitats are in the proximity of human settlements. The upper Prahova Valley in south-central Romania provides habitat to several native charismatic wildlife species. Since 1990 this area has become a major tourist destination now characterized by a high density of major transport infrastructures and homes. To assess how the changes in land use have affected HWIs, from September 2018 to August 2019, we discussed with 370 local citizens from the cities of Sinaia, Bușteni and Predeal. We developed maps of wildlife habitat fragmentation and connectivity and superimposed them over the locations with the HWI hotspots. According to the results, over 50% of the hotspots identified where located in areas exhibiting the greatest level of fragmentation, and the species frequently involved in interactions were the brown bear (Ursus arctos), wild boar (Sus scrofa), and red fox (Vulpes vulpes). The maps suggest that habitat fragmentation could represent a primal cause for the regions’ high frequency of animal descents into settlements, since wildlife habitats and ecological corridors are affected by the fragmentation of anthropogenic infrastructures especially in the proximity of major settlements. As a conclusion, our results highlight the need for sustainable landscape planning in order to optimize biodiversity management and diminish interactions between humans and wild animals, based on: a) mapping the areas characterized by high quality wildlife habitats and including them into a system of strict protection, b) developing or enhancing wildlife ecological corridors to favour animal movement between intact ecosystems, and c) regulating the expansion of human infrastructures in the proximity of strictly protected habitats.

The future of plant diversity within a Mediterranean endemism centre: Modelling the synergistic effects of climate and land-use change in Peloponnese, Greece

Climate- and land-use change stand as primary threats to terrestrial biodiversity. Yet, their synergistic impacts on species distributions remain poorly understood. To address this knowledge gap, we conducted the first-ever comprehensive species distribution analysis on an entire regional endemism centre within an eastern Mediterranean country, incorporating dynamic land-use/land-cover change data together with climate change scenarios. Specifically, we apply species distribution modelling and spatial data analysis techniques to compare the individual and synergistic effects of these environmental drivers on the endemic vascular flora of Peloponnese, focusing on potential range contractions, altitudinal shifts, and habitat fragmentation levels. Moreover, we identify fine-scale present and potential future endemism hotspots within our study area, incorporating taxonomic and phylogenetic information. Overall, we aim to enhance our current understanding of endemism patterns and contribute to the development of future-proof conservation strategies for safeguarding Greece's endangered endemic flora. The integration of land-use change projections with climate change yielded less severe impacts compared to the effects anticipated when considering climatic variables alone. Most taxa are expected to undergo significant range declines and nearly half might experience increased habitat fragmentation, due to the synergistic effects of climate- and land-use change. We identified endemism hotspots, which are concentrated in or along the main Peloponnesian mountain massifs. However, our predictions indicate that areas presently recognized as endemism hotspots will undergo a concerning area decline, across all future scenarios considered in this study. Our findings highlight the importance of including dynamic land-use variables alongside climatic predictors when projecting species distributions under global change. Moreover, we showed that endemism hotspots are not static and considering their potential geographic shifts is paramount to delineate effective forward-looking conservation strategies.

Evaluation of Selective Culling as a Containment Strategy for African Swine Fever at a Vietnamese Sow Farm.

Selective culling, also known as the "tooth extraction approach", is a strategy for controlling African swine fever (ASF) by removing only sick and suspect animals instead of the entire herd in Vietnam. This method prioritizes preserving healthy animals, particularly valuable breeding pigs. Despite its implementation in various forms, no standardized protocol based on scientific principles has been established. Farms typically adapt this strategy based on their understanding, which can vary significantly. In implementing of selective culling that is not based on scientific principles, there is a significant risk of spreading the disease. The aim of this study is to evaluate the consequences of selective culling as currently implemented in Vietnam. Our analysis on a large sow farm revealed that current practices rely heavily on clinical observations without laboratory confirmations. This approach allows ASF-infected animals to remain on the farm longer, potentially exacerbating the spread of the virus. Thus, selective culling poses a substantial risk by potentially exacerbating the spread of disease. Our findings emphasize that early diagnosis of ASF and systematic removal of infected pigs are critical components for the effective implementation of selective culling strategies and that a high level of fragmentation to minimize contact between animals plays a key role. The optimal approach is to test conspicuous animals and separate them. Under no circumstances should suspect animals be left in the herd for several days before they become severely ill and succumb to the disease.

Influence of surgical treatment of varicocele on the pathospermia and the level of sperm DNA fragmentation

Introduction. Varicocele is a varicose vein in the testicle caused by pathological venous reflux. Clinically, varicocele is found in 15% of the male population, with 35% of men with primary infertility and 70-81% of men with secondary infertility. Uncontrolled, excessive amounts of reactive oxygen species (ROS) can cause single- and/ or double-strand breaks in sperm DNA, commonly referred to as sperm DNA fragmentation. The purpose of the study was to determine the effect of surgical treatment of varicocele on the level of DNA fragmentation and to develop prognostic criteria for restoring the fertility of patients.Materials and methods. The study is based on a comparative analysis of the results of examination and treatment of 93 patients without concomitant pathology, with pathospermia, increased levels of DNA fragmentation and varicocele, who received surgical treatment. All patients were followed up for 3–9 months after surgery. The control group did not receive postoperative therapy – 34 people, the main group (59 patients) received drug rehabilitation – a complex of spermatoprotectors and antioxidants – for 3 months after surgical treatment.Results of the study and their discussion. In patients who received surgical treatment, there was a normalization of the level of DNA fragmentation after 3 months in the postoperative period, on average the positive dynamics was 21% (from an average fragmentation level of 22.82% to 15.84%, p <0.05), the prognosis for pregnancy in the couple became favorable. An improvement in DNA fragmentation indicators was observed in parallel with an improvement in spermogram indicators in the postoperative period. Progression of left testicular hypotrophy in the postoperative period was not detected. No relapses or other complications were noted.Conclusions. Treatment of varicocele improves fertility in patients with pathospermia, increased levels of DNA fragmentation, which is consistent with research data from other authors, which confirmed the fact of a significant improvement in ejaculate parameters (sperm concentration, motility and morphology) However, the dynamics of normalization of sperm DNA fragmentation occurs, according to our data, more dynamically (within 3 months in the postoperative period) than the improvement of pathospermia (OAT syndrome) and does not depend on the presence of postoperative drug rehabilitation.

P-086 Consequences of elevated levels of DNA fragmentation and its influence on pregnancy – A multicentric cohort analysis

Abstract Study question Does an abnormal DNA Fragmentation Index impact the clinical outcomes of patients undergoing IVF treatment? Summary answer Statistical differences were seen in biochemical, implantation, clinical, and ongoing pregnancy rate in sperms with high and low DFI scores with autologous and donated oocytes. What is known already Sperm chromatin integrity is associated with embryo development and clinical outcomes of an IVF cycle. DNA Fragmentation Index (DFI) is a diagnostic tool for the detection of spermatozoa with damaged chromosomes in the ejaculate. DNA fragmentation is a significant cause of day 3 embryo arrest and the development of poor-quality embryos. The interventions associated with elevated levels of DFI are ICSI, lifestyle changes and antioxidant therapies. However, the published data regarding the potential of embryo development with high DFI is limited to the sample size thus, encouraging a need for more research in this area. Study design, size, duration The study is population-based and included 768 male patients who underwent DFI score detection across 44 centers at Indira IVF Hospital Private Limited from January 2023 to September 2023. The abstinence period for all the patients was recorded during the DFI detection. After the DFI assessment patients were classified into normal DFI score ( < =15%) (n = 203) and abnormal DFI score (>15%) (n = 565). Participants/materials, setting, methods The retrospective cohort analysis was performed on our databank on the male patients regardless of their age (21-50), ethnicity, and medical history. DFI was calculated using a Sperm Chromatin Dispersion assay. Patients were categorized into two groups: Normal DFI ( < =15%) and Abnormal DFI (>15%). The characteristics of population along with clinical outcomes were evaluated after transferring embryos fertilized with normal DFI sperm and abnormal DFI sperm. The data was analyzed and estimated by SPSS software. Main results and the role of chance Our analyses discovered that embryos fertilized with sperms of normal DFI score have significantly enhanced the likelihood of implantation compared with embryos fertilized with sperms of abnormal DFI (50.27% v/s 44.40%; p = 0.047). Simultaneously, early pregnancy was evaluated by determining the values of beta-HCG in blood, also known as biochemical pregnancy rate (BPR). The embryos fertilized with sperms of normal DFI score significantly enhanced the BPR compared with embryos fertilised with sperms of abnormal DFI (67% v/s 59.1%; p = 0.048). Clinical Pregnancy Rate (CPR) defined as presence of a heartbeat of gestational sac observed during the 6-week scan. Furthermore, it was evaluated that the CPR showed a significant enhancement in embryos fertilized with sperms of normal DFI score in comparison with embryos fertilized with sperms of abnormal DFI (65.5% v/s 57.9%; p = 0.034). However, it was noticed that male DFI score had no impact on the miscarriage rate of embryos inseminated with normal DFI score and abnormal DFI score (7.52% v/s 7.34%; p = 0.687). Moreover, the pregnancy was tracked, and it was revealed that embryos inseminated with sperms of normal DFI score significantly enhanced the ongoing pregnancy till 12 weeks compared with embryos inseminated with sperms of abnormal DFI score (60.6% v/s 53.6%; p = 0.039). Limitations, reasons for caution Embryos are known to have a self-correct mechanism to correct the presence of mosaicism in the embryos. Hence, the oocyte quality also affects the outcomes of the IVF cycle. Therefore, using a high grade of the oocyte for the patients with abnormal DFI score can be useful. Wider implications of the findings Our data suggest that abnormal DFI have an impact on the clinical outcomes of an IVF cycle. Therefore, examining the levels of DNA Fragmentation is utmost imperative for a successful pregnancy. Trial registration number not applicable

P-092 PGT-A Evaluates Advanced Sperm Selection in Embryos from High Teratozoospermia Males; a prospective randomized controlled trial

Abstract Study question Can sperm selection techniques improve the reproductive outcomes of ICSI patients, particularly in males with high teratozoospermia index (TZI) and sperm DNA fragmentation (SDF)? Summary answer Sperm selection techniques significantly increase fertilization and blastocyst development rates in males with high TZI and SDF, achieving a euploidy rate comparable to normal cases. What is known already Abnormal sperm morphology negative effects begin at the fertilization and reach the blastocyst quality and euploidy status. Men with abnormal teratozoospermic index (TZI) exhibit high levels of chromosomal abnormalities and sperm DNA fragmentation (SDF). SDF adversely affects pre and post-implantation embryo development events. Blastocyst aneuploidy isn’t solely due to the oocyte; but also, sperm contribution should be taken into consideration. Previous studies reported that advanced sperm selection techniques (PICSI and MACS) can efficiently enhance reproductive outcomes in abnormal SDF ICSI cases. The efficiency of these techniques in improving ICSI outcomes of patients with high TZI plus SDF remains unexplored. Study design, size, duration A prospective randomized controlled trial included a total of 264 couples, divided into three groups: (1) Control-1 group: males having normal TZI and no sperm selection techniques were used (n = 111), (2) Control-2 group (NO-SS): males having high TZI and no sperm selection techniques were used (n = 63), and (3) Sperm selection (SS) group: males having high TZI and SDF, and sperm selection techniques were used (n = 90). The study started from January till October 2023. Participants/materials, setting, methods Eligibility: females had ≥5MII oocytes, males had abnormal (>1.8) TZI, and all blastocysts (≥4BB) had trophoectoderm biopsy. Cases were randomized on the ICSI day to (control-2) and (SS) groups, along with (control-1) with normal TZI for comparison. Females were categorized into 2 subgroups (<35, and ≥35 years). SS techniques included physiological ICSI and magnetic-activated cell sorting. PGT-A was done using Illumina platform (Miseq). Statistical analysis (SPSS) of pre-implantation embryo development and PGT-A results was done. Main results and the role of chance We found no significant differences between the female, male ages and number of mature oocytes between the studied groups. Limitations, reasons for caution The results included all pre-implantation embryo parameters but, subsequent post-implantation events like implantation, pregnancy, and miscarriage rates will add more data about the competency of the resulting embryos. A Small sample size of females aged from 36 to 40 years old. Wider implications of the findings Sperm selection techniques appear to be efficient in improving fertilization, blastocyst development rates, and euploidy rates in males with high TZI and sperm DNA fragmentation, suggesting potential benefits for couples undergoing infertility treatment. Trial registration number NCT06231589

P-017 DNA fragmentation Index before and after density gradient centrifugation using flow cytometry, its reflection in sperm ultrastructure and its association with clinical outcome

Abstract Study question Is DNA fragmentation Index (DFI) affected by Density Gradient Centrifugation (DGC), reflected in sperm ultrastructure and associated with clinical outcome? Summary answer DFI was not affected by DGC, nor associated with clinical outcome, but it was negatively correlated with sperm concentration/motility and positively correlated with ultrastructural anomalies. What is known already Oxidative stress induces DNA fragmentation during sperm transit through the male genital tract and it is suggested to be the main mechanism causing DNA breaks after ejaculation during in vitro manipulation and sperm selection for injection/insemination. High levels of sperm DNA fragmentation have been linked to lower fertilization rates, poor embryo quality and delayed cleavage. Limited studies, have investigated in detail the effects of DGC on DFI and the reflection in sperm ultrastructure, while the predictive value of DFI on clinical outcome still remains controversial. Study design, size, duration Semen samples were collected on the day of the female partener’s oocyte collection and analysed before and after DGC for standard characteristics by light microscopy, according to World Health Organization (WHO) criteria (n = 120), for DNA Fragmentation by Flow Cytometry and for ultrastructure by Transmission Electron Microscopy (TEM). The predictive value of DFI for clinical outcome was also assessed. Participants/materials, setting, methods Standard semen analysis and DGC using the Sil-Select 40%/ 80 % layers were performed at an IVF Laboratory of an academic Hospital. Sperm DFI before and after DGC was assessed by Flow Cytometry at the Department of Immunology and Histocompatibility of the same academic Hospital. Ultrastructure analysis by TEM was carried out at an academic Histology/Embryology Laboratory following fixation in 3% glutaraldehyde, 1% osmium tetroxide, washes in PBS and staining with 1% aqueous uranyl acetate. Main results and the role of chance DGC did not alter DFI significantly (p = 0.052), but lead to a statistically significant increase in Progressive Motility (p < 0.001). DFI was higher in men with a male factor infertility compared to men without a male factor (p = 0.032) and Oligoasthenoteratospermia (OAT) exhibited the highest DFI levels (36.38% ± 5.36). DFI was negatively correlated with sperm concentration (rho = -0.361, p < 0.01) and sperm motility (rho = -0.301, p < 0.05). No significant association was observed between DFI and clinical outcome, but statistically significant differences were observed both in the age of men (37.54±1.33 vs 42.8±1.27 p = 0.017) and the age of women (35.77±1.25 vs 40.17±0.78 p = 0.007) in the group of patients who achieved a pregmamcy and those that did not. TEM showed that DGC did not lead to an increase in ultrastructural anomalies of spermatozoa, but DFI was positively correlated with ultrastructural anomalies. In teratozoospermic men TEM revealed head, neck and tail defects, including elongated forms, axonemal defects and premature initiation of the acrosome reaction. Limitations, reasons for caution The ultrastructure analysis was not possible to be performed in severely oligospermic samples. Wider implications of the findings This study shows that sperm ultrastructure and DFI are not adversely affected by DGC confirming the safety of the preparation method for sperm selection prior to insemination/injection. Larger-scale studies are needed, to elucidate any potential correlations between DFI and the probability of pregnancy. Trial registration number not applicable

O-015 Enabling laboratory-quality male fertility analysis with 72-hour post ejaculation sample viability

Abstract Study question Can SPX72-treated semen samples sustain diagnostic viability and integrity for up to 72 hours, enabling comparable analysis of mail-in samples to fresh samples? Summary answer SPX72 preserves sperm viability, motility, and integrity up to 72 hours post-collection, equalling the diagnostic precision of fresh samples and enhancing at-home testing capabilities. What is known already Challenges with laboratory-based fertility assessments stem from logistical burdens and the necessity for timely sample delivery, often deterring male patient participation. Semen samples typically need to be assessed within 60 minutes of collection to ensure accuracy. Pre-existing solutions have failed to sufficiently prolong sample viability while matching the diagnostic accuracy of immediate laboratory analyses, an issue Sapyen's SPX72 medium addresses effectively. Study design, size, duration An exhaustive controlled study with over 100 semen samples compared SPX72-treated samples at 72 hours against fresh controls. The study incorporated actual postal logistics to emulate mail-in scenarios. Participants/materials, setting, methods A predetermined quantity of SPX72 per millilitre was added to the collected semen samples, with key indicators, including progressive and total motility, morphology, and DNA fragmentation, measured at intervals of 0, 24, 48, and 72 hours, contrasted with untreated, fresh samples. Main results and the role of chance SPX72 application preserved notable semen motility, evidenced by a mean progressive motility of 37.12% after 72 hours, a slight decrease from the initial 41.14%. This minor 4.02% reduction underscores SPX72's proficiency in maintaining diagnostic-grade motility over an extended duration. In terms of DNA integrity, SPX72 maintained DNA fragmentation at 14.78%, juxtaposed with 12.44% in control samples, a statistically insignificant variance (p-value of 1.15037E-08). This consistency in DNA fragmentation levels is pivotal for the accuracy of fertility diagnostic metrics. Additionally, SPX72 demonstrated its efficacy in stabilising the pH level of semen samples throughout the 72-hour period, ensuring an optimal environment for sperm preservation. This maintenance of pH is crucial as deviations can significantly affect sperm functionality and overall sample integrity. These results substantiate SPX72's capacity to preserve diagnostic sperm quality for up to three days, potentially redefining the logistics of male fertility testing by enabling accurate, laboratory-level assessment through a mail-in format. Limitations, reasons for caution Despite the controlled study and real-world postal validation, ongoing surveillance of sample stability in diverse global postal environments will be crucial to ensure consistent results across all geographies. Wider implications of the findings The data indicate that SPX72 sets a new standard for at-home fertility testing, matching clinical quality without the immediacy constraint. For the first time, home-based sperm diagnostics do not compromise clinical quality, revolutionising how male fertility is approached and managed. Trial registration number not applicable

P-083 Short ejaculatory abstinence time of 2 hours results in higher pregnancy outcomes compared to the recommended abstinence period of 2-7 days

Abstract Study question What is the impact of a short abstinence (2h) compared to the recommended (2-7 days) ejaculatory abstinence time on embryo utilization rate and pregnancy outcome? Summary answer A short abstinence of 2 hours results in similar embryo utilization rates, but in reduced sperm DNA fragmentation and higher pregnancy rates. What is known already Semen parameters exhibit variability among and within individuals, and ejaculatory abstinence time (EA) is a known factor contributing to intra-individual variation. Despite the challenge of providing a specific recommendation for the optimal timeframe, recent studies suggested that a shorter abstinence is likely to decrease DNA fragmentation levels in semen and improve pregnancy and live birth rates following assisted reproductive technology. At present, ESHRE recommends a more limited range than the WHO manual, advising an ejaculatory abstinence of 3 to 4 days. Study design, size, duration This prospective proof-of-concept sibling oocyte study (n = 100 cycles), conducted between January 2018 and June 2023, required two sperm samples per patient. The initial sperm sample was produced on the day of oocyte retrieval following the requested abstinence period of 2-7 days. The second sperm sample was collected 2 hours later. Sibling oocytes were randomly allocated to injection with the first or second sample. Sibling oocytes of each patient were cultured under the same conditions. Participants/materials, setting, methods Inclusion criteria were ≥ 6 mature oocytes (MII), fresh ejaculates with concentration >1x106/ml, ICSI with subsequent blastocyst culture. Endpoints were pregnancy outcome, fertilization, blastocyst development, utilization rate, sperm concentration/motility and DNA fragmentation index (DFI) measured by TUNEL. Statistical analysis was done taking into account female age and clustering. A total of 104 couples consented to provide two samples on the day of oocyte retrieval and were enrolled. Main results and the role of chance Average abstinence periods of 98:48h±20:01 vs 02:05h±00:18 were observed. Sperm concentration (x106/ml), progressive forward motility and DFI, were 46.53±39.6 vs 37.0±37.1 (p < 0.001), 52.14%±20.1 vs 49.79%±20.4 (p = 0.152) and 6.76%±7.0 vs 5.37%±7.2 (p = 0.024), respectively. Of the 1282 collected oocytes, 634 were injected with the first sperm sample and 648 with the ‘2h EA sample’. Fertilization rates per MII (79.3% vs 80.7%, p = 0.154) and good-quality day 3 embryos (76.7% vs 77.6%, p = 0.276) were similar between both groups. From the 93 patients with blastocyst culture (with average female age of 33.9±5.0 years; average male age of 36.4±6.6 years), 1173 oocytes were stratified for injection: 580 (first sample) and 593 (second sample), respectively. Fertilization rates (81.4% vs 82.3%, p = 0.074) and good-quality day 5 blastocysts (46.2% vs 49.5%, p = 0.534) did not differ between the groups. Utilization rate per inseminated (30.5% vs 35.8%, p = 0.126) or fertilized (37.5% vs 43.4%, p = 0.153) oocyte was similar between the groups. Single fresh and frozen embryo transfers were compared between embryos generated after the requested (2-7 days; n = 91) and short (2h; n = 94) abstinence. Pregnancy rate was significantly higher after short abstinence: 49.5% (45/91) vs 62.8% (59/94); p = 0.01. Ongoing pregnancy rate was 31.9% (29/91) and 40.4% (38/94); p = 0.10. Limitations, reasons for caution This is a prospective proof-of-concept study, not powered to detect differences in utilization and pregnancy outcomes. Not all patient are capable of producing two sperm samples within 2 hours from each other. Wider implications of the findings The results indicate a significant, yet not relevant difference in DNA fragmentation index (DFI) but a relevant difference in pregnancy rate between the two groups. If increased live birth rates are obtained after short abstinence, recommendations concerning EA might need adaptation. Trial registration number NCT03323892

P-016 Prevalence of high oxidative stress and poor sperm DNA fragmentation levels in different groups of patients

Abstract Study question What is the prevalence of patients with high oxidation-reduction potential (ORP) and high sperm DNA fragmentation (SDF) in different patient groups with and without varicocele? Summary answer A large percentage of normozoospermic patients and patients who are normally considered of manageable in assisted reproduction show high SDF and/or high ORP. What is known already Standard semen analysis, even according to WHO criteria, is very limited in predicting male fertility potential. Oxidative stress is widely recognized as a major contributor to male infertility. Therefore, sperm DNA fragmentation (SDF) is increasingly recommended as additional test in diagnostics. High levels of oxidative stress and SDF have also been reported in patients with varicocele. Yet, SDF and oxidative stress tests are not routinely tested in ART laboratories. Apart from the cost and/or complexity of these tests, a reason for the lack of usage is the lack of knowledge of the prevalence. Study design, size, duration This is a retrospective analysis of data obtained from 2,946 patients attending the Department of Urology, Hamad Medical Corporation, Doha, Qatar, for fertility diagnosis from January to December 2018. For this analysis only patients (n = 773) who had a full data set for sperm DNA fragmentation (SDF) and seminal oxidative stress as measured by means of the oxidation-reduction potential (ORP) were included. Participants/materials, setting, methods Apart from SDF and ORP, standard semen parameters (ejaculate volume, sperm concentration, total and progressive motility, and normal sperm morphology), and the presence of varicocele were determined. Semen parameters were determined according to WHO 2010 criteria. SDF was tested using the Halosperm G2 test (Halotech DNA SL, Madrid, Spain), while ORP was determined using the MiOXSYS® (Aytu BioScience, Englewood, CO). Patients were categorized as normo-, oligo-, astheno-, and teratozoospermic. Main results and the role of chance The mean sperm concentration of the patients was 33.5 × 106 sperm/mL with a mean total motility of 50.3%. The overall percentage of patients with high (>20% SDF) SDF was 57.4% and those with high (≥1.34 mV/106 sperm/mL) ORP 63.4%. Varicocele was diagnosed in 14.0% of the patients. In normozoospermic patients, 26.8% had high SDF, whereas in the non-normozoospermic group this was 59.8% (P = 0.0108). In non-oligozoospermic (53.7%), non-asthenozoospermic (34.7%) and non-teratozoospermic (43.4%) groups, SDF was significantly higher. High ORP values were recorded in 35.7% of normozoospermic, 53.4% non-oligozoospermic, 47.2% of non-asthenozoospermic, and 49.7% of teratozoospermic patients. The difference between the prevalence of high ORP in the normozoospermic (35.7%) and the non-normozoospermic group is significant (P = 0.0063). When both tests (high SDF or high ORP) were considered, 50.0% of normozoospermic, 76.9% of non-oligozoospermic, 59.7% of non-asthenozoospermic, and 71.2% of non-teratozoospermic patients showed seminal pathology. High SDF was detected in 59.3% of varicocele patients, while 50.0% of varicocele patients had high ORP values. In patients without varicocele, 57.1%, 65.6%, and 82.7% had high SDF, high ORP, and high SDF or high ORP, respectively. These percentages were significantly (P < 0.0003) higher than for the corresponding normal groups. Limitations, reasons for caution Although the overall sample size of 773 is reasonably high, after sub-grouping the sample size is much smaller. Since this study only analyzed the SDF and ORP in patients attending and andrological outpatient clinic, the results could be different if patients in IVF and/or ICSI treatment cycles were analyzed. Wider implications of the findings This study highlights that complementation of semen analysis with SDF and oxidative stress testing will identify a large percentage of patients who would benefit from andrological treatment before assisted reproductive techniques (ART). This approach would result in better chances of these patients for successful ART. Trial registration number not applicable

PENGARUH LAMA WAKTU KRIOPRESERVASI TERHADAP KARAKTERISTIK SPERMATOZOA KAMBING SETELAH SEMEN DI-THAWING

Cryopreservation is a technique utilized to store spermatozoa with the aim of preserving the quality of frozen semen. The quality of spermatozoa after thawing is a critical factor influencing the success of Artificial Insemination (AI). This decline in quality can be attributed to factors such as exposure to extreme temperatures, formation of ice crystals, reduced membrane permeability and the generation of Reactive Oxygen Species (ROS). Given this decline in quality, it becomes imperative to assess spermatozoa quality at the cellular level. This study was aimed to investigate the impact of storage time on frozen goat semen on spermatozoa membrane integrity, levels of Malondialdehyde (MDA), and sperm DNA fragmentation. The samples utilized included frozen goat semen stored for less than two years and six years, subjected to a thawing process at 37°C for 20 seconds. Data pertaining to the percentage of spermatozoa with intact membrane integrity, sperm DNA fragmentation, and MDA levels (nmol/L) were analyzed using a paired t-test with a 95% confidence level (? = 0.05). The findings of this study revealed that the duration of cryopreservation may not significantly impact post-thawing DNA fragmentation and MDA levels (p>0.05). However, a moderate decrease in sperm viability and intact plasma membrane (IPM) was observed with increasing cryopreservation duration (p<0.05). Furthermore, the IPM parameter emerged as a potentially suitable indicator for assessing the quality of post-thawing frozen semen, alongside sperm viability and motility.

Impact of Fragmentation on Carbon Uptake in Subtropical Forest Landscapes in Zhejiang Province, China

Global changes cause widespread forest fragmentation, which, in turn, has given rise to many ecological problems; this is especially true if the forest carbon stock is profoundly impacted by fragmentation levels. However, the way in which forest carbon uptake changes with different fragmentation levels and the main pathway through which fragmentation affects forest carbon uptake are still unclear. Remote sensing data, vegetation photosynthesis models, and fragmentation models were employed to generate a time series GPP (gross primary productivity) dataset, as well as forest fragmentation levels for forest landscapes in Zhejiang province, China. We analyzed GPP variation with forest fragmentation levels and identified the relative importance of the phenology (carbon uptake period—CUP) and physiology (maximum daily GPP—GPPmax) control pathways of GPP under different fragmentation levels. The results showed that the normalized mean annual GPP data of highly fragmented forests during the period from 2000 to 2018 were significantly higher than those of other fragmentation levels, while there was almost no significant difference in the annual GPP trend of forest landscapes with all fragmentation levels. Moreover, the percentage area of the control variable, GPPmax, gradually increased with fragmentation levels; the mean GPPmax between 2000 and 2018 of high-level fragmentation was higher than that of other fragmentation levels. Our results demonstrate that the carbon uptake capacity per unit area was enhanced in highly fragmented forest areas, and the maximum photosynthetic capacity (physiology-based process) played an important role in controlling carbon uptake, especially in highly fragmented forest landscapes. Our study calls for a better and deeper understanding of the potential of forest carbon uptake, and it is necessary to explore the mechanism by which forest fragmentation changes the vegetation photosynthetic process.

Analysis and prediction of the Horizon 2020 R&D&I collaboration network

This study presents a novel link prediction model for the analysis and prediction of collaboration networks in the context of H2020 projects. This model incorporates various databases and expands the list of variables used in the analysis. In this study, the accuracy and interpretability of both generic and nongeneric link prediction algorithms are examined through a comprehensive analysis. Additionally, collaborative groups that exhibit stronger linkages than initially anticipated are examined, and their prevalence across different regions is evaluated. In this study, it was found that the collaboration network within H2020 has significant concentration and fragmentation levels. Moreover, several key factors that strongly influence collaboration, including prior and concurrent project involvement, earned contribution, and the economic and technological characteristics of participating businesses, were identified. This study additionally demonstrated that cooperative communities exhibit geographical concentration in western Europe, whereas nongeneric models offer improved predictive accuracy for identifying relationships. This research contributes to the field of cooperation network analysis and offers guidance to decision-makers on strategies to promote future collaborations in the context of FP.

Антиоксидантная терапия в коррекции ДНК-фрагментации сперматозоидов.

Introduction. The progressive deterioration of reproductive function, observed throughout the world in recent decades, is an extremely alarming trend, while the incidence of male infertility is increasing. The universal pathogenetic mechanism of spermatogenesis disorders, which underlies almost all forms of male infertility, is oxidative stress of sperm. It has various manifestations, the essence of which, as a rule, boils down to the formation of mitochondrial dysfunction and fragmentation of sperm DNA. It has been found that consuming antioxidants and microelements helps increase the chances of conception for subfertile couples, and also reduces the risk of reproductive losses. Of greatest and justifiable interest are medicinal complexes designed to reduce the level of oxidative stress and sperm DNA fragmentation. The purpose of the study was to evaluate the effect of antioxidant therapy on the degree of sperm DNA fragmentation in men suffering from infertility. Materials and methods. The study involved 42 men aged 25 to 37 years with an established diagnosis of male infertility who were treated at the Center for Family Health and Reproduction. The patients were divided into 2 groups, comparable according to the main criteria assessed. Patients in the control group (12 people, 29%) received standard antioxidant therapy. Patients of the main group (30 people, 71%) received the BESTFertil-DHA® complex. The course of treatment for patients in both groups was 3 months. Control examinations were carried out at 1.5 and 3 months from the start of therapy. Spermogram data and assessment of the degree of DNA fragmentation in sperm using the SCD (Sperm chromatin dispersion test) method were used as the main criterion for assessing the effectiveness of therapy. Results. After three months of taking BESTFertil-DHA®, in 16 patients (53%), the degree of sperm DNA fragmentation decreased to less than 15%. Against this background, 5 couples became pregnant naturally, and 4 couples became pregnant as a result of the in vitro fertilization procedure. The average level of sperm DNA fragmentation was less than 13% (p < 0.05). A decrease in the level of DNA fragmentation was noted in 53% of patients after 1.5 months of taking BESTFertil-DHA®, which is especially important when preparing patients for assisted reproductive technology programs. In patients taking BESTFertil-DHA®, the total progressive motility of spermatozoa «A+B» increased by 28%, and the concentration of spermatozoa in 1 ml of ejaculate – by 61%. In the control group, the degree of sperm DNA fragmentation did not undergo statistically significant changes during treatment. Conclusion. BESTFertil-DHA® statistically significantly helps to reduce the degree of DNA fragmentation in sperm, increase the number of actively motile forms of sperm and reduce the degree of teratozoospermia.

Is sleep affected after microgravity and hypergravity exposure? A pilot study.

Sleep is known to be affected in space travel and in residents of the international space station. But little is known about the direct effects of gravity changes on sleep, if other factors, such as sleep conditions, are kept constant. Here, as a first exploration, we investigated sleep before and after exposure to short bouts of microgravity and hypergravity during parabolic flights. Sleep was measured through actigraphy and self-report questionnaires in 20 healthy men and women before and after parabolic flight. Higher sleep fragmentation and more awakenings were found in the night after the flight as compared with the night before, which was discrepant from participants' reports showing better and longer sleep after the parabolic flight. Variable levels of experience with parabolic flights did not affect the results, nor did levels of scopolamine, a medication typically taken against motion sickness. Pre-existing sleep problems were related to sleep fragmentation and wake after sleep onset by a quadratic function such that participants with more sleep problems showed lower levels of sleep fragmentation and nighttime awakenings than those with few sleep problems. These novel findings, though preliminary, have important implications for future research, directed at prevention and treatment of sleep problems and their daytime consequences in situations of altered gravity, and possibly in the context of other daytime vestibular challenges as well.

Spatiotemporal evolution of landscape stability in World Heritage Karst Sites: a case study of Shibing Karst and Libo-Huanjiang Karst

Landscape stability is a paramount concern within the field of landscape ecology. Indices of landscape patterns not only facilitate an effective analysis of land use transformations but also delve into the mechanisms of landscape disturbances across various spatial and temporal dimensions. Utilizing land use data spanning from 2014 to 2022 for the Shibing and Libo-Huanjiang South China Karst (SCK) World Heritage Sites (WHSs), this study delves into the landscape dynamics of these areas over the past 8 years. This investigation employs landscape pattern indices and a moving window technique to construct a landscape stability evaluation model, incorporating indices such as the Contagion Index, Patch Density, and Total Edge Contrast. Moreover, the study employs Moran's I, a spatial autocorrelation index, to scrutinize the shifts in geographical heterogeneity of landscape stability within the said period. The findings reveal: (1) Between 2014 and 2022, the landscape patterns of the Shibing and Libo-Huanjiang WHSs have undergone significant transformations, with Woodland emerging as the predominant landscape type and its area exhibiting an upward trend in recent years. (2) The level of fragmentation within the research area has decreased, landscape diversity has diminished, and the aggregation index has risen, according to the landscape pattern indices from 2014 to 2022, indicating that conservation measures have significantly influenced the evolution of landscape patterns. (3) Throughout most of the study period, landscape stability predominantly remained at a relatively stable level, albeit with noticeable fluctuations in years heavily impacted by human activities. (4) The areas of high-high agglomeration and the hotspots of the heritage sites were primarily concentrated within the core zones of these sites, suggesting focused areas of conservation and landscape integrity.

High seminal BPA in IVF patients correlates with lower sperm count and up-regulated miR-21 and miR-130a

Bisphenol A (BPA) is a widespread industrial chemical, used as the key monomer of polycarbonate plastics and epoxy resins. BPA has been detected in human seminal fluid and has been correlated with changes in sperm parameters, crucial in determining male fertility. In this study, semen samples were collected from 100 patients aged 29–47 years undergoing fertility assessment between 2021 and 2023 and analyzed according to WHO guidelines. BPA levels in the seminal plasma were then measured through an enzyme-linked immunosorbent assay (ELISA) and compared to sperm quality metrics. The relative mRNA/miRNA expression of key genes associated to male reproduction, including androgen receptor, miR-34c, miR-21, miR-130a, was then quantified and compared between groups with high or low BPA content. Our results revealed that BPA levels were increased with age and were negatively correlated with sperm counts (p<0.05). The negative correlation remained significant when patients were age-matched. No other relationships between seminal BPA and motility, morphology or DNA fragmentation levels were observed. qPCR analysis showed that androgen receptor mRNA expression was significantly greater in sperm with high seminal BPA (p<0.05). Moreover, we found that the expression of miR-21 and miR-130a was also upregulated in the higher BPA group (p<0.05). These results display a relationship between BPA content in the semen and male fertility parameters, and provide insights into the molecular mechanisms through which BPA may be affecting male reproductive capability. Ultimately, this research can potentially drive changes to guidelines and exposure limits for BPA exposure.

Evaluasi Kerusakan DNA Spermatozoa Semen Cair Babi Menggunakan Pewarnaan Toluidine Blue

The integrity of the deoxyribonucleic acid (DNA) in sperm plays a crucial role in the fertilization process and embryonic development. This study aimed to evaluate the level of the DNA fragmentation of boar sperm in tris-citrate-fructose extender preserved at temperatures between 16-18°C for 3 days. The level of sperm DNA damage was analysed using toluidine blue (TB) staining. Pig semen collected from 4 male pigs aged 1.3-1.8 years was used as the research sample. Semen collected was conducted twice a week. The experimental method using a Completely Randomized Design with 4 storage duration groups as follows: Group I (0 hours of storage), Group II (24 hours of storage), Group III (48 hours of storage), and Group IV (72 hours of storage), each with 5 replications. The data were analysed using analysis of variance (ANOVA) and, if significant differences were found, Duncan's test was performed. The results of the evaluation of the percentage of DNA damage during the storage process at 16-180C at 0 hours, 24 hours, 48 hours and 72 hours was: 0.33 ± 1.66%, 0.80 ± 0.447%, 1.60 ± 1.14% and 2.80 ± 0.447%, respectively. These results demonstrate that liquid boar semen preserved with tris citrate fructose extender at 16-18 °C for 3 days has spermatozoa DNA damage levels within the normal range, making it suitable for artificial insemination and ensuring optimal AI success.