Fragmentation Rate Research Articles

Possibilities for implementing of anti-fibrotic and anti-inflammatory effects of metabolic therapy in acute alcoholic disorders under experimental conditions

The aim of the work was to study antifibrotic and anti-inflammatory effects of metabolic therapy and mechanisms of regeneration in acute alcoholic liver damage (AALD) in rats under experimental conditions. Materials and methods. The experiment involved 66 white non-linear male rats with a mass of 120–130 g, which were divided into 5 groups: 1 – intact animals (n = 10); 2 – animals with AALD (n = 20), 3 – animals (n = 12) with AALD and intraperitoneally injected with Corvitin, 4 – animals (n = 12) with AALD and injected with Glutargin, 5 – аnimals with AALD (n = 12) and injected with Corvitin and Glutargin. The pro-inflammatory cytokines and cell cycle phases were analyzed. Results. The level of IGF-1 was significantly 24.1 % higher in group 2 compared to the control. In animals of group 3, the level of IGF-1 was 20.2 % decreased compared with group 2. The level of IGF-1 was significantly 9.7 % decreased in group 5 animals compared with group 2. There was a 31.6% increase in the level of TGF-β in animals of group 2 in comparison with the control ones. The level of TGF-β was 22.8 % decreased in group 3 compared with group 2. In group 5 animals, the value was 12.0 % lower than in group 2. The percentage of cell nuclei in the presynthetic phase in group 2 rats was 8.3 % higher than in controls. In animals of group 2, the number of cell nuclei in the phase of DNA synthesis were 33.3 % larger than in group 1. The rate of DNA fragmentation in AALD exceeded the corresponding value in control group by 27.5 %. Conclusions. AALD was accompanied by an increase in the serum IGF-1 and TGF-β in animals. The administration of corvitin decreased the level of IGF-1 in rats with AALD, and the use of glutargine mainly decreased the level of TGF-β. Combined use of the drugs did not show significant effectiveness. Compensatory regeneration mechanisms were activated in AALD processes and apoptotic cell death was evidenced by the increased indicators of nuclear DNA fragmentation.

Alterations in TRN-anterodorsal thalamocortical circuits affect sleep architecture and homeostatic processes in oxidative stress vulnerable Gclm−/− mice

Schizophrenia is associated with alterations of sensory integration, cognitive processing and both sleep architecture and sleep oscillations in mouse models and human subjects, possibly through changes in thalamocortical dynamics. Oxidative stress (OxS) damage, including inflammation and the impairment of fast-spiking gamma-aminobutyric acid neurons have been hypothesized as a potential mechanism responsible for the onset and development of schizophrenia. Yet, the link between OxS and perturbation of thalamocortical dynamics and sleep remains unclear. Here, we sought to investigate the effects of OxS on sleep regulation by characterizing the dynamics of thalamocortical networks across sleep-wake states in a mouse model with a genetic deletion of the modifier subunit of glutamate-cysteine ligase (Gclm knockout, KO) using high-density electrophysiology in freely-moving mice. We found that Gcml KO mice exhibited a fragmented sleep architecture and impaired sleep homeostasis responses as revealed by the increased NREM sleep latencies, decreased slow-wave activities and spindle rate after sleep deprivation. These changes were associated with altered bursting activity and firing dynamics of neurons from the thalamic reticularis nucleus, anterior cingulate and anterodorsal thalamus. Administration of N-acetylcysteine (NAC), a clinically relevant antioxidant, rescued the sleep fragmentation and spindle rate through a renormalization of local neuronal dynamics in Gclm KO mice. Collectively, these findings provide novel evidence for a link between OxS and the deficits of frontal TC network dynamics as a possible mechanism underlying sleep abnormalities and impaired homeostatic responses observed in schizophrenia.

Environmental Degradation of Microplastics: How to Measure Fragmentation Rates to Secondary Micro- and Nanoplastic Fragments and Dissociation into Dissolved Organics.

Understanding the environmental fate of microplastics is essential for their risk assessment. It is essential to differentiate size classes and degradation states. Still, insights into fragmentation and degradation mechanisms of primary and secondary microplastics into micro- and nanoplastic fragments and other degradation products are limited. Here, we present an adapted NanoRelease protocol for a UV-dose-dependent assessment and size-selective quantification of the release of micro- and nanoplastic fragments down to 10 nm and demonstrate its applicability for polyamide and thermoplastic polyurethanes. The tested cryo-milled polymers do not originate from actual consumer products but are handled in industry and are therefore representative of polydisperse microplastics occurring in the environment. The protocol is suitable for various types of microplastic polymers, and the measured rates can serve to parameterize mechanistic fragmentation models. We also found that primary microplastics matched the same ranking of weathering stability as their corresponding macroplastics and that dissolved organics constitute a major rate of microplastic mass loss. The results imply that previously formed micro- and nanoplastic fragments can further degrade into water-soluble organics with measurable rates that enable modeling approaches for all environmental compartments accessible to UV light.

Effect of Respite Time before Live Transportation on Muscle Quality of Blunt Snout (Wuchang) Bream.

To provide scientific support for improving the muscle quality of blunt snout bream, ultrahigh performance liquid chromatography, texture analyzer, and optical electron microscopy were applied to explore the effects of respite time (0, 24, 48, and 72 h) on the muscle quality of blunt snout bream before live transportation. The energy compounds (ATP and glycogen) of muscle significantly decreased with the respite time (p < 0.05). Lactic acid content declined and then increased, leading to a rise and then a fall in pH (p < 0.05). Water-holding capacity of fish muscle increased progressively when the respite time was increased to 48 h and then dramatically decreased when the respite time was further increased to 72 h (p < 0.05). Shear force gradually increased (p < 0.05), while the whiteness and lightness values remained stable (p > 0.05). Both the content of umami compounds (IMP) and bitter compounds (HxR, Hx), and the calculated K value decreased steadily with the respite time (p < 0.05). The extracellular gap of the fish muscles gradually decreased with the respite time. The most uniform and intact cellular structure occurred at 48 h. However, when the respite time was extended to 72 h, the extracellular gap and muscle fragmentation rate of the muscle increased considerably. The findings indicated that a 48 h respite time was suitable to improve the muscle quality of blunt snout bream after live transportation.

Characterizing Fragmentation of Polystyrene Foam Debris by Isopods Oniscus asellus (Isopoda: Oniscidae) and Trachelipus rathkii (Isopoda: Trachelipodidae).

Microplastics present a novel and potentially unique threat to soil ecosystems, one whose effects may be mediated by soil organisms themselves. We investigated fragmentation of polystyrene (PS) foam into microplastic particles by two isopods, Oniscus asellus L. and Trachelipus rathkii Brandt, in laboratory arena experiments. First, we examined the temporal dynamics of fragmentation across a time span of 96 h. O. asellus produced more fragments than T. rathkii, and neither species significantly fragmented the PS foam until 48 h had passed. Second, we asked whether O. asellus would still fragment PS foam in the presence of an alternate, more natural substrate like wood. Wood did not significantly affect fragmentation rates, in line with the few other studies examining the effect of alternate food on soil invertebrates' propensity to consume and/or fragment plastics. Our results provide additional characterization of PS foam fragmentation by isopods and indicate that laboratory experiments involving soil invertebrates and plastic debris can take place over relatively short timespans of four or fewer days, but do not necessarily need to provide alternate food to prove that plastic consumption would still occur in its presence.

Mini-Percutaneous Nephrolithotomy With an Endoscopic Surgical Monitoring System for the Management of Renal Stones: A Retrospective Evaluation.

PurposeTo compare the outcomes and postoperative quality of life of patients with renal calculi who underwent standard percutaneous nephrolithotomy (sPNL), mini-invasive percutaneous nephrolithotomy (mPNL) or mPNL with an endoscopic surgical monitoring system (ESMS) using a retrospective clinical trial.MethodsEighty-six adult patients with renal stones who were treated with sPNL were retrospectively compared to ninety-two patients who were treated with mPNL between July 2014 and December 2017. Next, further studies were retrospectively conducted using a matched paired method. The ninety-two patients treated with mPNL were divided into two groups based on whether the endoscopic surgical monitoring system (ESMS) was used (ESMS-mPNL vs. non-ESMS-mPNL). The ESMS used strain gauge transducers to measure the inflow and outflow of irrigation solution. Bleeding and fluid absorption during endoscopic surgery could be accurately calculated by computer program in ESMS.ResultsThe fluoroscopy time, complication rate, stone-free status and clinically insignificant residual fragment (CIRF) rate were not significantly different between the two groups (sPNL vs. mPNL). The mPNL group had a significantly longer operation time than the sPNL group, and the mPNL group exhibited a markedly reduced 12-h postoperative visual analogue pain scale (VAS) score, mean hospitalization time, and return to work time, had slightly reduced haemoglobin loss, and underwent more tubeless operations. Moreover, among the 92 patients who underwent mPNL, the operation time (P = 0.090), complication rate (P = 0.996), stone-free status (P = 0.731), CIRF rates (P = 0.125) and number of tubeless operations (P = 0.760) were not significantly different between the two subgroups (non-ESMS-mPNL vs. ESMS-mPNL); however, the patients in the ESMS-mPNL group had significantly longer irrigation times than those in the non-ESMS-mPNL subgroup, along with marked reductions in irrigation fluid, blood loss, haemoglobin loss, 12 h postoperative VAS score, mean hospitalization time, and return to work time.ConclusionsmPNL is less painful than sPNL in patients undergoing treatment for 20–40 mm renal stones. Similar stone-free rates were achieved by the two procedures, but mPNL was superior to sPNL in terms of blood loss, discomfort, hospitalization time and return to work time. We think that ESMS-mPNL is less painful for patients and more efficacious than non-ESMS-mPNL, and ESMS-mPNL achieves a stone-free rate that is similar to non-ESMS-mPNL in patients receiving treatment for 20–40 mm kidney stones.

Effect of Chronic Oral Exposure to Overdose of Cough Syrups on Rate of DNA Fragmentation in Liver and Brain of Wistar Rats

This study was designed to determine the effect of chronic oral exposure of overdose of cough syrups containing codeine (CSC) and dextromethorphan (DXM) on the rate of deoxyribonucleic acid (DNA) fragmentation in the liver and brain of Wistar rats. Forty-five rats divided into 9 groups of 5 rats were used. Groups 1, 2, 3, and 4 were treated with 0.1, 0.2, 0.4, and 0.6 mg/kg b/w of CSC, and Groups 5, 6, 7, and 8 were treated with the same doses of DXM, respectively for 21 days. Group 9 (control) received distilled water once daily and all the rats were sacrificed 24 hours after the last treatment. DNA analysis was done on the harvested liver and brain. Significant reductions (p &lt; 0.05) in the rate of DNA fragmentation of the liver tissues were observed in all the groups treated with the overdose of cough syrups when compared to the control. However, there was no significant difference in the rate of DNA fragmentation of the brain in all the groups treated with cough syrup as compared to the control. The result indicated that overdose of cough syrup may cause suppressed DNA fragmentation of the liver thereby predisposing the organ to dysfunctions and untimely aging.

DNA fragmentation of equine cumulus cells from Cumulus-Oocyte complexes submitted to vitrification and its relationship to the developmental competence of the oocyte.

The objectives of this study were to evaluate the effect of vitrification on the DNA fragmentation rate of equine cumulus cells and to assess its relationship to oocyte in vitro maturation (IVM) after vitrification. Cumulus cells (CC) from 14 mares were recovered from COCs, previously submitted to vitrification (VIT) and IVM. The DNA fragmentation rate of the cumulus cells (CC-DF) was assessed using a chromatin dispersion test. CC-DF rates between vitrified and control COCs were statistically compared by Student's t-test. The rates of CC-DF from control COCs were lower than in vitrified COCs. The percentage of CC-DF was not significantly different (p > .05) between groups of COCs able to reach metaphase II (MII > 0) and those in which oocyte maturation was not achieved (MII=0). In conclusion, vitrification has a deleterious effect on the DNA fragmentation of equine cumulus cells; however, this parameter cannot be used as a predictor for IVM success after COCs vitrification.

Field investigation- and dietary metabarcoding-based screening of arthropods that prey on primary tea pests.

Predatory natural enemies play key functional roles in biological control. Abundant predatory arthropod species have been recorded in tea plantation ecosystems. However, few studies have comprehensively evaluated the control effect of predatory arthropods on tea pests in the field. We performed a 1‐year field investigation and collected predatory arthropods and pests in the tea canopy. A total of 7931 predatory arthropod individuals were collected, and Coleosoma blandum (Araneae, Theridiidae) was the most abundant species in the studied tea plantation. The population dynamics between C. blandum and four main tea pest species (Aleurocanthus spiniferus, Empoasca onukii, Ectropis grisescens, and Scopula subpunctaria) were established using the individual number of predators and pests in each month. The results showed that C. blandum appeared to co‐occur in the tea canopy with A. spiniferus, Em. onukii, and Ec. grisescens in a longer period. The prey spectrum of C. blandum was further analyzed using DNA metabarcoding. Among prey species, A. spiniferus, Em. onukii, and Ec. grisescens were included, and the relative abundance and positive rates of target DNA fragments of A. spiniferus were greater than that of other two pests. Combined with the high dominance index of C. blandum, co‐occurrence between C. blandum and A. spiniferus in time and space and high positive rate and relative abundance of target DNA fragments of A. spiniferus, C. blandum was identified to prey on A. spiniferus, and C. blandum may be an important predator of A. spiniferus. Thus, C. blandum has potential as a biological control agent of A. spiniferus in an integrated pest management strategy.

Effects of Different Energy Diets on FSHR mRNA Expression and DNA Methylation in Promoter Region of Duolang Sheep During Diestrus.

The aim of this study was to study the relationship between the methylation level of the promoter region of follicle-stimulating hormone receptor (FSHR) gene and the mRNA expression of Duolang sheep fed different energy diets. In this experiment, three groups of diets with different energy levels (dietary nutrient level reference (NY/T816 - 2004)) were designed to obtain medium energy level diets with metabolic energy of 10.88 MJ/d. The treatments with high and low energy levels increased and decreased by 15%, 12.51, and 9.25 MJ/d, respectively, on the basis of medium energy level. Total RNA and genomic DNA were extracted from the ovaries of Duolang sheep, and qPCR was performed. Bisulfite genomic sequencing PCR and sequence alignment were used to detect the relationship between the relative expression level of FSHR and methylation. The results showed that the expression of FSHR in high-energy group and medium-energy group was significantly higher than that in low-energy group (p < 0.01), and there was no significant difference between high-energy group and medium-energy group (p > 0.05). The methylation rate of the target fragment in the promoter region of FSHR gene was 41.67% in the high-energy group, 75.00% in the medium-energy group, and 83.33% in the low-energy group. This study revealed that different dietary energy levels had certain effects on DNA methylation level of FSHR gene and then affected gene expression, and the expression level was negatively correlated with methylation level.

A Classy Memory Management System (CyM2S) using an Isolated Dynamic Two-Level Memory Allocation (ID2LMA) Algorithm for the Real Time Embedded Systems

Due to an increased scalability, flexibility, and reduced cost complexity, the dynamic memory allocation models are highly preferred for the real-time embedded systems. For this purpose, the different types of dynamic models have been developed in the conventional works, which are highly focused on allocating the memory blocks with increased searching capability. However, it faced some of the problems and issues related to the factors of complex operations, high time consumption, memory overhead, and reduced speed of processing. Thus, this research work objects to design an advanced and intelligent dynamic memory allocation mechanism for the real-time embedded systems. Here, a Classy Memory Management System (CyM2S) is developed by using an Isolated Dynamic Two-Level Memory Allocation (ID2LMA) algorithm for efficiently allocating the memory blocks with simple searching. The CyM2S helps to reduce the fragmentation rate and time consumption by optimally allocating the memory blocks. In this model, the small buffer has been maintained for surplus pointers, and the allocated blocks comprise the metadata and payload data. During evaluation, the performance of the proposed CyM2S- ID2LMA technique is validated and compared by using the measures of memory allocation time, release time, execution, and processing speed.

P-278 The presence of fragmentation during embryo cleavage does not reflect the chromosomal status of the blastocysts

Abstract Study question Does the presence of fragmentation at day-2 and day-3 of embryo development correlate with blastocyst formation and chromosomal status? Summary answer Embryos with high fragmentation have low chances to develop into blastocysts due to cytoplasmic incompetence. However, there is no correlation between aneuploidy and embryo fragmentation. What is known already The assessment of cleavage stage embryos is carried out with the evaluation of some morphological features such as fragmentation rate, irregularities in blastomeres, multinucleation and the blastomere number. It has been described that the fragments formed during cell division are responsible for the arrest and necrosis of the adjacent blastomeres; Moreover, they may hinder the blastomeres’ division planes that increase the change of abnormal compaction, cavitation, and blastocyst formation. Studies have searched for a link between aneuploidy and altered embryo morphology. However, it is unknown if the blastocyst with high fragmentation at the cleavage stage can lead to chromosomal abnormalities. Study design, size, duration From March 2019 to December 2022, couples (N = 175) underwent 190 ICSI cycles with donor oocytes and had their embryos tested through Pre-implantation Genetic Test for Aneuploidy (PGT-A). We evaluate the degree of fragmentation at their day-2 and day-3 of development of 1765 embryos and compare the blastocyst, euploidy, implantation and clinical pregnancy rate (CPR) between embryos with grade 1 and 2 (Grade 1-2) and those with grade 3 and 4 (Grade 3-4) of fragmentation. Participants/materials, setting, methods The embryo score was assigned according to the following cleavage grading system: grade-1 with 0% of fragmentation, grade-2 with 0-10%, grade-3 with &amp;gt;10-25% and grade-4 with 25-50%. Implantation, CPR and delivery rate were calculated per transferred embryo. Embryo quality was assessed with time-lapse technology (Geri®), PGT-A test was carried out on blastocyst’s trophectoderm using NGS (Illumina®). All analysis were conducted using SPSS software. Main results and the role of chance This retrospective study includes 175 couples who underwent 190 ICSI cycles using donor oocytes. A total of 2518 oocytes were injected with ICSI procedure achieving a fertilization rate of 73.8% (1858/2518). Of these, 93 fertilized oocytes were excluded from the study because they showed no cell division at day-1 of their development. Significant difference was seen in the number of embryos that reached the blastocyst stage. The blastocyst rate of grade 1-2 embryos was 60.4% (815/1348) compared to 16.8% (70/417) of those with grade 3-4 (P &amp;lt; 0.0001). Interestingly, comparable euploidy rate was found between grade 1-2 and grade 3-4 embryos being 69% (548/794) and 68.5 % (48/70), respectively (P=NS). Although Implantation and CPR decrease from grade 1-2 to grade 3-4, no significant difference was found. The implantation rate was 44.2% (104/235) and 36.8% (7/19) while CPR was 59.6% (93/156) and 36.8% (7/19) in grade 1-2 and grade 3-4 group, respectively (P=NS). Pregnancy loss was 16.1% (15/93) in grade 1-2 and 5.2% (1/19) in grade 3-4 (P=NS). Limitations, reasons for caution This study was carried out on donor oocytes to exclude any female factor that could have affected the euploidy rate. Therefore, the number of patients is limited. Moreover, Implantation parameters like receptivity or immunologic disorders weren’t addressed. No data have been included on perinatal and obstetrical outcomes for pregnancies. Wider implications of the findings This study demonstrated that although embryos with high fragmentation have a lower change of reaching the blastocyst stage, the fragmentation parameter doesn’t reflect the chromosomal status of the resulting blastocysts. Trial registration number N/A

P-320 The Effects of Endometriosis on Oocyte Morphology and Embryo Development

Abstract Study question What is the effect of the endometriosis on oocytes morphological abnormalities in intracytoplasmic sperm injection cycles.? Summary answer An increase in the detection rate of refractile body and polar body fragmentation was observed in the morphology of oocytes obtained from endometriosis patient. What is known already The quality of the oocyte is suspected for the lower clinical success in endometriosis patients compared to other patient groups. The oocyte quality is determined by its morphological, cellular, and molecular evaluations. However, there are limited studies on the association between endometriosis and oocyte morphology. Study design, size, duration This was a retrospective cohort study, conducted between January 2020 and August 2021, at a tertiary care hospital. The women aged 24-35 years and BMI 19-30 kg/m2 ranges who underwent fresh GnRH antagonist cycles and stimulated with 150-300 IU dose of recombinant FSH (rFSH) and then trigerred with 250 micrograms/0.5 ml of rhCG were included. Participants/materials, setting, methods The patients who were cycled with the etiology of endometriosis (n = 46 cycles, Group 1) and those who were cycled with the etiology of male infertility (n = 64 cycles, Group 2) were included in this study. In addition to the comparison of intracytoplasmic and extracytoplasmic (polar body, perivitelline space, zona pellisuda) anomalies observed in oocyte morphology between groups, morphology anomalies that affect fertilization and blastocyst development within the group were evaluated. Main results and the role of chance The patient age, BMI, antral follicle counts, duration of infertility, and basal endocrine parameters among patients who were cycled with the etiology of endometriosis (n = 46 cycles, Group 1) and those who were cycled with the etiology of male infertility (n = 64 cycles, Group 2) were not different (p&amp;gt;0.05). As a result of the comparison of intracytoplasmic anomalies of 800 oocytes in total (n = 263 Group 1; n = 537 Group 2) between the groups; the refractile body determination (OR:1.23; 95% CI 1.08 -1.4, p &amp;lt;0.00) was higher in the endometriosis group compared to the other group. As a result of the comparison of extracytoplasmic anomalies between the groups; the polar body fragmentation (OR:1.3; 95% CI 1.01 - 1.85, p&amp;gt; =0.03) determination was higher in the endometriosis group compared to the other group. When the effect of oocyte morphology anomalies on fertilization and blastocyst development within the groups was evaluated by logistic regression analysis, it was found that granulation of the perivitelline space had a negative effect on blastocyst development only in the endometriosis group (β = -0.69; p&amp;gt; =0.31). Limitations, reasons for caution The main limitation of our study was its retrospective design, which precludes us from reaching a more definitive conclusion on the relation between oocyte morphology of endometriosis an fertility outcomes. Wider implications of the findings The previous studies indicate that PVS and PBI abnormalities may be ascribed to over-mature oocytes and also be negatively correlated with fertilization rate and embryo quality. Our study suggest that endometriosis seems to negatively affect oocyte quality and maturation process. Trial registration number not applicable

P-093 A comparison of the effects of IMSI and HOST for sperm selection on morphokinetic parameters and ploidy status in sibling embryos

Abstract Study question Are hypo-osmotic swelling test (HOST) and intracytoplasmic morphologically selected sperm injection (IMSI) both effective alternative methods for sperm selection? Summary answer Similar results were obtained in embryo morphokinetics between HOST and IMSI. What is known already HOST is a simple, low-cost and rapid test based on the swelling of the tail of the sperm in a hypoosmotic environment. Less DNA fragmentation, membrane integrity and aneuploidy rates have been found in sperm selected by the HOST method. It also allows identification of viable sperm in a totally immotile sperm sample. IMSI, on the other hand, is a method for identifying the morphologically best sperm using high-magnification microscope. In studies, IMSI has been associated with high fertilization rates. However, it is a time-consuming method that requires expensive equipment. Study design, size, duration Time-Lapse Monitoring (TLM)-incubated sibling embryos of 25 male factor patients who applied for ART treatment to Istanbul, Sisli Memorial ART and Genetics Center in 2021 were included into the study. Sibling embryos were grouped according to their sperm selection method. In the IMSI group 170 embryos, in the HOST group 173 embryos were evaluated retrospectively in terms of the morphokinetic parameters, the rates of blastulation, Top- Quality (TQ) and Good Quality (GQ) embryos and euploidy. Participants/materials, setting, methods Blastocyst grading was performed according to Gardner’s classification. Blastocysts 3AA,4AA,4AB,4BA,4BB,5AA,5AB,5BA,5BB were accepted as TQ+GQ. Blastulation rates was determined including the embryos at least 2 expansion. PGT-A results were obtained by Next Generation Sequencing(NGS) method. The comparison of the TLM parameters and clinical features of the patients were performed using Student’s t-test. Chi-square test was used for comparing the rates of fertilization, blastulation, TQ+GQ embryo and euploidy in each group. Main results and the role of chance No statistical difference was observed between IMSI and HOST groups in terms of morphokinetic parameters (tPN, tPNf, t2, t3, t4, t5, t6, t7, t8, t9, tSC, tM, tSB, tB, tEB) (p &amp;gt; 0.05). There were higher rates of normal fertilization in the HOST group (90.8%) than in the IMSI group (82.4%) and of TQ+GQ embryo (HOST: 50%, IMSI: 47.3%). However, these diffrences were not statistically significant (p &amp;gt; 0.05). There was statistically lower abnormal fertilization rate was found in HOST group (2.3% vs 7.1%, p = 0.038). When PGT-A results were compared (HOST:19 embryos, IMSI: 19), slightly higher results were observed in IMSI group than HOST group ( 63%, 58% respectively) (p &amp;gt; 0.05). Limitations, reasons for caution This was a retrospective of a small number of cases. Wider implications of the findings The similarity of the outcome in these initial findings studying a small number of cases, suggest that this study should be extended. Trial registration number None

P-120 Microfluidic sperm selection device increases clinical pregnancy rate in IVF/ PGT-A cycles

Abstract Study question Evaluate the influence of sperm selection according to the sorting technique, and its effects in clinical and laboratory parameters in IVF/PGT-A cycles. Summary answer Cycles that used microfluidic sperm selection device presented an increase in clinical pregnancy rates. What is known already A common genomic anomaly detected in subfertile men is the Sperm DNA fragmentation. It is resulted of a high concentation of free radicals produced causing oxidative stress in sperm cells. In order to achieve a normal embryonic development is necessary good spermatozoa with intact genetic material. In fact, embryonic quality parameters as lower fertilization, blastocyst formation and pregnancy rates has been related with Sperm DNA fragmentation increased cases. The microfluidic sperm selection device has been commercialized promising to select spermatozoa with a low level of DNA fragmentation and suggest a relationship between embryo quality, euploidy and pregnancy rates. Study design, size, duration A case-control study involving 494 IVF/PGT-A cycles that resulted in embryo biopsy between June, 2020 and November, 2021 at a Brazilian private practice unit (Instituto Ideia Fertil) were included in this analysis. Participants/materials, setting, methods All cycles that would perform PGT-A were splitted in two groups considering the sperm sorting technique: case - Microfluidic sperm selection device (245 cycles) and control - Density gradient centrifugation + swim-up (249 cycles) were included. The variables considered in this analysis were patient age, sperm concentration, fertilization, D3 top quality, blastocyst top quality, blastulation, ploidy and pregnancy rates. This data were analysed between groups and female age (until 37; 38-40 and over 41). Main results and the role of chance The mean age of the female and male patients found were 38.05 and 38.2 respectively and there was not statistically significant different between groups. Fertilization rate compared between case (72.4%) and control (76.5%) groups (p &amp;lt; 0.05) demonstrated statistically significant difference, but when stratified accordingly to the age, only the female age until 37 years old group showed significant difference (73.25%; 76.43%; p &amp;lt; 0.05). The variables analyzed as D3 Top Quality, Blastocyst Top Quality, blastulation and ploidy did not present difference (p = 0.552; 0.699; 0.378). The clinical pregnancy rate analyzed revealed in case group 65.33% and control 56.38% (p &amp;lt; 0.05), demonstrating statistically significant difference. When groups were splitted according to the female age (until 37 years old; 38-40 and over 41) the findings behaved in the same characteristics, increasing only the clinical pregnancy rate. The tests used was Mann-Whitney and Kruskal-wallis. Limitations, reasons for caution The DNA fragmentation test was not evaluated because not all patients had this test performed. Besides that, another parameter that will be analysed is live birth rate. Wider implications of the findings Microfluidic sperm sorting device made possible to select high quality spermatozoa according to motility, morphology, and low rates of fragmentation, showing the importance of having a good sperm in clinical pregnancy rate. Trial registration number none

P-009 A novel sperm selection technique using a progesterone gradient

Abstract Study question Is it possible to select spermatozoa using progesterone gradients? Does chemotaxis improve sperm selection compared to a classical density gradient or swim up? Summary answer Spermatozoa selected using progesterone gradients yield higher percentages of high quality spermatozoa with lower DNA fragmentation and better morphology compared to classic sperm selection. What is known already There are few studies which aim at improving sperm selection, even though it is known to be a vital parameter for IVF success. DNA integrity is fundamental for embryo development although it is currently underestimated. Progesterone has been suggested to be a major chemoattractant for spermatozoa in the fallopian tubes and therefore using gradients in a clinical setting may lead to a more physiologically improved selection method. Study design, size, duration 19 normo spermic patients (according to W.H.O. parameters) were selected. After standard semen analysis the sample was split in two halves. One half was treated using classical swim up and the remaining half was treated on a progesterone gradient. The treated samples were then tested for fragmentation, motility and morphology. Sperm motility was analyzed using SCA computer system. DNA fragmentation using the TUNEL assay. Participants/materials, setting, methods The patients enrolled in the study were all normospermic. The results were compared using ANOVA one way. Progesterone gradients was obtaining using an horizontal density gradients with added progesterone. Main results and the role of chance The percentages of rapid progressive spermatozoa were 12,89 ± 4,50%; 56,37 ± 13,61% and 29,84±12,52% in the untreated sample (US), treated sample (TS) and control group (C)respectively; p &amp;lt; 0,005. Normal sperm morphology rates were 3,474 ± 1,46; 5,500 ±1,57 and 3,684 ± 1,15 in US, TS and C respectively. Sperm fragmentation rates were 9,77 ± 9,012,50 ± 2,66 and 7,94 ± 6,68 in US, TS and C respectively. Limitations, reasons for caution The number of normospermic patients enrolled in the study was limited. Moreover, the sperm concentration obtained in this kind of selection is lower than in the conventional techniques and therefore may be harder to use in patients with a very low sperm count. Wider implications of the findings Our data shows a significant improvement in sperm selection using chemotaxis gradient compared to the control group by all the parameters considered in this study. Chemotaxis gradients may be useful in a clinical setting to improve spermatozoa selection, lowering the rates of sperm with damaged DNA potentially increasing embryo quality. Trial registration number Not applicable

P-039 Male age is associated with sperm DNA integrity: Selection of high DNA integrity sperm by microfluidics sorting is critical to clinical outcomes in older patients

Abstract Study question Does sperm DNA integrity affect clinical outcomes of ICSI? Summary answer Use of high DNA integrity sperm selected by microfluidics sperm sorting results in lower miscarriage rates in the patients of 39-years old and more. What is known already High sperm DNA damage is associated with decreased normal fertilization, embryo development and pregnancy rates, and an increased miscarriage rate. On the other hand, oocytes from older women have decreased pregnancy rate, and increased miscarriage rate because of possibility of low ability to repair sperm with DNA fragmentation, and dramatical increases of aneuploidy as women age. A microfluidic sperm selection chamber (MSS, ZyMōt™; DxNow) is a device designed to collect sperm with higher chromatin integrity than density gradient centrifugation (DGC). Study design, size, duration Sperm analysis was performed by sperm chromatin dispersion (SCD) test and comet assay in the same sample of 15 cases between October 2020 and February 2021. ICSI outcomes by DGC and MSS were compared with blastocyst development, and pregnancy rates in vitrified-thawed single blastocyst transfers cycle for 518 cases between August 2018 and May 2021. Participants/materials, setting, methods SCD test was optimized as a rapid procedure, with sperm showing a halo deemed normal, and those without a halo abnormal. Comet assay results were analyzed using CometScore 2.0, with comparison of %Tail DNA. ICSI outcomes were analyzed using multiple logistics regressions of male and female ages. Main results and the role of chance We found a positive correlation between male age and sperm DNA fragmentation rates in raw semen using SCD test (r = 0.70) and Comet assay (r = 0.42). Higher DNA integrity sperm could select using MSS than DGC. In this study with ICSI outcomes, 170 of 318 (53.5%) blastocyst transfers resulted in pregnancy, and 49 (28.8%) subsequently miscarried. The data were classified according to less than or more than 39 years old of male age detected by multiple logistics regressions. In patients with ≥39 years of male age, the female age was significantly higher and blastocyst and pregnancy rates were significantly lower, and the miscarriage rate was significantly higher than &amp;lt;39 years of male age. Since sperm DNA fragmentation increased in accordance with male age, we compared MSS and DGC in the patients with male age ≥39 years. There was no significant difference in blastocyst, pregnancy, and miscarriage rates in female age &amp;lt;39 years. While in ≥ 39 years of female age, blastocyst and pregnancy rates in MSS were not significantly different from DGC, but the miscarriage rate in MSS was significantly lower than in DGC (27.3 vs. 57.1%). Limitations, reasons for caution The sample size for each study was small. Analysis of sperm DNA fragmentation and samples in ICSI outcomes were not the same. The retrospective nature of ICSI outcomes in this study does not allow controlling of unknown confounders. Wider implications of the findings Sperm DNA fragmentation depended on male age affected fertility outcomes. However, when male age is higher, masking the effect of male age by female age. In this study, we found out the improvement of ICSI outcome by using high DNA integrity sperm selected by MSS in both ≥39 years. Trial registration number Not applicable

Degradation Characteristics of Environment-Friendly Bamboo Fiber Lunch Box Buried in the Soil

The research on the development of lunch boxes made of clean, environment-friendly, and naturally degradable plant fibers has attracted enormous attention. A bamboo fiber lunch box prepared by the clean and efficient steam explosion method has the advantages of good stiffness, water and oil resistance, and easy degradation. The purpose of this study was to investigate the degradation behavior of the environment-friendly bamboo fiber lunch box under indoor soil burial, as represented by the changes in physical properties, mechanical strength, chemical components, morphological structure, and so on. The results showed that: with the extension of the burial time, the weight loss increased rapidly from slowly to quickly; the boxes were completely degraded in the soil on the 70th day; the microorganisms in the soil first decomposed the tapioca starch, hemicellulose, and cellulose in the lunch box, and finally decomposed the lignin; the residual debris in the soil was further decomposed into CO2, H2O, and inorganic salts. In short, the degradation process of the lunch box mainly included the following stages: stage I: the increase in apparent roughness, the generation of microcracks, the rapid increase in weight loss, and the breakdown of starch and hemicellulose; stage II: the slow increase in the weight loss rate of the box fragmentation, the rapid decay of the mechanical strength, and the cellulose decomposition; stage III: the decomposition of lignin, the complete degradation of the debris, and the integration with the soil.

The assessment of particle selection and blank correction to enhance the analysis of microplastics with Raman microspectroscopy

Although microplastics research has received enormous attention in the last decade, both the research practices and the quality of produced data should still be improved. In this study, the identification process of microplastics with Raman imaging microscope was improved by decreasing the time needed for the analysis. To do that, new features, including terrain mosaic and automatic particle selection, were utilized and various ways of handling the produced microplastics data were implemented and discussed. Furthermore, blank correction of microplastic concentrations was demonstrated and its effects on the recovery of spiked microplastics was assessed with aqueous and solid samples.Six types of microplastics, including fragments and fibers, were spiked in triplicates of ultra-pure water and reference sediment samples. The spiked samples were pretreated by a modified method of the universal enzymatic purification protocol. Microplastics were analyzed with Raman imaging microscope, using terrain mosaic combined with automatic particle selection. The microplastics data were subjected to different identification steps to estimate the potential overestimation and underestimation of microplastics counts.With the complete correction of Raman-based data, the average recovery rates of fragments (77–80%) were higher than fibers (20–33%). The decrease in recovery rates of spiked microplastics (49–57%) were observed when blank correction was applied (28–47%). The impact of the blank correction depended on the polymer, causing exclusion of PE, PET, and PP from sediment samples. For the completely corrected Raman-based data, the average recovery rates of microplastics were higher for water than sediment samples both with and without blank correction.The results demonstrated the impact of blank correction on the microplastics recovery rates. To our knowledge, this is the first study to explore the use of automatic particle selection of Raman imaging microscope for microplastics analysis. Hence, potential drawbacks and advantages of the new features of Raman imaging microscope were explicitly discussed.

Microbial translocation, toll-like and vitamin D receptor polymorphisms in blood and risk of recurrence in stage III colorectal cancer.

3531 Background: Microbial translocation from the intestinal lumen into the blood circulation is significantly linked to intestinal dysbiosis; thus, leading to colorectal cancer (CRC), disease progression and decreased survival. Toll-like (TLRs) and vitamin D receptors (VDRs) play essential role in immunity and gut microbiome determination. Polymorphisms in such receptors have been associated with increased CRC incidence risk and mortality. The aim was to evaluate the microbial translocation in the blood of stage III CRC patients and correlate the presence of TLR and VDR genetic variants with microbial DNA fragments at risk of CRC development and progression. Methods: A total of 132 stage III CRC patients and 100 healthy donors were enrolled in the study. Peripheral blood DNA was analyzed using PCR for the amplification of microbial DNA encoding 16S rRNA, β-galactosidase gene of Escherichia coli, glutamine synthase of Bacteroides fragilis, and 5.8S rRNA of Candida albicans. Moreover, DNA from patients and controls was analyzed using PCR and PCR-RFLP for genotyping functional polymorphisms of both TLR (TLR2, TLR4, TLR9) and VDR ( TaqI, ApaI, FokI and BsmI) genes. Results: Median age of patients was 62 years, 59.8% were males, 92.7% had a colon/sigmoid tumor, 24.4% had a right colon tumor and 99.2% had a good performance status. Microbial DNA fragments from 16S rRNA, E. coli, B. fragilis, and C. albicans were detected in 43.2%, 20.5%, 31.8% and 36.4% of patients, respectively. Significantly higher rates of all microbial fragments, but E. coli, were detected in the group of patients in comparison to healthy donors ( p &lt; 0.001). Similarly, higher rates of both TLR and VDR genetic variants were detected in CRC patients compared to healthy donors ( p &lt; 0.001). Moreover, individuals with homozygous mutant alleles of either TLR or VDR polymorphisms had significantly higher detection rates of microbial DNA fragments. KRAS, BRAF and MSI status were significantly correlated with TLR9 genetic variants ( p= 0.001, p= 0.013 and p= 0.011, respectively) and MSI status was significantly correlated with all four VDR polymorphisms ( TaqI, p= 0.044; ApaI, p= 0.037; FokI, p= 0.002 and BsmI, p&lt; 0.001). Cox regression analysis revealed that BRAF mutations, histology type and ApaI genetic variants are significantly associated with shorter disease-free survival (DFS). C. albicans detection is significantly associated with shorter overall survival, and B. fragilis is an independent poor prognostic factor for decreased DFS (HR = 33.85; p = 0.018). Conclusions: The detection of higher frequencies of the TLR/ VDR genetic variants was correlated with significantly higher detection rates of microbial DNA fragments. The detection of these TLR/VDR polymorphisms and microbial DNA fragments in CRC patients highlighted their role in cancer development, progression, and patients’ survival.