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Blast furnaces create iron and steel from pig iron, which in turn produces iron slag. Iron ore is the primary raw material for these transformations. Slag aggregate, a byproduct of the iron and steel industry, is a sustainable building material. In order to produce more environmentally friendly and cost-effective concrete, this study evaluated the effect of inclusion of two waste materials in concrete including, 10% treated cement kiln dust (TCKD) as cement weight replacement with different volumetric contents (15, 25, and 35%) of iron slag coarse aggregate (ISCA) as a replacement to natural coarse aggregate. Microstructure, static modulus of elasticity, splitting tensile strength, flexural strength, water absorption, and workability were among the many concrete qualities studied. There was an improvement in oven-dry, splitting tensile, flexural, compressive, ultrasonic pulse velocity, and static modulus of elasticity as ISCA content rose, as shown in the results. Increasing the ISCA concentration reduces thermal conductivity. Depending of the ISCA content, increasing the compressive strength by 1.43–12.4% and the splitting tensile strength by 0.4–5.34% were both possible. There was an additional increase of 1.3–9.15% in flexural strength. From the experimental results, it can be observed that innovative and potent method for producing structural sustainable concrete is provided in this study by inclusion of two waste materials, including TCKD and ISCA. The sustainable concrete produced has high strength and low thermal conductivity relative to concrete not containing these waste materials, which can be used in different construction projects. Moreover, the use of these waste materials in concrete has a benefit of reducing the environmental pollution. All the above-mentioned results conforms the goal of this study.

Geopolymer in recent concrete (GP) has gained significant attention due to its sustainability and environmental friendliness. Local metakaolin-based geopolymer is weaker than geopolymers made from other materials due to its low Si/Al ratio. A consistent mixture design is also lacking because geopolymers are affected by many variables. This study presents a method to find the optimal geopolymer mixture based on locally modified metakaolin as a source of aluminates and silicates using the Taguchi method. Metakaolin was modified using different contents of materials rich in silica, such as silica fume (SF), or materials rich in calcium, such as calcium oxide (CaO) of 5, 10, and 15%. The inclusion of 5% SF, 10% CaO, and a combination of 5% SF and 5% CaO as substitutes for metakaolin increases the compressive strength by 16.8, 6.9, and 22.8%, respectively, compared to the reference mixture without any modifications. In other words, adding 5% SF and CaO increased the molar ratio of SiO2/Al2O3 (R) from 1.782 to 1.914, resulting in the highest compressive strength of 53.3 MPa after 7 days of sun curing. To obtain the optimal mixture that achieves the highest compressive strength, the impact of three main variables, including the concentration of sodium hydroxide (SH), alkali solution-to-binder (Al/B) ratio, and sodium silicate-to-sodium hydroxide (SS/SH) ratio, must be considered using Taguchi method. A total of nine mixtures were investigated. It was found that 13 M of SH, 0.65 Al/B, and 2.5 SS/SH give a high compressive strength of 58.6 MPa at 7 days. Also, it was found that the concentration of SH plays a more important role in increasing the compressive strength than the alkali-to-binder ratio and SS/SH ratio. The scanning electron microscopy images show that the 5% weight replacement of metakaolin by silica and CaO could source fewer and smaller pores and reduce the microcracks’ width.

This study combines two types of waste materials to achieve sustainable lightweight concrete (LWC) including agricultural and industrial wastes that hold high potential to reduce global environmental pollution. A novel application of 35% treated date seed (DS) wastes as partially replaced by volume to natural coarse aggregate in producing sustainable LWC was investigated. Calcium hydroxide solution was utilized as a treatment to extract cellulose from the achieved hemicelluloses of the DS. Fourier transform infrared spectroscopy was utilized to analyze the structural characteristics of the isolated cellulosic samples before and after treatment. Then different volume fractions of steel fibers (0.5, 0.75, and 1%) recycled from scrap tires were added to the LWC with 35% DS. Several experimental tests, including slump test, compressive strength, oven-dry density, split tensile strength, flexural strength, and water penetration depth, were carried out in this investigation. The results demonstrate that concrete containing 35% DS and reinforced with 1% recycled steel fibers show enhancement in the compressive, tensile, and flexural strengths by about 15.2, 32.6, and 58.2%, respectively, and reduces the permeability by 16.1% relative to the reference concrete mixture.

Roller-compacted concrete pavement (RCCP) is one of the most durable, economical, and practical solutions for the construction of roads for various heavy-duty purposes. To make RCCP more sustainable, different waste materials have been utilized. These materials were densified silica fume (SF), ground granulated blast furnace slag (S), crumb rubber (CR), and recycled steel fibers (RSFs) from waste tires. The weight percentages of replacement for SF and S from cement were 5, and 27.5%, respectively. CR was utilized as a volumetric replacement of sand with 0, 2, 5, and 10%. As a volumetric addition of concrete, RSF with 0.2, 0.4, and 0.6% was utilized. Water content was 6% for all mixtures. The impact resistance test was performed to evaluate the behavior of RCCP to the repeated load on roads. Also, ultrasonic pulse velocity (UPV) (nondestructive) and abrasion resistance tests were performed to validate roller-compacted concrete (RCC) as pavement. There is a substantial increase in impact energy by using 10% of CR and 0.6% of RSF, compared with that of reference specimens. The use of CR and RSF can improve the abrasion resistance of RCC, and this can ensure its applications in pavements. The relationships between impact, abrasion, and UPV were established, and models have been proposed to predict these relationships.

This work presents a development of a new radiocarbon dating system based on CO2 and liquid scintillation counting. The system is flexible enough to be applied in different fields such as radiocarbon dating (up to about 32 000 years span), environmental radioactive monitoring, and determining the fraction of biological carbon in environmentally-ecologically safe biodegradable and bio-synthetic products, and help accreditation to customaries. The implemented system is developed following the standard method (ASTM D6866-06). It has been calibrated tested and verified for CO2 production, absorption, and counting (using appropriate reference materials for the present-day, fossil, and in between ages). These results are optimistic that the developed system can contribute to the accreditation of ongoing and upcoming environment- friendly productions.

This study investigated the types and profile of defects facing newly constructed houses through conducting a survey and analyzing defect records of data observed for 652 houses out of 1000 houses newly constructed for Salahaddin University academic staff in Erbil City. The result of analysis revealed that the overall of 6758 defects identified with the mean average of 10 defects per house. The overall percentage of defected houses for each type of defects and the location of the defect ranged from 10% to 67%. The most defected components found in the doors and windows, which comes in rank 1 with the highest percentage of defected houses reached to 76%, whereas coating and painting of doors come in rank 2 with percentage of 75%, and cracks in structural elements come in rank 3 with 73%. Analyzing the defects in terms of area and location showed that the finishing works representing the major defects area of 48%. While, the defects in the doors and window representing second highest defects of 42%. The results indicated that the quality performance in newly constructed houses is low due to poor workmanship and lack of experience and skills of construction staff and inadequate supervision

Short-term grassland set-asides (GLSA) have been incorporated into intensive annual crop rotations to improve soil quality. The legacy of the GLSA to subsequent annual crops, however, is not well understood. The objective of this study was to determine the impacts of 3-year-old GLSA on nitrogen (N) dynamics and the yield of the subsequent cash crop. A regional analysis was conducted over two years, utilizing eight production fields transitioning from GLSA, paired with fields in continuous annual crop rotation (ACR) with matching management. A controlled plot-scale experiment was also conducted on a single 3-year-old GLSA, comparing fertilizer types, rates, and timing of incorporation. In each experiment, soils were sampled every 10–14 days for ammonium (NH4+-N) and nitrate (NO3−-N), along with ion probes, installed near the rooting zone to track plant available nitrogen (PAN) throughout the season. The results from the regional analysis are confounding, in 2015 showing that GLSA supplied an additional 18 kg PAN ha−1 compared to ACR but showed no PAN benefits in 2016. The controlled plot-scale experiment highlighted the importance of fertilizer type to subsequent PAN, showing synthetic treatments consistently supplied more PAN than organic. The results from this study suggest that 3-year-old GLSAs can potentially improve PAN to subsequent crops depending on how they are managed.

The present study included detection of genetic variability, and identification of genetic relationship and finding a fingerprint of the ten clinical isolates related to Trichophyton rubrum using RAPD and ISSR markers. The experiment was carried out and the results performed using six primer of the RAPD markers these primers showed 239 amplified band , out of these band 90 of them was considered as a main band, and 149 was Polymorphic band , the largest number of bands was 30 in the isolate TR6 and less number of bands in the isolate TR7. The results clear the value of genetic diversity based on RAPD analysis the lowest value of genetic diversity (0.13005) between the isolate TR3 and TR9while the highest genetic diversity (0.55941) was between the isolates TR5 and TR8. The analysis of the relationship shows that there are three main groups: the first group include isolate TR8, while the second group included three isolates are isolate TR2, isolate TR10 and isolates TR3, The third group included three subgroups included the first isolate TR1,isolates TR4 and second isolate TR3 and isolate TR9 and the third included the isolate TR5 and isolates TR6.The results of ISSR experiments: the use six a primers in the of the ISSR showed 192 bands, in the isolate of Trichophyton rubrum, two of these primers showed monomorphic bands (in number and location) and six primers showed monomorphic and polymorphic bands, while one showed only polymorphic bands among Trichophyton rubrum isolates. And the largest number of bands was 24 in the TR5 and less number of bands 16 in isolate TR3 and isolate TR8 were finding DNA fingerprint to isolate TR1, isolate TR5, and isolate TR3. The ISSR markers showed lowest genetic polymorphism was (0.05561) between the isolates TR2 and isolate TR7 and the largest genetic distance was (0.40501) between the isolate TR4 and isolate TR8. The analysis of the relationship of genetic showed that there are three groups key first included isolates and only one isolate TR8 and the second involving isolates TR2 and isolate TR7, isolate TR4 and isolate TR3, while the group included three sub-groups, the first included isolate TR1 and isolate TR6, and the second involving isolate TR5, isolate TR10 and isolate TR9. The results confirmed the efficiency markers of the RAPD and ISSR in contrast, find a genetic fingerprint to the isolates of Trichophyton rubrum, and these markers differ in mechanical detection contrast, and coverage is Genome. Therefore Complementary to each other, although the ISSR markers were more efficient in terms of the number of binding sites in each type of the isolates of Trichophyton rubrum and so can the initiator of the discovery of a much larger area from Genome.

This investigation includes the use of 15% of glass wastes as a partial substitution to cement in combination with plastic wastes as volumetric replacement to natural coarse aggregate to produce sustainable concrete. Different volumetric replacements of plastic waste to natural coarse aggregate (25%, 50%, 75%, and 100%) were used in concrete containing 15% glass powder as a replacement by weight of cement. Generally, the results show that the inclusion of 15% glass powder improves the compressive strength, splitting tensile strength and flexural strength by about 13.3%, 36.3%, and 34.7%respectively at 60 day age in comparison with reference concrete without wastes, also the results show a decrease in water absorption and an increase in dry density. The inclusion of plastic waste aggregate in the presence of 15% glass powder leads to a decrease in the compressive strength, flexural strength, splitting tensile strength, dry density, ultrasonic pulse velocity, and thermal conductivity. The percentage reductions are 59.8%, 46.3%, 43.6%, 20.5%, 28.6%, and 54.4% respectively for concrete specimens that including 100% plastic waste coarse aggregate in comparison with concrete specimens without plastic waste aggregate.

In this investigation, sustainable High Performance Lightweight Aggregate Concrete (HPLWAC) containing artificial aggregate as coarse lightweight aggregate (LWA) and reinforced with mono fiber, double and triple hybrid fibers in different types and aspect ratios were produced. High performance artificial lightweight aggregate concrete mix with compressive strength of 47 MPa, oven dry density of 1828 kg/m3 at 28 days was prepared. The Fibers used included, macro hooked steel fiber with aspect ratio of 60 (type S1), macro crimped plastic fiber (P) with aspect ratio of 63, micro steel fiber with aspect ratio of 65 (type S), and micro polypropylene fiber (PP) with aspect ratio of 667. Four HPLWAC mixes were prepared including, one plain concrete mix (without fiber), one mono fiber reinforced concrete mixes (reinforced with plastic fiber with 0.75% volume fraction), one double hybrid fiber reinforced concrete mixes (0.5% plastic fiber + 0.25% steel fiber type S), and a mix with triple hybrid fiber (0.25% steel fiber type S1+ 0.25% polypropylene fiber + 0.25% steel fiber type S). Fresh (workability and fresh density) and hardened concrete properties (oven dry density, compressive strength, ultrasonic pulse velocity, splitting tensile strength, flexural strength, static modules of elasticity, thermal conductively, and water absorption) were studied. Generally, mono and hybrid (double and triple) fiber reinforced HPLWAC specimens give a significant increase in splitting tensile strength and flexural strength compared with plain HPLWAC specimens. The percentage increases in splitting tensile strength for specimens with mono plastic fiber are, 20.8%, 31.9%, 36.4% and 41%, while the percentage increases in flexure strength are 19.5%, 37%, 33.9% and 34.2% at 7, 28, 60, 90 days age respectively relative to the plain concrete. The maximum splitting tensile and flexure strengths were recorded for triple hybrid fiber reinforced HPLWAC specimens. The percentage increases in splitting tensile strength for triple hybrid fiber reinforced specimens are 19.5%, 37%, 33.9% and 34.2%, while the percentage increases in flexure strength are 50.5%, 62.4. %, 66.8% and 62.2% at 7, 28, 60 and 90 days age respectively relative to the plain concrete specimens.

Due to the rapid depletion of natural resources, the use of waste materials and by-products from different industries of building construction has been gaining increased attention. Geopolymer concrete based on Pozzolana is a new material that does not need the presence of Portland cement as a binder. The main focus of this research is to produce lightweight geopolymer concrete (LWGPC) using artificial coarse lightweight aggregate which produced from locally available bentonite clays. In this investigation, the binder is low calcium fly ash (FA) and the alkali activator is sodium hydroxide and sodium silicate in different molarities. The experimental tests including workability, fresh density, also, the compressive strength, splitting tensile strength, flexural strength, water absorption and ultrasonic pulse velocity at the age of 7, 28 and 56 days were studied. The oven dry density and thermal conductivity at 28 days age are investigated. The results show that it is possible to produce high strength lightweight geopolymer concrete successfully used as insulated structural lightweight concrete. The 28-day compressive strength, tensile strength, flexural strength, dry density, and thermal conductivity of the produced LWGPC are 35.8 MPa, 2.6MPa, 5.5 MPa, 1835kg/m3, and 0.9567 W/ (m. K), respectively.

The main aim of this investigation is to study the combined effect of hot-dry weather conditions on the plastic and drying shrinkage of high performance lightweight aggregate concrete (HPLWAC) specimens, along with the other properties including, workability, setting time compressive and splitting strength. The experimental program including the use of fixed mix proportions and was carried out in a typical Iraqi summer day (under actual conditions) of different times during the day. The results indicate that the use of lightweight aggregate play a main role in decreasing the effect of hot dry weather on plastic shrinkage, as well as the plastic shrinkage strain of HPLWC specimens cast at 12:00 pm. less than that of normal weight aggregate concrete specimens by about 36.7%. While drying shrinkage of HPLWC specimens at initial ages up to 7 days was low and it increases with a higher rate at later ages.

Epidermal growth factor (EGF) plays a crucial role in maintaining male reproductive capacity in mammals, however, its protective effects on cryopreserved dromedary camel epididymal spermatozoa have not been thoroughly investigated. This study aims to investigate the potential protective role of EGF on cryopreserved camel epididymal spermatozoa, supported by evidence from a molecular docking study. We assessed sperm motility, kinematics parameters, oxidative stress, ultrastructural changes, apoptosis, and molecular docking markers in camel epididymal spermatozoa following cryopreservation. Camel epididymal spermatozoa (n = 30 pairs of testes) were collected from local slaughterhouses. The epididymal spermatozoa were diluted with a freezing medium (SHOTOR extender) supplemented with different concentrations of EGF; 0 (EGF0), 50 (EGF50), 100 (EGF100), 200 (EGF200), and 400 (EGF400) ng/mL in SHOTOR extender and cryopreserved using a standard protocol. All EGF groups showed significant improvements in sperm progressive motility, viability, and sperm membrane function after equilibration at 5 °C for 24 hours. Regarding frozen-thawed samples, sperm progressive motility and some kinematic parameters (DAP, VSL, VCL and AHL) were significantly higher in the EFG400 group compared to the other groups (P < 0.01). A significant increase in the percentage of live/acrosome-intact sperm was observed, accompanied by a significant decrease in malondialdehyde levels in all EGF groups (P < 0.05). Both the EGF200 and EGF400 groups showed significantly higher sperm viability and significantly lower percentages of apoptotic and necrotic sperm compared to the other groups (P < 0.05). EGF supplementation preserved the ultrastructural integrity and cryotolerance of epididymal camel spermatozoa. The docking analysis indicated that EGF exhibited higher binding affinity with apoptosis sperm markers, including caspase-3 and bcl-2-associated X (Bax) proteins, with binding energies of -502.0 and -621.0 kcal/mol, respectively. In conclusion, the addition of EGF to SHOTOR extender was found to have beneficial effects on sperm motility, kinematics parameters, sperm viability, acrosome integrity, sperm ultrastructural features, and reduced oxidative stress and apoptosis-like changes in cryopreserved epididymal camel spermatozoa., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This experiment was conducted to determine the most suitable glycerol concentration (3 or 6%) and/or non-penetrating cryoprotectants (trehalose and sucrose) for the cryopreservation of buffalo semen, with the aim of enhancing the cryopreservation protocol. Semen of Egyptian buffalo were pooled and diluted with eight Tris extenders supplemented with either 6% glycerol (control group, GL6), 3% (low level, GL3), sucrose (SU, 50 mM), trehalose (TR, 50 mM), 6% glycerol together with 50 mM of sucrose (GL6SU) or 50 mM of trehalose (GL6TR), and 3% of glycerol together with 50 mM of sucrose (GL3SU) or 50 mM of trehalose (GL3TR), then frozen following the standard protocol. Findings indicated that GL3 extender resulted in the highest values of progressive motility, sperm kinematics, sperm membrane integrity, and viability of post-thawed semen (37 °C for 30 s). On the contrary, the Tris extender enriched only with SU and TR groups had the lowest values of sperm quality compared to the other groups (p < 0.05). All GL supplemented groups showed higher intact acrosome levels and lower detached acrosome and dead sperm with intact acrosome compared to those with TR and SU alone (p < 0.05). A significant increase in viable sperm was observed in the GL3, GL6, and GL3SU groups compared to the other groups (p < 0.05). The Tris extender supplemented with low glycerol (3%) significantly reduced the levels of MDA. In the in vivo fertility trial, it was shown that the pregnancy rate was higher in the GL6SU group (72%) than in the GL3SU group (68%; p > 0.05). Collectively, these results suggest that there is potential in using low glycerol (3%) as a cryoprotective agent in the medium for buffalo sperm cryopreservation without significant adverse effects compared to the addition of 6% glycerol. This study supported the sustainability of materials used in assisted reproductive technology by reducing the glycerol content in the freezing medium. Further research is needed to confirm this hypothesis., Competing Interests: Declarations. Ethics approval and consent to participate: All investigational techniques applied in this research were approved by the Animal Ethics Committee of Zagazig University, based on IACUC (Institutional Animal Use and Care Committee) rules (Approval No: ZU IACUC/2/F/173/2022). The study complies with the European Directive 2010/63/EU regarding husbandry and experimental handling Consent for publication: Not applicable Competing interests: The authors declare no conflict of interest., (© 2024. The Author(s).)

Nanomaterials offer several promising prospects in the field of farm animal reproduction, encompassing a broad range of applications such as transgenesis and the precise delivery of substances to sperm cells, antimicrobial, antioxidants properties as well as their potent role in improving cryopreservation methods. The aim of the present study is to explore the effect of supplementing the semen extender with 10 µg/mL nano gold (Au-NPs10), 10 µg/mL nano silver (Ag-NPs10), 1 µg/mL nano selenium (Se-NPs1), and 100 µg/mL nano zinc oxide (ZnO-NPs100) on sperm characteristics and kinematics parameters, acrosome integrity, oxidative biomarkers, morphological and apoptosis-like changes of frozen-thawed buffalo bull sperm, and, ultimately, their fertilizing capacity. The results revealed that all aforementioned nano materials significantly improved viability, progressive motility, membrane integrity, acrosome integrity, and kinematic parameters as well as apoptosis-like changes of post-thawed buffalo bull sperm compared to the control (p < 0.05). No discernible effects were observed on sperm ultrastructure morphology measures as a response to the addition of these metallic nanoparticles to the extender. The values of caspase 3 significantly decreased by 64.22, 45.99, 75.59, and 49.39% in Au-NPs10, Ag-NPs10, Se-NPs1, and ZnO-NPs100 treated groups, respectively, compared to the control. The addition of 100 µg ZnO-NPs to the extender significantly decreased the total count of bacteria, fungi, and yeast compared to the control (p < 0.05). The AuNPs10 and SeNPs1 treated groups showed lower content of malondialdehyde, hydrogen peroxide, and nitric oxide concentrations and higher values of total antioxidant capacity of post-thawed extended semen (p < 0.05). Pregnancy rates increased by 17.5, 20, and 30% in buffalo cows inseminated with sperm treated with ZnO-NPs100, Se-NPs1, and Au-NPs10, respectively, compared to the control group. The present results indicate that the freezing extender supplemented with metallic nanoparticles can be an effective strategy to enhance the cryotolerance and fertility potential of buffalo bull sperm., (© 2024. The Author(s).)

Sperm cryopreservation can lead to subfertility due to potential damage to sperm DNA, membranes, and overall motility caused by the freeze-thaw process. Interleukin-6 (IL-6) is a versatile cytokine with various roles in reproductive processes. However, the impacts of IL-6 supplementation on cryopreserved ram sperm have not been thoroughly investigated. Therefore, this study aims to assess the influence of IL-6 on the sperm quality of cryopreserved ram sperm. Ram semen was collected, pooled, and extended with tris-citrate soybean lecithin extender supplemented with 0, 50, 100, and 200 ng/mL of IL-6. The samples experienced a standard freezing protocol, and sperm quality, kinematic parameters, ultrastructure, and molecular docking of cryopreserved ram spermatozoa were evaluated. The results showed that sperm kinematics, viability, progressive motility, and membrane integrity were significantly enhanced by the addition of 100 or 200 ng of IL-6/mL (p < 0.05). Semen supplemented with 100 or 200 ng/mL of IL-6 also exhibited higher percentages of sperm kinematics, including DAP, DCL, DSL, VSL, VAP, VCL, and ALH, compared to other groups (p < 0.05). IL-6 supplementation enhanced acrosome integrity, and reduced caspase-3 activity in post-thawed ram spermatozoa (p < 0.05) compared to untreated group. Supplementation with IL-6 (200 ng/mL) significantly decreased oxidative biomarkers (NO, MDA, and H 2 O 2 ) (p < 0.001) and improved total antioxidant capacity (p < 0.05). The percentage of sperm damage (tail, head, and midpiece) was significantly reduced by IL-6 supplementation (p < 0.05). Electron micrographs showed that supplementation with 100 or 200 ng/mL IL-6 protected acrosome stability, plasma membrane integrity, and sustained the ultrastructure integrity of cryopreserved ram spermatozoa. The docking exploration indicates a higher binding affinity with sperm function biomarkers, including caspase 3, BCL2, and PSMA6, with binding energies of - 52.30 kcal/mol, - 56.04 kcal/mol, and - 57.06 kcal/mol, respectively. In conclusion, the addition of IL-6 to the freezing extender can enhance the post-thaw quality of cryopreserved ram spermatozoa., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.)

Kisspeptins are neuropeptides encoded by the Kiss1 gene that was discovered as a metastasis suppressor gene in melanoma and breast cancer. Kisspeptin has pivotal functions for gonadotropin-releasing hormone secretion and plays integrated roles in the hypothalamic-pituitary-gonadal axis. However, little is known about the peripheral expression of kisspeptin in ruminants, especially in the female reproductive tract. Here, the objectives of the current study were to investigate the spatial localization of kisspeptin and mRNA expression of Kiss1 and its receptor (Kiss1r) in the fallopian tubes (FT) and uterus of goats at varied reproductive activity (cyclic versus true anoestrous goats, n=6, each). Specimens of the uterus and FT were collected and fixed using paraformaldehyde to investigate the localizations of kisspeptin in the selected tissues by immunohistochemistry. Another set of samples was snape-frozen to identify the expressions of mRNAs encoding Kiss1 and Kiss1r using real-time PCR. Results revealed immunolocalizations of kisspeptin in the uterus and the FT. The staining of kisspeptin was found mainly in the mucosal epithelium of the uterus the FT, and the endometrial glands. Very intense staining of kisspeptin was found in the uterine and FT specimens in the true anoestrous goats compared to that in cyclic ones. The expression of mRNA encoding Kiss1 gene was significantly higher in the uterine specimen of cyclic goats (1.00±0.09) compared to that in the true anoestrous goats (0.62±0.08) (P ˂0.05), while the expression of mRNA encoding Kiss1r was significantly (P ˂0.001) higher in the uterine tissues of true anoestrous goats (1.78±0.17) compared to that in cyclic ones (1.00±0.11). In conclusion, immunohistochemical localization of kisspeptin and the expression of mRNA encoding Kiss1/Kiss1r revealed spatial changes in the uterus and FT of goats according to the reproductive potential of goats (cyclic versus true anoestrous goats). However, the definitive local role of kisspeptin in the uterus and FT need further investigation., Competing Interests: Declaration of competing interest The authors have declared no conflict of interest for this paper., (Copyright © 2024. Published by Elsevier Inc.)

Introduction: Atrial arrhythmia is the most common complication of patent foramen ovale (PFO) closure. The real incidence of post-PFO closure atrial arrhytmia and whether this complication can be prevented is unknown., Methods/design: The Assessment of Flecainide to Lower the PFO closure risk of Atrial fibrillation or Tachycardia (AFLOAT) trial is a prospective, national, multicentre, randomized, open-label, superiority trial with a blind evaluation of all the endpoints (PROBE design). A total of 186 patients are randomized in a 1:1:1 ratio immediately after PFO closure to receive Flecainide (150 mg per day in a single sustained-release (SR) dose) for 6 months (Group 1), Flecainide (150 mg per day in a single SR dose) for 3 months (Group 2), or no additional treatment (standard of care) for 6 months (Group 3). The primary endpoint is the percentage of patients with at least one episode of symptomatic or asymptomatic atrial arrhythmia episode (≥30 s) recorded within 3 months after PFO closure on long-term monitoring with an insertable cardiac monitor. Whether 3 months of treatment is sufficient compared to 6 months will be analysed as a secondary objective of the study., Conclusion: AFLOAT is the first trial to test the hypothesis that a short treatment with oral Flecainide can prevent the new-onset of atrial arrhythmia after PFO closure., Clinical Trial Registration: NCT05213104 (clinicaltrials.gov)., (© The Author(s) 2024. Published by Oxford University Press on behalf of the European Society of Cardiology.)

Although cryopreservation is a reliable method used in assisted reproduction to preserve genetic materials, it can stimulate the occurrence of oxidative stress, which affects sperm structure and function. This research was conducted to explore the effects of quinoa seed extracts (QSE) on ram sperm quality, oxidative biomarkers, and the gene expression of frozen-thawed ram sperm. Semen samples were diluted in extenders supplemented with 0 (QSE0), 250 (QSE1), 500 (QSE2), 750 (QSE3), and 1000 (QSE4) µg of QSE /mL, and then frozen according to the typical procedure. The findings indicate that the QSE3 and QSE4 groups provided the optimal results in terms of sperm viability and progressive motility. Sperm kinematics were considerably enhanced in the QSE3 group compared to the other groups (P<0.01). QSE (500-1000 µg/mL) significantly decreased the apoptosis-like changes (higher viable and lower apoptotic sperm) in ram sperm (P<0.001). The percentage of live sperm with intact acrosomes was significantly increased, while the percentage of detached and intact acrosomes in live and dead sperm were significantly decreased respectively by the QSE addition (P<0.001). All QSE groups had higher TAC and lower MDA and H 2 O 2 levels than the control group (P<0.001). The expressions of SOD1, CAT, GABPB1, and GPX1 genes in sperm samples were significantly increased, while the CASP3 gene was significantly decreased in all QSE-supplemented samples. Our data suggest that QSE has beneficial effects on sperm quality of cryopreserved ram semen, which are achieved by promoting sperm antioxidant-related genes and reducing apoptosis-related gene., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sperm cryopreservation technology significantly contributes to the safeguarding of genetic resources, particularly for endangered species, and supports the use of artificial insemination in domestic animals. Therefore, cryopreservation can negatively affect sperm health and function leading to reduce the freezing ability and fertility potential. Therefore, it is essential to prioritize the improvement of cryotolerance in cryopreserved sperm to enhance reproductive efficiency and ensure sustainability in livestock herds. The main reason for sperm dysfunction after thawing may be related to the excessive amount of oxidative stress (OS) produced during cryopreservation. Scientists have different ways for counteracting this OS including the use of plant extracts, enzymes, minerals, anti-freezing proteins, and amino acids. Recently, one such amino acid is L-proline (LP), which has multiple roles such as osmotic and OS defense, nitrogen, and carbon metabolism, as well as cell survival and signaling. LP has been found in seminal plasma and has recently been added to the freezing extender to improve the various post-thaw parameters of sperm. This improvement is related to the ability of LP to reduce the OS, sustain the plasma membrane and to act as an osmoregulatory agent. Moreover, LP can suppress cell apoptosis by modulating intracellular redox in sperm. This review addresses the ongoing research on the addition of L-proline as an osmoregulatory agent in freezing extenders to increase the cryotolerance of animal spermatozoa to freeze-thaw., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Coronary slow flow (CSF) often links to inflammation and endothelial function disturbance. While conventional ejection fraction measurements fall short in identifying myocardial dysfunction, left ventricular global longitudinal strain (LV GLS) has shown superior efficacy in this regard. Our study aimed to explore subclinical left ventricular systolic dysfunction by assessing LV GLS in patients diagnosed with coronary slow flow (CSF)., Methods: The study included sixty patients with CSF and sixty control individuals without CSF. Coronary angiography employed the Thrombolysis in Myocardial Infarction (TIMI) frame count (TFC) to identify CSF. LV GLS values were evaluated and compared between the two groups., Results: Significantly reduced LV GLS was evident in the CSF group compared to the control group (- 16.18 ± 1.25 vs. - 19.34 ± 1.33, p < 0.001). A notable correlation (r = 0.492, p < 0.001) between LV GLS and TFC was observed in the CSF group. Multivariate logistic regression analysis highlighted reduced LV-GLS (OR 2.2, 95% CI 1.57-3.09, p < 0.001) and smoking (OR 11.55, 95% CI 3.24-41.2, p < 0.001) as significant predictors for CSF presence. The receiver operating characteristic curve established that an LV GLS value of ≥ - 17.8% accurately predicted the presence of CSF (AUC: 0.958, 95% CI: 0.924-0.991, p < 0.001) with 90% specificity and 91.7% sensitivity., Conclusion: Our study indicates that reduced LV GLS is associated with CSF presence, offering a valuable means to early detect subclinical left ventricular systolic dysfunction in high-risk patients susceptible to heart failure., Trial Registration: ZU-IRB#7038/12-7-2021 Registered 12 July 2021, email: IRB&#95;123@medicine.zu.edu.eg., (© 2023. The Author(s).)

The current study aimed to investigate the impact of nano-formulations of clove bud ethanolic extract (CBENF) in the extender on sperm characteristics, antioxidant capacity, oxidative biomarkers, enzymatic activity, apoptosis and fertility of post-thawed rabbit semen. Twelve mature male rabbits semen samples were pooled and cryopreserved in a Tris-egg yolk-based extender containing varying concentrations of CBENF (0, 25, 50, 75 and 100 μg/mL). After the equilibration and freezing-thawing process, CBENF (100 μg /mL) significantly enhanced progressive motility, viability and membrane integrity. Conversely, sperm abnormality was significantly reduced by CBENF supplementation. Total antioxidant capacity was increased in the post-thawed sperm medium, while nitric oxide and malondialdehyde were decreased in all CBENF concentrations. The lactic dehydrogenase and caspase-3 activities were decreased, whereas the number of live spermatozoa with an intact acrosome was increased in all CBENF concentrations. Conception rate and litter size per doe were higher in doe rabbits inseminated with semen supplemented with 100 μg CBENF/mL than un-supplemented group (76% vs. 52% and 8.4 vs. 7.7/doe), with no statistical differences. These findings suggest that supplementing rabbit extenders with 100 μg of CBENF/mL could be an effective strategy for enhancing freeze-thawing rabbit sperm attributes and fertility., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

The existing treatise targeted to compare the effects of adding different nano-emulsions essential oils (olive, flaxseed, and grapeseed oils) in freezing extender on semen quality and freezability in buffalo. Nano-emulsions were prepared from olive, flaxseed, and grapeseed oils and characterized for their sizes and shapes. Semen extended in four tubes were supplemented with 0 (control) and 3.5% nanoemulsion oils, including olive (NEO), flaxseed (NEFO) and grape seed oils (NEGSO) respectively. NEGSO resulted in the highest (p < 0.05) membrane integrity, vitality, progressive motility (P-motility) of sperm compared to the other groups in post-thawed buffalo bull semen (at 37 °C for 30 s). The addition of NEGSO had the best results for membrane integrity, progressive motility, and vitality of sperm after incubation (at 37 °C and 5% CO 2 for 2 h). A superior (p < 0.05) value of total antioxidant capacity in frozen-thawed spermatozoa was monitored in all supplemented groups as relative to the control. The values of malondialdehyde (MDA) and nitric oxide (NO) were lower (p < 0.05) in NEGSO group compared with other groups. Both NEO and NEFO exhibited the same results for MDA, and NO levels (p > 0.05). All supplemented groups exhibited lower hydrogen peroxide levels (p < 0.05) as relative to the un-treated group. The lowest (p < 0.05) caspase 3 levels were verified in NEGSO treatment, followed by NEFO and NEO treatments. Post-thawed sperm showed ultrastructural damages in the control group, and theses damages were attenuated or resorted by the NEGSO, NEFO and NEO supplemented to freezing extender. In consequences with in vitro results regarding the sperm attribute, a greater pregnancy rate (92%) was observed in NEGSO group as compared with NEFO (88%), NEO (76%) and CON (68%) groups. Our findings demonstrate that NEGSO (3.5%) could be used as a new strategy in enhancing sperm functionality, potential fertility and reducing the oxidative damage and apoptosis markers. This could be significantly applicable for sperm physiology cryopreservation in the milieu of assisted reproduction systems., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Sperm preservation is a well-established technique in reproductive biotechnology that is widely used to maintain the genetic quality of male individuals. However, there are several factors during the preservation process that can affect the vitality, functionality, and quality of sperm, thereby reducing their fertility potential after thawing. One of these factors is the synthesis of high levels of oxidative stress (OS) during semen preservation, which can have detrimental effects on sperm health and functionality. To counter the negative impact of OS on sperm, researchers have explored the supplementation of several exogenous antioxidants in the extenders used to preserve ram sperm. This approach has shown promising results in improving sperm health, functionality, and fertility potential in ram. Additionally, the preservation process can induce modifications in the ram sperm proteome. By employing targeted proteomics techniques, researchers have been able to identify and modify specific proteins in cryopreserved ram sperm, potentially offering further improvements in the quality of the cryopreserved ram sperm. In summary, this review provides a comprehensive overview of the antioxidants and targeted proteomics modifications that have been investigated for enhancing ram sperm preservation. These advancements aim to mitigate the negative effects of OS and optimize the techniques used in preserving ram sperm., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

Thymoquinone nanoparticles (TQNPs) are broadly utilized in numerous pharmaceutical applications. In the present study, we tested the effects of TQNP supplementation on sperm quality and kinematics, acrosome exocytosis, oxidative biomarkers, apoptosis-like and morphological changes of frozen-thawed buffalo sperm, as well as the fertilizing capacity. Semen was collected from buffalo bulls, diluted (1:10; semen/extender), and divided into five aliquots comprising various concentrations of TQNP 0 (CON), 12.5 (TQNP12.5), 25 (TQNP25), 37.5 (TQNP37.5), and 50 (TQNP50) µg/mL, and then cryopreserved and stored in liquid nitrogen (-196 °C). The results revealed that TQNPs (25 to 50 µg/mL) provided the most optimal results in terms of membrane integrity ( p < 0.001) and progressive motility ( p < 0.01). In contrast, TQNP50 resulted in a greater post-thawed sperm viability ( p = 0.02) compared with other groups. The addition of TQNPs to the extender had no discernible effects on sperm morphology measures. Sperm kinematic motion was significantly improved in the TQNP50 group compared to the control group ( p < 0.01). TQNPs effectively reduced the content of H 2 O 2 and MDA levels and improved the total antioxidant capacity of post-thawed extended semen ( p < 0.01). The addition of TQNP significantly increased the number of intact acrosomes ( p < 0.0001) and decreased the number of exocytosed acrosomes ( p < 0.0001). A significant reduction in apoptosis-like changes was observed in TQNP groups. The non-return rates of buffalo cows inseminated with TQNP50-treated spermatozoa were higher than those in the control group ( p < 0.05; 88% vs. 72%). These findings suggested that the freezing extender supplemented with TQNPs could effectively enhance the cryotolerance and fertility of buffalo sperm.

Buffalo sperm is sensitive to cryoinjuries, thus improving sperm cryoresistance is a critical approach for wide spreading the assisted reproductive technologies in buffalo. The intention of this work was to assess the effect of propolis-loaded in nanoliposomes (PRNL) supplementation of semen extender on semen quality, antioxidant status and some apoptotic genes of cryopreserved buffalo semen. PRNL were prepared using cholesterol (Chol) as well as soybean lecithin and their physicochemical properties were characterized. Egyptian buffalo bulls (4-6 years) were involved, and the semen samples were collected using the artificial vagina method. Buffalo semen was pooled (n = 25 ejaculates) and cryopreserved in tris extender containing PRNL at 0 (PRNL0), 2 (PRNL2), 4 (PRNL4) and 6 µg/mL (PRNL6), respectively. The PRNL had a size of 113.13 nm and a negative zeta potential (- 56.83 mV). Sperm progressive motility, viability, membrane integrity, abnormalities, chromatin damage, redox status, apoptosis status, and apoptotic genes were investigated after post-thawed buffalo semen. Using 2 or 4 µg/mL PRNL significantly increased sperm progressive motility, viability, and membrane integrity, while sperm abnormalities and the percentage of chromatin damages were the lowest in PRNL2 group. Moreover, the PRNL2 group exhibited the best results for all antioxidative activities (TAC, SOD, GPx and CAT) with significantly higher levels than the other groups (P < 0.05). The levels of ROS and MDA were significantly lower in the PRLN2 compared with other groups. The sperm caspase 3 enzyme activities showed the lowest values in PRNL2 groups followed by PRNL4 and PRNL6 groups with significant differences compared with the control. Adding 2 µg/mL PRNL to freezing media significantly reduced apoptotic genes such as Bax and Caspase 3 in sperm, while significantly increase in Bcl2 expression compared with the control (P < 0.001). The expression of Bcl2, Caspase 3 and Bax genes in sperm were not affected by the 6 µg/mL PRNL addition (P > 0.05). The electron micrography descriptions exemplified that the fortification of 2 or 4 µg/mL PRNL maintained the acrosomal and plasma membrane integrities as well as sustained the ultrastructure integrity of the cryopreserved buffalo spermatozoa when compared with control group, whereas the 6 µg/mL of PRNL demonstrated highest injury to the acrosome and plasma membranes. Results show supplementation of the buffalo freezing extender with 2 or 4 µg/mL of PRNL enhanced post-thawed sperm quality via boosting the antioxidant indices, diminishing the oxidative stress and apoptosis as well as maintained the ultrastructure integrity of frozen-thawed buffalo sperm., (© 2023. The Author(s).)

This research was designed to explore the protective effect of alpha-lipoic acid-loaded nanoliposomes (ALAN) during cryopreservation of buffalo sperm. Buffalo semen was cryopreserved in a tris-citrate egg yolk extender without any supplement (ALAN0, control group) or with ALAN at levels of 25, 50, 75 or 150 µg (ALAN25, ALAN50, ALAN75 and ALAN150, respectively). The ALAN had a size of 171.80 nm and a negative zeta potential (- 43.40 mV). The progressive motility, vitality and membrane integrity significantly improved in all ALAN groups (except ALAN25 for membrane integrity). ALAN150 group exhibited the best values of progressive sperm motility, vitality and membrane integrity after thawing at 37 °C for 30 s or incubated for 2 h at 37 °C and 5% CO 2 compared with those in other groups. Both ALAN75 and ALAN150 groups significantly improved the TAC, GR and catalase, while lipid peroxidation and early apoptotic spermatozoa significantly decreased in ALAN150 group followed by ALAN75 group. Collectively, the adding ALAN to buffalo semen freezing extender plays a substantial shielding function against cryodamage by preserving the sperm functional parameters., (© 2022. The Author(s).)

Bee bread has numerous nutritional benefits and bioactive compounds. Other bee byproducts have been used as extender additives to improve semen cryopreservation. Here, we examined the effects of supplementing egg yolk extender (EYE) or soybean lecithin extender (SBLE) with bee bread extract (BBE) on the quality of cryopreserved ram semen. Semen was collected from five adult Rahmani rams once a week for 7 weeks. EYE and SBLE were supplemented with BBE. Antioxidant capacity and total phenolic compound, total flavonoid compound, and total soluble carbohydrate levels of BBE were measured. Sperm characteristics, including progressive motility, viability, abnormalities, membrane integrity, and acrosome integrity, were analyzed after equilibration, thawing, and thawing followed by a 2-h incubation. The total antioxidant capacity and malondialdehyde, hydrogen peroxide, aspartate transaminase, alanine transaminase, alkaline phosphatase, and total acid phosphatase levels in extenders were determined after thawing. Sperm apoptosis was analyzed using annexin V assays. SBLE was more effective than EYE for cryopreserving ram semen. Extender supplementation with BBE improved ram semen quality during freezing in a concentration-dependent pattern. Motility, vitality, and membrane integrity were particularly enhanced in BBE-treated semen. Additionally, BBE promoted antioxidant and enzymatic activities and reduced apoptosis in semen. Thus, extender supplementation with BBE improved sperm cryopreservation.

Objectives: The current study aims to investigate the morphology and symmetry of root canals of mandibular second premolar teeth via Cone-Beam Computed Tomography (CBCT) among a sample of the Saudi population., Methods: A total of 1200 CBCT images were examined in the study. The number of roots in the mandibular second premolars was identified, and canal configurations were classified according to Vertucci classification. Bilateral symmetry and association between gender and number of roots and root canal configuration were investigated using a chi-square test was utilized for data analysis and p value was set at 0.05 for significance., Results: Of the total 2400 Mandibular second premolars, 98.33% were single rooted and 1.67% were double rooted. The prevalence of double roots in females on the right side was significantly higher than that on the left side (p = 0.017), but the opposite was found among males. Only Type I and II canal configurations showed significant differences regarding gender and sides. 98.0% of teeth showed the symmetrical number of roots in both genders. The symmetry in canal configuration was 96.7% in males and 97.3% in females and 99.1% was Type I in both sides, 0.5% Type III, and 0.3% Type IV in both sides., Conclusions: The root morphology of mandibular second premolars in the Saudi population demonstrated a high degree of variability. The vast majority of mandibular premolars have a single root with one canal and a low prevalence of two rooted teeth. Contralateral mandibular second premolars exhibited great symmetry of roots and root canal configuration., (© 2021. The Author(s), under exclusive licence to Japanese Society for Oral and Maxillofacial Radiology and Springer Nature Singapore Pte Ltd.)

The aim of the present study was to investigate the effect of supplementing rabbit semen extender with zeolite loaded with different charges (Z + or Z - , Z ± ) on sperm cryopreservation. Semen was collected from six healthy, fertile New Zealand rabbit bucks using an artificial vagina. The collected ejaculates were pooled and diluted with a tris-yolk fructose (TYF) extender supplemented with Z ± (+16, +12, +8, -16, -12, and -8) at a concentration of 1% for a final sperm concentration of 25 × 10 6 sperm cells/mL. The diluted semen samples were then cryopreserved in 0.25 mL straws and stored in liquid nitrogen for 1 month. To evaluate sperm quality, we examined sperm progressive motility, vitality, morphological abnormalities, and plasma membrane integrity. In addition, apoptotic rates were determined using flow cytometry and by examining sperm ultrastructure under a transmission electron microscope (TEM). Moreover, total antioxidant capacity and markers of lipid peroxidation were measured in the extender after thawing. Addition of Z ± had a positive effect on progressive motility, vitality, and membrane integrity after an equilibration period and post-thawing as compared with the controls (P < 0.05). Z ± supplementation, particularly with a strong negative charge, also decreased the percentages of apoptotic and necrotic sperm cells compared to controls (P < 0.05), as shown both by flow cytometry and TEM. This was not associated with any marked effects on the oxidative biomarkers in the extender. In conclusion, addition of Z ± to semen extender improved post-thawing sperm quality by improving sperm characteristics, decreasing apoptosis, and minimizing sperm damage during cryopreservation., (Copyright © 2021 Elsevier Inc. All rights reserved.)

This study examined the effects of zinc chloride (ZnCl2) and sodium selenite (Na2SeO3) supplementation in maturation medium on in vitro maturation (IVM) rate, oxidative biomarkers and gene expression in buffalo oocytes. Ovaries from a slaughterhouse were aspirated and good quality cumulus-oocyte complexes (COCs) with at least four layers of compact cumulus cells and evenly granulated dark ooplasm were selected. COCs were randomly allocated during IVM (22 h) to one of four treatment groups: (1) control maturation medium (basic medium), or basic medium supplemented with (2) ZnCl2 (1.5 µg/ml), (3) Na2SeO3 (5 µg/l), or (4) ZnCl2 + Na2SeO3 (1.5 µg/ml + 5 µg/l, respectively). Oocytes were denuded after 22 h of IVM in the first four replicates. Specimens were fixed and stained to evaluate the stage of nuclear maturation. The spent medium was collected for biochemical assays of total antioxidant capacity (TAC), malondialdehyde (MDA) and hydrogen peroxide concentrations. A second four replicates were used for COCs for RNA extraction. The expression levels of antioxidant (SOD1, GPX4, CAT and PRDX1), antiapoptotic (BCL2 and BCL-XL) and proapoptotic (BAX and BID) genes were measured. Supplementation with ZnCl2 and Na2SeO3 during IVM increased the ratio of oocytes reaching metaphase II at 22 h, increased TAC and decreased MDA and H2O2 concentrations in the maturation medium (P < 0.05). Moreover, beneficial effects were associated with complementary changes in expression patterns of antioxidative, antiapoptotic and proapoptotic genes, suggesting lower oxidative stress and apoptosis. Supplementation medium with zinc chloride and sodium selenite improves the maturation rate, reduces oxidative stress and increases expression levels of antioxidative and antiapoptotic genes.

This study evaluated the efficacy of Turraea fischeri leaf extract for maintaining the viability of cryopreserved goat sperm. Ejaculated semen was collected from 5 mature Baladi bucks (50-60 kg, 2-4 years of age) and those samples with mass motility ≥ 70% and sperm concentration ≥ 2.5 × 10 9 /mL were selected, pooled, and divided into 4 aliquots. Each aliquot was diluted in Tris-citric-soybean lecithin extender containing a different concentration of T. fischeri leaf extract (0, 125, 250, or 375 µg/mL). Treated semen samples were cooled to 5 °C, transferred to 0.25-mL French straws, and stored in liquid nitrogen (LN 2 ) at -196 °C. After thawing, membrane integrity was examined by transmission electron microscopy, apoptotic activity by Annexin/propidium iodide staining and flow cytometry, and both enzyme activities and antioxidant capacity by spectroscopic assays. The leaf extract at 375 µg/mL significantly improved semen quality as indicated by enhanced total antioxidant capacity, reduced H 2 O 2 concentration, a greater proportion of structurally intact motile sperm, and concomitant reductions in apoptosis and necrosis. The extract also significantly increased the proportion of sperm with a contiguous plasma membrane and intact acrosome ( p < 0.05). Furthermore, LC-MS revealed numerous secondary metabolites in the extract that may contribute to sperm cryopreservation.

This study determined the effects of antioxidant supplementation and storage time at cool temperatures on the characteristics of epididymal camel spermatozoa. Camel testes were collected at the abattoir after animal slaughtering and kept at 4 °C during transportation and until processing (max 6 h). Spermatozoa were retrieved and diluted with SHOTOR extender, split in aliquots, supplemented with the following antioxidants: 200 μm/mL vitamin E, 1.0 g/L vitamin C, 1 μg/mL selenium nanoparticles, 50 μg/mL zinc nanoparticles, 2 μg/mL sodium selenite, and 100 μg/mL zinc sulfate, and stored at 4 °C for 2, 48, 96, and 144 h. The storage time significantly affected (P < 0.05) the sperms' motility and livability, the sperms' membrane integrity, and the percentages of cytoplasmic droplets as well as the percentage of morphologically normal spermatozoa. Epididymal sperm characteristics (progressive motility, livability, membrane integrity, and abnormalities) were significantly improved (P < 0.05) when the spermatozoa were diluted with antioxidants as compared with the control group, and the best additives were identified as nano-selenium, sodium selenite, nano-zinc, and zinc sulfate. In conclusion, adding nano-sized minerals or inorganic trace elements and vitamins maintained the progressive motility, livability, and membrane integrity, and decreased abnormalities and cytoplasmic droplet percentages of epididymal camel spermatozoa stored at 4 °C up to 144 h.

Cryopreservation is an essential tool to preserve sperm cells for zootechnical management and artificial insemination purposes. Cryopreservation is associated with sperm damage via different levels of plasma membrane injury and oxidative stress. Nanoparticles are often used to defend against free radicals and oxidative stress generated through the entire process of cryopreservation. Recently, artificial or natural nanovesicles including liposomes and exosomes, respectively, have shown regenerative capabilities to repair damaged sperm during the freeze-thaw process. Exosomes possess a potential pleiotropic effect because they contain antioxidants, lipids, and other bioactive molecules regulating and repairing spermatozoa. In this review, we highlight the current strategies of using nanoparticles and nanovesicles (liposomes and exosomes) to combat the cryoinjuries associated with semen cryopreservation.

The Splinter bean, Entada abyssinica , is widely used in folk medicine. In the current work, we profiled the secondary metabolites from E. abyssinica bark extract using LC-MS and investigated its effect on cryopreserved ram semen. Twenty-eight compounds, including tannins and gallic acid derivatives that prevailed in the extract, were tentatively identified. Results showed that the quality of the post-thawed semen showed a significant improvement when the extract was added to the extender at a concentration of 375 μg/mL. The progressive motility and plasma membrane integrity of sperm cells were significantly increased in the post-thawed semen; however, the total antioxidant capacity (TAC) was insignificantly increased. A significant decrease in the concentration of hydrogen peroxide was detected as well. No significant changes were observed in activities of lactate dehydrogenase (LDH), alanine aminotransaminase (ALT), and aspartate transaminase (AST) within the treated samples. Intact sperm percentage was significantly increased, while apoptotic and necrotic sperm percentages were reduced significantly. Molecular docking of some individual components from the extract revealed their potential to interfere with the apoptosis cascade in which Bcl-2 is involved. In conclusion, Entada abyssinica appears to be useful for cryopreservation presumably owing to its polyphenol content that has potent antioxidant capacity scavenging reactive oxygen species (ROS), enhancing the endogenous antioxidant system and inhibiting lipid peroxidation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Sobeh, Hassan, Hassan, Khalil, Abdelfattah, Wink and Yasri.)

MicroRNA-208a is a cardiac specific oligo-nucleotide. We aimed at investigating the ability of microRNA-208a to diagnose myocardial infarction and predict the outcome of primary percutaneuos coronary angiography (PCI). Patients (n = 75) presented by chest pain were recruited into two groups. Group 1 (n = 40) had ST elevation myocardial infarction (STEMI) and underwent primary PCI: 21 patients had sufficient reperfusion and 19 had no-reflow. Group 2 (n = 35) had negative cardiac troponins (cTns). Plasma microRNA-208a expression was assessed using quantitative polymerase chain reaction and patients were followed for occurrence of in-hospital major adverse cardiac events (MACE). MicroRNA-208a could diagnose of MI (AUC of 0.926). After primary PCI, it was superior to cTnT in prediction of no-reflow (AUC difference of 0.231, P = 0.0233) and MACE (AUC difference of 0.367, P = 0.0053). Accordingly, circulating levels of miR-208a can be used as a diagnostic marker of MI and a predictor of no-reflow and in-hospital MACE. Graphical abstract Receiver operating curve analysis of no-reflow prediction of miRNA208a, CK-MB and hs-Troponin T. MicroRNA-208a shows significantly higher prediction of no-reflow as compared to routine cardiac biomarkers.

Goat semen cryopreservation is a challenging process as it results in reduced motility, vitality, and fertility of spermatozoa after freezing. In this study, we evaluated the effects of different herbal extract nanoformulations (NFs) [mint (MENFs), thyme (TENFs), and curcumin (CENFs)], supplemented at either 50 or 100 μg into Tris-extender on the cryopreserved goat semen quality. The hydrothermal squeezing method was used for the preparation of the NFs extracts. The morphological evaluation of the NFs extracts was conducted by transmission electron microscopy. All NFs supplements improved (p < 0.05) the progressive motility, vitality, and plasma membrane integrity of sperm compared with the control extender after equilibration (5 °C for 2 h) and thawing (37 °C for 30 s), but had no effect on sperm abnormality and acrosome integrity. All NFs supplements decreased (p < 0.05) the apoptosis, malondialdehyde level, and chromatin decondensation of sperm cells, while increased (p < 0.05) the total antioxidant capacity and catalase activity in the frozen/thawed extender. Particularly, CENFs at a level of 100 μg showed improvement of sperm parameters and antioxidant status during cryopreservation of goat semen more than TENFs and MENFs. The CENFs improved the quality of goat spermatozoa in post-thawed semen in terms of preventing cryodamage and promoting the cryotolerance of spermatozoa when compared with TENFs and MENFs. Therefore, supplementation of Tris-extender with CENFs could enhance goat semen processing during cryopreservation., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Purpose: This work investigates the possible renoprotective effects of date palm fruits and seeds extract against renal ischemia and their underlying mechanisms., Methods: 108-Sprague Dawle male rats were randomly allocated into 6 equal groups differently receiving aqueous or methanolic fruit and seed extracts. Assay of serum creatinine, BUN and TNF-α, morphological examination of the left kidney, markers of the redox state (MDA, CAT, and GSH), the expression of TNFα and Nrf2 genes at the level of mRNA, the expression of caspase-3 and TGF-β proteins by immunohistochemistry were performed., Results: 45-min renal I/R caused significant deterioration of kidney functions (increase in serum creatinine and BUN) and morphology (P < 0.001) and significant reduction in CAT activity and GSH levels with significant increase in serum TNF-α and MDA concentration and the expression of Nrf2, caspase-3, TNF-α, and TGF-β in kidney tissues. Pre-treatment with either date palm fruit or seed extracts significantly improved kidney functions and morphology (P ≤ 0.001) with a significant increase in the expression of Nrf2 and CAT activity, and GSH concentration and a reduction in serum TNF-α and expression of caspase-3, TNF-α, and TGF-β (P < 0.001)., Conclusions: Administration of date palm extracts exhibited a renoprotective effect against renal I/R injury.This renoprotective action might be due to their antioxidants, anti-apoptotic and anti-inflammatory actions. Moreover, aqueous fruit extracts offered powerful renoprotective effect than aqueous seed extracts, and aqueous fruit and seed extracts were generally more effective than methanolic extracts., (Copyright © 2020. Published by Elsevier Masson SAS.)

New blend films based on polyethylene terephthalate (PET) with different concentrations (50, 100 and 200 µL) of chamomile oil (CAO) were prepared. The effect of oil on the dielectric properties, structural and surface properties of PET was studied. The wettability of the blend films was evaluated by contact angle measurements. In vitro platelet adhesion on the surface and coagulation assessment were conducted to evaluate the behavior of the new blends for blood contact applications. Results of the study indicate that the wettability of PET-CAO blends up to 100 µL has been enhanced relative to the pure PET as indicated by the decrease in contact angle measurements. The attenuation total reflection-Fourier transform infrared spectra of the blends confirmed the presence of chamomile oil in the polymer matrix and suggested the presence of interaction between them. The permittivity ε' values decreased by increasing oil content upto 100 µL. On the other hand, the values of dielectric loss ε″ were found to increase by increasing oil content to 100 µL after which it decreased. The delay in partial thromboplastin time (PTT) of the blood would validate the anti-coagulant property of PET-CAO blends. The results demonstrated that the PET-CAO blends with concentration of 100 µL could be considered as a promising candidate material in blood contact application.

Surface properties and morphology of the biomaterial play an essential role in the polymer-material interaction. In this work, laser surface modification of polyethylene terephthalate as a polymer with distinguished mechanical properties was carried out using (neodymium-doped yttrium aluminum garnet) Nd:YAG laser (1.064 µm) with different output power (0.3, 3, and 6 W). The structural, surface, and dielectric properties of PET before and after laser irradiation have been studied using attenuation total reflection-Fourier transform infrared (ATR-FTIR), dielectric spectroscopy (DS), scanning electron microscope (SEM), and contact angle measurements. Moreover, the anticoagulant properties of the laser-irradiated PET was determined through measuring the prothrombin time (PT), partial thromboplastin time (PTT), and international normalized ratio (INR). In vitro platelet adhesion test was used to assess the platelets adhered to the surface of the samples; in addition to hematological study. It was found that contact angle (θ) measurements of laser-irradiated PET samples decreased compared to the unirradiated PET. The irradiated samples at 0.3 W have the lowest contact angle which is a clear indication that surface treatment with Nd:YAG laser brought about improving the wettability of the polymer. From the dielectric measurements, both values of permittivity and dielectric loss decrease by increasing the laser power. The electrical conductivity decreases with increasing laser power, but still in the same order 10 -14  S/cm. The decrease in electrical conductivity σ may be due to the cross-linking of the polymeric matrix which led to a decrease in the total polarity and consequently decrease in electrical conductivity. The magnitude of σ obtained is highly recommended to be used for insulator purposes in addition to the main purpose that is blood contact.

Increased reactive oxygen species (ROS) generation may disrupt the oocytes function and their competence. In this study, we introduced BTZO-1, a new supplement that can regulate the oxidative stress. Addition of BTZO-1 during IVM of bovine oocytes improved their developmental competence in the term of improvement of blastocyst rates. In addition, the quality of the produced embryos was improved by decreasing the apoptosis level by showing a decreased number of TUNNEL positive cells., (© 2020 Blackwell Verlag GmbH.)

Background: The reinforcement of teeth with internal root resorption is essential to prevent their fracture., Objectives: The aim of this study was to assess the fracture resistance of the premolar teeth with internal root resorption cavities (IRCs), repaired with glass-ionomer cement (GIC), gutta-percha (GP) or EndoSequence® Root Repair MaterialTM (RRM)., Material and Methods: Forty lower premolars, instrumented to size 50, were used. Ten teeth were assigned to the control group, which received the full obturation of the root canals. In the remaining 30, IRCs were prepared with Gates-Glidden burs. The apical 8 mm was obturated to the level of IRC using the single-cone technique. Then, the teeth were divided into 3 groups according to the material used for repairing the cavities (n = 10): GIC; GP; and RRM. The canals were filled with respective materials and backfilled with GP. All of the specimens were scanned at the level of IRC with a micro-computed tomography (micro-CT) system, and the volume of the IRCs and the percentages of voids in the filling materials were measured. The specimens were subjected to fracture testing. The force recorded at the time of fracture was analyzed with the Kruskal-Wallis test and the independent t-test., Results: The control group showed a significantly higher mean value of fracture resistance as compared to the groups with IRCs (p < 0.05). No significant difference was found between GIC and RRM, whereas the GP group had a significantly lower fracture resistance than other tested IRC groups (p < 0.05). The percentage of voids was significantly higher in the GIC group as compared to the GP and RRM groups (p < 0.05)., Conclusions: EndoSequence Root Repair Material provides more strength to the teeth than the GP/sealer technique when both are used to fill a resorption cavity. The fracture resistance of the teeth filled with RRM was close to that obtained with GIC.

There are several obstacles in camel semen cryopreservation; such as increasing semen viscosity and the reduction in motile spermatozoa after ejaculation. Epididymal spermatozoa offer an efficient alternative to overcome these problems and are well-suited for artificial insemination in camels. In the current study, we compared the effects of supplementation with vitamin C, E, inorganic trace elements of selenium (Na 2 SeO 3 ) and zinc (ZnSO 4 ), and zinc and selenium nanoparticles (ZnONPs and SeNPs, respectively) on the cryopreservation of dromedary camel epididymal spermatozoa. When the SHOTOR extender was supplemented with ZnONPs and SeNPs; the sperm showed increased progressive motility; vitality; and membrane integrity after cooling at 5 °C for 2 h; when compared to the control and vitamin-supplemented groups. Moreover, the ZnONPs and SeNPs supplementation improved the progressive motility, vitality, sperm membrane integrity, ultrastructural morphology, and decreased apoptosis when frozen and thawed. SeNPs significantly increased reduced glutathione (GSH), superoxide dismutase (SOD), and decreased lipid peroxide malondialdehyde (MDA) levels. The advantageous effects of the trace elements were potentiated by reduction into a nano-sized particle, which could increase bioavailability and reduce the undesired liberation of toxic concentrations. We recommend the inclusion of SeNPs or ZnONPs to SHOTOR extenders to improve the cryotolerance of camel epididymal spermatozoa., Competing Interests: The authors declare no conflict of interest.

Several materials such as silver are used to enhance graphene oxide (GO) sheets antimicrobial activity. However, these toxic materials decrease its biocompatibility and hinder its usage in many biological applications. Therefore, there is an urgent need to develop nanocomposites that can preserve both the antimicrobial activity and biocompatibility simultaneously. This work highlights the importance of functionalisation of GO sheets using Polyvinylpyrrolidone (PVP) and decorating them with silver nanoparticles (AgNPs) in order to enhance their antimicrobial activity and biocompatibility at the same time. The structural and morphological characterisations were performed by UV-Visible, Fourier transform infrared (FTIR), and Raman spectroscopic techniques, X-ray diffraction (XRD), and high-resolution transmission electron microscopy (HR-TEM). The antimicrobial activities of the prepared samples against Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans were studied. The cytotoxicity of prepared materials was tested against BJ1 normal skin fibroblasts. The results indicated that the decoration with AgNPs showed a significant increase in the antimicrobial activity of GO and FGO sheets, and functionalisation of GO sheets and GO-Ag nanocomposite with PVP improved the cell viability about 40 and 35%, respectively.

Objectives: The purpose of this study was to evaluate the cyclic fatigue, bending resistance, and surface roughness of EdgeEvolve (EdgeEndo) and ProTaper Gold (Dentsply Tulsa Dental Specialties) nickel-titanium (NiTi) rotary files., Materials and Methods: The instruments ( n = 15/each) were tested for cyclic fatigue in single- (60° curvature, 5-mm radius) and double-curved (coronal curvature 60°, 5-mm radius, and apical curvature of 30° and 2-mm radius) artificial canals. The number of cycles to fracture was calculated. The bending resistance of both files were tested using a universal testing machine where the files were bent until reach 45°. Scanning electron microscopy and x-ray energy-dispersive spectrometric analysis were used for imaging the fractured segments, while the atomic force microscope was used to quantify the surface roughness average (Ra)., Results: EdgeEvolve files exhibited higher cyclic fatigue resistance than ProTaper Gold files in single- and double-curved canals ( p < 0.05) and both files were more resistant to cyclic fatigue in single-curved canals than double-curved canals ( p < 0.05). EdgeEvolve files exhibited significantly more flexibility than did ProTaper Gold files ( p < 0.05). Both files had approximately similar Ni and Ti contents ( p > 0.05). EdgeEvolve files showed significantly lower Ra values than ProTaper Gold files ( p < 0.05)., Conclusions: Within the limitation of this study, EdgeEvolve files exhibited significantly higher cyclic fatigue resistance than ProTaper Gold files in both single- and double-curved canals., Competing Interests: Conflict of Interest: No potential conflict of interest relevant to this article was reported.

The aim of this study was to evaluate the effect of adding different concentrations of selenium nano-particles (Se-NPs) in semen extender on bull sperm cryopreservation. Five healthy, fertile Friesian bulls were used, and the ejaculates were obtained using an artificial vagina. Semen of all bulls were pooled and diluted in a tris-yolk fructose (TYF) extender supplemented with Se-NPs at concentrations of 0 (T1, control), 0.5 (T2), 1.0 (T3) and 1.5 (T4) μg/ml for a final sperm concentration of 80 × 10 6 sperm cells/ml. Diluted semen was packed in straws (0.25 ml) and stored in liquid nitrogen (-196 °C) for one month. After thawing, semen of each treatment was evaluated for sperm quality parameters, including sperm progressive motility, livability, morphological abnormalities, plasma membrane integrity and chromatin integrity. Apoptosis and sperm ultrastructure were also examined. Total antioxidant capacity and lipid peroxidation markers were determined in seminal plasma of semen in each treatment. Finally, the effect of Se-NPs on fertilization capacity was checked in vivo using n = 81cows. Results showed that T2 and T3 had a positive effect (P < 0.05) on post-thawing sperm progressive motility, livability and membrane integrity as compared with the control. Percentage of viable sperm increased (P < 0.05), while percentages of early apoptotic, apoptotic and necrotic sperm cells decreased (P < 0.05) in T3 as compared to T1. Total antioxidants capacity (TAC) in seminal plasma increased (P < 0.05) and malondialdhyde (MDA) concentration decreased (P < 0.01) in T3 as compared to T1, but did not differ in T4 from that in T1. In vivo fertility rate was higher in T3 (90%) than in T1 (59%). In conclusion, enrichment of semen extender with Se-NPs at a concentration of 1.0 μg/ml improved post-thaw sperm quality of Holstein bulls, and consequently in vivo fertility rate by reducing apoptosis, lipid peroxidation and sperm damage occurring by cryopreservation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Recent advances in nanotechnology have tremendously expanded its possible applications in biomedicine. Although, the effects of nanoparticles (NPs) at cellular and tissue levels have not been fully understood, some of these biological effects might be employed in assisted reproduction to improve male fertility particularly by enhancing sperm cell quality either in vivo or in vitro . This review summarises the available literature regarding the potential applications of nanomaterials in farm animal reproduction, with a specific focus on the male gamete and on different strategies to improve breeding performances, transgenesis and targeted delivery of substances to a sperm cell. Antioxidant, antimicrobial properties and special surface binding ligand functionalization and their applications for sperm processing and cryopreservation have been reviewed. In addition, nanotoxicity and detrimental effects of NPs on sperm cells are also discussed due to the increasing concerns regarding the environmental impact of the expanding use of nanotechnologies on reproduction.

Dietary polyunsaturated fatty acids (PUFAs) can influence fertility in farm animals. Some evidence in mice and sheep have suggested that PUFAs may influence offspring sex ratio, which may have significant value for cattle production. To test this hypothesis, three groups of Holstein cows were supplemented with either 0%, 3% or 5% protected fat (PF) in the form of calcium salt of fatty acids (rich in omega-6) from 14-21 days pre-partum until conception. Proven-fertile frozen semen from the same ejaculate was used for insemination. Calf sex recorded at birth was 8/19 (42.1%) male offspring in the control group, increasing to 14/20 (70%, P  > 0.05) and 17/20 (85%, P  < 0.05) in 3% and 5% PF, respectively. To test if this effect was caused by a direct influence on the oocyte, we supplemented bovine cumulus oocyte complexes during in vitro maturation with either omega-3 alpha-linolenic acid (ALA), omega-6 linoleic acid (LA) or trans-10, cis-12 conjugated linoleic acid (CLA). Sex ratio of the produced transferable embryos was determined using PCR of SRY gene. Similar to the in vivo results, sex ratio was skewed to the male side in the embryos derived from LA- and CLA-treated oocytes (79% and 71%) compared to control and ALA-treated oocytes (44% and 54%, respectively). These results indicate that both dietary and in vitro supplementation of omega-6 PUFAs can skew the sex ratio towards the male side in cattle. Further experiments are required to confirm this effect on a larger scale and to study the mechanisms of action that might be involved.