Effect of breeding season and age on follicular dynamics and hemodynamics in embryo donor mares subjected to luteolysis after embryo flushing.

Background: Mares are the only companion animals simulating women in the large diameter of their follicles. Horses start reproduction at the age of three years, and some of them live for >30 years, so aging influences their reproductive capacity. Mares are sensitive to summer heat stress as they can sweat like humans. Aim: The current work aimed to study the effects of age (young versus senile), season (cold versus hot), and the hormonal treatments during embryo collection on the dominant and subordinate follicular dynamics and hemodynamics and circulating ovarian hormones in embryo donor mares ovulated twice spontaneously before inducing ovulation for flushing embryos. Methods: Spontaneous oestrous cycles were studied for young mares (<10 years; N = 6) or senile (>20 years; N = 5) during months of the cold season (November to April) and hot season (May to August). In young embryo donor mares, oestrous cycles after inducing ovulation and luteolysis were studied using Doppler ultrasound. Estradiol (E2), progesterone (P4), nitric oxide (NO), total cholesterol, and lactate dehydrogenase (LDH) were measured in blood serum. Results: A decrease in the dominant follicle antrum diameter (p > 0.05) and LDH (p = 0.016) was observed after inducing luteolysis in young embryo donor mares. Both human chorionic gonadotropin (hCG) and PGF2α treatments increased dominant follicle area (p = 0.0001), antrum area (p = 0.001), perimeter (p = 0.001), granulosa area (p = 0.0001), cholesterol (p = 0.0001), NO (p = 0.0001), and E2 (p = 0.0001). The dominant follicle area, antrum area, perimeter, color area, granulosa area, LDH, cholesterol, NO, and E2 increased (p = 0.0001) during the oestrous cycles of the hot season, but the circulatory % (p = 0.0001) declined. Senile mares had lower dominant follicle area (p = 0.002), antrum area (p = 0.0001), granulosa area (p > 0.05), LDH (p = 0.001), cholesterol (p = 0.0001), NO (p = 0.0001), and E2 (p = 0.0001) but higher circulatory % (p = 0.0001) and color area % (p = 0.023). The dominant follicle possesses the largest diameter, area, perimeter, granulosa area, and color area but the lowest circulatory % during spontaneous oestrous cycles, after inducing ovulation, or luteolysis with significant effects of the day of the spontaneous oestrous cycles on their dynamics and hemodynamics. Conclusion: During hot months, mares treated with hCG ovulated 24 hours later and prostaglandin-induced luteolysis was followed by new ovulation five days later. Follicles ovulated during the hot months were larger than those ovulated during the cold months and both had nearly the same color area %. Senile mares ovulated follicles with a lower area and antrum area but a higher color area %, so senile mares can be used as embryo or oocyte donors during the hot season.

Evaluation of non-thyroidal illness syndrome in shock patients admitted to pediatric intensive care unit in a developing country.

During critical illness, children my experience various changes in their thyroid hormone levels. Such changes are termed non-thyroidal illness syndrome (NTI). The extent of change correlates with the severity of the illness and its outcomes in critically ill patients. This study aimed to investigate the correlation between the severity of shock and thyroid hormone derangement. This prospective observational study included forty patients aged one month to five years who were admitted to the pediatric intensive care unit (PICU) with shock. Thyroid function tests were conducted on admission, after shock reversal, and five days later. NTI patterns were observed in 70% of patients. The PIM2 score showed a significant negative correlation with T3 (r = - 0.353, p = 0.026) and FT3 levels on admission (r = - 0.417, p = 0.007). Furthermore, after shock reversal, the PIM2 score continued to exhibit significant negative correlations with T4 (r = - 0.444, p = 0.004), T3 (r = - 0.329, p = 0.038), FT3 (r = - 0.355, p = 0.025), and FT4 levels (r = - 0.379, p = 0.016).    Conclusion: This study underscores the high prevalence of NTI in PICU shock patients and suggests monitoring thyroid hormone levels for outcome prediction and treatment guidance. Further research is needed to optimize NTI management in critically ill children. What is Known: • Non-thyroidal illness syndrome (NTIS) is a condition observed in critically ill patients. • There has been limited research on NTI in children, and existing studies have generated conflicting results regarding the relationship between thyroid hormones and clinical outcomes in cases of sepsis and septic shock. What is New: • The study has revealed dynamic changes in free triiodothyronine (FT3) levels during the process of shock reversal and recovery in children who experienced shock. • A significant negative correlation was found between the Pediatric Index of Mortality 2 (PIM2) score and several thyroid hormone levels, including FT3 on admission and T4, FT3, and FT4 on shock reversal.

Accelerating wound healing: Unveiling synergistic effects of P25/SWCNT/Ag and P25/rGO/Ag nanocomposites within PRP-gelatin scaffold, highlighting the synergistic antimicrobial activity.

Wound healing is a multifaceted biological process requiring innovative strategies to enhance efficiency and counter infections. In this groundbreaking study, we investigate the regenerative potential of platelet-rich plasma (PRP) integrated into a gelatin (GLT) scaffold along with nanocomposites of titanium dioxide (TiO2) (P25)/single-walled carbon nanotubes (SWCNTs)/Ag and P25/reduced graphene oxide (rGO)/Ag. Incorporating these advanced materials into the PRP/GLT delivery system aims to optimize the controlled release of growth factors (GFs) and leverage the exceptional properties of nanomaterials for enhanced tissue repair and wound healing outcomes. Antioxidant activity assessment using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity reveals the superior performance of P25/SWCNTs/Ag compared to P25/rGO/Ag. Their synergistic effects are evaluated in conjunction with antibacterial and antifungal antibiotics. Furthermore, the wound healing potential of P25/SWCNTs/Ag and P25/rGO/Ag, combined with PRP/GLT, is examined. Notably, both nanocomposites exhibit promising synergistic effects with gentamicin and fluconazole against pathogenic strains. Significantly, the inclusion of non-activated PRP substantially augments the wound healing efficacy of P25/SWCNTs/Ag on days 3 (p < 0.01) and 15 (p < 0.05). These findings pave the way for advanced wound dressing and therapeutic interventions, capitalizing on the synergistic effects of PRP and nanomaterials, thus ultimately benefiting patients and advancing regenerative medicine.

Curcumin-infused nanostructured lipid carriers: a promising strategy for enhancing skin regeneration and combating microbial infection.

BACKGROUND: Curcumin is a biomolecule that can be extracted from the Curcuma longa that has been shown to have the potential to aid skin wound healing. It has been studied for its anti-inflammatory and antioxidant properties, which may help to reduce swelling and promote tissue repair. However, curcumin has low solubility in water, which can limit its absorption and bioavailability. Encapsulating it in lipid nanoparticles may help to increase its absorption, leading to improved bioavailability. METHODS: Curcumin-loaded nanostructure lipid nanocarriers (CURC-NLCs) were prepared and characterized. Also, the phenolic, flavonoid contents, antioxidant and antimicrobial efficacy against gram-positive and gram-negative bacteria were investigated. Furthermore, in vivo rabbit animal model was used to test its regenerative capacity and wound-healing efficiency. RESULTS: The CURC-NLCs significantly increased the content of phenolic and flavonoid compounds compared to curcumin, resulting in a dramatic increase in antioxidant activity. CURC-NLCs also showed a potent inhibitory effect on Gram-positive, Gram-negative, and fungi, two times higher than curcumin. CURC-NLCs showed a higher potential to fasten the wound healing of full-thickness skin injuries as it resulted in 1.15- and 1.9-fold higher wound closure at the first week of injury compared to curcumin and control, respectively (p < 0.0001). CONCLUSION: These results suggest that CURC-NLCs have an excellent potential to promote skin regeneration, which could be attributed to its antioxidant and broad-spectrum antimicrobial effect.

Comparative evaluation of propolis nanostructured lipid carriers and its crude extract for antioxidants, antimicrobial activity, and skin regeneration potential.

BACKGROUND: Propolis extracted from beehives has been conferred with natural antimicrobial and antioxidant properties. Hence, it has been recommended as a wound healing therapy. This study investigated the additive value of nanotechnology to the herbal extract, (propolis rebuts), after which we examined its efficacy in wound healing. METHODS: Propolis nanostructured lipid carriers (NLCs) were first prepared using the emulsion-evaporation-solidification method at three concentrations. Then, we compared their flavonoid and phenolic contents and phenolic contents. Their antioxidant, antibacterial, and antifungal effects were also investigated after which, the skin regenerative capacity of propolis-NLCs was assessed using full-thickness skin wounds in rabbits. RESULTS: This study showed that propolis-NLCs had increased the phenolic and flavonoid contents compared to the raw propolis extract (EXTR) (9-fold and 2-fold, respectively). This increase was reflected in their antioxidant activities, which dramatically increased by 25-fold higher than the propolis-EXTR. Also, propolis-NLCs exhibited a 2-fold higher potent inhibitory effect than propolis-EXTR on Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), Gram-negative bacterium (Salmonella spp.), and fungus (Candida albicans) microbes (p < 0.0001). Investigations also revealed that treatment of full-thickness skin injuries with propolis-NLCs resulted in significantly higher wound closure compared to propolis-EXTR and the control after two weeks (p < 0.0001). CONCLUSION: With a prominent broad-spectrum antibacterial effect propolis-NLCs exhibited higher skin regenerative potency than propolis-EXTR. We also highlighted the additive impact of nanotechnology on herbal extract, which accounted for the increased flavonoid content and hence a better antioxidant and antimicrobial effect and propose it as a potential therapy for wound healing.

Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa.

Melatonin is known to protect sperm against freezing-inflicted damage in different domestic species. The aim of the study was to evaluate the effect of supplementation of semen extender with melatonin on the quality and DNA integrity of cooled and frozen/thawed rabbit spermatozoa. We also investigated whether the addition of melatonin to the semen extender could improve the fertility of rabbit does artificially inseminated with frozen/thawed semen. Semen samples collected from eight rabbit bucks were pooled and then diluted in INRA-82 supplemented either with (0.5, 1.0 or 1.5 mM) or without (0.0 mM) melatonin. Diluted semen was cooled at 5°C for 24 hr. For cryopreservation and based on the first experiment's best result, semen samples were diluted in INRA-82 in the presence or absence of 1.0 mM melatonin and then frozen in 0.25 ml straws. Following cooling or thawing, sperm quality and DNA integrity were evaluated. Furthermore, the fertility of frozen/thawed semen was investigated after artificial insemination. Supplementation of semen extender with 1.0 mM melatonin improved (p < .05) motility, viability, membrane and acrosome integrities in cooled semen compared with other groups. Sperm quality and DNA integrity were higher (p < .05) in frozen/thawed semen diluted in 1.0 mM melatonin-supplemented extender than in the control group. Conception and birth rates were higher in does inseminated with 1.0 mM melatonin treated semen compared with the controls. In conclusion, supplementation of semen extender with 1.0 mM melatonin improved the quality of cooled and frozen/thawed rabbit spermatozoa. Melatonin can preserve DNA integrity and enhance the fertility of frozen/thawed rabbit spermatozoa.

Gelatin Loaded Titanium Dioxide and Silver Oxide Nanoparticles: Implication for Skin Tissue Regeneration.

Treatment of burn wounds has many requirements to ensure wound closure with healthy tissue, increased vascularization, guarantee edema resolution, and control bacterial infection. We propose that titanium oxide (TiO2) nanoparticles (NPs) will be more efficient than silver dioxide (Ag2O) in the treatment of burn wounds. Herein, gelatin loaded NPs (GLT-NPs) were evaluated for their efficacy to regenerate second-degree burn wound in rabbit skin. TEM results revealed that the average particle sizes were ⁓ 7.5 and 17 nm for Ag2O and TiO2 NPs, respectively. The results of the in vivo application of GLT-NPs on burn wound in the rabbit revealed that both Ag2O and TiO2 NPs were efficient than the control none treated (CTRL) and GLT group. In terms of the healing rate, the GLT-TiO2 did not show any significant difference than GLT-Ag2O (99.57% vs. 99.85%, p = 0.2). Meanwhile, the healing rate was significantly higher in both NPs' treated groups than CTRL (94.16%, p < 0.01) and GLT group (95.07%, p < 0.05). Also, the histological analysis using H&E staining showed re-epithelization, less edema, and enhanced vascularization in both GLT-NPs than CTRL and GLT groups. Furthermore, immunohistochemical analysis of TGF-ß1 and α-SMA revealed significantly a higher expression in both GLT-NPs groups than CTRL and GLT groups at weeks 1 and 2 (p < 0.05). Interestingly, TGF-ß1 and α-SMA were substantially higher in GLT- TiO2 than GLT-Ag2O at weeks 1 and 2 (p < 0.05), but the expression was not significant at week 3. In conclusion, GLT-NPs showed higher regenerative capacity and enhanced the healing quality after burn wound compared to CTRL and GLT. Graphical abstract.

Passive immunization against inhibin increases testicular blood flow in male goats.

The present study investigated whether or not passive immunization against inhibin modulates testicular blood flow in goats. Male Shiba goats were injected with either 10 ml of inhibin antiserum (INH group; n = 5) or 10 ml of normal castrated goat serum (NGS group; n = 4). Concentrations of FSH, LH, testosterone (T), and estradiol (E2) in the plasma were measured by radioimmunoassay. Blood flow within the supratesticular (STA) and marginal testicular arteries (MTA) were measured by color pulsed-Doppler ultrasonography, and Doppler indices (resistive index; RI and pulsatility index; PI) were recorded. Results revealed significant increases in concentrations of FSH and E2 in the INH group compared to those in the NGS group (P < 0.05). Animals in the INH group had greater (P < 0.05) FSH concentrations than those in the NGS group in the period between 60 h and 144 h after treatment than at any other time. Estradiol concentrations were greater (P < 0.05) in the INH group than in the NGS group at 6 h (12.15 ± 2.09 pg/ml vs 5.49 ± 1.17 pg/mL), 12 h (8.27 ± 1.29 pg/mL vs 3.05 ± 0.38 pg/mL), and 36 h (9.35 ± 1.31 pg/mL vs 5.09 ± 0.46 pg/mL) after treatment than at any other time. Concentrations of LH and T did not significantly change between the two groups. Goats in the INH group had lesser (P < 0.05) RI of the STA than those in the NGS group and RI values were lesser at 24 h (0.37 ± 0.031 vs 0.49 ± 0.004) and 120 h (0.38 ± 0.028 vs 0.55 ± 0.048) after treatment than at any other time. Furthermore, values of RI and PI of the MTA were significantly lesser (P < 0.05) in the INH group compared to those in the control group at 48 h (RI of MTA: 0.21 ± 0.014 vs 0.37 ± 0.039; PI of MTA: 0.24 ± 0.016 vs 0.46 ± 0.058) after treatment than at any other time. In conclusion, passive immunization against inhibin has a stimulatory effect on testicular blood flow in goats by inducing decreases in the RI values of the STA and MTA.

Optimization of the Protocol for Cryopreservation of Arabian Stallion Spermatozoa: Effects of Centrifugation, Semen Extenders and Cryoprotectants.

BACKGROUND: Cryopreservation of Arabian stallion semen is important in order to improve the function and fertility of frozen/thawed semen in this breed. OBJECTIVE: To investigate the effects of centrifugation, type of semen extenders, and type of cryoprotectants on the quality of frozen/thawed Arabian stallion spermatozoa. MATERIALS AND METHODS: Semen samples collected from four adult Arabian stallions (one ejaculate per week for 10 consecutive weeks) were either processed directly without centrifugation (no centrifugation; NC) or subjected to centrifugation on the gel-free fraction. Centrifugation protocols were divided into six categories; 600 x g for 10 min (C1), 600 x g for 15 min (C2), 900 x g for 10 min (C3), 900 x g for 15 min (C4), 1200 x g for 10 min (C5), or 1200 x g for 15 min (C6) (Experiment 1). Two semen extenders, INRA-82 and modified Kenney's were compared (Experiment 2). Three different cryoprotectants, [namely 5% glycerol, 5% dimethylformamide (DMF) and 2.5% glycerol] plus 2.5% DMF were used (Experiment 3). Following freezing and thawing, motility, viability, plasma membrane integrity, acrosome status and viability index were evaluated. RESULTS: Centrifugation at 600 x g for 15 min before cryopreservation increased (P< 0.05) sperm motility, viability, membrane integrity and percentage of spermatozoa with intact acrosome compared to other centrifugation protocols. Dilution of Arabian stallion semen with INRA-82 before cryopreservation improved (P< 0.05) sperm quality after freezing and thawing compared to modified Kenney's extender. Supplementation of semen diluent INRA-82 with 5% DMF improved (P< 0.05) the quality of frozen/thawed Arabian stallion spermatozoa compared to 5% glycerol. CONCLUSION: These findings suggest that optimized conditions such as centrifugation, types of semen extenders and cryoprotectants play an important role in processing Arabian stallion spermatozoa for cryopreservation.

Development of a cobalt(II) phthalocyanine- MWCNT modified carbon paste electrode for the detection of polyunsaturated fatty acids.

In this work the development of an electrochemical sensor for the determination of polyunsaturated fatty acids (PUFAs), in particular linoleic acid, in commercially available safflower oil as complex matrix is described. The sensor consists of a carbon paste electrode with cobalt(II) phthalocyanine, Co(II)Pc, as mediator and multiwalled carbon-nanotubes (MWCNT) as nanomaterial. As carrier medium a sodium borate buffer (0.1 M, pH 9) was used. PUFAs were detected at a working voltage of 0.35-0.45 V (vs. Ag/AgCl). The sensor development was carried out in a batch system with differential pulse voltammetry (DPV) and cyclic voltammetry (CV). Sensor specification was tested by using various fatty acids (stearic acid, oleic acid, linoleic acid, linolenic acid and arachidonic acid) as well as esterified fatty acids, resulting in a specific applicability towards PUFAs, especially linoleic acid (LAH). The optimized sensor was applied in a flow injection analysis system (FIA) for the analysis of PUFAs in complex matrix. Linoleic acid was used as standard substrate to determine the analytical parameters. The linearity ranges between 7.5 and 200 µg mL-1 LAH, while the limit of detection was determined to be 2.5 µg mL-1 and the limit of quantification is approximately 7.5 µg mL-1 LAH. The LAH content was successfully detected in commercially available safflower oil via standard addition method and the results could be confirmed by a reference method. The PUFA content was calculated as LAH-equivalent.

High frequency oscillatory ventilation versus conventional mechanical ventilation in pediatric acute respiratory distress syndrome: A randomized controlled study.

El-Nawawy A, Moustafa A, Heshmat H, Abouahmed A. High frequency oscillatory ventilation versus conventional mechanical ventilation in pediatric acute respiratory distress syndrome: A randomized controlled study. Turk J Pediatr 2017; 59: 130-143. The aim of this prospective randomized study is to compare the outcomes of the early use of either high frequency oscillation (HFO) or conventional mechanical ventilation (CMV) in patients with pediatric acute respiratory distress syndrome (PARDS). We allocated two hundred PARDS patients over 5 years in 1:1 ratio to either mode. The HFO group showed a significantly higher median partial arterial oxygen pressure to fraction of inspired oxygen (PaO2/FiO2) values after 24 hours of enrollment (p=0.011), higher oxygenation index (OI) decrease percent (p=0.004) and lower cross-over rates (p < 0.001), whereas no differences in 30-day mortality, length of stay (LOS) or ventilation days (p=0.77, p=0.28, p=0.65 respectively). The second day values (after 24 hours) of both OI and PaO < sub > 2 < /sub > /FiO < sub > 2 < /sub > were found to be more significant discriminators for mortality when compared to the baseline values (cutoff values > 8.5, ≤139 respectively). PARDS patients with baseline OI > 16 had a better chance of survival if initially ventilated with the HFO (p=0.004). Although the HFO mode appeared to be a safe mode with a significant better oxygenation improvement (after the first 24 hours) and fewer cross-over rates, it failed to show differences as regards mortality or LOS when compared to the CMV adopting protective lung strategy. In PARDS, HFO had a superior advantage in improving oxygenation, yet with no significant mortality improvement, as multi-organ dysfunction syndrome (MODS) was the most common cause of death in our study and not refractory hypoxemia which is the main problem in PARDS; highlighting that mortality in PARDS is multi-factorial and may not depend only on how fast oxygenation improves.

The carpal joint of the donkey (Equus asinus): morphological investigation / Articulación del carpo del asno (Equus asinus): investigación morfológica

The current study has been achieved to be an essential resource for all veterinary practitioners that deal with the anatomy of the carpal joint of the donkey. Ten adult donkeys of both sexes were used in the current study. The topographical approach to the carpal joint was investigated in this study. Radiography and computed tomography (CT) of the carpus delineated the articulations of the carpal joint: radiocarpal, intercarpal, and carpometacarpal. The carpal ligaments were well delineated and the carpal canal was demonstrated with its content such as superficial digital flexor tendon (SDFT) and deep digital flexor tendon (DDFT). The contrast radiography visualized that the radiocarpal joint outpouched proximal to the accessory carpal bone by large palmarolateral pouch and small palmaromedial pouch, however the intercarpal joint outpouched distal to the accessory carpal bone by two small palmarolateral and palmaromedial pouches. The carpometacarpal joint showed medial and lateral palmarodistal outpouchings in distal direction between the corresponding 2nd and 4th metacarpal bones and the 3rd metacarpal bone. Scanning electron microscopy (SEM) displayed two types of synoviocytes macrophages type A cells and fibroblast like type B cells at the cellular lining of the synovial membrane of the joint capsule.

Este estudio constituye un recurso esencial para todos los veterinarios que se ocupan de la anatomía de la articulación del carpo del asno. Fueron utilizados diez asnos adultos, de ambos sexos. Se analizó la articulación del carpo desde un enfoque topográfico. Se realizaron estudios por radiografía y tomografía computarizada para identificar las articulaciones del carpo en su conjunto: radiocarpiana, intercarpiana y carpometacarpiana. Los ligamentos del carpo se encontraron bien delineados y en el canal carpiano se identificó su contenido, como el tendón del flexor digital superficial y el tendón del flexor digital profundo. La radiografía de contraste permitió visualizar que la articulación radiocarpiana se articula proximalmente con el hueso accesorio del carpo a través una larga evaginación palmarolateral y una pequeña evaginación palmaromedial; sin embargo, la articulación intercarpiana se relaciona distalmente con el hueso accesorio del carpo por medio de dos pequeñas evaginaciones, palmarolateral y palmaromedial. La articulación carpometacarpiana mostró evaginaciones palmarodistales, medial y lateral, en dirección distal entre los correspondiente 2 y 4 huesos metacarpianos y el 3 hueso metacarpiano. La microscopía electrónica de barrido muestra dos tipos de células: células A, correspondientes a sinoviocitos macrófagos y células B, correspondientes a fibroblastos, en el revestimiento celular de la membrana sinovial de la cápsula articular.