Ultrasound Neuromodulation of an Anti-Inflammatory Pathway at the Spleen Improves Experimental Pulmonary Hypertension.

BACKGROUND: Inflammation is pathogenically implicated in pulmonary arterial hypertension; however, it has not been adequately targeted therapeutically. We investigated whether neuromodulation of an anti-inflammatory neuroimmune pathway involving the splenic nerve using noninvasive, focused ultrasound stimulation of the spleen (sFUS) can improve experimental pulmonary hypertension. METHODS: Pulmonary hypertension was induced in rats either by Sugen 5416 (20 mg/kg SQ) injection, followed by 21 (or 35) days of hypoxia (sugen/hypoxia model), or by monocrotaline (60 mg/kg IP) injection (monocrotaline model). Animals were randomized to receive either 12-minute-long sessions of sFUS daily or sham stimulation for 14 days. Catheterizations, echocardiography, indices of autonomic function, lung and heart histology and immunohistochemistry, spleen flow cytometry, and lung single-cell RNA sequencing were performed after treatment to assess the effects of sFUS. RESULTS: Splenic denervation right before induction of pulmonary hypertension results in a more severe disease phenotype. In both sugen/hypoxia and monocrotaline models, sFUS treatment reduces right ventricular systolic pressure by 25% to 30% compared with sham treatment, without affecting systemic pressure, and improves right ventricular function and autonomic indices. sFUS reduces wall thickness, apoptosis, and proliferation in small pulmonary arterioles, suppresses CD3+ and CD68+ cell infiltration in lungs and right ventricular fibrosis and hypertrophy and lowers BNP (brain natriuretic peptide). Beneficial effects persist for weeks after sFUS discontinuation and are more robust with early and longer treatment. Splenic denervation abolishes sFUS therapeutic benefits. sFUS partially normalizes CD68+ and CD8+ T-cell counts in the spleen and downregulates several inflammatory genes and pathways in nonclassical and classical monocytes and macrophages in the lung. Differentially expressed genes in those cell types are significantly enriched for human pulmonary arterial hypertension-associated genes. CONCLUSIONS: sFUS causes dose-dependent, sustained improvement of hemodynamic, autonomic, laboratory, and pathological manifestations in 2 models of experimental pulmonary hypertension. Mechanistically, sFUS normalizes immune cell populations in the spleen and downregulates inflammatory genes and pathways in the lung, many of which are relevant in human disease.

Dissecting the Interplay Mechanism among Process Parameters toward the Biofabrication of High-Quality Shapes in Embedded Bioprinting.

Embedded bioprinting overcomes the barriers associated with the conventional extrusion-based bioprinting process as it enables the direct deposition of bioinks in 3D inside a support bath by providing in situ self-support for deposited bioinks during bioprinting to prevent their collapse and deformation. Embedded bioprinting improves the shape quality of bioprinted constructs made up of soft materials and low-viscosity bioinks, leading to a promising strategy for better anatomical mimicry of tissues or organs. Herein, the interplay mechanism among the printing process parameters toward improved shape quality is critically reviewed. The impact of material properties of the support bath and bioink, printing conditions, cross-linking mechanisms, and post-printing treatment methods, on the printing fidelity, stability, and resolution of the structures is meticulously dissected and thoroughly discussed. Further, the potential scope and applications of this technology in the fields of bioprinting and regenerative medicine are presented. Finally, outstanding challenges and opportunities of embedded bioprinting as well as its promise for fabricating functional solid organs in the future are discussed.

Bee venom enhances performance and immune function in thinlip mullet: A promising approach for sustainable aquaculture.

As an environmentally friendly alternative to antibiotics, bee venom holds promise for aquaculture due to its diverse health advantages, including immune-amplifying and anti-inflammatory features. This study investigated the effects of dietary bee venom (BV) on the growth and physiological performance of Thinlip mullet (Liza ramada) with an initial body weight of 40.04 ± 0.11 g for 60 days. Fish were distributed to five dietary treatments (0, 2, 4, 6, and 8 mg BV/kg diet) with three replicates. Growth traits, gut enzyme ability (lipase, protease, amylase), intestinal and liver histology, blood biochemistry, immune responses [lysozyme activity (LYZ), bactericidal activity (BA), nitroblue tetrazolium (NBT%)], and antioxidant status [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), malondialdehyde (MDA)] were evaluated. BV supplementation significantly improved growth performance, digestive enzyme activity, histological integrity of organs, immune responses (LYZ, BA), and antioxidant status (SOD, CAT, GPx), while declining MDA levels. Optimal BV levels were identified between 4.2 and 5.8 mg/kg diet for different parameters. Overall, the findings suggest that BV supplementation can enhance growth and physiological performance in Thinlip mullet, highlighting its potential as a beneficial dietary supplement for fish health and aquaculture management.

Cas9-expressing chickens and pigs as resources for genome editing in livestock.

Genetically modified animals continue to provide important insights into the molecular basis of health and disease. Research has focused mostly on genetically modified mice, although other species like pigs resemble the human physiology more closely. In addition, cross-species comparisons with phylogenetically distant species such as chickens provide powerful insights into fundamental biological and biomedical processes. One of the most versatile genetic methods applicable across species is CRISPR-Cas9. Here, we report the generation of transgenic chickens and pigs that constitutively express Cas9 in all organs. These animals are healthy and fertile. Functionality of Cas9 was confirmed in both species for a number of different target genes, for a variety of cell types and in vivo by targeted gene disruption in lymphocytes and the developing brain, and by precise excision of a 12.7-kb DNA fragment in the heart. The Cas9 transgenic animals will provide a powerful resource for in vivo genome editing for both agricultural and translational biomedical research, and will facilitate reverse genetics as well as cross-species comparisons.

Influence of nano-formulations of clove bud (Syzygium aromaticum) extract on freezing ability, antioxidant capacity, caspase-3 activity, acrosome reaction and fertility of frozen rabbit semen.

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.

Investigating the impact of nanoemulsion of curcumin-loaded olive oil on growth performance, feed utilization, immunological responses, and redox status of Litopenaeus vannamei shrimp with emphasis on economic efficiency of supplementation.

The trail aimed to explore the effect of dietary supplementation of curcumin loaded olive oil nanoemulsion (CUR-OLNE) on growth performance, feed utilization, blood biochemical, redox status, and immune response of Litopenaeus vannamei shrimp, considering the economic efficiency of supplementation. A total of 280 healthy shrimps (3.42 ± 0.02 g) were randomly distributed into five equal groups and were fed diets containing 0 (CUR-OLNE0), 5(CUR-OLNE5), 10(CUR-OLNE10), 15(CUR-OLNE15) and 20 (CUR-OLNE20) mg CUR-OLNE/kg diet, respectively for 16 weeks. Among CUR-OLNE treated groups, CUR-OLNE20 showed the highest growth performance and feed utilization traits, including final body weight, specific growth rate, feed conversion ratio, and protein efficiency ratio. Notably, the photomicrographs provided further compelling evidence regarding the potential effect of CUR-OLNE supplementation on muscle structure and integrity. Compared to the control, the levels of blood protein significantly induced in CUR-OLNE15 and CUR-OLNE20 treated groups (p < 0.05). All CUR-OLNE -supplemented groups possessed lower activities of liver enzymes as well as the levels of urea and creatinine compared to the control (p < 0.05). The addition of 20 mg CUR-OLNE/kg diet decreased the concentrations of cortisol, glucose and triglycerides. The dietary treatment significantly improved the secretion of digestive enzymes, including amylase, lipase, and protease. The lowest levels of Malondialdehyde and the highest levels of total antioxidant capacity, super oxide dismutase, catalase, lysozyme and immunoglobulin M were detected in both of CUR-OLNE15, and CUR-OLNE20 treated groups compared to the control (p < 0.05). There were considerable significant effects of dietary supplementation of CUR-OLNE on economic efficiency. In conclusion, the application of nanocarriers for the delivery of dietary immune stimulants such as CUR-OLNE to Litopenaeus vannamei shrimp is a promising strategy for improving shrimp nutrition. The addition of 20 mg CUR-OLNE/kg to the diets of can be recommended as an affective intervention to improve growth performance, feed utilization, and health status of shrimp. Implementing this intervention can maximize the economic efficiency of shrimp farming while promoting sustainable practices in the industry.

Respirable dust exposure impact on respiratory symptoms among cleaners in a Selangor Public University, Malaysia.

INTRODUCTION: Cleaners perform a vital role in environmental health by keeping the place clean, but they are also exposed to various hazards. Yet, there is a lack of effective and accessible occupational safety standard measures, thus making this to be difficult to monitor the long-term health effects of cleaners. This study aims to determine the respirable dust exposure on respiratory symptoms among cleaners in a public university in Selangor. MATERIALS AND METHODS: A cross-sectional study was carried out among 51 cleaners. The respondents' background information and respiratory symptoms were gathered using a series of standardised questionnaires validated by the American Thoracic Society (ATS-DLD-78-A). The 8- hour respirable dust exposure to cleaners was measured using an air sampling pump (Gillian & Sensodyne Gil Air 3). RESULTS: The mean of respirable dust was lower than permissible exposure limit with 0.63±0.57mg/m3. The respiratory symptoms among the cleaners showed no significant association between cough, phlegm, and breathing difficulties with working tenure. Meanwhile, wheezing and coughing with phlegm have an almost significant association with working tenure among cleaners with (Χ2=1.00, p=0.08) and (Χ2=1.00, p=0.07) respectively. Exposure to respirable dust has exhibited 6 times the prevalence of coughing with phlegm among cleaners (PR=6.28, 95% CI: 0.44, 89.38). CONCLUSION: The findings of this study demonstrated that the cleaners were significantly affected by the respirable dust. The cleaners' working environment has caused them to be exposed to respirable dust.

From tap to table: An assessment of drinking water quality in Perak, Malaysia.

INTRODUCTION: A study on the quality of drinking water was conducted at Air Kuning Treatment Plant In Perak, Malaysia, based on a sanitary survey in 14 sampling points stations from the intake area to the auxiliary points. This was to ensure the continuous supply of clean and safe drinking water to the consumers for public health protection. The objective was to examine the physical, microbiological, and chemical parameters of the water, classification at each site based on National Drinking Water Standards (NDWQS) and to understand the spatial variation using environmetric technique; principal component analysis (PCA). MATERIALS AND METHODS: Water samples were subjected to in situ and laboratory water quality analyses and focused on pH, turbidity, chlorine, Escherichia coli, total coliform, total hardness, iron (Fe), aluminium (Al), zinc (Zn), magnesium (Mg) and sodium (Na). All procedures followed the American Public Health Association (APHA) testing procedures. RESULTS: Based on the results obtained, the values of each parameter were found to be within the safe limits set by the NDWQS except for total coliform and iron (Fe). PCA has indicated that turbidity, total coliform, E. coli, Na, and Al were the major factors that contributed to the drinking water contamination in river water intake. CONCLUSION: Overall, the water from all sampling point stations after undergoing water treatment process was found to be safe as drinking water. It is important to evaluate the drinking water quality of the treatment plant to ensure that consumers have access to safe and clean drinking water as well as community awareness on drinking water quality is essential to promote public health and environmental protection.

Voltammetry in the spleen assesses real-time immunomodulatory norepinephrine release elicited by autonomic neurostimulation.

BACKGROUND: The noradrenergic innervation of the spleen is implicated in the autonomic control of inflammation and has been the target of neurostimulation therapies for inflammatory diseases. However, there is no real-time marker of its successful activation, which hinders the development of anti-inflammatory neurostimulation therapies and mechanistic studies in anti-inflammatory neural circuits. METHODS: In mice, we performed fast-scan cyclic voltammetry (FSCV) in the spleen during intravenous injections of norepinephrine (NE), and during stimulation of the vagus, splanchnic, or splenic nerves. We defined the stimulus-elicited charge generated at the oxidation potential for NE (~ 0.88 V) as the "NE voltammetry signal" and quantified the dependence of the signal on NE dose and intensity of neurostimulation. We correlated the NE voltammetry signal with the anti-inflammatory effect of splenic nerve stimulation (SpNS) in a model of lipopolysaccharide- (LPS) induced endotoxemia, quantified as suppression of TNF release. RESULTS: The NE voltammetry signal is proportional to the estimated peak NE blood concentration, with 0.1 µg/mL detection threshold. In response to SpNS, the signal increases within seconds, returns to baseline minutes later, and is blocked by interventions that deplete NE or inhibit NE release. The signal is elicited by efferent, but not afferent, electrical or optogenetic vagus nerve stimulation, and by splanchnic nerve stimulation. The magnitude of the signal during SpNS is inversely correlated with subsequent TNF suppression in endotoxemia and explains 40% of the variance in TNF measurements. CONCLUSIONS: FSCV in the spleen provides a marker for real-time monitoring of anti-inflammatory activation of the splenic innervation during autonomic stimulation.

IDH mutations in G2-3 conventional central bone chondrosarcoma: a mono institutional experience.

BACKGROUND: Heterozygous isocitrate dehydrogenase (IDH) mutations occur in about half of conventional central bone chondrosarcomas (CCBC). Aim of this study was to assess the frequency and prognostic impact of IDH mutations in high grade CCBC patients. METHODS: 64 patients with G2 and G3 CCBC were included. DNA extraction, PCR amplification of IDH1/2 exon 4s, and sequencing analysis with Sanger were performed. RESULTS: IDH mutations were detected in 24/54 patients (44%): IDH1 in 18, IDH2 in 4, and both IDH1/2 in 2 patients. The frequency of mutations was 37% in G2 vs. 69% in G3 (p = 0.039), and 100% in three Ollier disease associated chondrosarcoma. 5-year overall survival (OS) at 124 months (range 1-166) was 51%, with no significant difference based on the IDH mutational status: 61% in IDHmut vs. 44% in IDH wild type (IDHwt). The 5-year relapse free survival (RFS) was 33% (95% CI:10-57) for IDHmut vs. 57% (95%CI: 30-77) for IDHwt. Progression free survival (PFS) was 25% (95%CI:1-65) IDHmut vs. 16% (95%CI: 0.7-52) IDHwt. 55% (5/9) of IDHmut G2 became higher grade at the recurrence, as compared with 25% (3/12) of G2 IDHwt. CONCLUSIONS: This study shows a higher frequency of IDH mutations in G3 CCBC as compared with G2. No significant differences in OS, RFS, and PFS by mutational status were detected. After relapse, a higher rate of G3 for IDH mutated CCBC was observed.

Acaricide resistance and novel photosensitizing approach as alternative acaricides against the camel tick, Hyalomma dromedarii.

The control of the camel tick, Hyalomma dromedarii is very crucial. This study evaluated the novel toxicity of photosensitizers and Phoxim insecticide against H. dromedarii males using the adult immersion tests. Ticks were subjected to sunlight for 10 min post-treatment (PT). The optical characters of the applied materials were determined by UV-Vis spectroscopy (250-900 nm wavelengths). The intensity of spectra decreased as dye concentration decreased. The optical bandgap energies of the dyes at different concentrations were not changed as the concentration changed and decreased as the absorption peak of individual dyes red-shifted. The mortalities 72 h PT reached 42.2%, 44.4%, 51.1%, 71.1%, 46.7%, 48.9%, 44.4%, and 55.6% for chlorophyllin, echinochrome, field stain, methylene blue, phthalocyanine, rhodamine 6G, riboflavin, and safranin, respectively. Methylene blue recorded the highest median lethal concentration (LC50 = 127 ppm) followed by safranin, field stain, rhodamine 6G, phthalocyanine, echinochrome riboflavin, and chlorophyllin (LC50 = 209, 251, 271, 303, 324, 332, and 362 ppm, respectively, 72 h PT). Their median lethal time, LT50, values PT with 240 ppm were 45, 87, 96, 72, 129, 115, 131, and 137 h, respectively. The relative toxicities of the LC50 values 72 h PT showed that chlorophyllin, echinochrome, field stain, methylene blue, phthalocyanine, rhodamine 6G, riboflavin, and safranin were 3.2, 3.6, 4.6, 9.1, 3.8, 4.3, 3.5, and 5.6 times, respectively, more effective than Phoxim. Methylene blue, safranin, and field stain showed a broad absorbance area indicating a large photoactivity and better phototoxicity and could be used as alternative agents to synthetic acaricides.

Impacts of selenium nanoparticles and spirulina alga to alleviate the deleterious effects of heat stress on reproductive efficiency, oxidative capacity and immunity of doe rabbits.

Effects of dietary inclusion of spirulina platensis (SP) and selenium nanoparticles (SeNPs) combination (SP-SeNPs) on the reproductive performance in vivo and in vitro, reproductive and metabolic hormones, hemato-bichemical parameters, oxidative stress, and immunity of heat-stressed doe rabbis were evaluated. All supplements significantly increased live litter size at birth and weaning, viability rate at birth, hemoglobin and red blood cells, and plasma T3, T4, insulin, total proteins and albumin compared with control. Plasma estradiol 17-ß (pre-mating), progesterone (mid-pregnancy), and prolactin (day -7 postpartum) were significantly increased only by SeNPs (0.3, 0.4, and 0.5 mg/kg). All dietary supplements significantly reduced WBCs, cortisol, lipid profile, and improved liver and kidney functions. Immunoglobulins levels, antioxidants capacity were significantly increased, superoxide dismutase was increased by SeNPs (0.4 and 0.5 mg/kg), while malondialdehyde was reduced by 0.3, 0.4 and 0.5 SeNPs mg/kg. Sexual receptivity, pregnancy rate, viability rate at weaning, ovulation rate, and embryo quality were significantly increased by increasing SeNPs above 0.1 mg, while embryo yield was increased by >0.2 mg SeNPs/kg. A combination of SP and SeNPs, could be potentially used as a strong antioxidant to enhance heat regulation and doe rabbit reproduction via improving reproductive and metabolic hormones, antioxidant status and immunological parameters.

Ameliorative impacts of sodium humate on heat-stressed laying Japanese quail (Coturnix coturnix Japonica).

A total of 300 laying Japanese quails (230.10 ± 20 g body weight) divided into four groups (15 birds in 5 replicates/group) were used to examine the impacts of dietary sodium humate (SH) supplementation at levels of 0% (control diet), 0.2%, 0.4% and 0.6% on egg variables and physiological merits of laying quails for 10 weeks under heat stress conditions (15 June and 23 August 2021). Results showed 0.4% SH increased (p < 0.05) weight (12.27 vs. 11.91 g), production (79.84% vs. 69.20%), mass (597.13 vs. 510.48 g) and brokenness (2.8% vs. 5.4%) of eggs as compared to control. Egg shape, shell thickness, shell strength and cholesterol content as well as feed conversion ratio were higher (80.2, 295.8 µm, 1.468 kg/cm,2 11.08 mg/g and 2.69, p < 0.05) in 0.4% SH than in control group (75.2, 279.0 µm, 1.304 kg/cm,2  14.94 mg/g and 2.76). Feed intake, percentages of eggs' shells, yolk, albumen and serum biochemistry (total protein, albumin, AST and HDL) were not altered with the dietary SH. Birds fed on SH diets showed higher levels of globulin, calcium and phosphorus, as well as lower contents of albumin/globulin ratio, triglycerides, cholesterol, corticosterone compared with the control. Regression analysis of antioxidants expected higher total antioxidant capacity (TAC), superoxide dismutase, glutathione peroxidase at 0.35%, and glutathione at 0.40% SH, while the lowest concentration of malondialdehyde was computed at 0.45%. Similarly, immunoglobulins (IgG and IgM) maximum values were determined at 0.35% and 0.40% levels. Moreover, the concentration of tumour necrosis factor-alpha increased (p < 0.05) in all SH levels as compared to the control group. It is conceivable to conclude that the dietary implementation of SH at a level of 0.4% improved egg variables and well-being aspects of laying quail exposed to heat stress conditions.

Analysis of the interrelationships of stakeholders involved in the management of transhumance in southern Benin.

The management of pastoral mobility is a stakeholder-centered approach for the integration of resource conservation and agricultural development. This study aimed to characterize the stakeholders of transhumance and to analyze their influence in the municipality of Djidja in southern Benin. For this purpose, semi-structured interviews were conducted with 300 stakeholders involved in transhumance and pastoral resource management. The Likert scale (1 to 5) was used to assess the levels of influence and focus groups were conducted. The results showed that several stakeholders (transhumant herders, agro-pastoralists, farmers, hunters, fishermen, loggers, gendarmerie, Garso, CTAF, cattle farmers' associations, farmers' associations, SCDA, and communal transhumance committee) were involved in transhumance with diverse interests, backgrounds, knowledge, and power (P < 0.05). More than half of the farmers (72%) blame transhumant herders whose practices are source of multiple conflicts among (please mention what kind of conflicts and with whom by providing example). Statistical analysis indicated a strong influence with significant differences (P < 0.001) in the pastoral resources by four stakeholders including the communal transhumance committee, the association of herders, the Garso (scout and intermediary for transhumant herders), and the transhumant herder. This research demonstrates how the systematic analysis of the activities carried out by the stakeholders, the interconnected activities between them, and their relationships can offer insights for a better coordination of transhumance. For effective pastoral management, it is therefore important to build a dialogue between the different stakeholders involved in transhumance in southern Benin.

Galantamine attenuates autoinflammation in a mouse model of familial mediterranean fever.

BACKGROUND: Autoinflammatory diseases, a diverse group of inherited conditions characterized by excessive innate immune activation, have limited therapeutic options. Neuroimmune circuits of the inflammatory reflex control innate immune overactivation and can be stimulated to treat disease using the acetylcholinesterase inhibitor galantamine. METHODS: We tested the efficacy of galantamine in a rodent model of the prototypical autoinflammatory disease familial Mediterranean fever (FMF). Multiple chronic disease markers were evaluated in animals that received long-term galantamine treatment compared to vehicle. RESULTS: Long-term treatment with galantamine attenuated the associated splenomegaly and anemia which are characteristic features of this disease. Further, treatment reduced inflammatory cell infiltration into affected organs and a subcutaneous air pouch. CONCLUSIONS: These findings suggest that galantamine attenuates chronic inflammation in this mouse model of FMF. Further research is warranted to explore the therapeutic potential of galantamine in FMF and other autoinflammatory diseases.

Chemotherapeutics and CAR-T Cell-Based Immunotherapeutics Screening on a 3D Bioprinted Vascularized Breast Tumor Model.

Despite substantial advancements in development of cancer treatments, lack of standardized and physiologically-relevant in vitro testing platforms limit the early screening of anticancer agents. A major barrier is the complex interplay between the tumor microenvironment and immune response. To tackle this, a dynamic-flow based 3D bioprinted multi-scale vascularized breast tumor model, responding to chemo and immunotherapeutics is developed. Heterotypic tumors are precisely bioprinted at pre-defined distances from a perfused vasculature, exhibit tumor angiogenesis and cancer cell invasion into the perfused vasculature. Bioprinted tumors treated with varying dosages of doxorubicin for 72 h portray a dose-dependent drug response behavior. More importantly, a cell based immune therapy approach is explored by perfusing HER2-targeting chimeric antigen receptor (CAR) modified CD8+ T cells for 24 or 72 h. Extensive CAR-T cell recruitment to the endothelium, substantial T cell activation and infiltration to the tumor site, resulted in up to ≈70% reduction in tumor volumes. The presented platform paves the way for a robust, precisely fabricated, and physiologically-relevant tumor model for future translation of anti-cancer therapies to personalized medicine.

Effect of addition of l-carnitine to cryopreservation extender on rabbit post-thaw semen parameters, antioxidant capacity, mitochondrial function, apoptosis and ultrastructure changes.

l-Carnitine (LC) is considered to be a natural antioxidant agent that could be used to improve the efficiency of reproduction. However, the precise machinery of the effect of LC supplementation on frozen-thawed rabbit sperm has not been evaluated. Thus, the aim of this study was to evaluate the effect of the addition of LC to a freezing medium on parameters and ultrastructure changes in frozen-thawed rabbit sperm. Rabbit bucks (7 months of age) were involved, and semen was collected using the artificial vagina method. Pooled rabbit semen was cryopreserved in a tris yolk fructose extender without any supplement (LC0, control group) or with LC at levels of 1, 2 or 4 mM (LC1, LC2 and LC4, respectively). The samples were then loaded into 0.25-ml straws and frozen over liquid nitrogen vapours before being plunged into the liquid nitrogen and stored at -196°C until evaluation. Data showed that the addition of LC significantly increased sperm motility, viability and membrane function, while sperm abnormalities decreased (p < .001). Sperm-like apoptosis (early, late and necrosis spermatozoa) was lower in the LC4 group compared with the other groups. l-Carnitine addition significantly enhanced the total antioxidant capacity and superoxide dismutase and glutathione peroxide activities and significantly reduced the protein carbonyl and malondialdehyde levels compared with the control group. Moreover, electron microscopy images demonstrated that LC addition (2 or 4 mM) preserved the acrosome and plasma membrane and protected the ultrastructure integrity of the cryopreserved spermatozoa in relation to the control group. Spermatozoa treated with LC exhibited higher mitochondria membrane potential (MMP) values compared with the control group. We conclude that the addition of LC (4 mM) to the freezing extender enhanced the quality, increased the antioxidant capabilities, preserved the ultrastructure integrity and reduced lipid and protein peroxidation as well as increased MMP activity of frozen-thawed rabbit sperm.

Impacts of different antioxidants sources on semen quality and sperm fertilizing ability of Muscovy ducks under high ambient temperature.

The potentiality of coenzyme Q10 (CoQ10), D-Aspartic acids (D-Asp), Maca or vitamin C, as antioxidant agents, to reduce negative impacts of high ambient temperature on semen quality, oxidative capacity and fertility of Muscovy ducks was investigated. Seventy-five Muscovy males (34-wk of age) were distributed randomly into five experimental groups of fifteen ducks each. The first group was fed a basal diet without supplementation and served as a control. The other four groups were fed a basal diet supplemented with 400 mg CoQ10, 400 mg D-Asp, 500 mg Maca and 200 mg vitamin C (ascorbic acid) per kg diet for 17 consecutive weeks under high ambient temperature conditions. The dietary inclusion of antioxidants significantly maintains better semen variables and a higher fertility rate either for fresh or preserved semen. Among the tested antioxidants, the Maca group showed the best status and outperformed the others in terms of motility, viability, sperm cell concentration, intact acrosome and membrane integrity percentages, total proteins, total antioxidants capacity, glutathione peroxidase, superoxide dismutase (SOD), malondialdehyde (MDA), testosterone, and the fertility rate for the fresh semen, as well as, forward motility, SOD and MDA for the preserved semen. The CoQ10 showed similar results to Maca in some measurements. Conversely, the basal diet had the poorest performance in all examined variables. The dietary incorporation of antioxidants (Maca or CoQ10) enhances fresh and preserved semen quantity and quality, as well as the fertility rate of Muscovy males under high ambient temperature conditions.

Molecular surveillance revealed no SARS-CoV-2 spillovers to raccoons (Procyon lotor) in four German federal states.

Raccoons (Procyon lotor), which are closely related to the family Mustelidae, might be susceptible to natural infection by SARS-CoV-2. This assumption is based on experimental evidence that confirmed the vulnerability of farmed fur-carnivore species, including Procyon lotor to SARS-CoV-2. To date, there are no reports of natural SARS-CoV-2 infections of raccoons in Germany. Here, we use RT-PCR to analyze 820 samples from raccoons hunted in Germany with a focus on 4 German federal states (Saxony-Anhalt, Thuringia, Hesse, North Rhine-Westphalia). Lung tissues were homogenized and processed for RNA extraction and RT-qPCR for detecting SARS-CoV-2 was performed. No viral RNA was detected in any samples (0/820). Next, we compared raccoons and human ACE-2 residues that are known to serve for binding with SARS-CoV-2 receptor binding domain (RBD). Interestingly, we found only 60% identity on amino acid level, which may have contributed to the absence of SARS-CoV-2 infections in raccoons. In conclusion, the chance of raccoons being intermediate reservoir hosts for SARS-CoV-2 seems to be very low.

Intra-Operative Bioprinting of Hard, Soft, and Hard/Soft Composite Tissues for Craniomaxillofacial Reconstruction.

Reconstruction of complex craniomaxillofacial (CMF) defects is challenging due to the highly organized layering of multiple tissue types. Such compartmentalization necessitates the precise and effective use of cells and other biologics to recapitulate the native tissue anatomy. In this study, intra-operative bioprinting (IOB) of different CMF tissues, including bone, skin, and composite (hard/soft) tissues, is demonstrated directly on rats in a surgical setting. A novel extrudable osteogenic hard tissue ink is introduced, which induced substantial bone regeneration, with ≈80% bone coverage area of calvarial defects in 6 weeks. Using droplet-based bioprinting, the soft tissue ink accelerated the reconstruction of full-thickness skin defects and facilitated up to 60% wound closure in 6 days. Most importantly, the use of a hybrid IOB approach is unveiled to reconstitute hard/soft composite tissues in a stratified arrangement with controlled spatial bioink deposition conforming the shape of a new composite defect model, which resulted in ≈80% skin wound closure in 10 days and 50% bone coverage area at Week 6. The presented approach will be absolutely unique in the clinical realm of CMF defects and will have a significant impact on translating bioprinting technologies into the clinic in the future.