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Mitofusin-2 (MFN2), a large GTPase residing in the mitochondrial outer membrane and mutated in Charcot-Marie-Tooth type 2 disease (CMT2A), is a regulator of mitochondrial fusion and tethering with the ER. The role of MFN2 in mitochondrial transport has however remained elusive. Like MFN2, acetylated microtubules play key roles in mitochondria dynamics. Nevertheless, it is unknown if the α-tubulin acetylation cycle functionally interacts with MFN2. Here, we show that mitochondrial contacts with microtubules are sites of α-tubulin acetylation, which occurs through MFN2-mediated recruitment of α-tubulin acetyltransferase 1 (ATAT1). This activity is critical for MFN2-dependent regulation of mitochondria transport, and axonal degeneration caused by CMT2A MFN2 associated R94W and T105M mutations may depend on the inability to release ATAT1 at sites of mitochondrial contacts with microtubules. Our findings reveal a function for mitochondria in α-tubulin acetylation and suggest that disruption of this activity plays a role in the onset of MFN2-dependent CMT2A., Competing Interests: The authors declare no competing interests., (© 2024 The Authors.)

The primary goal of this study was to explore the social buffering effect that humans offer to goats and dogs with limited exposure to human socialization, particularly in situations involving interactions with unfamiliar humans. A total of 13 dogs and 14 goats were selected for the study, all of which had limited prior socialization with humans. Each animal was placed in a testing room with unfamiliar humans for 15 min. Three experimenters aimed to establish a comfortable environment, encouraging social interaction by offering food to the animals and assessing the animals' willingness to accept food and their response to being approached and petted. If both conditions were satisfied, the animals were classified as "social". If one or none of the conditions were met, the animals were classified as "not social". Cortisol levels were measured by collecting blood samples before and after the test. Non-parametric tests together with a GzLM showed that the effect of human social buffering in goats was different in comparison to dogs: goats exhibited higher cortisol levels after the test, while dogs did not show a significant change. Further analysis demonstrated that "social" goats had a lower likelihood of experiencing significant changes in cortisol levels than dogs. Thus, once human interactions are accepted, both species could benefit from social buffering. In summary, this study enhances our understanding of how dogs and goats respond to social interactions with humans in the social buffering effect., (© 2024. The Author(s).)

The study aimed to explore how limited human socialization affects the socio-cognitive abilities and interactions with unfamiliar individuals of a selected group of domesticated dogs and goats. These animals were raised and kept under conditions characterized by limited human socialization, and their behavior was assessed using the "impossible task" paradigm. The study found that dogs, with a history of cooperative interactions and human companionship, exhibited more frequent social engagement with human experimenters in the experimental setting than goats, traditionally domesticated for utilitarian purposes. However, differences in interaction duration and latency were not significant, highlighting the complexity of these interactions. The results suggest that domestication history and behavioral ecology play significant roles in shaping animals' willingness to engage with humans. However, this study acknowledges limitations, such as the specific population studied, and calls for further research with larger and more diverse samples to generalize these findings. Understanding the interplay between domestication history, behavioral ecology, and human socialization could provide insights into the complex factors influencing animal-human interactions and cognitive behaviors, with implications for animal welfare and human-animal relationships.

In this study, we explored the correlations between circulating levels of oxytocin, cortisol, and different social behaviors toward humans in 26 Italian Red Pied calves (all females, with an average age of 174 ± 24 days) using the impossible task paradigm. This paradigm has proved fruitful in highlighting the effect of socialization on the willingness to interact with humans in several domesticated species. The test consists of the violation of an expectation (recovering food from an experimental apparatus) while a caregiver and a stranger are present. Immediately after the end of the test (less than one minute), blood was collected from the coccygeal vein. Statistics were performed by the Spearman's rank correlation; significant differences were adjusted according to Bonferroni's correction. Cortisol correlates positively (ρ = 0.565; p < 0.05) with the latency of behaviors directed at the caregiver, and the duration of behaviors directed at the apparatus correlates negatively with both the caregiver (ρ = -0.654; p < 0.05) and a stranger (ρ = -0.644; p < 0.05). Contrary to what is reported in the literature on cows, no correlations were found between oxytocin levels and direct behaviors toward the caregiver. This highlights a different behavioral strategy between calves and cows when placed in front of an impossible task.

Peripheral neuropathy is a common disorder that results from nerve damage in the periphery. The degeneration of sensory axon terminals leads to changes or loss of sensory functions, often manifesting as debilitating pain, weakness, numbness, tingling, and disability. The pathogenesis of most peripheral neuropathies remains to be fully elucidated. Cumulative evidence from both early and recent studies indicates that tubulin damage may provide a common underlying mechanism of axonal injury in various peripheral neuropathies. In particular, tubulin post-translational modifications have been recently implicated in both toxic and inherited forms of peripheral neuropathy through regulation of axonal transport and mitochondria dynamics. This knowledge forms a new area of investigation with the potential for developing therapeutic strategies to prevent or delay peripheral neuropathy by restoring tubulin homeostasis., Competing Interests: Declaration of Competing Interest The authors report no conflict of interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Microtubules play fundamental roles in the maintenance of neuronal processes and in synaptic function and plasticity. While dynamic microtubules are mainly composed of tyrosinated tubulin, long-lived microtubules contain detyrosinated tubulin, suggesting that the tubulin tyrosination/detyrosination cycle is a key player in the maintenance of microtubule dynamics and neuronal homeostasis, conditions that go awry in neurodegenerative diseases. In the tyrosination/detyrosination cycle, the C-terminal tyrosine of α-tubulin is removed by tubulin carboxypeptidases and re-added by tubulin tyrosine ligase (TTL). Here we show that TTL heterozygous mice exhibit decreased tyrosinated microtubules, reduced dendritic spine density and both synaptic plasticity and memory deficits. We further report decreased TTL expression in sporadic and familial Alzheimer's disease, and reduced microtubule dynamics in human neurons harbouring the familial APP-V717I mutation. Finally, we show that synapses visited by dynamic microtubules are more resistant to oligomeric amyloid-β peptide toxicity and that expression of TTL, by restoring microtubule entry into spines, suppresses the loss of synapses induced by amyloid-β peptide. Together, our results demonstrate that a balanced tyrosination/detyrosination tubulin cycle is necessary for the maintenance of synaptic plasticity, is protective against amyloid-β peptide-induced synaptic damage and that this balance is lost in Alzheimer's disease, providing evidence that defective tubulin retyrosination may contribute to circuit dysfunction during neurodegeneration in Alzheimer's disease., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Guarantors of Brain.)

This study aimed to investigate the efficacy of DìRelax TM , a nutraceutical formulated to reduce anxiety in dogs, using a randomized controlled trial (RCT) design. The C-BARQ questionnaire, some clinical investigations, and the impossible task test were performed in dogs before and after treatment. The C-BARQ questionnaire is particularly useful for assessing the frequency and severity of problematic behaviors. The impossible task paradigm provides insight into the decision-making processes in the realm of expectancy frustration. Results showed an ameliorative effect on the performances of treated dogs during the solvable phases, with a significant decrease in the time needed to solve the task. No behavioral difference was found between treated and untreated anxious dogs during the unsolvable phase. According to the results from the C-BARQ questionnaire, some of the behaviors appeared to improve. Clinical investigations, including a complete blood cell count and blood chemistry, showed no difference between groups, thus suggesting the safety of the product. In general, this study suggests that DìRelax TM can be safely administered with no adverse effects and can exercise a beneficial effect on anxious dogs by enhancing their cognitive abilities, but further studies should investigate the best method of administration.

This study explored a possible relationship between the circulating oxytocin, cortisol, and the willingness of dairy cows to engage in social behaviors with humans in an experimental context. The behaviors of twenty-nine cows were recorded during the impossible task paradigm, a procedure aimed at creating a violation of expectancy, in the presence of the caregiver and a stranger. The results showed that serum oxytocin levels were positively correlated with duration and negatively correlated with the latency of the cows' social interactions with the caregiver. This research provides a clear correlation between circulating oxytocin and a willingness to engage in social contact with the caregiver, excluding the possible effect of different cortisol levels on such behavior.

Transthyretin (TTR), a plasma and cerebrospinal fluid protein, increases axon growth and organelle transport in sensory neurons. While neurons extend their axons, the microtubule (MT) cytoskeleton is crucial for the segregation of functional compartments and axonal outgrowth. Herein, we investigated whether TTR promotes axon elongation by modulating MT dynamics. We found that TTR KO mice have an intrinsic increase in dynamic MTs and reduced levels of acetylated α-tubulin in peripheral axons. In addition, they failed to modulate MT dynamics in response to sciatic nerve injury, leading to decreased regenerative capacity. Importantly, restoring acetylated α-tubulin levels of TTR KO dorsal root ganglia (DRG) neurons using an HDAC6 inhibitor is sufficient to completely revert defective MT dynamics and neurite outgrowth. In summary, our results reveal a new role for TTR in the modulation of MT dynamics by regulating α-tubulin acetylation via modulation of the acetylase ATAT1, and suggest that this activity underlies TTR neuritogenic function., 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 © 2021 Eira, Magalhães, Macedo, Pero, Misgeld, Sousa, Bartolini and Liz.)

Antimicrobial peptides (AMPs), α- and β-defensins, possess antiviral properties. These AMPs achieve viral inhibition through different mechanisms of action. For example, they can: (i) bind directly to virions; (ii) bind to and modulate host cell-surface receptors, disrupting intracellular signaling; (iii) function as chemokines to augment and alter adaptive immune responses. Given their antiviral properties and the fact that the development of an effective coronavirus disease 2019 (COVID-19) treatment is an urgent public health priority, they and their derivatives are being explored as potential therapies against COVID-19. These explorations using various strategies, range from their direct interaction with the virus to using them as vaccine adjuvants. However, AMPs do not work in isolation, specifically in their role as potent immune modulators, where they interact with toll-like receptors (TLRs) and chemokine receptors. Both of these receptors have been shown to play roles in COVID-19 pathogenesis. In addition, it is known that a healthy lifestyle accompanied by controlled physical activity can represent a natural weapon against COVID-19. In competitive athletes, an increase in serum defensins has been shown to function as self-protection from the attack of microorganisms, consequently a controlled physical activity could act as a support to any therapies in fighting COVID-19. Therefore, including information on all these players' interactions would produce a complete picture of AMP-based therapies' response.

Chemotherapy-induced peripheral neuropathy (CIPN) is a major side effect from cancer treatment with no known method for prevention or cure in clinics. CIPN often affects unmyelinated nociceptive sensory terminals. Despite the high prevalence, molecular and cellular mechanisms that lead to CIPN are still poorly understood. Here, we used a genetically tractable Drosophila model and primary sensory neurons isolated from adult mouse to examine the mechanisms underlying CIPN and identify protective pathways. We found that chronic treatment of Drosophila larvae with paclitaxel caused degeneration and altered the branching pattern of nociceptive neurons, and reduced thermal nociceptive responses. We further found that nociceptive neuron-specific overexpression of integrins, which are known to support neuronal maintenance in several systems, conferred protection from paclitaxel-induced cellular and behavioral phenotypes. Live imaging and superresolution approaches provide evidence that paclitaxel treatment causes cellular changes that are consistent with alterations in endosome-mediated trafficking of integrins. Paclitaxel-induced changes in recycling endosomes precede morphological degeneration of nociceptive neuron arbors, which could be prevented by integrin overexpression. We used primary dorsal root ganglia (DRG) neuron cultures to test conservation of integrin-mediated protection. We show that transduction of a human integrin β-subunit 1 also prevented degeneration following paclitaxel treatment. Furthermore, endogenous levels of surface integrins were decreased in paclitaxel-treated mouse DRG neurons, suggesting that paclitaxel disrupts recycling in vertebrate sensory neurons. Altogether, our study supports conserved mechanisms of paclitaxel-induced perturbation of integrin trafficking and a therapeutic potential of restoring neuronal interactions with the extracellular environment to antagonize paclitaxel-induced toxicity in sensory neurons., Competing Interests: The authors declare no competing interest.

The pathogenesis of chemotherapy-induced peripheral neuropathy (CIPN) is poorly understood. Here, we report that the CIPN-causing drug bortezomib (Bort) promotes delta 2 tubulin (D2) accumulation while affecting microtubule stability and dynamics in sensory neurons in vitro and in vivo and that the accumulation of D2 is predominant in unmyelinated fibers and a hallmark of bortezomib-induced peripheral neuropathy (BIPN) in humans. Furthermore, while D2 overexpression was sufficient to cause axonopathy and inhibit mitochondria motility, reduction of D2 levels alleviated both axonal degeneration and the loss of mitochondria motility induced by Bort. Together, our data demonstrate that Bort, a compound structurally unrelated to tubulin poisons, affects the tubulin cytoskeleton in sensory neurons in vitro, in vivo, and in human tissue, indicating that the pathogenic mechanisms of seemingly unrelated CIPN drugs may converge on tubulin damage. The results reveal a previously unrecognized pathogenic role for D2 in BIPN that may occur through altered regulation of mitochondria motility., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

Alzheimer's disease (AD), a chronic multifactorial and complex neurodegenerative disorder is a leading cause of dementia. Recently, neuroinflammation has been hypothesized as a contributing factor to AD pathogenesis. The role of adaptive immune responses against neuronal antigens, which can either confer protection or induce damage in AD, has not been fully characterized. Here, we measured T cell responses to several potential antigens of neural origin including amyloid precursor protein (APP), amyloid beta (Aβ), tau, α-synuclein, and transactive response DNA binding protein (TDP-43) in patients with AD and age-matched healthy controls (HC). Antigen-specific T cell reactivity was detected for all tested antigens, and response to tau-derived epitopes was particularly strong, but no significant differences between individuals with AD and age-matched HC were identified. We also did not observe any correlation between the antigen-specific T cell responses and clinical variables including age, gender, years since diagnosis and cognitive score. Additionally, further characterization did not reveal any differences in the relative frequency of major Peripheral Blood Mononuclear Cells (PBMC) subsets, or in the expression of genes between AD patients and HC. These observations have not identified a key role of neuronal antigen-specific T cell responses in AD., (Copyright © 2020 Dhanwani, Pham, Premlal, Frazier, Kumar, Pero, Bartolini, Dutra, Marder, Peters, Sulzer, Sette and Lindestam Arlehamn.)

Aim of the present study was to evaluate the potential beneficial and/or adverse effects of Capsicum annuum L. (cv. Fiesta) extracts at two stage of ripening (immature and mature), and at two dosages (low and high) by evaluation of biochemical profile and oxidative status in CD-1 mice. The extracts were daily administered to mice by oral gavage for 20 days. At the end of the trial, the animals were euthanatized and blood was collected. Evidence of liver damage (increase of AST, ALT and bilirubin) in the group receiving the higher dosage of immature peppers extract were observed. Even if no adverse effects were seen at the lower doses, also no signs of beneficial effects in term of health status, biochemical profile and oxidative status were detected. These results are in contrast with in vitro findings and raise doubts about the possible use of Capsicum annuum L. (cv. Fiesta) as a nutritional supplement.

The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100 μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1 hr with 0, 20 and 100 µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (p < .05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8 ± 1.5 and 69.1 ± 1.4, respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (p < .05), while the percentage of spermatozoa with high ΔΨm increased (p < .05). The results of Experiment 2 showed that 1-hr treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4 ± 5.8 and 83.7.1 ± 1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (p < .05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (p < .05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0 ± 11.4). In conclusion, the treatment with 100 µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100 µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation., (© 2020 Blackwell Verlag GmbH.)

ARFGEF1 encodes a guanine exchange factor involved in intracellular vesicle trafficking, and is a candidate gene for childhood genetic epilepsies. To model ARFGEF1 haploinsufficiency observed in a recent Lennox Gastaut Syndrome patient, we studied a frameshift mutation (Arfgef1 fs ) in mice. Arfgef1 fs/+ pups exhibit signs of developmental delay, and Arfgef1 fs/+ adults have a significantly decreased threshold to induced seizures but do not experience spontaneous seizures. Histologically, the Arfgef1 fs/+ brain exhibits a disruption in the apical lining of the dentate gyrus and altered spine morphology of deep layer neurons. In primary hippocampal neuron culture, dendritic surface and synaptic but not total GABA A receptors (GABA A R) are reduced in Arfgef1 fs/+ neurons with an accompanying decrease in the number of GABA A R-containing recycling endosomes in cell body. Arfgef1 fs/+ neurons also display differences in the relative ratio of Arf6 + :Rab11 + :TrfR + recycling endosomes. Although the GABA A R-containing early endosomes in Arfgef1 fs/+ neurons are comparable to wildtype, Arfgef1 fs/+ neurons show an increase in the number of GABA A R-containing lysosomes in dendrite and cell body. Together, the altered endosome composition and decreased neuronal surface GABA A R results suggests a mechanism whereby impaired neuronal inhibition leads to seizure susceptibility., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The aim of this research was to investigate the influence of hydrogenated palm oil (HPO) added to a dairy goat diet on serum biochemistry and progesterone levels. Thirty pregnant Cilentana dairy goats were equally divided into 2 groups (control [CTR] and HPO groups). After kidding, concentrated feed for both groups was gradually increased up to 400 g/(animal·d), and the HPO group received 50 g/(animal·d) of HPO. Supplementation with HPO significantly increased cholesterol levels (mg/dL, 63.80 vs. 54.68 at 30 d, P  ≤ 0.05; 78.20 vs. 58.00 at 60 d, P  ≤ 0.05; 83.80 vs. 57.83 at 120 d, P  ≤ 0.01) compared with the CTR group although no significant differences were detected for liver and kidney function indicators. Moreover, other biochemical parameters were not affected by HPO supplementation thus suggesting no change occurred in lipid and protein metabolism. Furthermore, a significant correlation was found between progesterone levels and serum cholesterol ( r  = 0.65, P  ≤ 0.01) although these were not significantly higher in HPO supplemented goats. The dose and time of HPO supplementation appears critical as regards assessing the limits between the risks and benefits of HPO supplementation in dairy goats. At the tested dose, HPO was well tolerated by the animals and may represent a useful tool to increase energy availability during highly demanding periods.

The aim of this work was to propose a model of free-range raising for rabbit able to maximize the animal welfare and at the same time the productive performances through the use of mirrors. A total of 81 rabbits were allocated into free-range areas and divided into three groups (nine replicates per group): in the first group (face to face, F2F), the rabbits of each replicate could see each other. In the second group (blind) each replicate was isolated from the others; in the third group (mirrors), the replicates were divided as for the Blind group but two mirrors were placed in a corner of the perimeter. The blind group rabbits showed the lowest final weight ( p < 0.05), while rabbits from the mirrors groups showed the best FCR and net dressing out values. The blind group showed the highest production of total short chain fatty acids, acetate ( p < 0.05) and propionate ( p < 0.01). The F2F rabbits showed higher levels of creatine phosphokinase and lactate dehydrogenase and lower values of blood glucose than those of the other groups, due to the higher locomotion activity. The use of mirrors can improve rabbit's growth performance and carcass traits by lowering the rabbit's locomotion activity in comparison to the other tested systems., Competing Interests: The authors declare no conflict of interest.

The effect of pasture on the stearoyl-CoA desaturase (SCD) and miRNA 103 expression was evaluated on dairy goats divided into two homogeneous groups (G, grazing, and S, stable). Group S was housed in a stall and received alfalfa hay as forage, while group G was led to pasture. The goats of both the groups received the same amount of concentrate. Milk yield did not differ statistically between the groups. Group G showed significantly higher fat (4.10% vs. 2.94%, p < 0.01) and protein percentage (3.43% vs. 3.25%; p < 0.05) than group S. Among milk fatty acids, group S showed significantly higher levels of saturated fatty acids (SFA) and lower values of mono-unsaturated fatty acid (MUFA). The percentages of polyunsaturated fatty acid (PUFA) were not different between groups even if pasture significantly affected the percentages of C18:3 and total omega 3. In group G, total CLAs were twice than in group S (0.646% vs. 0.311%; p < 0.01) mainly due to the differences in CLA cis9 trans 11 (0.623% vs. 0.304%; p < 0.01). Milk total CLA in grazing group was significantly ( p < 0.01) higher in August according to the highest value of both linoleic and α-linolenic acids in the pasture. In grazing animals, SCD expression decreased from April to June, increased in July and decreased again in August, while it was almost unvaried along the trial in group S. By contrast, the expression of miRNA 103 showed a similar trend for both groups, decreasing from April to June, increasing in July and falling down in August. To our knowledge, this is the first observation of the effects of pasture on miRNA expression in milk from ruminant species.

The objective of this research was to investigate the efficacy of DìSenior TM , a nutraceutical formulated to improve cognitive functions in elderly dogs. To this purpose, some clinical and metabolic investigations and a spatial navigation test were performed in treated and untreated dogs. Moreover, the nutraceutical was also tested on primary hippocampal neuron cultures. Results showed no adverse effects on the dogs' health and a positive effect on learning. In vitro effects on neuron cultures showed an increase in the level of cFOS in treated neurons compared with the vehicle, suggesting that DiSenior TM has also a positive effect on neuronal functions. Overall, this study suggests that DiSenior TM can exert a beneficial effect on aged dogs by preventing the negative effects of aging on cognition. Further studies are needed to assess the mechanisms by which it acts on neurons and the specific effect of the different components alone or combined.

Given probable the increment in the nutritional needs of both humans and animals, animal production will have increased dramatically by 2050. Insect meals could be an alternative protein source for livestock, and they would also be able to reduce the environmental problems related to intensive animal production system. The aim of this study was to evaluate productive performance, blood analysis, nutrient digestibility, and changes in the internal organs of laying hens fed Hermetia illucens larvae meal (HI) at two different levels in substitution (25 or 50%) of soybean meal (SBM). A total of 162 Hy-line Brown hens (sixteen weeks old) were equally divided into three experimental groups and fed isoprotein and isoenergetic diets. Egg weight, feed intake, and feed conversion rate were not affected by the soybean meal substitution at both inclusion levels of insect meal. Egg mass was positively affected by the insect meal diets, as was the lay percentage, although only at the lowest inclusion level. Dry matter, organic matter, and crude protein digestibility coefficients were lower for the HI50 diet, probably due to the negative effect of chitin. A reduction in serum cholesterol and triglycerides was observed in both insect-meal fed groups, while serum globulin level increased only at the highest level of insect meal inclusion, and, consequently, the albumin to globulin ratio decreased. Overall, a protein replacement of 25% with an insect meal from Hermetia illucens larvae in the diet of laying hens seems to be more suitable and closer to the optimal level., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The aim of this work was to evaluate whether the treatment with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK) during cryopreservation and post-warming in vitro culture improves cryotolerance of bovine in vitro produced (IVP) embryos. Abattoir derived bovine oocytes were in vitro matured, fertilized and cultured according to standard procedure. On Day 7, embryo yields were assessed and blastocysts randomly divided in 2 groups: vitrification and post-warming culture in the absence (n = 184) or presence (n = 156) of 20 μM Z-VAD-FMK. Resistance to cryopreservation was evaluated post-warming culture by assessing the survival rate and hatching rate. Differential staining combined with in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) technique was performed to evaluate total cells number, cell allocation into inner cell mass (ICM) and trophectoderm (TE) lineages, as well as the DNA fragmentation rate of vitrified blastocysts, while immunohystochemical staining was used to assess the level of cleaved-caspase 3. It was demonstrated that inhibition of caspase activity by Z-VAD-FMK increases embryo cryotolerance, as indicated by higher survival (76.1 vs 51.1%; P < 0.01) and hatching rates (26.5 vs 17.6%; P < 0.05) after 48 h of post-warming culture. Furthermore, Z-VAD-FMK decreased both the average number (4.7 ± 0.3 vs 7.7 ± 0.5; P < 0.01) and the percentage (3.4 ± 0.2 vs 6.1 ± 0.5; P < 0.01) of DNA fragmented cells in blastocysts compared to the control. No differences were recorded in the average number of ICM, TE and total cells between groups. The level of cleaved-caspase-3, the downstream effector of apoptosis, and its relative percentage on total area of blastocysts was reduced (P < 0.01) in the presence of Z-VAD-FMK both at thawing (1.29 ± 0.17 vs 3.24 ± 0.46) and after 48 h post-warming culture (1.46 ± 0.17 vs 5.06 ± 0.41). In conclusion, the addition of 20 μM Z-VAD-FMK during vitrification/warming and post-warming culture partially inhibits cryopreservation-induced apoptosis by reducing the level of active caspase 3, suggesting a potential use as an additive to ameliorate the efficiency of embryo cryopreservation in cattle, critical for a further diffusion of IVEP technology in the field. Further studies are though needed to evaluate the effect of Z-VAD-FMK on post-transfer embryo development before considering a commercial application., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Oligomeric Amyloid β 1-42 (Aβ) plays a crucial synaptotoxic role in Alzheimer's disease, and hyperphosphorylated tau facilitates Aβ toxicity. The link between Aβ and tau, however, remains controversial. In this study, we find that in hippocampal neurons, Aβ acutely induces tubulin posttranslational modifications (PTMs) and stabilizes dynamic microtubules (MTs) by reducing their catastrophe frequency. Silencing or acute inhibition of the formin mDia1 suppresses these activities and corrects the synaptotoxicity and deficits of axonal transport induced by Aβ. We explored the mechanism of rescue and found that stabilization of dynamic MTs promotes tau-dependent loss of dendritic spines and tau hyperphosphorylation. Collectively, these results uncover a novel role for mDia1 in Aβ-mediated synaptotoxicity and demonstrate that inhibition of MT dynamics and accumulation of PTMs are driving factors for the induction of tau-mediated neuronal damage., (© 2017 Qu et al.)

The aim of the research was to study the effects of an insect meal from Hermetia illucens larvae (HILM) as complete replacement of soybean meal (SBM) on productive performance and blood profiles of laying hens, from 24 to 45 wk of age. A total of 108 24-week-old Lohmann Brown Classic laying hens was equally divided into 2 groups (54 hens/group, 9 replicates of 6 hens/group). From 24 to 45 wk of age, the groups were fed 2 different isoproteic and isoenergetic diets: the control group (SBM) was fed a corn-soybean meal based diet, while in the HILM group the soybean meal was completely replaced by Hermetia illucens larvae meal. Feed intake, number of eggs produced, and egg weight were recorded weekly along the trial. At 45 wk of age, blood samples were collected from 2 hens per replicate. The use of HIML led to a more favorable (P < 0.01) feed conversion ratio in hens but lay percentage, feed intake, average egg weight, and egg mass were higher (P < 0.01) in hens fed the SBM diet. Hens fed insect meal produced a higher percentage of eggs from small (S), medium (M), and extra-large (XL) classes (P < 0.01) than SBM, while the SBM group had a higher percentage of eggs from the large (L) class (P < 0.01). The levels of globulin and albumin to globulin ratio were, respectively, higher and lower (P < 0.05) in HILM than the SBM group. Cholesterol and triglycerides were higher (P < 0.05 and P < 0.01, respectively) in hens from SBM than in the HILM group. Blood levels of Ca were higher (P < 0.01) in hens fed insect meal, while creatinine was higher (P < 0.01) in blood of hens fed SBM. Hermetia illucens larvae meal can be a suitable alternative protein source for laying hens even if the complete replacement of soybean meal needs further investigation to avoid the negative effects on feed intake., (© 2017 Poultry Science Association Inc.)

Aquaporins (AQPs) are membrane channel proteins that play a role in regulating water permeability in many tissues. To date, seven isoforms of AQPs have been reported in the gastrointestinal tract in different mammalian species. In contrast, both tissue distribution and expression of AQPs are unknown in the buffalo. The purpose of this study was to investigate the expression of both AQP4 and AQP5 mRNAs and their relative proteins in the large intestinal tracts of buffalo calves after colostrum suckling using reverse transcriptase polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry. Our results revealed a diversified tissue AQP4 and AQP5 immunolocalization accompanied by their highest expression in the tissues of colostrum-suckling buffalo calves confirmed by Western blotting. In particular, AQP4 was distributed along the endothelium and enterocytes while AQP5 in the endocrine cells. These findings provide direct evidence for AQP4 and AQP5 expression in the large intestine, suggesting that different AQPs collaborate functionally and distinctively in water handling during intestinal development, especially during the first period after delivery., (© 2015 Blackwell Verlag GmbH.)

The aim of this work was to assess the effect of supplementation of bovine culture medium with the natural antioxidant crocetin on in vitro blastocyst development and quality. This was evaluated as cryotolerance, apoptosis index, and total cells number and allocation. Abattoir-derived oocytes were matured and fertilized in vitro according to standard procedure. Twenty hours after IVF, presumptive zygotes were cultured in synthetic oviduct fluid medium, supplemented with 0, 1, 2.5, and 5 μM crocetin (experiment 1) at 39 °C under humidified air with 5% CO2, 7% O2, and 88% N2. On Day 7, embryo yields were assessed and the blastocysts were vitrified by Cryotop method in 16.5% ethylene glycol, 16.5% DMSO, and 0.5 M sucrose. Finally, blastocysts produced on Day 8 in the absence (control) and presence of 1 μM crocetin were used for terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling and differential staining to evaluate, respectively, the apoptotic rate and the allocation of cells into inner cell mass (ICM) and trophectoderm (TE) lineages (experiment 2). Embryo development was higher in the 1 μM crocetin group compared to the control, both in terms of total embryo output (37.7 ± 4.2%, 52.9 ± 6.3%, 40.9 ± 7.6%, and 42.4 ± 8.7%, respectively, with 0, 1, 2.5, and 5 μM; P < 0.01) and grade 1 and 2 blastocysts (33.6 ± 4.9%, 46.1 ± 7.3%, 37.8 ± 7.9%, and 39.4 ± 7.9%, respectively, with 0, 1, 2.5, and 5 μM; P < 0.05). Moreover, the percentage of fast-developing embryos increased in 1 μM crocetin group compared to the control (23.4 ± 4.7%, 32.7 ± 6.6%, 27.2 ± 6.6%, and 30.1 ± 7.2%, respectively, with 0, 1, 2.5, and 5 μM; P < 0.05). In addition, the enrichment of culture medium with 1 μM crocetin improved embryo cryotolerance compared to the control, as indicated by higher hatching rates recorded after 48 hours postwarming culture (46.5% vs. 60.4%; P < 0.05). Furthermore, 1 μM crocetin decreased both the average number (9.9 ± 0.4 vs. 7.1 ± 0.3) and the percentage of apoptotic cells (7.1 ± 0.4 vs. 4.2 ± 0.2) in blastocysts compared to the control (P < 0.01). However, no differences were recorded in the average number of ICM, TE, and total cells between 1 μM crocetin and control groups. In conclusion, the enrichment of bovine culture medium with 1 μM crocetin increased both blastocyst yield and quality, as indicated by the improved chronology of embryo development, increased resistance to cryopreservation, and reduced incidence of apoptosis., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Functional studies indicate differences in newborn gastrointestinal morphology and physiology after a meal. Both water and solutes transfer across the intestinal epithelial membrane appear to occur via aquaporins (AQPs). Given that the physiological roles of AQP4 and AQP5 in the developing intestine have not been fully established, the objective of this investigation was to determine their distribution, expression and respective mRNA in the small intestine of colostrums-suckling buffalo calves by using immunohistochemistry, Western blot, and reverse transcriptase-PCR analysis. Results showed different tissue distribution between AQP4 and AQP5 with the presence of the former along the enteric neurons and the latter in the endocrine cells. Moreover, their expression levels were high in the ileum of colostrum-suckling buffalo calves. The data present a link between feeding, intestinal development and water homeostasis, suggesting the involvement of these channel proteins in intestinal permeability and fluid secretion/absorption during this stage of development after birth., (Published by Elsevier Ltd.)

Aquaporin-1 (AQP1), a six-transmembrane domain protein, belongs to a highly conserved group of proteins called aquaporins known to regulate permeability across cell membranes. Although the role of AQP1 has been extensively studied, its specific activity along the gastrointestinal tract in animals during early postnatal development is poorly known. This study investigates the expression of AQP1 mRNA and protein in the small and large intestine of water buffalo calves after colostrum ingestion using reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and cellular localization of AQP1 by immunohistochemistry. Our results revealed AQP1 immunoreactivity and the presence of the corresponding mRNA in all the examined tracts of the intestine but with a different cellular localization. Western blotting confirmed the presence of AQP1, with a more intense band in colostrum-suckling animals. These findings offer insights into AQP1 expression in the small and large intestine, suggesting its involvement in osmoregulation in gastrointestinal physiology particularly during the first week after birth in relation to specific maturation of intestinal structures., (© 2014 Blackwell Verlag GmbH.)

Neuronal apoptotic death generally requires de novo transcription, and activation of the transcription factor c-Jun has been shown to be necessary in multiple neuronal death paradigms. Caspase-2 has been implicated in death of neuronal and non-neuronal cells, but its relationship to transcriptional activation has not been clearly elucidated. In the present study, using two different neuronal apoptotic paradigms, β-amyloid treatment and NGF (nerve growth factor) withdrawal, we examined the hierarchical role of caspase-2 activation in the transcriptional control of neuron death. Both paradigms induce rapid activation of caspase-2 as well as activation of the transcription factor c-Jun and subsequent induction of the pro-apoptotic BH3 (Bcl-homology domain 3)-only protein Bim (Bcl-2-interacting mediator of cell death). Caspase-2 activation is dependent on the adaptor protein RAIDD {RIP (receptor-interacting protein)-associated ICH-1 [ICE (interleukin-1β-converting enzyme)/CED-3 (cell-death determining 3) homologue 1] protein with a death domain}, and both caspase-2 and RAIDD are required for c-Jun activation and Bim induction. The present study thus shows that rapid caspase-2 activation is essential for c-Jun activation and Bim induction in neurons subjected to apoptotic stimuli. This places caspase-2 at an apical position in the apoptotic cascade and demonstrates for the first time that caspase-2 can regulate transcription.

Canine platelets have been extensively studied but little is known about specific aspects such as adhesion. Platelet adhesion is a critical step during haemostasis and thrombosis as well as during inflammatory and immunopathogenic responses. The aim of this study was to evaluate the adhesive properties of canine platelets using fibrinogen and collagen as substrates immobilized on plates. Adhesion was monitored for 120 min and the effect of adenosine 5'-diphosphate (ADP) was assayed. The results showed that canine platelets displayed good adhesion activity that was significantly time-dependent. Moreover, ADP was able to enhance platelet adhesion in a dose-dependent manner. The findings aid knowledge of the adhesion process and suggest a specific role of surface platelet receptors in mediating the interaction with extracellular matrix proteins., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Evidence is presented that the buffalo mammary gland contains enzymes that catalyse the synthesis and utilization of glutathione. A significant, inverse correlation (r = 0.79) was detected between colostrum gamma-glutamyltransferase (GGT) and glutathione (GSH), suggesting that the enzyme uses GSH as a substrate for its activity. A similar trend was shown in mammary gland homogenates (r = 0.75). Our results show that GSH is secreted into buffalo colostrum and suggest that the enzyme GGT degrades it. To the authors' knowledge, this is the first demonstration of the involvement of GGT-mediated GSH metabolism in the synthesis of colostrums, which elucidates the role of the enzyme that has always been reported very high in colostrum.

Urocortin (UCN) is a 40 aminoacid peptide which belongs to corticotropin-releasing factor (CRF) family. This family of peptides stimulates the secretion of proopiomelanocortin (POMC)-derived peptides, adrenocorticotropic hormone (ACTH), beta-endorphin and melanocyte-stimulating hormone (MSH) in the pituitary gland. In the present study, using Western blotting and immunohistochemistry, the distribution of UCN in the primary lymphoid organs of the duck was investigated at different ages. In the cloacal burse and thymus, Western blot demonstrated the presence of a peptide having a molecular weight compatible with that of the mammalian UCN. In the cloacal burse, immunoreactivity was located in the medullary epithelial cells and in the follicular associated and cortico-medullary epithelium. In the thymus, immunoreactivity was located in single epithelial cells. Double labelling immunofluorescence studies showed that UCN immunoreactivity completely colocalised with cytokeratin immunoreactivity in both the thymus and cloacal burse. Statistically significant differences in the percentage of UCN immunoreactivity were observed between different age periods in the cloacal burse. The results suggest that, in birds, urocortin has an important role in regulating the function of the immune system.

Forty-four dogs naturally infected by Leishmania infantum were divided into two groups: 20 thrombocytopenic dogs with fewer than 150 x 10(9) platelets/l, and 24 non-thrombocytopenic dogs with more than 200 x 10(9) platelets/l. Ten clinically healthy dogs were used as controls. A haematological profile was obtained and the dogs' serum was used to assess the presence of platelet-binding IgM and IgG antibodies using a flow cytometry technique. Nineteen of the 20 thrombocytopenic dogs, and 13 of the 24 non-thrombocytopenic dogs had detectable levels of platelet-binding immunoglobulins, but none of the control dogs did so. The differences were significantly different for both IgM and IgG platelet-binding antibodies.

Leishmania-infected dogs, which represent an important reservoir of infection in many parts of the world, frequently suffer from haematological disorders, including thrombocytopenia. In this study, the ability of platelets from healthy (control) dogs (n = 11) and from dogs with naturally acquired clinical leishmaniasis (n = 24) to aggregate in the presence of two different agonists (adenosine 5'-diphosphate [ADP] and collagen) was assayed. Haematological parameters examined consisted of the platelet count, prothrombin time, activated partial thromboplastin time (aPTT), fibrinogen concentration and D-dimer concentration. In dogs with leishmaniasis, a significant decrease in ADP- and collagen-induced platelet aggregation was observed. Compared with platelets from the control dogs, those from leishmania-infected dogs showed a higher sensitivity to collagen, as demonstrated by a reduction in platelet aggregation of up to 20.4%, and a significant (P < 0.0001) difference for all the doses tested. With ADP the reduction was up to 10.4%, the difference reaching a significant level of P < 0.0001 only at the maximum dose used. The nature of this response, which was not accompanied by any clinical signs of bleeding other than an increase in aPTT, emphasizes the role of platelets in the parasite-host cell interaction.