Proteomic signature profiling in the cortex of dairy cattle unravels the physiology of brain aging.

Introduction: Aging is a physiological process occurring in all living organisms. It is characterized by a progressive deterioration of the physiological and cognitive functions of the organism, accompanied by a gradual impairment of mechanisms involved in the regulation of tissue and organ homeostasis, thus exacerbating the risk of developing pathologies, including cancer and neurodegenerative disorders. Methods: In the present work, for the first time, the influence of aging has been investigated in the brain cortex of the Podolica cattle breed, through LC-MS/MS-based differential proteomics and the bioinformatic analysis approach (data are available via ProteomeXchange with identifier PXD044108), with the aim of identifying potential aging or longevity markers, also associated with a specific lifestyle. Results and discussion: We found a significant down-regulation of proteins involved in cellular respiration, dendric spine development, synaptic vesicle transport, and myelination. On the other hand, together with a reduction of the neurofilament light chain, we observed an up-regulation of both GFAP and vimentin in the aged samples. In conclusion, our data pave the way for a better understanding of molecular mechanisms underlying brain aging in grazing cattle, which could allow strategies to be developed that are aimed at improving animal welfare and husbandry practices of dairy cattle from intensive livestock.

Preliminary evaluation of the proteomic profiling in the hippocampus of aged grazing cattle.

Brain aging is a physiological process associated with physical and cognitive decline; however, in both humans and animals, it can be regarded as a risk factor for neurodegenerative disorders, such as Alzheimer's disease. Among several brain regions, hippocampus appears to be more susceptible to detrimental effects of aging. Hippocampus belongs to limbic system and is mainly involved in declarative memories and context-dependent spatial-learning, whose integrity is compromised in an age-dependent manner. In the present work, taking advantage of liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomics, we sought to identify proteins differentially expressed in the hippocampus of the aged grazing milk cows. Our exploratory findings showed that, out of 707 identified proteins, 112 were significantly altered in old cattle, when compared to the adult controls, and functional clusterization highlighted their involvement in myelination, synaptic vesicle, metabolism, and calcium-related biological pathways. Overall, our preliminary data pave the way for the future studies, aimed at better characterizing the role of such a subcortical brain region in the age-dependent cognitive decline, as well as identifying early aging markers to improve animal welfare and husbandry practices of dairy cattle from intensive livestock.

Description of Twenty-Nine Animal Hoarding Cases in Italy: The Impact on Animal Welfare.

The hoarding of animals is a psychiatric disease, characterized by a compulsive collection of animals, with a relevant impact upon the care and welfare of animals, as well as on human society. In Italy, there are neither substantial reports nor information shared about such a phenomenon, making it difficult to draw a clear picture of the hoarder profile. Therefore, in the present work, we sought to detail 29 cases of animal accumulators in Italy, who lived within two areas of the Lazio region, and accumulated a total of 1080 animals from 2019 to 2022. In line with other international studies, we observed a prevalence of middle-aged (in their fifties) women, who lived mainly alone in a high level of social and health degradation. Most of the hoarded animals exhibited severe signs of dehydration and malnutrition, muscle hypotrophy, dermatological injuries, and behavioral disorders. Animal hoarding is not yet fully understood nor recognized as a psychosocial disorder, although it produces a deep suffering for the hoarder themselves, as well as corresponding family members, and the animals accumulated. Therefore, given the crucial impact of animal hoarding upon human and animal welfare, cross-cultural networks aimed at properly raising awareness of the problem could be established.

Combining α-s1 Casozepine and Fluoxetine Treatment with a Behavioral Therapy Improves Symptoms in an Aggressive Dog: An Italian Case Report.

Behavioral dysfunctions in dogs represent a critical issue of the human-animal relationship. In particular, aggression can make interspecific coexistence quite complicated within family units, thus exposing all members to greater health risks. In this present study, we documented multiple aggression episodes against one of the two family members caused by a 4-year-old neutered male pure breed Lagotto Romagnolo dog. To minimize impulsivity and anxiety-like behaviors of the patient as much as possible and improve his relationship with the adopting family, we used an interdisciplinary approach, employing specific skilled personnel, including a veterinary behaviorist and a rehabilitating dog instructor. Nine months after fluoxetine treatment (0.8 mg/kg, SID), in combination with oral α-s1 casozepine administration, and behavioral rehabilitation, the owners reported a significant reduction in aggressive events in terms of intensity and frequency. Collectively, our promising data pave the way toward a more detailed characterization of α-s1-casozepine to better evaluate the potential involvement of such a compound in the modulation of aggressive behaviors in dogs affected by relational dysfunctions.

Combining cannabis and melatonin treatment with a rehabilitation program improved symptoms in a dog with compulsive disorder: A case report.

Compulsive disorder in dogs (CD) is characterized by constant and time-consuming repetition of behaviors, emancipated from the environment, that definitely compromise their everyday life activities. Here, we documented the efficacy of a novel approach to counteract the negative symptoms of CD in a 5-year-old mongrel dog, previously found to be resistant to the conventional antidepressant. The patient underwent an integrated and interdisciplinary approach, based on the cannabis and melatonin co-administration, together with a tailored 5-month-lasting behavioral program. Observational findings showed a lower rate of compulsive episodes and better management of the dog as well, when compared to the previous paroxetine treatment. We followed him for an additional four months of therapy, and the owners reported easier management of the dog, as reduction of abnormal behaviors to a level acceptable to the owners. Overall, our data so far collected in the CD dog may allow us to test more deeply the feasibility and safety of such an off-label approach, at both preclinical and clinical levels.

Puppies Raised during the COVID-19 Lockdown Showed Fearful and Aggressive Behaviors in Adulthood: An Italian Survey.

During the COVID-19 pandemic, the Prime Minister's decree issued by the Italian government on 9 March 2020, known as "#Iorestoacasa" (I stay at home), required everyone to stay indoors, with a few exceptions, from 11 March to 3 May 2020. This decree had a substantial impact on the mental health of both people and dogs. We carried out a national survey to compare the personalities of adult dogs who were puppies in lockdown (from March to May 2020) with those of adult dogs born after that time (from June 2020 to February 2021). Our results highlighted a significant increase in personality traits related to fear and aggression in dogs who experienced lockdown restrictions during their socialization period, further confirming that the COVID-19 pandemic strongly impacted the behavioral development of dogs. Therefore, it may be advantageous for these dogs to be closely monitored by veterinary behaviorists and receive specialized behavioral rehabilitation therapy to lower the risk of episodes of aggression and fearfulness as well as to increase the wellbeing of dogs raised under social restrictions.

The Key Role of Lysosomal Protease Cathepsins in Viral Infections.

Cathepsins encompass a family of lysosomal proteases that mediate protein degradation and turnover. Although mainly localized in the endolysosomal compartment, cathepsins are also found in the cytoplasm, nucleus, and extracellular space, where they are involved in cell signaling, extracellular matrix assembly/disassembly, and protein processing and trafficking through the plasma and nuclear membrane and between intracellular organelles. Ubiquitously expressed in the body, cathepsins play regulatory roles in a wide range of physiological processes including coagulation, hormone secretion, immune responses, and others. A dysregulation of cathepsin expression and/or activity has been associated with many human diseases, including cancer, diabetes, obesity, cardiovascular and inflammatory diseases, kidney dysfunctions, and neurodegenerative disorders, as well as infectious diseases. In viral infections, cathepsins may promote (1) activation of the viral attachment glycoproteins and entry of the virus into target cells; (2) antigen processing and presentation, enabling the virus to replicate in infected cells; (3) up-regulation and processing of heparanase that facilitates the release of viral progeny and the spread of infection; and (4) activation of cell death that may either favor viral clearance or assist viral propagation. In this review, we report the most relevant findings on the molecular mechanisms underlying cathepsin involvement in viral infection physiopathology, and we discuss the potential of cathepsin inhibitors for therapeutical applications in viral infectious diseases.

The Potential Impact of a Dog Training Program on the Animal Adoptions in an Italian Shelter.

One of the main concerns of the human-dog relationship is today associated with the quality life inside the kennels, which are very often regarded as animal dump where dogs are exiled, representing a burden on society. In the present study we sought to investigate the importance of performing an appropriate behavioral program on the adoption chances within an Italian shelter, near Naples (Ottaviano). In this respect, we enrolled 555 adopted dogs of different ages, who followed a tailored-4-month lasting training program between 2018 and 2020. Once entered there, they were carefully examined by the veterinary behaviorist, and directed towards a suited training program, to improve living conditions. We documented a higher number of both adult and senior dogs who left the kennel and were adopted, compared to the age-matched untrained animals (n = 479), housed in the same kennel from 2015 to 2017. Taken together, the present data highlight an important role for training in improving the natural attitudes of the companion dogs, thus pointing towards a better human-animal bond.

An Interdisciplinary Approach for Compulsive Behavior in Dogs: A Case Report.

Compulsive disorder is a debilitating condition affecting both humans and animals, characterized by intrusive thoughts and recurring out-of-place behaviors. Among them, tail chasing might represent one of the most common traits in compulsive dogs. Herein, we reported the case of a 7-year-old intact male German Shepherd mixed-breed dog, presenting with tail chasing behavior. He underwent a first behavioral evaluation 1 year before (at the age of 6), when he injured himself with severe wounds at level of the tail and left thigh. To avoid any specific suffering and increase his physical health, of course, the study was carried out through an interdisciplinary approach, employing a veterinary behaviorist and a rehabilitating dog instructor. Three months after pharmacological treatment with fluoxetine and α-s1 casozepine, associated with a behavioral recovery program, the owner reported an improvement of compulsive events in his dog, in terms of intensity and frequency. Interestingly, over the following 3 months, the dog did not experience any new tail chasing episodes.

Human-Dog Relationship during the First COVID-19 Lockdown in Italy.

The SARS-CoV2 pandemic forced an abrupt interruption of social contacts and interpersonal affective relationships all over the world, according to national directives. Many considerable inconveniences occurred with important repercussions also on the emotional state of people and their pets. We carried out a national survey to evaluate the human-dog relationship in a social isolation context using an adapted version of Monash Dog Owner Relationship Scale, the perception of the dogs' discomfort by their human owners, and the resilience of the dog through the quantification of symptoms, in time of the first lockdown of the COVID-19 pandemic. The results highlighted that the human-dog interaction was similar during quarantine; however, there was lower owner's perception of a dog's cost during the quarantine than before it.

Heparan Sulfate Proteoglycans in Viral Infection and Treatment: A Special Focus on SARS-CoV-2.

Heparan sulfate proteoglycans (HSPGs) encompass a group of glycoproteins composed of unbranched negatively charged heparan sulfate (HS) chains covalently attached to a core protein. The complex HSPG biosynthetic machinery generates an extraordinary structural variety of HS chains that enable them to bind a plethora of ligands, including growth factors, morphogens, cytokines, chemokines, enzymes, matrix proteins, and bacterial and viral pathogens. These interactions translate into key regulatory activity of HSPGs on a wide range of cellular processes such as receptor activation and signaling, cytoskeleton assembly, extracellular matrix remodeling, endocytosis, cell-cell crosstalk, and others. Due to their ubiquitous expression within tissues and their large functional repertoire, HSPGs are involved in many physiopathological processes; thus, they have emerged as valuable targets for the therapy of many human diseases. Among their functions, HSPGs assist many viruses in invading host cells at various steps of their life cycle. Viruses utilize HSPGs for the attachment to the host cell, internalization, intracellular trafficking, egress, and spread. Recently, HSPG involvement in the pathogenesis of SARS-CoV-2 infection has been established. Here, we summarize the current knowledge on the molecular mechanisms underlying HSPG/SARS-CoV-2 interaction and downstream effects, and we provide an overview of the HSPG-based therapeutic strategies that could be used to combat such a fearsome virus.

Involvement of the Protein Ras Homolog Enriched in the Striatum, Rhes, in Dopaminergic Neurons' Degeneration: Link to Parkinson's Disease.

Rhes is one of the most interesting genes regulated by thyroid hormones that, through the inhibition of the striatal cAMP/PKA pathway, acts as a modulator of dopamine neurotransmission. Rhes mRNA is expressed at high levels in the dorsal striatum, with a medial-to-lateral expression gradient reflecting that of both dopamine D2 and adenosine A2A receptors. Rhes transcript is also present in the hippocampus, cerebral cortex, olfactory tubercle and bulb, substantia nigra pars compacta (SNc) and ventral tegmental area of the rodent brain. In line with Rhes-dependent regulation of dopaminergic transmission, data showed that lack of Rhes enhanced cocaine- and amphetamine-induced motor stimulation in mice. Previous studies showed that pharmacological depletion of dopamine significantly reduces Rhes mRNA levels in rodents, non-human primates and Parkinson's disease (PD) patients, suggesting a link between dopaminergic innervation and physiological Rhes mRNA expression. Rhes protein binds to and activates striatal mTORC1, and modulates L-DOPA-induced dyskinesia in PD rodent models. Finally, Rhes is involved in the survival of mouse midbrain dopaminergic neurons of SNc, thus pointing towards a Rhes-dependent modulation of autophagy and mitophagy processes, and encouraging further investigations about mechanisms underlying dysfunctions of the nigrostriatal system.

Antioxidants in assisted reproductive technologies: An overview on dog, cat, and horse.

Assisted reproductive technologies (ARTs) are widely used as a tool to improve reproductive performance in both humans and animals. In particular, in the veterinary field, ARTs are used to improve animal genetics, recover endangered animals, and produce offspring in the event of subfertility or infertility in males or females. However, the use of ARTs did not improve the fertilization rate in some animals due to various factors such as the difficulty in reproducing an anatomical and humoral substrate typical of the natural condition or due to the increase in catabolites and their difficult elimination. The in vitro environment allows the production and increase in the concentration of substances, including reactive oxygen species (ROS), which could be harmful to gametes. If produced in high concentration, the ROS becomes deleterious, both in vitro and in vivo systems. It has been seen that the use of antioxidants can help neutralize or counteract the production of ROS. The present study aims to report the latest findings regarding the use of antioxidants in ARTs of some domestic species, such as dogs, cats, and horses, compared to other animal species, such as cattle, in which ARTs have instead developed more widely.

Cortisol Levels of Shelter Dogs in Animal Assisted Interventions in a Prison: An Exploratory Study.

Previous studies regarding the Animal Assisted Interventions (AAI) have mainly focused on the beneficial effects of human-animal interactions on human health; whereas the impact of such activities on the welfare of the animals involved has received limited attention. So far, few studies have addressed this issue by evaluating the physiological and behavioral reactions of therapy dogs during the interventions. The aim of this study was to evaluate the potential effect of AAI on the cortisol levels of shelter dogs. Five dogs participated in weekly AAI working activities with adult inmates held at a prison of the South of Italy for two months. Saliva samples were collected every two weeks in three conditions: at the kennel (baseline), after transportation and at the end of the working sessions. The results revealed a significant decrease in the cortisol baseline at the end of the AAI program, suggesting that the activities carried out with humans and in a different environment could improve the welfare of dogs housed in kennels. Moreover, we found that transportation significantly increased subjects' cortisol levels, suggesting that it is a critical phase that deserves particular care.

Id(entifying) the inhibitor of DNA binding 3 in the brain of Nothobranchius furzeri upon aging.

Inhibitors of DNA (Id) are key transcription factors (TFs) regulating neurogenic processes. They belong to the helix-loop-helix (HLH) TF family and are dominant negative regulators of basic HLH proteins (bHLHs). Specifically, they inhibit cell differentiation and enhance cell proliferation and motility. The Id family includes four members, Id1, Id2, Id3, and Id4, which have been identified in nearly all vertebrates. The transcript catalog of the African turquoise killifish, Nothobranchius furzeri, contains all four TFs and has evolved showing positive selection for Id3. N. furzeri, a teleost, is the short-lived vertebrate and is gaining increasing scientific interest as a new model organism in aging research. It is characterized by embryonic diapause, explosive sexual maturation, and rapid aging. In this study, we investigated both the expression and the role of Id3 in the brain of this model organism. Interestingly, Id3 was upregulated age-dependently along with a distribution pattern resembling that of other vertebrates. Additionally, the gene has undergone positive selection during evolution and shows a high degree of conservation relative to that of other vertebrates. These features make N. furzeri a valid tool for aging studies and a potential model in translational research.

Lepidium meyenii (Maca) in male reproduction.

Lepidium meyenii (Maca) is an edible root plant that grows in the Andean region of Peru. For centuries, the plant has been used as a dietary supplement for its nutritional and therapeutic properties. Maca are rich in high value nutritional elements and secondary metabolites (macaridine, macamides and glucosinolates) with high biological activity. Several studies demonstrated various biological effects of Maca mainly in the field of fertility. The aim of this review is to summarize the state of knowledge on the properties of Maca on male reproduction. Literature data was performed in PubMed with researches published from 2000 to 2019. The research showed results related to the effects of Maca on the quality and quantity of the semen, sexual behaviour and disorders of the male genital tract. Despite the numerous studies carried out on different animal species, further research is needed to clarify the mechanisms of action of Maca.

Triple-Negative Breast Cancer Comparison With Canine Mammary Tumors From Light Microscopy to Molecular Pathology.

Many similar characteristics in human and dog cancers including, spontaneous development, clinical presentation, tumor heterogeneity, disease progression, and response to standard therapies have promoted the approval of this comparative model as an alternative to mice. Breast cancer represents the second most frequent neoplasm in humans after lung cancer. Triple-negative breast cancers (TNBC) constitute around 15% of all cases of breast cancer and do not express estrogen receptor (ER), progesterone receptor (PR), and do not overexpress human epidermal growth factor receptor 2 (HER2). As a result, they do not benefit from hormonal or trastuzumab-based therapy. Patients with TNBC have worse overall survival than patients with non-TNBC. Lehmann and collaborators described six different molecular subtypes of TNBC which further demonstrated its transcriptional heterogeneity. This six TNBC subtype classification has therapeutic implications. Breast cancer is the second most frequent neoplasm in sexually intact female dogs after skin cancer. Canine mammary tumors are a naturally occurring heterogeneous group of cancers that have several features in common with human breast cancer (HBC). These similarities include etiology, signaling pathway activation, and histological classification. Molecularly CMTs are more like TNBCs, and therefore dogs are powerful spontaneous models of cancer to test new therapeutic approaches, particularly for human TNBCs. More malignant tumors of the breast are more often ER and PR negative in both humans and dogs. Promising breast cancer biomarkers in both humans and canines are cancer-associated stroma (CAS), circulating tumor cells and tumor DNA (ctDNA), exosomes and miRNAs, and metabolites.

Role of Aquaporins in the Physiological Functions of Mesenchymal Stem Cells.

Aquaporins (AQPs) are a family of membrane water channel proteins that control osmotically-driven water transport across cell membranes. Recent studies have focused on the assessment of fluid flux regulation in relation to the biological processes that maintain mesenchymal stem cell (MSC) physiology. In particular, AQPs seem to regulate MSC proliferation through rapid regulation of the cell volume. Furthermore, several reports have shown that AQPs play a crucial role in modulating MSC attachment to the extracellular matrix, their spread, and migration. Shedding light on how AQPs are able to regulate MSC physiological functions can increase our knowledge of their biological behaviours and improve their application in regenerative and reparative medicine.

Rapamycin, by Inhibiting mTORC1 Signaling, Prevents the Loss of Striatal Bidirectional Synaptic Plasticity in a Rat Model of L-DOPA-Induced Dyskinesia.

Levodopa (L-DOPA) treatment is the main gold-standard therapy for Parkinson disease (PD). Besides good antiparkinsonian effects, prolonged use of this drug is associated to the development of involuntary movements known as L-DOPA-induced dyskinesia (LID). L-DOPA-induced dyskinesia is linked to a sensitization of dopamine (DA) D1 receptors located on spiny projection neurons (SPNs) of the dorsal striatum. Several evidences have shown that the emergence of LID can be related to striatal D1/cAMP/PKA/DARPP-32 and extracellular signal-regulated kinases (ERK1/2) pathway overactivation associated to aberrant N-methyl-d-aspartate (NMDA) receptor function. In addition, within striatum, ERK1/2 is also able to modulate in a D1 receptor-dependent manner the activity of the mammalian target of rapamycin complex 1 (mTORC1) pathway under DA depletion and L-DOPA therapy. Consistently, increased mTORC1 signaling appears during chronic administration of L-DOPA and shows a high correlation with the severity of dyskinesia. Furthermore, the abnormal activation of the D1/PKA/DARPP-32 cascade is paralleled by increased phosphorylation of the GluA1 subunit of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor at the PKA Ser845 site. The GluA1 promotes excitatory AMPA receptor-mediated transmission and may be implicated in the alterations found at the corticostriatal synapses of dyskinetic animals. In our study, we investigated the role of mTORC1 pathway activation in modulating bidirectional striatal synaptic plasticity in L-DOPA-treated parkinsonian rats. Inhibition of mTORC1 by coadministration of rapamycin to L-DOPA was able to limit the magnitude of LID expression, accounting for a therapeutic effect of this drug. In particular, behavioral data showed that, in L-DOPA-treated rats, rapamycin administration induced a selective decrease of distinct components of abnormal involuntary movements (i.e., axial and orolingual dyskinesia). Furthermore, ex vivo patch clamp and intracellular recordings of SPNs revealed that pharmacological inhibition of mTORC1 also resulted associated with a physiological bidirectional plasticity, when compared to dyskinetic rats treated with L-DOPA alone. This study uncovers the important role of mTORC1 inhibition to prevent the loss of striatal bidirectional plasticity under chronic L-DOPA treatment in rodent models of PD.

Human-Animal Relationship Dysfunction: A Case Study of Animal Hoarding in Italy.

"Animal hoarding" or "compulsive hoarding of animals" is a psychiatric disease, which has important social implications and a profound influence on animal welfare. To date, this phenomenon has been little investigated and largely unexplored. The present study aims to systematically describe a case of animal hoarding, which remains unresolved. The report refers to a case of a woman suffering from animal hoarding that emerged in 2005. From March 2014 to December 2019, 450 animals were seized over nine different occasions. This disease had significant implications on the welfare of the animals collected, which lived in poor housing and hygiene conditions that frequently led to their death. Since animal hoarding cases involve sanitary, legal, and veterinary aspects, we believe that a multidisciplinary approach is necessary in order to prevent a recurrence and a new accumulation of animals. A holistic approach should be taken according to the One Health principle that involves different stakeholders at every level in order to adopt an efficient solution.