Aged Animals Research Articles

Piezo1 Activation Drives Enhanced Collagen Synthesis in Aged Animal Skin Induced by Poly L-Lactic Acid Fillers.

Poly L-lactic acid (PLLA) fillers stimulate collagen synthesis by activating various immune cells and fibroblasts. Piezo1, an ion channel, responds to mechanical stimuli, including changes in extracellular matrix stiffness, by mediating Ca2+ influx. Given that elevated intracellular Ca2+ levels trigger signaling pathways associated with fibroblast proliferation, Piezo1 is a pivotal regulator of collagen synthesis and tissue fibrosis. The aim of the present study was to investigate the impact of PLLA on dermal collagen synthesis by activating Piezo1 in both an H2O2-induced cellular senescence model in vitro and aged animal skin in vivo. PLLA elevated intracellular Ca2+ levels in senescent fibroblasts, which was attenuated by the Piezo1 inhibitor GsMTx4. Furthermore, PLLA treatment increased the expression of phosphorylated ERK1/2 to total ERK1/2 (pERK1/2/ERK1/2) and phosphorylated AKT to total AKT (pAKT/AKT), indicating enhanced pathway activation. This was accompanied by upregulation of cell cycle-regulating proteins (CDK4 and cyclin D1), promoting the proliferation of senescent fibroblasts. Additionally, PLLA promoted the expression of phosphorylated mTOR/S6K1/4EBP1, TGF-β, and Collagen I/III in senescent fibroblasts, with GsMTx4 treatment mitigating these effects. In aged skin, PLLA treatment similarly upregulated the expression of pERK1/2/ERK1/2, pAKT/AKT, CDK4, cyclin D1, mTOR/S6K1/4EBP1, TGF-β, and Collagen I/III. In summary, our findings suggest Piezo1's involvement in PLLA-induced collagen synthesis, mediated by heightened activation of cell proliferation signaling pathways such as pERK1/2/ERK1/2, pAKT/AKT, and phosphorylated mTOR/S6K1/4EBP1, underscoring the therapeutic potential of PLLA in tissue regeneration.

Individual flexibility in group foraging behaviour of reef manta rays (Mobula alfredi)

Flexibility in animal foraging strategies can increase overall feeding efficiency for individuals. For example, group foraging can increase the efficiency of resource exploitation; conversely solo foraging can reduce intraspecific competition, particularly at low resource densities. The cost–benefit trade-off of such flexibility is likely to differ within and among individuals. Reef manta rays (Mobula alfredi) are large filter-feeding elasmobranchs that often aggregate to feed on ephemeral upwellings of zooplankton. Over three years in the Maldives, we free-dived to film 3106 foraging events involving 343 individually identifiable M. alfredi. Individuals fed either solo or in groups with a clear leader plus between one and eight followers. M. alfredi were significantly more likely to forage in groups than solo at high just prior to high tide and when aggregations were larger. Within aggregations, individuals foraged in larger groups when more food was available, and when the overall aggregations were relatively large suggesting that foraging in large groups was more beneficial when food is abundant, and the costs of intraspecific competition were outweighed by the efficiency resulting from group foraging strategies. Females, the larger sex, were more likely to lead foraging groups than males. The high within-individual variance (over 70%), suggested individuals were unpredictable across all foraging behaviours, thus individual M. alfredi cannot be classified into foraging types or specialists. Instead, each individual was capable of considerable behavioural flexibility, as predicted for a species reliant on spatially and temporally ephemeral resources.Significance statementReef manta rays (Mobula alfredi), listed as “Vulnerable” on the IUCN Red List, are at risk from targeted and by-catch fisheries due to their slow life history and aggregative behaviour. M. alfredi feed together in aggregations on short-lived glut of microscopic zooplankton. Over three years in the Maldives, we filmed 3106 foraging events involving 343 individually identifiable M. alfredi. Manta rays were more likely to forage in groups than solo just prior to high tide and when aggregations were larger, attracted by the influx of zooplankton. Foraging groups included more individuals when plankton was more abundant. However, individuals flipped between solo and group foraging and did not specialise. Foraging groups were most often led by females, the larger sex. Individuals were very flexible in how they foraged, which makes sense for a species that relies on a food source that varies enormously in when, where and for how long it is available. Understanding manta ray foraging behaviour will help conservation management efforts and predict their responses to climate change.

Оцінка доз опромінення нориці рудої 137Cs на дослідних полігонах Чорнобильської зони відчуження з урахуванням вік-залежної динаміки зміни фізіологічних параметрів

Methods for dose estimation of 137Cs in mouse-like rodents, specifically bank voles (Myodes glareolus), commonly found within the research sites in the Chornobyl Exclusion Zone, are being proposed. The International Commission on Radiological Protection recommends using the specially developed "BiotaDC" software, but it needs to be adapted to solve specific applied issues. It is shown that the "BiotaDC" approach adequately describes dose accumulation of 137Cs for external exposure inside nest chambers if the mean integral value of soil activity concentration at a depth of 0.5 m is used, for external exposure above soil – if the mean integral value of soil activity concentration at a depth of 0.2 m is used. However, the authors propose a method considering the age of animals at the time of capture and the age-related dynamics of changes in such parameters as excretion and accumulation of radioisotopes for internal dose estimation using the "BiotaDC" software. A detailed dose estimation algorithm is being proposed. Dose estimates for internal irradiation according to the authors' method and the method of the International Commission on Radiological Protection are being compared.

Dynamics of mercury stable isotope compounds in Arctic seals: New insights from a controlled feeding trial on hooded seals Cystophora cristata

Accurate interpretation of mercury (Hg) isotopic data requires the consideration of several biotic factors such as age, diet, geographical range, and tissue metabolic turnover. A priori knowledge of prey-predator isotopic incorporation rates and Hg biomagnification is essential. This study aims to assess Hg stable isotopes incorporation in an Arctic species of Phocidae, the hooded seal Cystophora cristata, kept in human care for 24 months (2012–2014) and fed on a constant diet of Norwegian Spring Spawning herring Clupea harengus. We measured THg, MMHg and iHg levels, as well as Hg stable isotope composition with both mass dependent (MDF) and mass independent (MIF) fractionation (e.g. δ202Hg and Δ199,200,201,204Hg) in hooded seal kidney, liver, hair and muscle, in addition to herring muscle. We then calculated Hg MDF and MIF isotopic fractionation between hooded seals and their prey. We found a significant shift in δ202Hg between hooded seal hair (+0.80‰) and kidney (−0.78‰), and herring muscle. In hooded seals tissues δ202Hg correlated positively with MMHg percentage. These findings suggest that tissue-specific Hg speciation is the major driver of changes in Hg isotopic fractionation rates in this Arctic predator. Δ199Hg, Δ200Hg, Δ201Hg and Δ204Hg values did not vary between herring and hooded seal tissues, confirming their utility as tracers of Hg marine and atmospheric sources in top predators. To our knowledge, this represents the first attempt to assess complex Hg isotope dynamics in the internal system of Arctic Phocidae, controlling the effects of age, diet, and distribution. Our results confirm the validity of Hg stable isotopes as tracers of environmental Hg sources even in top predators, but emphasize the importance of animal age and tissue selection for inter-study and inter-species comparisons.

Prevalence and Risk Factors of Bone and Dental Lesions in Neotropical Deer.

Bone and dental lesions have been documented in various deer species globally, affecting the efficiency of ingestion and digestion, consequently influencing their general health and leading to a decline in survival and reproductive performance. The present study aimed to characterize bone and dental lesions in the dry skulls of individual deer, estimate the prevalence of these lesions, and assess potential risk factors associated with the development of bone and dental alterations. This study assessed bone and dental lesions in 180 dry skulls of eleven neotropical deer species, originating from both captivity and wildlife conditions, through direct visual inspection. A high prevalence of bone and dental lesions was observed in all analyzed species. Dental calculus was the most common alteration (96.7%), followed by dental wear (71.1%). Animal age positively correlated with most bone and dental alterations, indicating that older animals showed more lesions. Additionally, the prevalence of these alterations was similar between sexes. Moreover, all lesions were more common in captive-bred animals, likely attributed to their older age and a less diverse diet. Blastocerus dichotomus and Mazama americana were most affected by bone resorption and dental trauma and had the highest dental calculus prevalence, along with Subulo gouazoubira and Passalites nemorivagus. All eleven species evaluated in the present study were susceptible to the occurrence of bone and dental lesions. Therefore, monitoring oral health and diet in captivity are fundamental practices for the conservation of these species.

The neuron-specific IIS/FOXO transcriptome in aged animals reveals regulatory mechanisms of cognitive aging

Cognitive decline is a significant health concern in our aging society. Here, we used the model organism C. elegans to investigate the impact of the IIS/FOXO pathway on age-related cognitive decline. The daf-2 Insulin/IGF-1 receptor mutant exhibits a significant extension of learning and memory span with age compared to wild-type worms, an effect that is dependent on the DAF-16 transcription factor. To identify possible mechanisms by which aging daf-2 mutants maintain learning and memory with age while wild-type worms lose neuronal function, we carried out neuron-specific transcriptomic analysis in aged animals. We observed downregulation of neuronal genes and upregulation of transcriptional regulation genes in aging wild-type neurons. By contrast, IIS/FOXO pathway mutants exhibit distinct neuronal transcriptomic alterations in response to cognitive aging, including upregulation of stress response genes and downregulation of specific insulin signaling genes. We tested the roles of significantly transcriptionally-changed genes in regulating cognitive functions, identifying novel regulators of learning and memory. In addition to other mechanistic insights, a comparison of the aged vs young daf-2 neuronal transcriptome revealed that a new set of potentially neuroprotective genes is upregulated; instead of simply mimicking a young state, daf-2 may enhance neuronal resilience to accumulation of harm and take a more active approach to combat aging. These findings suggest a potential mechanism for regulating cognitive function with age and offer insights into novel therapeutic targets for age-related cognitive decline.

The neuron-specific IIS/FOXO transcriptome in aged animals reveals regulatory mechanisms of cognitive aging.

Cognitive decline is a significant health concern in our aging society. Here, we used the model organism C. elegans to investigate the impact of the IIS/FOXO pathway on age-related cognitive decline. The daf-2 Insulin/IGF-1 receptor mutant exhibits a significant extension of learning and memory span with age compared to wild-type worms, an effect that is dependent on the DAF-16 transcription factor. To identify possible mechanisms by which aging daf-2 mutants maintain learning and memory with age while wild-type worms lose neuronal function, we carried out neuron-specific transcriptomic analysis in aged animals. We observed downregulation of neuronal genes and upregulation of transcriptional regulation genes in aging wild-type neurons. By contrast, IIS/FOXO pathway mutants exhibit distinct neuronal transcriptomic alterations in response to cognitive aging, including upregulation of stress response genes and downregulation of specific insulin signaling genes. We tested the roles of significantly transcriptionally-changed genes in regulating cognitive functions, identifying novel regulators of learning and memory. In addition to other mechanistic insights, a comparison of the aged vs young daf-2 neuronal transcriptome revealed that a new set of potentially neuroprotective genes is upregulated; instead of simply mimicking a young state, daf-2 may enhance neuronal resilience to accumulation of harm and take a more active approach to combat aging. These findings suggest a potential mechanism for regulating cognitive function with age and offer insights into novel therapeutic targets for age-related cognitive decline.

Overexpression of the receptor for resolvin E1 (ERV1) prevents early alveolar bone loss in leptin receptor deficiency-induced diabetes.

This study was designed to test the hypothesis that the leptin receptor (LepR) regulates changes in periodontal tissues and that the overexpression of the receptor for resolvin E1 (ERV1) prevents age- and diabetes-associated alveolar bone loss. LepR-deficient transgenic (TG) mice were cross-bred with those overexpressing ERV1 (TG) to generate double-TG mice. In total, 95 mice were divided into four experimental groups: wild type (WT), TG, LepR deficient (db/db), and double transgenic (db/db TG). The groups were followed from 4weeks up to 16weeks of age. The natural progression of periodontal disease without any additional method of periodontitis induction was assessed by macroscopic and histomorphometric analyses. Osteoclastic activity was measured by tartrate-resistant acid phosphatase (TRAP) staining. At 4weeks, ERV1 overexpression prevented weight gain. From Week 8 onward, there was a significant increase in the weight of db/db mice with or without ERV1 overexpression compared to the WT mice, accompanied by an increase in glucose levels. By 8weeks of age, the percentage of bone loss in the LepR deficiency groups was significantly greater compared to WT mice. ERV1 overexpression in the db/db TG mice prevented early alveolar bone loss; however, it did not impact the development of diabetic bone loss in aging mice after the onset of weight gain and diabetes. The findings suggest that the overexpression of ERV1 prevents LepR-associated alveolar bone loss during the early phases of periodontal disease by delaying weight gain, diabetes onset, and associated inflammation; however, LepR deficiency increases susceptibility to naturally occurring inflammatory alveolar bone loss as the animal ages, associated with excess weight gain, onset of diabetes, and excess inflammation.

Improved resilience and proteostasis mediate longevity upon DAF-2 degradation in old age

Little is known about the possibility of reversing age-related biological changes when they have already occurred. To explore this, we have characterized the effects of reducing insulin/IGF-1 signaling (IIS) during old age. Reduction of IIS throughout life slows age-related decline in diverse species, most strikingly in the nematode Caenorhabditis elegans. Here we show that even at advanced ages, auxin-induced degradation of DAF-2 in single tissues, including neurons and the intestine, is still able to markedly increase C. elegans lifespan. We describe how reversibility varies among senescent changes. While senescent pathologies that develop in mid-life were not reversed, there was a rejuvenation of the proteostasis network, manifesting as a restoration of the capacity to eliminate otherwise intractable protein aggregates that accumulate with age. Moreover, resistance to several stressors was restored. These results support several new conclusions. (1) Loss of resilience is not solely a consequence of pathologies that develop in earlier life. (2) Restoration of proteostasis and resilience by inhibiting IIS is a plausible cause of the increase in lifespan. And (3), most interestingly, some aspects of the age-related transition from resilience to frailty can be reversed to a certain extent. This raises the possibility that the effect of IIS and related pathways on resilience and frailty during aging in higher animals might possess some degree of reversibility.

Induction of Nitric Oxide and Its Role in Facial Nerve Regeneration According to the Method of Facial Nerve Injury.

Nitric oxide (NO) is an important molecule in cell communication that also plays an important role in many biological processes. Given the dual role of NO in nerve degeneration and regeneration after facial nerve injury, we sought to delve deeper into its role through a systematic literature review. A comprehensive review of the literature employing SCOPUS, PubMed, Cochrane Library, EMBASE, and Google Scholar databases was conducted to evaluate the induction and role of NO in neurodegeneration and regeneration after facial nerve injury. From the 20 papers ultimately reviewed, the central findings were that neuronal nitric oxide synthase(nNOS), endothelial nitric oxide synthase (eNOS), and induced nitric oxide synthase (iNOS) increased or decreased depending on the method of facial nerve damage, damaged area, harvested area, and animal age, and were correlated with degeneration and regeneration of the facial nerve. Research conducted on rats and mice demonstrated that NO, nNOS, eNOS, and iNOS play significant roles in nerve regeneration and degeneration. However, the relationship between nerve damage and NO could not be defined by a simple causal relationship. Instead, the involvement of NOS depends on the type of nerve cell, source of NO, timing, and location of expression, age of the target animal, and proximity of the damage location to the brainstem. Consequently, nNOS, eNOS, and iNOS expression levels and functions may vary significantly.

Creating a Lactation Model for 305-Day Milk Yield with Different Resampling Techniques (Bagging Mars) in Mars Modeling

The main purpose of this research is to obtain a prediction model for milk yield by using Multivariate Adaptive Regression Splines (MARS) and Bagging MARS algorithms as a non-parametric regression technique. For this purpose, the effects on milk yield of 305 days were investigated by using lactation parameters in dairy cattle. In the study, 9337 lactation milk yield records belonging to 37 animals belonging to the 2022-2023 period were used and the data set was created by randomly ordering the animals. Data on milk yield results were analyzed with MARS and Bagging MARS algorithms. For dairy cattle; it was modeled with explanatory variables such as lactation month (month), service period (SP), last 7 days average milk yield (L7DMMY), animal's first birth age (FP), animal's age (Age), number of lactations (LN).Correlation coefficient (r), coefficient of determination (R2), Adjusted R2, Root of Square Mean Error (RMSE), standard deviation ratio (SD ratio), mean absolute percent error (MAPE), mean absolute for MARS algorithm estimating total average milk yield deviation (MAD) and Akaike Information Criteria (AIC) values are 0.9986, 0.997, 0.977, 0.142, 0.052, 0.2389, 0.086 and -88, respectively. Similar statistics for the Bagging MARS algorithm are 0.754, 0.556, 0.453, 1.8, 0.666, 3.96, 1.47, and 115, respectively. It has been observed that MARS and Bagging MARS algorithms provide correct results according to the goodness of fit statistics. In this study, it was revealed that MARS algorithm gave better results in milk yield modeling of 305-day lactation.

Effects of methylphenidate on mitochondrial dynamics and bioenergetics in the prefrontal cortex of juvenile rats are sex-dependent

Methylphenidate (MPH) is a central nervous system stimulant drug and a first order prescription in the treatment of Attention Deficit Hyperactivity Disorder (ADHD). Although MPH biochemistry in neurodevelopment is not completely understood, studies showed it alters energy metabolism in rat brains. ADHD prevalence during neurodevelopment is related to males and the investigation has been mainly done in these subjects, therefore, little is known about MPH action in females and, consequently, about sexual dimorphism. In the present study we evaluated markers of mitochondrial dynamics (DRP1 and MFN2, fission and fusion, respectively), biogenesis (mtTFA) and bioenergetics (respiratory chain complexes) in prefrontal cortex of male and female juvenile rats submitted to exposure to MPH to better understand MPH effect during postnatal neurodevelopment. ATP and oxidative stress levels were also evaluated. Wistar rats received intraperitoneal injection of MPH (2.0 mg/kg) or control (saline), once a day, from 15th to 45th day of age. Results showed that MPH increased DRP1 and decreased MFN2, as well as increased mtTFA in prefrontal cortex of male rats. In female, MPH decreased NRF1 and increased Parkin, which are mitochondrial regulatory proteins. Respiratory chain complexes (complex I, SDH, complexes III and IV), ATP production and oxidative stress parameters were altered and shown to be sex-dependent. Taken together, results suggest that chronic MPH exposure at an early age in healthy animals changes mitochondrial dynamics, biogenesis and bioenergetics differently depending on the sex of the subjects.

Aging in animals – Individuals decline and the impacts on toxicity – Hazard of Cd in Enchytraeus crypticus

Aging is a natural process for all living beings and there is often a decline of overall performance after a certain age tipping point. The response to stressors, e.g. exposure to chemicals can differ depending on the individuals’ age. Standard guidelines do not always include synchronized age animals and are often optimized based on healthy animals of homogeneous populations, not aiming to cover age aspects. Hence, in the present study we aimed to assess the effects of aging on Enchytraeus crypticus (Oligochaeta) response to cadmium (Cd) exposure. For that, cultures with 3 synchronized ages [young: 18–20 days old, old: 91–93 days old and oldest: 127–129 days old] plus the standard (non-synchronized adults), were exposed to CdCl2 (0, 1, 3.2, 10, 32, 100, 320 mg Cd/kg) in LUFA 2.2 soil. Results showed that Cd toxicity varied depending on the animal’s age, with the young and oldest being most sensitive to Cd (reproduction EC50_old = 138 > EC50_young = 69 > EC50_oldest = 46 mg Cd/kg soil). This highlights the importance of age when assessing toxicity. Testing animals with synchronized age should improve results comparability but will not necessarily capture the most sensitive age. Toxicity to Cd has been shown to be higher in younger animals compared to standard cultures, although this may not be a general pattern with other chemicals or stressors. When using standardized guidelines, if cultures are not synchronized results will suffer from variability due to various ages of the animals. For a standard context we recommend the start with young (18–20 days) animals instead of non-synchronized adults. On the other hand, the study of populations on the edge window is highly relevant and can be much more explored in an ecotoxicology frame, as this can also enlighten impacts of aging in a wider species context.

Gastric mill ossicles record chronological age in the Caribbean spiny lobster (Panulirus argus)

Information on individual age and size relationships is crucial for sustainable management of fished populations. Previous research has indicated it may be possible to directly age crustaceans by counting annular bands deposited within gastric mill ossicles. The method has been applied to multiple crustacean species; however, its applicability has been controversial because studies have shown gastric mill ossicles are reworked and/or replaced during ecdysis. Over the last 8 years, we have refined and improved upon methods of processing gastric mill ossicles and investigated the validity of this method for ageing Caribbean spiny lobster (Panulirus argus) using captive-raised animals of known age from the Florida Keys (USA). Although the biological process that creates annular bands within gastric mill ossicles remains unclear, validation of the method for P. argus based on known-age lobsters from 10 months–5.5 years old (n = 33) confirms that the deposition of bands correlates with chronological age. We compared precision and accuracy of age estimates for ossicles processed using two imaging techniques (transmitted vs. reflected light) and found that transmitted light provided the best images for ageing P. argus. The mean coefficient of variation (CV) of age estimates between two readers was 20.2 %. The consensus age estimates provided by the readers were, on average, within 0.5 years of the known age and overall, 94 % of consensus age estimates from readers were within 1 year of the known age. We also compared lobsters of the same size but different age as well as those of the same age but different size to address whether the number of endocuticular bands in the ossicle is a measure of true chronological age or simply related to the size of the animal. Our results suggest that the gastric mill ossicles record chronological age in P. argus, providing a reliable ageing method with which to inform age-based management of the Caribbean spiny lobster population. Future research is needed to understand the mechanism that controls the deposition of endocuticular bands within the crustacean gastric mill ossicles and to determine the applicability of the method for other crustaceans.

Organic light-emitting diode therapy promotes longevity through the upregulation of SIRT1 in senescence-accelerated mouse prone 8 mice

Phototherapy has been extensively used to prevent and treat signs of aging and stimulate wound healing, and phototherapy through light-emitting diodes (LEDs). In contrast to LED, organic LED (OLED) devices are composed of organic semiconductors that possess novel characteristics. We investigated the regenerative potential of OLED for restoring cellular potential from senescence and thus delaying animal aging. Bone marrow–derived stem cells (BMSCs) and adipose-derived stem cells (ADSCs) were isolated from the control and OLED- treated groups to evaluate their proliferation, migration, and differentiation potentials. Cellular senescence was evaluated using a senescence-associated β-galactosidase (SA-β-gal) activity assay and gene expression biomarker assessment. OLED treatment significantly increased the cell proliferation, colony formation, and migration abilities of stem cells. SA-β-gal activity was significantly decreased in both ADSCs and BMSCs in the OLED-treated group. Gene expression biomarkers from treated mice indicated a significant upregulation of IGF-1 (insulin growthfactor-1). The upregulation of the SIRT1 gene inhibited the p16 and p19 genes then to downregulate the p53 expressions for regeneration of stem cells in the OLED-treated group. Our findings indicated that the survival rates of 10-month aging senescence-accelerated mouse prone 8 mice were prolonged and that their gross appearance improved markedly after OLED treatment. Histological analysis of skin and brain tissue also indicated significantly greater collagen fibers density, which prevents ocular abnormalities and β-amyloid accumulation. Lordokyphosis and bone characteristics were observed to resemble those of younger mice after OLED treatment.In conclusion, OLED therapy reduced the signs of aging and enhanced stem-cell senescence recovery and then could be used for tissue regeneration.

Most genetic roots of fungal and animal aging are hundreds of millions of years old according to phylostratigraphy analyses of aging networks.

Few studies have systematically analyzed how old aging is. Gaining a more accurate knowledge about the natural history of aging could however have several payoffs. This knowledge could unveil lineages with dated genetic hardware, possibly maladapted to current environmental challenges, and also uncover "phylogenetic modules of aging," i.e., naturally evolved pathways associated with aging or longevity from a single ancestry, with translational interest for anti-aging therapies. Here, we approximated the natural history of the genetic hardware of aging for five model fungal and animal species. We propose a lower-bound estimate of the phylogenetic age of origination for their protein-encoding gene families and protein-protein interactions. Most aging-associated gene families are hundreds of million years old, older than the other gene families from these genomes. Moreover, we observed a form of punctuated evolution of the aging hardware in all species, as aging-associated families born at specific phylogenetic times accumulate preferentially in genomes. Most protein-protein interactions between aging genes are also old, and old aging-associated proteins showed a reduced potential to contribute to novel interactions associated with aging, suggesting that aging networks are at risk of losing in evolvability over long evolutionary periods. Finally, due to reshuffling events, aging networks presented a very limited phylogenetic structure that challenges the detection of "maladaptive" or "adaptative" phylogenetic modules of aging in present-day genomes.

Effect of sainfoin (Onobrychis viciifolia Scop.) seed-based diet on rats: A comprehensive evaluation of hemogram, biochemistry, and histopathology.

Sainfoin species (Onobrychis spp.) have been employed for centuries as an essential forage for ruminant animals, both for grazing and as hay. The seeds produced by sainfoin have also been investigated as an animal feed source and were indicated to be a particularly protein-rich supplement for monogastric animals. This study explores the effects of two sainfoin seed inclusion rates in rat diets compared to a control diet, focusing on blood biochemical parameters and a comprehensive histopathological evaluation of multiple organ systems. Thus, we provide a novel contribution to the body of evidence investigating sainfoin seeds as a protein supplement in monogastric animal diets. In this 21-day experiment, seven rats each were assigned to the control group, a 5% sainfoin seed group, and a 10% sainfoin seed group. The control group received standard feed and water; the second group received feed with 5% sainfoin seeds; and the third group received feed with 10% sainfoin seeds. At the experiment's end, necropsies and evaluations were conducted. Histopathological exams revealed normal organ structures in all 21 samples, regardless of the group. Blood analysis showed statistically significant decreases in creatine, ALT, P, Ca, and Mg levels in the sainfoin seed groups compared to the control group, with most values nearing reference levels, suggesting potential benefits. Notably, no adverse effects were observed when sainfoin seeds were included at 5% and 10% in the rat feed. These findings contribute to a growing body of research investigating the inclusion of sainfoin seeds in monogastric animal diets, which is a foundational component of assessing sainfoin's potential as a novel pulse crop for human consumption.

Increased expression of SLC25A18 is associated with Alzheimer’s disease and is involved in Aβ42-induced mitochondrial dysfunction and apoptosis in neuronal cells

Alzheimer’s disease (AD) is currently one of the most serious public health concerns in the world. However, the best approach to treat AD has yet to be discovered, implying that we must continue to work hard to find new AD target genes. In this study, we further analysed Gene Expression Omnibus (GEO) data and discovered that the expression of the Mitochondria glutamate carrier SLC25A18 is associated with AD by screening the differentially expressed genes in different regions of the brains of Alzheimer’s disease patients. To verify the expression of SLC25A18 during Alzheimer’s disease development, we analysed animal models (5×FAD transgenic AD animal model, chemically induced AD animal model, natural ageing animal model), and the results showed that the expression of SLC25A18 was increased in animal models of AD. Further investigation of the different regions found that SLC25A18 expression was elevated in the EC, TeA, and CA3, and expressed in neurons. Next, We found that Aβ42 treatment elevated SLC25A18 expression in Neuro 2A cells. Reducing SLC25A18 expression attenuated mitochondrial dysfunction and neuronal apoptosis caused by Aβ42. Overexpression of SLC25A18 increased ATP and intracellular superoxide anions but decreased mitochondrial membrane potential. The results indicate that SLC25A18 affects mitochondrial function and neuronal apoptosis, and is related to AD, which makes it a potential target for treating brain dysfunction.

Extracellular vesicles derived from mesenchymal stem cells ameliorate cognitive impairment caused by neuroinflammation in young but not aged mice

Aim: The aim of this work was to study the effects of extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) on inflammation-impaired cognitive functions and the brain of mice. Methods: Young mice (~3-month-old) and aged mice (~18-month-old) were injected with bacterial lipopolysaccharide (LPS) and obtained intravenously donor 106 human umbilical cord MSCs, EVs isolated from a similar amount of MSCs or conditioned medium (CM) of MSCs. Subsequently, the mice were examined in behavioral tests and the mouse brains were analyzed for the levels of pro-inflammatory cytokines, α7 nicotinic acetylcholine receptors (α7 nAChRs) and amyloid beta 1-42 (Aβ1-42). Results: EVs prevented LPS-induced memory impairment in mice, whereas CM provided a weaker and temporal effect. Both EVs and MSCs injected once after regular injections of LPS stably improved memory of young mice. In contrast, both cells and EVs provided only transient effect in aged mice injected with LPS. The brains of aged LPS-treated mice contained elevated amounts of IL-1β and IL-6; both MSCs and EVs decreased them significantly. The brains of non-treated aged mice contained decreased levels of α7 nAChRs and increased levels of Aβ1-42 and α7-bound Aβ1-42 compared to the brains of young mice. LPS treatment decreased α7 nAChRs in both young and aged mice, while both MSCs and EVs restored them up to the control level. In young mice, LPS treatment increased the level of Aβ1-42 and α7-bound Aβ1-42, whereas MSCs and EVs decreased it. In contrast, neither LPS nor MSCs/EVs influenced the elevated level of Aβ1-42 but increased α7-bound Aβ1-42 in the brains of aged mice. Conclusions: Regenerative potential of MSCs and MSC-derived EVs is sufficient to support cognitive functions of LPS-treated young mice but is quite poor for aged animals, possibly, due to decreased levels of α7 nAChRs and accumulated Aβ1-42 in their brains.

Seroprevalence and Endemic Status of Babesia ovis by Enzyme-Linked Immunosorbent Assay in East Azerbaijan Province, North-West of Iran.

Babesia ovis, an intraerythrocytic parasite carried by ticks and one of the most common subclinical ovine illnesses, was studied to ascertain its seroprevalence and endemic status in ram and ewe populations in East Azerbaijan Province, Iran, in lambs, yearlings, and adults of over two years of age. A total of 960 sheep from 10 cities were selected from Jan 2018 to Nov 2019. Blood samples were collected from each animal and tested for the presence of B. ovis antibodies by applying a developed enzyme-linked immunosorbent assay (ELISA) technique. Checkerboard titrations were used to determine the optimal dilution of the antigen using negative and positive control sera. To determine whether the disease is endemically stable, inoculation rates for each age group were also calculated. Correlation coefficients were calculated between age and infection rates and also between age and inoculation rates. The results revealed an average infection rate of 49.4% in East Azerbaijan Province. There was a positive correlation between the age of animals and susceptibility to infection except for lambs and yearlings, whereas there was no meaningful difference in exposure to B. ovis between rams and ewes. The negative correlation between age and inoculation rates indicates increased disease instability with age. Inoculation rate results revealed the endemically instable status of B. ovis in the studied area. High prevalence rates and endemically instable status of the disease suggest demand for vaccine development and implementation of appropriate control measures for ovine babesiosis to mitigate the associated economic losses.