HDAC2 dysregulation in the nucleus basalis of Meynert during the progression of Alzheimer's disease.

AIMS: Alzheimer's disease (AD) is characterized by degeneration of cholinergic basal forebrain (CBF) neurons in the nucleus basalis of Meynert (nbM), which provides the major cholinergic input to the cortical mantle and is related to cognitive decline in patients with AD. Cortical histone deacetylase (HDAC) dysregulation has been associated with neuronal degeneration during AD progression. However, whether HDAC alterations play a role in CBF degeneration during AD onset is unknown. We investigated global HDAC protein levels and nuclear HDAC2 immunoreactivity in tissue containing the nbM, changes and their association with neurofibrillary tangles (NFTs) during the progression of AD. METHODS: We used semi-quantitative western blotting and immunohistochemistry to evaluate HDAC and sirtuin (SIRT) levels in individuals that died with a premortem clinical diagnosis of no cognitive impairment (NCI), mild cognitive impairment (MCI), mild/moderate AD (mAD) or severe AD (sAD). Quantitative immunohistochemistry was used to identify HDAC2 protein levels in individual cholinergic nbM nuclei and their colocalization with the early phosphorylated tau marker AT8, the late-stage apoptotic tau marker TauC3 and Thioflavin-S, a marker of ß-pleated sheet structures in NFTs. RESULTS: In AD patients, HDAC2 protein levels were dysregulated in the basal forebrain region containing cholinergic neurons of the nbM. HDAC2 nuclear immunoreactivity was reduced in individual cholinergic nbM neurons across disease stages. HDAC2 nuclear reactivity correlated with multiple cognitive domains and with NFT formation. CONCLUSIONS: These findings suggest that HDAC2 dysregulation contributes to cholinergic nbM neuronal dysfunction, NFT pathology, and cognitive decline during clinical progression of AD.

Training-related improvements in musculoskeletal health and balance: a 13-week pilot study of female cancer survivors.

Cancer survivors often experience poor post-treatment musculoskeletal health. This study examined the feasibility of combined aerobic and resistant training (CART) for improving strength, skeletal health and balance. Cancer survivors (n = 24) were identified by convenience sampling in Los Angeles County with 11 survivors consenting to 13 weeks of CART. Pre- and post-intervention assessments of bone mineral density (BMD), strength, flexibility and biomarker analysis were performed. Paired t-test analysis suggested increases in lower and upper body strength. The average T-score for BMD at the femoral neck improved from -1.46 to -1.36 and whole body BMD improved from -1.65 to -1.55. From baseline to follow-up, participants also displayed decreases in sway velocity on the eyes open (7%) and eyes closed (27%) conditions. Improvement in lower body strength was associated with increases in lean body mass (LBM) (r = 0.721) and an inverse association was observed between sway velocity and LBM (r = 0.838). Age and time since last treatment were related with biomarkers of anabolic growth (IGF-1, IGFbp-3) and bone (DPD, BAP). In summary, observed physiological changes were consistent with functional improvements, suggesting that isometric and dynamic exercise prescription may reduce the risk for falls and fall-related fractures among survivors.

Comparative study on the in vitro replication and genomic variability of Argentinean field isolates of bovine herpesvirus type 4 (BoHV-4).

Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus, belonging to the Rhadinovirus genus, which is increasingly associated with various problems of the reproductive tract of cattle. In Argentina, analysis of BoHV-4 strains isolated from cervico-vaginal mucus of aborted cows revealed a high genetic divergence among strains, which could be classified in three different groups: Genotype 1 comprises Movar-like strains (European prototype), Genotype 2 includes DN599-like strains (American prototype) and Genotype 3 corresponds to a novel genotype group. Understanding the replication behavior in cell cultures and the molecular characteristics of this pathogen of cattle is critical for the rational design of in vitro experiments. The aim of this work was to quantitatively evaluate the replication properties of different Argentinean BoHV-4 strains and to characterize their phylogenetic relationships. Significant differences were evident among the virus titers of the different BoHV-4 isolates in vitro. The most conserved gene was the major capsid protein (ORF25). The glycoprotein B (gB), glycoprotein H (gH), and thymidine kinsase (TK) genes displayed both synonymous and non-synonymous substitutions, with the highest diversity observed for gB, which displayed amino acid substitutions in 24 out of the 178 positions examined. Strains 09/759, 12/512, and 07/568 presented a deletion encompassing amino acid position 27 to 35, whereas strains 07/435 and 09/227 had a deletion from position 28 to 35. Two strains, 07/435 and 09/227, also displayed the highest divergence compared to the other strains analyzed. This study provides information about the in vitro replication and behavior of nine field isolates of BoHV-4. These findings are relevant since available information on the in vitro growth characteristics of BoHV-4 strains is scarce. The results from this study may also be useful for establishing comparisons with other related viruses.

Resistance training suppresses intra-abdominal fatty acid synthesis in ovariectomized rats.

Ovarian hormone loss is associated with a shift in fat distribution to intra-abdomin al adipose tissue (intra-AAT) depots and with lipid metabolism disorders, which predisposes individuals to developing insulin resistance. Resistance training (RT) prevents increases in intra-AAT after ovarian hormone loss. However, the molecular mechanisms underlying these changes remain unclear. We investigated the effects of ovariectomy and RT on gene expression related to lipogenesis and fat oxidation in the intra-AAT of ovariectomized rats. Sprague-Dawley rats (n=6/group) were divided into the groups: sham-sedentary, ovariectomized-sedentary, sham-RT and ovariectomized-RT. RT groups performed a 10-week climbing program on a ladder with progressive overload. Intra-AAT was subjected to morphometric and mRNA analysis. Ovariectomized-sedentary group had larger adipocytes and higher expression of peroxisome proliferator-activated receptor-γ (PPAR-γ), sterol regulatory element-binding protein-1c (SREBP-1c), stearoyl-CoA desaturase-1 (SCD-1), acetyl-CoA carboxylase (ACC), hormone-sensitive lipase (HSL) and lower expression of the oxidative carnitinepalmitoyltransferase-I (CPT-1). RT counteracted OVX-induced increases in PPAR-γ and SCD-1 and decreased SREBP-1c. ACC and HSL were downregulated in ovariectomized-RT compared with the ovariectomized-sedentary group. Ovariectomized-RT group had the highest CPT-1 gene expression. Adipocyte size decreased in ovariectomized-RT group. Results suggest that RT reduces intra-AAT adipocyte size in ovariectomized rats by suppressing intra-AAT fatty acid synthesis and enhancing fatty acid ß-oxidation.

Effects of long-term periodized resistance training on body composition, leptin, resistin and muscle strength in elderly post-menopausal women.

AIM: Objective of the study was to determine the effects of a periodized resistance training program on body composition, plasmatic levels of leptin and resistin, and muscle strength in elderly post-menopausal women. METHODS: Twenty-three post-menopausal women (age= 63.02±4.42 years; height 1.55±0.06 m; body mass 67.56±2.26 kg) were submitted to 12 months of periodized resistance training twice a week. The training program consisted of 3 sets of 6-14 repetitions maximal (RM). Body composition (DXA), muscle strength (bench press, leg press 45º and arm curl), plasmatic levels of resistin and leptin (ELISA method) were assessed before and after the training program. Paired Student's t test was used for comparison between pre- and post-training values. RESULTS: There was a significant increase in muscle strength and lean body mass; decrease in body mass, body fat percentage and fat mass after 12 months of resistance training, a part from the decrease in leptin and resistin levels. CONCLUSION: Long-term periodized resistance training prevents aging sarcopenia, decreases body fat and systemic markers of inflammation in postmenopausal elderly women.

Acute resistance training affects cell surface markers for apoptosis and migration in CD4⁺ and CD8⁺ lymphocytes.

The purpose of the present study was to examine the acute effects of resistance training (RT) on CD4⁺ and CD8⁺ T lymphocytes apoptosis (annexin V⁺) and migration (CX3CR1). Twelve subjects performed two RT sessions (3 sets of 9 exercises) with 1 min (Hyper-1) and 3 min (Hyper-3) of rest-interval length between sets and exercises. CD4⁺ and CD8⁺ cells count displayed no change following Hyper-1 and Hyper-3. There was an increase in the percentage of CD4⁺ positive for annexin V⁺ and CX3CR1⁺ immediately after and 24 h post Hyper-1. Percentage of CD4⁺ positive for annexin V⁺ increased 2 and 24 h post Hyper-3, and decreased after CXCR1⁺ for the same time-points. There was an increase in CD8⁺ positive for annexin V⁺ and CX3CR1⁺ immediately after, 2 and 24 h post Hyper-1 and Hyper-3, while no differences were found between Hyper-1 and Hyper-3. Acute RT increase the apoptosis and migration of CD4⁺ and CD8⁺ lymphocytes even 24h after exercise, with minimal effects of rest-interval length.

Effects of resistance training on matrix metalloproteinase-2 activity and biomechanical and physical properties of bone in ovariectomized and intact rats.

The purpose of this study was to investigate the influence of resistance training on the activity of matrix metalloproteinase (MMP)-2 and bone biomechanical properties in ovariectomized and intact rats. Forty-eight female rats were divided into two distinct groups, ovariectomized (OVX) and intact (Int), which were subdivided into three similar subgroups: sedentary, acute exercise and chronic exercise. Rats performed a resistance training for 12 weeks in which animals climbed a vertical ladder of 1.1 m with weights attached to their tails. Sessions were performed with an interval of 3, 4-9 and 8-12 days scaled dynamic movements of climbing. Biomechanical and physical analyses were performed using a universal testing machine, and MMP-2 activity analysis by zymography. Bone density (BD), mineral density (MD), maximum load and fracture load was reduced in sedentary and acute exercise OVX groups compared with the sedentary intact group (P<0.05); in contrast, chronically trained groups (OVX and Int) showed a significant increase in BD, MD and fracture load compared with all the other groups. MMP-2 activity in chronically trained groups also showed a significant increase, while the sedentary OVX group showed a decrease in MMP-2 activity compared with the intact sedentary group (P<0.05). Our results suggest that the resistance training proposed in our work was efficient in reverting the deleterious effects of ovariectomy on bone tissue, and also produced modeling effects in intact rats. On the other hand, ovariectomy reduced the activity of MMP-2 and produced deleterious effects on bone tissue, mimicking menopause intrinsically.

Acute cardiorespiratory and metabolic responses during resistance exercise in the lactate threshold intensity.

The aims were both to determine lactate and ventilatory threshold during incremental resistance training and to analyze the acute cardiorespiratory and metabolic responses during constant-load resistance exercise at lactate threshold (LT) intensity. Ten healthy men performed 2 protocols on leg press machine. The incremental test was performed to determine the lactate and ventilatory thresholds through an algorithmic adjustment method. After 48 h, a constant-load exercise at LT intensity was executed. The intensity of LT and ventilatory threshold was 27.1±3.7 and 30.3±7.9% of 1RM, respectively (P=0.142). During the constant-load resistance exercise, no significant variation was observed between set 9 and set 15 for blood lactate concentration (3.3±0.9 and 4.1±1.4 mmol x L(-1), respectively. P=0.166) and BORG scale (11.5±2.9 and 13.0±3.5, respectively, P=0.783). No significant variation was observed between set 6 and set 15 for minute ventilation (19.4±4.9 and 22.4±5.5 L x min(-1), respectively, P=0.091) and between S3 and S15 for VO2 (0.77±0.18 and 0.83±0.16 L x min(-1), respectively, P=1.0). Constant-load resistance exercise at LT intensity corresponds to a steady state of ventilatory, cardio-metabolic parameters and ratings of perceived exertion.

Resistance training and glycogen content in ovariectomized rats.

The aim of the present study was to investigate the effects of resistance training on glycogen content and muscle cross-sectional area (CSA) in ovariectomized rats. Wistar rats were divided into: sedentary; ovariectomized sedentary; resistance trained; and ovariectomized resistance trained. In the 12-week resistance training, the animals climbed a 1.1 m vertical ladder, 3 days per week, with 4-8 climbs. Cardiac, liver and muscle glycogen content was determined. After the 12-week resistance training period there was a higher hepatic and muscle glycogen content in the resistance training group compared with the other groups (p<0.01). CSA was higher in soleus for the resistance trained, ovariectomized resistance trained and sedentary compared with ovariectomized sedentary (p<0.05). Ovariectomy attenuated the increase in liver and muscle glycogen content, while soleus muscle cross-sectional area increased with resistance training, even in ovariectomized rats. Resistance training could be an important exercise to increase muscle function in situations of reduced estrogen and progesterone.

Infection by bovine alphaherpesvirus types 1 and 5 induces IFN-λ3 expression in neuronal-type cells and bovine neural tissues.

Type III interferons (IFNs) are components of the innate immunity, with IFN lambda- (λ)3 having the most potent bioactivity in humans. IFN-λ has a predominant role in epithelial cells. However, antiviral function in certain infections of the central nervous system has also been demonstrated. IFN-λ3 expression in neural tissues of cattle has not been investigated. Thus, the aim of this study was to analyze whether an antiviral IFN-λ3 response is mounted after infection with bovine alphaherpesviruses (BoHV-1 and BoHV-5) in vitro, in neuronal-type cells, and in neural tissues from experimentally-infected calves. This study demonstrated that there is a strong IFN-λ3 response early after BoHV-1infection of undifferentiated neuroblastoma cells. During acute BoHV-1 and BoHV-5 infection of calves, low levels of IFN-λ3 expression were detected in the brain, which would favor virus spread within this tissue. Striking differences in the transcriptional levels of IFN-λ3 were observed in trigeminal ganglion, particularly in BoHV-1-acutely- and latently-infected calves. During reactivation, IFN-λ3 expression was down-regulated, which may be a requirement for virus replication and spread. Overall, different patterns of IFN-λ3 expression were detected during BoHV-1 and BoHV-5 infection, particularly during latency.

Biomechanical responses of different rat tendons to nandrolone decanoate and load exercise.

Androgenic-anabolic steroids (AAS) have been associated with an increased incidence of tendon rupture. The aim of this study was to compare the biomechanical properties of the rat calcaneal tendon (CT), superficial flexor tendon (SFT), and deep flexor tendon (DFT), and to determine the effect of jump training in association with AAS. Animals were separated into four groups: sedentary, trained, AAS-treated sedentary rats (AAS), and AAS-treated and trained animals. Mechanical testing showed that the CT differed from the DFT and SFT, which showed similar mechanical properties. Jump caused the CT to exhibit an extended toe region, an increased resistance to tensional load, and a decreased elastic modulus, characteristics of an elastic tendon capable of storing energy. AAS caused the tendons to be less compliant, and the effects were reinforced by simultaneous training. The DFT was the most affected by training, AAS, and the interaction of both, likely because of its involvement in the toe-off step of jumping, which we suggest is related to the rapid transmission of force as opposed to energy storage. In conclusion, tendons are differently adapted to exercise, but responded equally to AAS, showing reduced flexibility, which is suggested to increase the risk of tendon rupture in AAS consumers.

Menopause: highlighting the effects of resistance training.

The increase in lifespan and in the proportion of elderly women has increased the focus on menopause induced physiological alterations. These modifications are associated with the elevated risk of several pathologies such as cardiovascular disease, diabetes, obesity, hypertension, dyslipidemia, non-alcoholic fat liver disease, among others. Because of estrogen levels decline, many tissue and organs (muscular, bone, adipose tissue and liver) are affected. Additionally, body composition suffers important modifications. In this sense, there is a growing body of concern in understanding the physiological mechanisms involved and establishing strategies to prevent and reverse the effects of menopause. The hormone reposition therapy, diet and physical exercise have been recommended. Among the diverse exercise modalities, resistance training is not commonly used as a therapeutic intervention in the treatment of menopause. Thus, the aim of this review was to analyze the physiological alterations on several organs and systems induced by menopause and ovariectomy (experimental model to reproduce menopause), as well as, to study the effects of resistance training in preventing and reverting these modifications. In conclusion, resistance training promotes beneficial effects on several organs and systems, mainly, on muscular, bone and adipose tissue, allowing for a better quality of life in this population.

Cathelicidin bovine myeloid antimicrobial peptide (BMAP) 28 is involved in the inflammatory response against alpha-herpesviruses in the bovine nervous system.

The role of the local innate immune response in the neuropathogenesis of bovine herpesvirus (BoHV) type 1 and 5 remains largely unknown. This study determined the gene transcriptional expression of relevant bovine cathelicidins, TNFα and IFNß in the nervous system of experimentally-infected cattle during the different stages of BoHV-1 and BoHV-5 infectious cycle. We studied the modulation of bovine myeloid antimicrobial peptide (BMAP) 27 and 28 by alpha-herpesviruses during acute infection of the central nervous system (CNS). However, BMAP28 was the main cathelicidin modulated. BoHV-5 supressed BMAP28 expression mainly in frontal cortex and cervical medulla whereas BoHV-1 slightly induced the expression of cathelicidins in the olfactory and posterior cortex. The differences in the regulation of the innate response are likely related to distinct replication rates of both alpha-herpesviruses in the CNS. During latency and reactivation, BoHV-1 and -5 decreased BMAP28 and BMAP27 expression, accompanied by high levels of TNFα and IFNß transcripts in the posterior brain region and medulla during BoHV reactivation. In terms of cytokines, a remarkably overexpression of IFNß was induced by BoHV-5 (133.8-fold). In trigeminal ganglion (TG) both alpha-herpesviruses induced cathelidicins gene expression at all stages of the infection cycle, while only acute BoHV-5 infection increased TNFα (129-fold) mRNA levels. This study suggests that the pronounced downregulation of BMAP28 in BoHV-5-acutely-infected CNS is due to a decreased immune stimulation during viral infection, favouring its establishment in the CNS with a low replication rate until latency. Thus, cathelicidins, together with IFNß and TNFα, are differentially regulated by BoHV-5 and BoHV-1 infections and this regulation is dependent on the stage of virus infection in the bovine nervous system.

Regional binding of tau and amyloid PET tracers in Down syndrome autopsy brain tissue.

INTRODUCTION: Tau pathology is a major age-related event in Down syndrome with Alzheimer's disease (DS-AD). Although recently, several different Tau PET tracers have been developed as biomarkers for AD, these tracers showed different binding properties in Alzheimer disease and other non-AD tauopathies. They have not been yet investigated in tissue obtained postmortem for DS-AD cases. Here, we evaluated the binding characteristics of two Tau PET tracers (3H-MK6240 and 3H-THK5117) and one amyloid (3H-PIB) ligand in the medial frontal gyrus (MFG) and hippocampus (HIPP) in tissue from adults with DS-AD and DS cases with mild cognitive impairment (MCI) compared to sporadic AD. METHODS: Tau and amyloid autoradiography were performed on paraffin-embedded sections. To confirm respective ligand targets, adjacent sections were immunoreacted for phospho-Tau (AT8) and stained for amyloid staining using Amylo-Glo. RESULTS: The two Tau tracers showed a significant correlation with each other and with AT8, suggesting that both tracers were binding to Tau deposits. 3H-MK6240 Tau binding correlated with AT8 immunostaining but to a lesser degree than the 3H-THK5117 tracer, suggesting differences in binding sites between the two Tau tracers. 3H-THK5117, 3H-MK6240 and 3H-PIB displayed dense laminar binding in the HIPP and MFG in adult DS brains. A regional difference in Tau binding between adult DS and AD was observed suggesting differential regional Tau deposition in adult DS compared to AD, with higher THK binding density in the MFG in adult with DS compared to AD. No significant correlation was found between 3H-PIB and Amylo-Glo staining in adult DS brains suggesting that the amyloid PIB tracer binds to additional sites. CONCLUSIONS: This study provides new insights into the regional binding distribution of a first-generation and a second-generation Tau tracer in limbic and neocortical regions in adults with DS, as well as regional differences in Tau binding in adult with DS vs. those with AD. These findings provide new information about the binding properties of two Tau radiotracers for the detection of Tau pathology in adults with DS in vivo and provide valuable data regarding Tau vs. amyloid binding in adult DS compared to AD.

Kinesin-II is required for axonal transport of choline acetyltransferase in Drosophila.

KLP64D and KLP68D are members of the kinesin-II family of proteins in Drosophila. Immunostaining for KLP68D and ribonucleic acid in situ hybridization for KLP64D demonstrated their preferential expression in cholinergic neurons. KLP68D was also found to accumulate in cholinergic neurons in axonal obstructions caused by the loss of kinesin light chain. Mutations in the KLP64D gene cause uncoordinated sluggish movement and death, and reduce transport of choline acetyltransferase from cell bodies to the synapse. The inviability of KLP64D mutations can be rescued by expression of mammalian KIF3A. Together, these data suggest that kinesin-II is required for the axonal transport of a soluble enzyme, choline acetyltransferase, in a specific subset of neurons in Drosophila. Furthermore, the data lead to the conclusion that the cargo transport requirements of different classes of neurons may lead to upregulation of specific pathways of axonal transport.

Modulation of cathelicidins, IFNß and TNFα by bovine alpha-herpesviruses is dependent on the stage of the infectious cycle.

Production of antimicrobial peptides cathelicidins, interferons and cytokines is an important feature in airway epithelial host defense. The innate immune response to alpha-herpesvirus infection at the sites of primary replication has not been fully studied. Thus, the aim of this study was to determine the expression of innate immune components, cathelicidins, IFNß, TNFα and TNF receptors (TNFRI and TNFRII) during acute infection and reactivation of bovine herpesvirus type 1 (BoHV-1) and 5 (BoHV-5) in the respiratory tract and lymphoid tissue of their natural host. We found that BoHV infection modulates mainly the expression of BMAP28, a key cathelicidin in cattle. It was downregulated by both viruses in retropharyngeal lymph nodes of acutely infected-calves, and it was accompanied by a lower expression of IFNß, TNFα and TNFRI. BoHV-5 showed a pronounced role in the downregulation of BMAP28, even in nasal mucosa and lung. However, during reactivation, BoHV-5 upregulated both BMAP28 and IFNß in retropharyngeal lymph nodes. Acute replication induced also TNFα mRNA and protein synthesis, and expression of TNFRI and II was positively regulated during both acute infection and reactivation, particularly in the trachea. Moreover, BMAP27 was detected during BoHV-1 reactivation suggesting a potential role at this stage. Thus, cathelicidins are implicated in alpha-herpesvirus infections of the bovine respiratory system and the response is distinct during BoHV-1 and BoHV-5 acute infection and reactivation. This demonstrates that these viruses modulate differentially the components of innate immune response, possibly influencing their pathogenesis. This study provides an initial pilot analysis of factors that might be implicated in alpha-herpesvirus infection of the bovine respiratory system.

Effect of magnetic ion exchange (MIEX®) on removal of emerging organic contaminants.

In this study, the removal of nine emerging organic contaminants was investigated by using anion exchange resins. The selected compounds were carbamazepine, atrazine, simazine, estrone, bisphenol A, methylparaben, ethylparaben, propylparaben and butylparaben. Two different magnetic anionic exchanger resins were tested: MIEX® DOC and MIEX® GOLD. The optimal resin dose (40 mL/L) and contact time (20 min) had been previously determined. Once these optimum parameters were set, the effect of the initial concentration of contaminants on the removal efficiency of the contaminants by the resins was studied. The study was carried out using mono and multicomponent systems, with distilled water and natural waters, to which contaminants had been previously added, in order to evaluate the competitive and matrix effects. Results showed that the average removal percentages obtained with the MIEX® DOC resin were: 51%, 61%, 68% and 80% for methyl-, ethyl-, propyl-, and butylparaben, respectively. For bisphenol A the result was similar, i.e., 66%, whereas for the rest of the compounds studied, removal efficiencies lower than 15% were obtained. The MIEX® GOLD resin achieved lower elimination rates than the MIEX® DOC resin in all cases.

Immunization with inactivated antigens of Neospora caninum induces toll-like receptors 3, 7, 8 and 9 in maternal-fetal interface of infected pregnant heifers.

Neospora caninum is an obligate parasite and a major cause of abortion in cattle. Pregnancy failures appear to be associated with weak innate defences on the maternal-fetal interface during infection with N. caninum. Herein, we studied the gene expression of Toll-like receptors (TLRs) in pregnant heifers immunized with different vaccine formulations against N. caninum before mating and then challenged the heifers with live N. caninum on day 70 of gestation. TLR7 and TLR8 expression was upregulated in the placental caruncle of infected-pregnant heifers previously exposed to live N. caninum as immunogen. However, TLR7 and 8 expression in both placenta and caruncle as well as, TLR3 and 9 expression in caruncle were upregulated when heifers were previously immunized with inactivated soluble whole antigens and recombinant NcSAG1, NcHSP20 and NcGRA7 proteins. All dams were carrying viable fetuses when they were culled at day 104 of gestation. Upregulation of TLR7 and IFNγ expression was detected in fetal spleen when their mothers where previously vaccinated with soluble antigens and recombinant NcSAG1, NcHSP20 and NcGRA7 proteins. These studies demonstrate that soluble or recombinant NcSAG1, NcHSP20 and NcGRA7 antigens induce key TLRs expression at the maternal-fetal interface, probably triggering damaging inflammatory cellular immune responses associated with abortion. Previous infection with N. caninum seems to attenuate the innate immune response at the maternal-fetal interface, which could favour pregnancy maintenance and perpetuation of the disease. This finding represents novel information on how N. caninum vaccination and infection modulate TLRs expression at the placenta and fetal spleen, the possible role in the pregnancy outcomes and transplacental transmission of the protozoa.

Toll-like receptors 3, 7 and 8 are upregulated in the placental caruncle and fetal spleen of Neospora caninum experimentally infected cattle.

Innate immune responses at the maternal-fetal interface are key in the pathogenesis of Neospora caninum, an obligate parasite that causes abortion in cattle. Herein, we determined the gene expression of endosomal Toll-like receptors (TLRs) in the placenta and fetuses from both non-infected pregnant heifers and pregnant heifers intravenously challenged with live tachyzoites of N. caninum on day 70 of gestation. On day 104 of pregnancy, mRNA expression of TLRs 3 and 8, as well as that of TLRs 7 and 9, was high in the spleen of fetuses from N. caninum-infected heifers. Gene expression levels of endosomal TLRs were also detectable in the placenta and the maternal caruncle from infected heifers, being TLRs 3, 7 and 8 particularly upregulated, mostly in the caruncle. Basal TLR levels were higher in fetal spleen than in placental tissues. This study provides novel information on how innate TLR responses are induced at the maternal-fetal interface of cattle in response to intracellular N. caninum.