Parturition is a challenging physiological process with perfect timing dictated by the events leading to the end of pregnancy in the female, and by the maturation of the fetus(es). The process of ...parturition remains an intricate interaction of hormones in a fine-tuned timing that remains to be better elucidated in the dog. In the dog pregnancy is maintained by the progesterone production by the corpora lutea, in which some hormones play a luteotrophic action. At term of pregnancy, the pre-parturient luteolytic cascade is the most apparent event, characterized by a rapid decline in plasma progesterone concentrations and useful in predicting the onset of spontaneous parturition. In contrast, cortisol plasma concentrations measurement showed high variability and suggested to be related to the stress condition instead of the onset of parturition. Both prostaglandin F2α and E2 concur in the process of parturition in the dog. The measurement of oxytocin plasma concentrations, also very variable, is implicated in uterine contractions. The measurement of plasma oxytocin concentrations showed to be useful to distinguish between normal and disturbed parturition, allowing the prompt recognition of dystocia and the immediate obstetrical intervention. In contrast to other species, no significant roles of estrogens for the initiating of parturition were demonstrated. Relaxin, the main pregnancy hormonal marker in the dog, beside an endocrine action, is also supposed to play a paracrine/autocrine role at the utero-placental unit and to support the maintenance of high plasma progesterone concentrations through a luteotrophic action, concurring to the main action of prolactin. Although important information has been provided, some aspects in the understanding the hormonal interactions and action timing implications in the process of parturition in the dog remains to be better investigated and represent intriguing topic for basic knowledge and applied research.
Contents
Perinatal mortality is relatively high in dogs, with deaths peaking around the time of birth and during the first week of age. Among the several causes of canine perinatal mortality, ...whelping is the greatest cause. Therefore, early neonatal assistance at birth should be mandatory with dogs. In comparison with human neonatology, knowledge and technological ability in canine neonatology is tremendously scarce. The Apgar score for the newborn viability assessment at birth represents a feasible method for the prompt recognition of newborns that will need special assistance immediately after birth. The five parameters of the Apgar score were adapted to the canine species by different studies. Advantages and limits, as well as clinical applications, are presented and discussed in further detail. It was concluded that the Apgar score represents the easiest and simplest, non‐invasive and reliable method, that could be performed under every clinical and practical condition, for newborn puppies viability evaluations and short‐term survival prognosis.
Parkinson's disease is a neurodegenerative disorder characterized by the death of dopaminergic neurons and by accumulation of alpha-synuclein (aS) aggregates in the surviving neurons. The dopamine ...catabolite 3,4-dihydroxyphenylacetaldehyde (DOPAL) is a highly reactive and toxic molecule that leads to aS oligomerization by covalent modifications to lysine residues. Here we show that DOPAL-induced aS oligomer formation in neurons is associated with damage of synaptic vesicles, and with alterations in the synaptic vesicles pools. To investigate the molecular mechanism that leads to synaptic impairment, we first aimed to characterize the biochemical and biophysical properties of the aS-DOPAL oligomers; heterogeneous ensembles of macromolecules able to permeabilise cholesterol-containing lipid membranes. aS-DOPAL oligomers can induce dopamine leak in an in vitro model of synaptic vesicles and in cellular models. The dopamine released, after conversion to DOPAL in the cytoplasm, could trigger a noxious cycle that further fuels the formation of aS-DOPAL oligomers, inducing neurodegeneration.
Hair steroid measurement has received increasing attention for monitoring hypothalamic-pituitary-adrenal axis function, as it offers the advantages of being noninvasive, fast, and able to indicate ...steroid concentrations over long periods. The objects of the study were to evaluate cortisol (C) and dehydroepiandrosterone sulfate (DHEA-S) hair concentrations and their ratio (C/DHEA-S) in beef cows from calving to 100 days (d) postpartum (pp) and to assess possible differences related to parity (primiparous vs multiparous) and conception outcome (pregnant vs not pregnant). Hair samples were collected from 6 primiparous and 5 multiparous pregnant beef cows by clipping the coat at calving (T0) and every 20 d for 5 times (T1-T5), collecting only the regrown hair. Starting from the 6th-week pp, cows were submitted to artificial insemination at spontaneous estrus; by 100 d pp, 7 cows were pregnant and 4 were not pregnant. Statistical analysis showed higher hair C concentrations in the 11 cows at calving (T0) compared to all the subsequent samplings except for T1, and higher C concentrations at T1 compared to T3, T4, and T5. These results indicate that hair C concentrations in beef cows are affected by sampling time, with a decrease from calving, as reported in other matrices. When exploring changes within parity groups, no differences were found in the multiparous among sampling times, while hair C concentrations at T0 and T1 tended to be higher than at T2 (0.01 ≤ p < 0.05) and were higher (p < 0.01) than in all the subsequent samplings (T3, T4 and T5) within the primiparous group. Higher hair C concentrations were found at T0 and T1 in the primiparous compared to multiparous (p < 0.01), suggesting that primiparous cows undergo a greater stress level before and around parturition compared to multiparous, probably due to the novelty of the calving experience. No differences were detected in C hair concentrations according to conception outcome (pregnant versus not pregnant) in each sampling time. Hair DHEA-S concentrations were neither affected by time nor by parity or conception outcome. Differences in the C/DHEA-S ratio were found at T1, with higher C/DHEA-S in the multiparous compared to primiparous cows (p < 0.001), and a tendency for higher ratio in the not pregnant compared to the pregnant (0.01 ≤ p < 0.05). These results support the choice of hair as a valuable biological matrix when investigating long-time periods such as postpartum in cows and suggest an enhanced immunoprotective effect of DHEA-S in the postpartum of primiparous cows, and in cows that get pregnant within 100 d postpartum.
Cortisol (C) and dehydroepiandrosterone (DHEA) are recognized as the main fetal steroids, and they are likely to influence fetal development and have long-term effects on newborn ...hypothalamic-pituitary-adrenal axis (HPA) function. DHEA is often measured as its sulfates and expressed as DHEA-S. Hair analysis represents a promising methodological approach for the non-invasive measurement of steroids, allowing for a retrospective analysis of the total exposure to steroids over time, and avoiding the influence of acute events or circadian fluctuations. Hair cortisol and DHEA concentrations have been investigated in cows, but no studies have been performed on calves. The object of this study was to evaluate hair cortisol (HC) and hair DHEA-S (HDHEA-S) concentrations in beef calves from birth to six months of age. Hair samples of 12 beef calves (seven males, five females) were firstly collected at birth (T1) and then every three weeks up to six months of age (T2-T10), collecting only the re-grown hair. HC and HDHEA-S were analyzed by radioimmunoassay (RIA). Calves sex, weight and APGAR score were registered immediately after birth. Statistical analysis revealed that both HC and HDHEA-S were influenced by sampling time (P < 0.001). HC concentrations were higher at T1 compared to all subsequent samplings (T2-T10, P < 0.01); HC concentrations were higher at T2 compared to T4-T10 (P < 0.01), while no further changes were detected from T3 onward. Higher HDHEA-S concentrations were registered at T1, T2 and T3 compared to all the other samplings (P < 0.01). No correlation was found between hair concentrations of both steroids and calf sex or birthweight. APGAR score was negatively correlated only with HC at birth (P < 0.05). These data demonstrate that C and DHEA-S are quantifiable in the hair of calves and are influenced by their age. The higher HC detected at birth (T1) probably reflects the high serum C concentrations present late in pregnancy and increased by the fetal HPA axis, by which parturition is initiated in cows. The highest HDHEA-S at birth (T1) in calves indicates that the largest amounts of DHEA and its sulfates are produced during fetal development. Moreover, the findings of higher HC at three weeks after birth and of higher HDHEA-S until six weeks after birth, suggest that C and DHEA secretion continues also beyond birth, and that these steroids could be involved in the events occurring during the challenging first weeks of age in the calf.
•The utility of hair matrix for retrospective hormonal changes was assessed in calves.•Cortisol and DHEA-S hair concentrations were investigated from birth to 6 months of age.•A significant effect of age was found on both hormones hair concentrations.•Stable hair C and DEHA-S concentrations were reached 3 and 6 weeks after birth respectively.•Both steroids could be involved in the events occurring in the first weeks of age in calves.
Objective
Attenuation of the growth supportive environment within the distal nerve stump after delayed peripheral nerve repair profoundly limits nerve regeneration. Levels of the potent Schwann cell ...mitogen neuregulin and its receptor ErbB2 decline during this period, but the regenerative impact of this change is not completely understood. Herein, the ErbB2 receptor pathway is inhibited with the selective monoclonal antibody Herceptin (trastuzumab) to determine its significance in regulating acute and chronic regeneration in a rat hindlimb.
Methods
The common peroneal nerve of Sprague–Dawley rats was transected and repaired immediately or after 4 months of chronic denervation, followed by administration of Herceptin or saline solution. Regenerated motor and sensory neurons were counted using a retrograde tracer 1, 2, or 4, weeks after repair. Distal myelinated axon outgrowth after 4 weeks was quantified using histomorphometry. Immunofluorescent imaging was used to evaluate Schwann cell proliferation and epidermal growth factor receptor (EGFR) activation in the regenerating nerves.
Results
Herceptin administration increased the rate of motor and sensory neuron regeneration and the number of proliferating Schwann cells in the distal stump after the first week. Herceptin also increased the number of myelinated axons that regenerated 4 weeks after immediate and delayed repair. Reduced EGFR activation was observed using immunofluorescent imaging.
Interpretation
Inhibition of the ErbB2 receptor with Herceptin unexpectedly enhances nerve regeneration after acute and delayed nerve repair. This finding raises the possibility of using targeted molecular therapies to improve outcomes of peripheral nerve injuries. The mechanism may involve a novel inhibitory association between ErbB2 and EGFR. Ann Neurol 2016;80:112–126
A proper canine neonatal assistance, required to reduce the high perinatal loss rate, imply a full knowledge about the fetal-to-neonatal physiology. Because fetal fluids play an important role ...throughout mammals pregnancy, influencing fetal growth and development, fetal well being, and contributing to guarantee the most suitable environment for the fetus, the knowledge about fetal fluids biochemical composition is of major importance. At first, the biochemical composition of fetal fluids collected by normal developed, healthy and viable newborns, is necessary to depict the normal features, and represent the first step for the further detection of abnormalities associated to fetal/neonatal distress and useful for the early identification of newborns needing special attention, immediately after birth. The present study was aimed to define the biochemical composition of amniotic and allantoic fluids collected from fetus delivered by caesarean section at term of pregnancy. To reduce the possible confounding effect of maternal labor or troubles at parturition, fetal fluids were collected only from puppies born by elective caesaeran section, at term of normal pregnancies. Fetal fluids from 76 puppies, 70 normal and six pathologic newborns, born by elective caesarean section were collected and analyzed for alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, lactate dehydrogenase (LDH), creatine-kinase (CK), alkaline phosphatase (ALP), creatinine, urea, amylase, lipase, gamma-glutamyl transferase (γ-GT), triglycerides, cholesterol, total proteins, albumin, globulins, glucose, magnesium, potassium, chloride, sodium, calcium, phosphorus and osmolarity. No significant differences were found between biochemical composition of amniotic or allantoic fluid in normal and pathologic newborns, maybe due to the small number of the pathologic puppies. Although some correlations between the two fluids were found (albumin, phosphorus, glucose and triglycerides), the results showed significant differences between the amniotic and allantoic biochemical composition (for all the parameters, except of alanine aminotransferase, triglycerides, cholesterol, albumin, amylase and glucose), suggesting that diverse sources could concur to the final composition of each fluid. A wide variability within and among litters was found for both amniotic and allantoic biochemical composition, and for some parameters an influence of breed body size (amniotic amylase, cholesterol, and allantoic calcium and glucose), maternal parity (amniotic and allantoic CK, glucose, LDH, chloride) and newborn gender (allantoic phosphorus) was found. Further investigations are needed for addressing the origin of each fetal fluid biochemical composition in the dog and also to indeep possible differences in fetal fluids biochemical composition between normal and pathologic puppies, providing potential markers for the quick identification of newborns that need special surveillance and cares immediately after birth.
•Biochemical canine fetal fluids composition at term.•Fetal fluids from 76 puppies, 70 normal and six pathologic newborns.•Several significant differences between the amniotic and allantoic biochemical composition.•Some parameters were influenced by breed body size, maternal parity and fetal gender.•No differences between biochemical fetal fluids composition in normal and pathologic newborns.
REV-ERBα and REV-ERBβ nuclear receptors regulate several physiological processes, including circadian rhythm and metabolism. A previous study reported the REV-ERBα gene to be co-overexpressed with ...ERBB2 in breast cancer cell lines. Surprisingly, we found that several tumor types, including a number of breast cancer cell lines, predominantly express the REV-ERBβ variant. This pattern was independent of ERBB2 and ER status, and opposite to that of non-cancer mammary epithelial HMEC cells, in which REV-ERBα was the major variant. Consistent with this molecular profile, REV-ERB target genes in both circadian and metabolic pathways were derepressed upon silencing of REV-ERBβ, but not REV-ERBα. Strikingly, we found that REV-ERBβ is a determinant of sensitivity to chloroquine, a clinically relevant lysosomotropic agent that suppresses autophagy. The cytoprotective function of REV-ERBβ appears to operate downstream of autophagy blockade. Through compound screening, we identified ARN5187, a novel lysosomotropic REV-ERBβ ligand with a dual inhibitory activity toward REV-ERB-mediated transcriptional regulation and autophagy. Remarkably, although ARN5187 and chloroquine share similar lysosomotropic potency and have a similar effect on autophagy inhibition, ARN5187 is significantly more cytotoxic. Collectively, our results reveal that dual inhibition of REV-ERBβ and autophagy is an effective strategy for eliciting cytotoxicity in cancer cells. Furthermore, our discovery of a novel inhibitor compound of both REV-ERB and autophagy may provide a scaffold for the discovery of new multifunctional anticancer agents.
The Apgar scoring system is an easy and reliable method for evaluating both human and animal neonates. However, its use is not widespread in veterinary medicine. The current study assessed a modified ...Apgar scoring system for routine evaluation of newborn puppies. Heart rate, respiratory effort, reflex irritability, motility, and mucus color have been evaluated in the score. Specifically, we used 5
min after birth Apgar score to assess newborn viability and short-term survival prognosis, as well as related characteristics, in 193 puppies from 42 litters, 65 born by spontaneous delivery, 66 by assisted delivery, and 62 by cesarean section. The percentage of puppies that were dead 2
h after birth was higher in the 4 to 6 Apgar score group versus that in the 7 to 10 score group (P
<
0.01) and in the 0 to 3 score group versus that in the 7 to 10 score group (P
<
0.0001). Delivery method did not affect survival. There was a marked reduction in the number of puppies searching for the mammary gland in the 0 to 3 and 4 to 6 Apgar score groups compared with that in the 7 to 10 score group (P
<
0.0001); there was a difference between the 0 to 3 and the 4 to 6 score groups as well (P
<
0.05). Suckling/swallowing reflexes were present in fewer puppies in the 0 to 3 and 4 to 6 score groups compared with that in the 7 to 10 group (P
<
0.0001), with no significant differences between the 0 to 3 and the 4 to 6 score groups.
Along the perinatal period, in mammals, cortisol (C) plays a pivotal role in the final intrauterine fetal maturation and in the early neonatal adaptation. Because of the scarce knowledge about canine ...perinatology, the present study was aimed to assess the C concentrations in amniotic and allantoic fluids collected, without invasiveness, from small-sized, purebred newborn puppies born by elective cesarean section, at term of pregnancy. Possible correlations between fetal fluid C concentrations and maternal parity, litter size, birth weight, Apgar score, were evaluated. In addition, the possible effect of fetal fluid C concentrations on newborn survival at 24 hours of age, and the effect of the litter or the newborn gender on fetal fluid C concentrations were also assessed. The results, obtained from 50 born alive, normal-weight puppies, without gross physical malformation, showed that C concentration was higher in allantoic than in amniotic fluid (P < 0.01), even if a strong positive correlation between the two fluids C concentration was found (P < 0.0001; R = 0.83). Neither amniotic nor allantoic C concentrations were correlated to maternal parity, litter size, birth weight, and Apgar score. Interestingly, higher amniotic (P < 0.05), but not allantoic, C concentrations were found in puppies not surviving at 24 hours after birth. Therefore, it could be suggested that this parameter may be useful for the recognition, at birth, of puppies needing special surveillance during the first day of age. A significant (P < 0.001) effect of the litter in both amniotic and allantoic C concentrations was found. In conclusion, the present results showed that in small-sized purebred puppies, born at term by elective cesarean section, the exact fetal, maternal, or placental source contributing to fetal fluid C concentrations remains to be clarified. From a clinical perspective, however, the evaluation of amniotic C concentration at birth seems useful for the detection of puppies that need special surveillance during the first 24 hours of age, and should be coupled to the early newborn evaluation by Apgar score. However, the small total number of newborns, and especially of the dead puppies enrolled in the present study, suggests that further, more-focused investigations on a large number of subjects are needed before the method could be considered for application in the clinical practices.
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