NON INVASIVE STUDY ON CANINE PERINATOLOGY

Al fine di migliorare le conoscenze riguardo la fisiologia perinatale nel cane e nel tentativo di fornire dei potenziali markers diagnostici e/o prognostici per la gestione dei neonati poco vitali o patologici, il progetto di dottorato si \ue8 concentrato su due aspetti principali: a) la definizione di alcune caratteristiche dei fluidi fetali nel cane in condizioni di normalit\ue0; b) il possibile uso di pelo e unghie come matrici per lo studio retrospettivo sulla concentrazione di deidroepiandrosterone (DHEA) e deidroepiandrosterone solfato (DHEAS), coinvolti nelle fasi finali della maturazione fetale e nell\u2019adattamento neonatale. Per quanto riguarda il primo argomento un primo studio ha indagato la concentrazione di cortisolo nei fluidi fetali del cane alla nascita, indagando il possibile effetto del cortisolo sulla sopravvivenza dei neonati a 24 ore di et\ue0, indagando inoltre l\u2019effetto di alcuni parametri materni e fetali sulla concentrazione di cortisolo nei fluidi fetali. Lo studio ha dimostrato una concentrazione maggiore di cortisolo nel fluido allantoideo rispetto al fluido amniotico, con una correlazione positiva nelle concentrazioni tra i due fluidi, \ue8 stato riscontrato infine un effetto della cucciolata sulle concentrazioni del cortisolo nei fluidi amniotico e allantoideo. La concentrazione di cortisolo nel fluido amniotico dei cuccioli morti nelle 24 ore dopo la nascita \ue8 risultata maggiore rispetto al fluido amniotico dei neonati normali. Un secondo studio ha indagato le concentrazioni di acido urico, lattato, glucosio e creatinina nel fluido amniotico appartenente a cuccioli di taglia piccola nati da taglio cesareo elettivo al termine di gravidanze normali, valutando la possibile influenza della parit\ue0 materna e del sesso dei neonati sulla concentrazione di questi metaboliti, cos\uec come sull\u2019effetto di questi metaboliti sull\u2019outcome neonatale. Da questo studio non sono emerse differenze significative sulla concentrazione di acido urico e creatinina in relazione a parit\ue0 materna e sesso dei neonati. Per quanto riguarda il glucosio \ue8 stata riscontrata un\u2019influenza del sesso dei neonati ma non della parit\ue0 materna sulla concentrazione nel liquido amniotico. La concentrazione del lattato nel fluido amniotico \ue8 risultata statisticamente pi\uf9 elevata nelle cagne multipare rispetto alle primipare. Un terzo studio ha indagato la composizione biochimica dei fluidi fetali del cane a termine di gravidanza. \uc8 stata effettuata una comparazione tra i fluidi di soggetti normali e patologici ma l\u2019analisi statistica non \ue8 stata in grado di evidenziare differenze significative tra i due gruppi, probabilmente anche per la scarsa numerosit\ue0 dei soggetti patologici rispetto ai sani. Per quanto riguarda le possibili differenze tra i fluidi nei soggetti sani, sono state evidenziate differenze significative nelle concentrazioni di alcuni dei parametri analizzati, suggerendo un diverso meccanismo di produzione e accumulo per i due fluidi nel cane. Lo studio ha rivelato inoltre l\u2019influenza di taglia materna e parit\ue0, cos\uec come del sesso dei neonati, su alcuni dei parametri indagati. Altri studi sono necessari per meglio indagare le possibili differenze nella composizione dei fluidi fetali tra soggetti sani e patologici al fine di individuare possibili markers per la rapida individuazione dei soggetti patologici che richiedono una pronta assistenza nelle prime ore di vita. In relazione al secondo aspetto indagato \ue8 emerso che il DHEA, prodotto dai surreni fetali, presenta una concentrazione maggiore nel pelo dei neonati prematuri rispetto ai neonati morti tra 1 e 30 giorni di vita, concentrazione che risulta tuttavia simile a quella dei neonati morti nelle prime 24 ore di vita, questi risultati suggeriscono una possibile produzione di questo ormone da parte del feto stesso durante l\u2019ultima fase di sviluppo intrauterino. Sebbene il pelo si sia rivelato una matrice utile per gli studi sulle concentrazioni del DHEA nei neonati, tuttavia il suo uso in vivo \ue8 limitato dal quantitativo richiesto per le analisi; \ue8 stata quindi indagata la concentrazione del DHEA e della sua forma di trasporto, DHEAS, anche nelle unghie dei cuccioli neonati. I risultati di questo studio hanno rivelato che le unghie sono una matrice ottimale per la determinazione di DHEA e DHEAS e che questa matrice potrebbe essere usata efficacemente anche per studi seriali in vivo. Le concentrazioni di DHEA e DHEAS nelle unghie sono risultate significativamente differenti ma fortemente e positivamente correlate. La concentrazione di DHEAS e non di DHEA \ue8 risultata statisticamente maggiore nei cani di tagli piccola rispetto ai soggetti di taglia grande, inoltre la concentrazione di DHEAS \ue8 risultata maggiore nei cuccioli prematuri rispetto ai nati morti o ai cuccioli morti tra 11 e 20 giorni di vita.In order to improve knowledge about perinatal physiology in dogs, and in the attempt to provide some potential diagnostic/prognostic markers for a better management of diseased and less viable newborn puppies, the PhD project was focused on two main topics: a) the definition of some fetal fluids characteristics under normal condition; b) the investigations of suitability of hair and nail single collection for the retrospective analysis of dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEAS), involved in the fetal final maturation and neonatal adaptation. In relation to the first topic a first study was aimed to measure the fetal fluids cortisol concentrations in puppies at birth, and to assess the possible effect of cortisol on newborn survival at 24 hours of age, and the effect of some neonatal or maternal parameters on fetal fluids cortisol concentrations. A significant higher cortisol concentration in the allantoic than in the amniotic fluid was found, but with significant high positive correlation between amniotic and allantoic cortisol concentrations. Significant higher amniotic cortisol concentrations were found in puppies dead at 24 hours, as well a significant effect of the litter on fetal fluids cortisol concentrations. The second study investigated the concentrations of uric acid, lactate, glucose and creatinine in amniotic fluid of small sized purebred newborn dogs born by elective cesarean section at term of normal pregnancies, in relation to newborn outcome and the possible effect played by maternal parity and newborn gender on uric acid, glucose, lactate and creatinine concentrations. When the statistical analysis was performed on fetal fluids belonging to normal puppies no significant difference on uric acid concentration were found in relation to maternal parity or newborn gender. Regarding amniotic glucose concentration a significant influence of newborn gender, but not of maternal parity, was found. Amniotic lactate concentration was higher in multiparous in comparison to primiparous bitches. Regarding creatinine no significant differences were found in relation to maternal parity or newborn gender. The third study investigated the biochemical composition of fetal fluids at term pregnancy in dogs. A comparison between fetal fluids characteristics of normal and pathologic puppies was done, but, the statistical analysis did not show significant results, due to the small number of pathologic puppies. When the statistical analysis was applied to the normal puppies, differences between the two fluids were found for many parameters, suggesting a different source and mechanism of production and accumulation of the two fluids in dogs. The study showed also the possible influence of breed body size and of maternal parity and newborn gender on some parameters. Further studies are needed in order to better investigate possible differences between fetal fluids belonging to normal and to pathological puppies, and therefore to detect potential markers of fetal/neonatal diseases or for a quick identification of newborns at risk, that need special surveillance and cares, immediately after birth. In relation to the second topic the present thesis highlighted that DHEA, recognized to be produced by the adrenals of offspring, is higher in the hair of premature as compared to puppies 1-30 days old, although not different from stillborn puppies, suggesting the possible production of this hormone by the fetus itself in the last period of intrauterine development. Although the hair is suitable for perinatal DHEA analysis in dead puppies, the hair necessary for the analysis still limits the use of this matrix for in vivo studies in newborn puppies. Therefore, the measurement of DHEA and of its transport form DHEAS, also in the nails of newborn puppies was assessed. The results showed that nails were suitable for DHEA and DHEAS measurement, allowing the use of this matrix for serial studies on alive newborn puppies. DHEA and DHEAS concentrations were significantly different in the overall concentration in nails of newborn puppies, with a high correlation between the concentrations of DHEA and DHEAS; DHEAS and not DHEA concentrations were significantly higher in small size breeds in comparison with large size breeds, while DHEAS was higher in premature puppies when compared to puppies born-dead or dead between 11 and 20 days of age. When the usefulness of these non-invasive matrices for the study of the dog perinatology was considered, the nails resulted more suitable in comparison to the hairs. In fact, in the perspective of using these matrices in alive newborn puppies, at present the relatively large amount of hair necessary for the analysis prevent its use on alive newborns

Apgar score or birthweight in Chihuahua dogs born by elective Caesarean section : which is the best predictor of the survival at 24 h after birth?

In the dog, the correct management of parturition and the prompt neonatal evaluation and assistance can reduce the perinatal mortality rates that are particularly high in toy breeds. Newborn evaluation and factors addressing prognosis are pivotal to guarantee the correct neonatal assistance. Assessment of the Apgar score with viability classification and birthweight are recognized as predictors for neonatal survival in dogs, but breed-specific data are needed for a more feasible application in the dog species, in which wide differences among breeds are known. The present study aimed therefore to: (a) assess the role of Apgar score and birthweight as predictors for the survival of Chihuahua newborn puppies in the first 24 h of life; (b) to assess a cut-off of the Apgar score and birthweight values that can predict the survival of Chihuahua newborn puppies in the first 24 h after birth; (c) to assess the possible effect played by maternal parity, newborn gender and litter-size on Apgar score in Chihuahua newborn puppies, in order to provide breed-specific data for a better neonatal assistance..Data obtained from 176 normal developed Chihuahua puppies born by elective Caesarean section, showed that 62%, 28% and 10% of puppies were classified in the Apgar score classes 7\u201310, 4\u20136 and 0\u20133, respectively, with survival at 24 h after birth of 97%, 96%, 39%, in the three Apgar classes of viability, respectively. Apgar score was a better predictor for survival at 24 h after birth than birthweight (AUC 0.93, P < 0.0001; AUC 0.69, P < 0.01, respectively). Litter-size of 7 puppies/litter plays a negative effect on Apgar score. Apgar score is a better predictor of survival at 24 h than birthweight, and the best cut-off of Apgar score for survival at 24 h after birth is 4, with 96% sensitivity and 77% specificity. The different proportion of \u201cnormal viable\u201d and \u201cless viable\u201d neonates in comparison to other studies highlights that Chihuahua puppies born by elective Caesarean section should be carefully evaluated at birth to provide correct assistance

Effects of Alfaxalone or Propofol on Giant-Breed Dog Neonates Viability During Elective Caesarean Sections

Nowadays, thanks to the increased awareness of owners and breeders and to the most recent techniques available to veterinarians, the management of parturition, especially of C-sections, has become a topic of greater importance. Anesthesia is crucial and must be targeted to both the mother and neonates. The present study aimed to evaluate the effect of the induction agent alfaxalone on the vitality of puppies born from elective C-section, in comparison to propofol. After inducing general anesthesia for elective C-section, puppies from the mothers induced with alfaxalone had higher 5-min Apgar scores than those induced with propofol. The concentration of cortisol in fetal fluids collected at birth is neither influenced by the anesthetic protocol used, nor does it differ between amniotic and allantoic fluids. Nevertheless, the cortisol concentration in fetal fluids affects the relationship between anesthesia and the Apgar score: the present study highlights a significant relationship between the anesthetic protocol used and Apgar score in puppies, and fetal fluids cortisol concentration acts as a covariate of this relationship. Alfaxalone is a safe and effective drug for the induction of anesthesia in dogs, and it can be successfully employed for elective C-section, with neonatal wellbeing improvements

Doxorubicin versus doxorubicin and cisplatin in endometrial carcinoma: definitive results of a randomised study (55872) by the EORTC Gynaecological Cancer Group

BACKGROUND: Combination chemotherapy yields better response rates which do not always lead to a survival advantage. The aim of this study was to investigate whether the reported differences in the efficacy and toxicity of monotherapy with doxorubicin (DOX) versus combination therapy with cisplatin (CDDP) in endometrial adenocarcinoma lead to significant advantage in favour of the combination. PATIENTS AND METHODS: Eligible patients had histologically-proven advanced and/or recurrent endometrial adenocarcinoma and were chemo-naïve. Treatment consisted of either DOX 60 mg/m(2) alone or CDDP 50 mg/m2 added to DOX 60 mg/m2, every 4 weeks. RESULTS: A total of 177 patients were entered and median follow-up is 7.1 years. The combination DOX-CDDP was more toxic than DOX alone. Haematological toxicity consisted mainly of white blood cell toxicity grade 3 and 4 (55% versus 30%). Non-haematological toxicity consisted mainly of grade 3 and 4 alopecia (72% versus 65%) and nausea/vomiting (36 % versus 12%). The combination DOX-CDDP provided a significantly higher response rate than single agent DOX (P <0.001). Thirty-nine patients (43%) responded on DOX-CDDP [13 complete responses (CRs) and 26 partial responses (PRs)], versus 15 patients (17%) on DOX alone (8 CR and 7 PR). The median overall survival (OS) was 9 months in the DOX-CDDP arm versus 7 months in the DOX alone arm (Wilcoxon P = 0.0654). Regression analysis showed that WHO performance status was statistically significant as a prognostic factor for survival, and stratifying for this factor, treatment effect reaches significance (hazard ratio = 1.46, 95% confidence interval 1.05-2.03, P = 0.024). CONCLUSIONS: In comparison to single agent DOX, the combination of DOX-CDDP results in higher but acceptable toxicity. The response rate produced is significantly higher, and a modest survival benefit is achieved with this combination regimen, especially in patients with a good performance status

Hodgkin's lymphoma: The pathologist's viewpoint

Despite its well known histological and clinical features, Hodgkin's lymphoma (HL) has recently been the object of intense research activity, leading to a better understanding of its phenotype, molecular characteristics, histogenesis, and possible mechanisms of lymphomagenesis. There is complete consensus on the B cell derivation of the tumour in most cases, and on the relevance of Epstein-Barr virus infection and defective cytokinesis in at least a proportion of patients. The REAL/WHO classification recognises a basic distinction between lymphocyte predominance HL (LP-HL) and classic HL (CHL), reflecting the differences in clinical presentation and behaviour, morphology, phenotype, and molecular features. CHL has been classified into four subtypes: lymphocyte rich, nodular sclerosing, with mixed cellularity, and lymphocyte depleted. The borders between CHL and anaplastic large cell lymphoma have become sharper, whereas those between LP-HL and T cell rich B cell lymphoma remain ill defined. Treatments adjusted to the pathobiological characteristics of the tumour in at risk patients have been proposed and are on the way to being applied

Activation of the JAK/STAT pathway leads to proliferation of ST14A central nervous system progenitor cells

We were interested in whether central nervous system progenitor cells possess the signal transduction machinery necessary to mediate cytokine functions and whether this machinery can become activated upon stable expression of a particular cytokine receptor. For this purpose we utilized a previously obtained conditionally immortalized striatum-derived nestin-positive cell line (ST14A). We found that ST14A cells express Jak2, but not Jak1 or Tyk2. An identical pattern of expression was found in embryonic striatal tissue. To evaluate the susceptibility of these cytokine specific cytoplasmic transducers to activation, ST14A cells were stably transfected with the alpha and beta (AIC2A) chains of the murine interleukin-3 receptor. Four independent lines expressing both the alpha and beta receptor subunits were obtained. We found that cells from each of these lines were induced to proliferate upon exposure to interleukin-3. Dose response curve, antibody blocking experiments and binding studies revealed that the response was mediated by the reconstituted high affinity interleukin-3 receptor. Immunoprecipitation studies on these cells showed that Jak2 and Stat5 were being phosphorylated after stimulation of the reconstituted receptor. These results indicate that members of the JAK/STAT family of proteins are expressed in central nervous system progenitor cells and are susceptible to activation through stimulation of an exogenously expressed cytokine receptor, ultimately leading to cell proliferation

Hydrophobic and ionic-interactions in bulk and confined water with implications for collapse and folding of proteins

Water and water-mediated interactions determine thermodynamic and kinetics of protein folding, protein aggregation and self-assembly in confined spaces. To obtain insights into the role of water in the context of folding problems, we describe computer simulations of a few related model systems. The dynamics of collapse of eicosane shows that upon expulsion of water the linear hydrocarbon chain adopts an ordered helical hairpin structure with 1.5 turns. The structure of dimer of eicosane molecules has two well ordered helical hairpins that are stacked perpendicular to each other. As a prelude to studying folding in confined spaces we used simulations to understand changes in hydrophobic and ionic interactions in nano droplets. Solvation of hydrophobic and charged species change drastically in nano water droplets. Hydrophobic species are localized at the boundary. The tendency of ions to be at the boundary where water density is low increases as the charge density decreases. Interaction between hydrophobic, polar, and charged residue are also profoundly altered in confined spaces. Using the results of computer simulations and accounting for loss of chain entropy upon confinement we argue and then demonstrate, using simulations in explicit water, that ordered states of generic amphiphilic peptide sequences should be stabilized in cylindrical nanopores

Direct Observation of Shock-Induced Disordering of Enstatite Below the Melting Temperature

We report in situ structural measurements of shock-compressed single crystal orthoenstatite up to 337 ± 55 GPa on the Hugoniot, obtained by coupling ultrafast X-ray diffraction to laser-driven shock compression. Shock compression induces a disordering of the crystalline structure evidenced by the appearance of a diffuse X-ray diffraction signal at nanosecond timescales at 80 ± 13 GPa on the Hugoniot, well below the equilibrium melting pressure (>170 GPa). The formation of bridgmanite and post-perovskite have been indirectly reported in microsecond-scale plate-impact experiments. Therefore, we interpret the high-pressure disordered state we observed at nanosecond scale as an intermediate structure from which bridgmanite and post-perovskite crystallize at longer timescales. This evidence of a disordered structure of MgSiO3 on the Hugoniot indicates that the degree of polymerization of silicates is a key parameter to constrain the actual thermodynamics of shocks in natural environments. © 2020. The Authors

Time evolution and asymmetry of a laser produced blast wave

Studies of a blast wave produced from carbon rods and plastic spheres in an argon background gas have been conducted using the Vulcan laser at the Rutherford Appleton Laboratory. A laser of 1500 J was focused onto these targets, and rear-side observations of an emission front were recorded using a fast-framing camera. The emission front is asymmetrical in shape and tends to a more symmetrical shape as it progresses due to the production of a second shock wave later in time, which pushes out the front of the blast wave. Plastic spheres produce faster blast waves, and the breakthrough of the second shock is visible before the shock stalls. The results are presented to demonstrate this trend, and similar evolution dynamics of experimental and simulation data from the FLASH radiation-hydrodynamics code are observed