Life history of a wide-ranging deepwater lantern shark in the north-east Atlantic, Etmopterus spinax (Chondrichthyes: Etmopteridae), with implications for conservation

In this paper, the population biology of the velvet belly lanternshark Etmopterus spinax was studied and life-history coefficients determined. Age was estimated from sections of the second dorsal spine and validated by marginal increment analysis. Males attained a maximum age of 8 years while 11 year-old females were found. Several growth models were fitted and compared for both size-at-age and mass-at-age data, showing that even though this is a small-sized species, it has a relatively slow growth rate. This species matures late, specifically at 49.6 and 42.5% of the maximum observed ages for males and females, respectively. It has a low fecundity, with a mean ovarian fecundity of 9.94 oocytes and a mean uterine fecundity of 7.59 embryos per reproductive cycle. This species seems to have a long reproductive cycle, and even though no conclusive data were obtained, a 2-3 year cycle is possible. The estimated coefficients indicate that this species has a vulnerable life cycle, typical of deepwater squalid sharks. Given the high fishing pressures that it is suffering in the north-east Atlantic, this fish may already be facing severe declines or in risk of facing them in the near future. (C) 2008 The Authors Journal compilation (C) 2008 The Fisheries Society of the British Isles

Reproductive aspects of the oceanic whitetip shark, Carcharhinus longimanus (Elasmobranchii: Carcharhinidae), in the equatorial and southwestern Atlantic Ocean

The present study sought to study the reproductive biology of the oceanic whitetip shark, Carcharhinus longimanus, in the equatorial and southwestern Atlantic Ocean. A total of 234 specimens were collected as bycatch during pelagic longline fisheries targeting tunas and swordfish, between December 2003 and December 2010. The fishing area was located between latitudes 10N and 35S and longitudes 3E and 40W. Of the 234 individuals sampled, 118 were females (with sizes ranging from 81 to 227 cm TL, total length) and 116 males (ranging from 80 to 242 cm TL). The reproductive stages of the females were classed as immature, mature, preovulatory and pregnant, while males were divided into immature, maturing and mature. The size at maturity for females was estimated at 170.0 cm TL, while that for males was between 170.0 and 190.0 cm TL. Ovarian fecundity ranged from 1 to 10 follicles and uterine fecundity from 1 to 10 embryos. The reproductive cycle of this species is most likely biennial, with parturition occurring once every two years.

The role of inflammation in vascular function and pregnancy outcome in the pregnant stroke prone spontaneously hypertensive rat

During pregnancy, the maternal cardiovascular system undergoes major adaptation. One of these changes is a 40-50 % increase in circulating blood volume which requires a systemic remodelling of the vasculature in order to regulate maternal blood pressure and maximise blood supply to the developing placenta and fetus. These changes are broadly conserved between humans and rats making them an appropriate pre-clinical model in which to study the underlying mechanisms of pregnancy-dependent cardiovascular remodelling. Whilst women are normally protected against cardiovascular disease; pregnancy marks a period of time where women are susceptible to cardiovascular complications. Cardiovascular disease is the leading cause of maternal mortality in the United Kingdom; in particular hypertensive conditions are among the most common complications of pregnancy. One of the main underlying pathologies of these pregnancy complications is thought to be a failure of the maternal cardiovascular system to adapt. The remodelling of the uterine arteries, which directly supply the maternal-fetal interface, is paramount to a healthy pregnancy. Failure of the uterine arteries to remodel sufficiently can result in a number of obstetric complications such as preeclampsia, fetal growth restriction and spontaneous pregnancy loss. At present, it is poorly understood whether this deficient vascular response is due to a predisposition from existing maternal cardiovascular risk factors, the physiological changes that occur during pregnancy or a combination of both. Previous work in our group employed the stroke prone spontaneously hypertensive rat (SHRSP) as a model to investigate pregnancy-dependent remodelling of the uterine arteries. The SHRSP develops hypertension from 6 weeks of age and can be contrasted with the control strain, the Wistar Kyoto (WKY) rat. The phenotype of the SHRSP is therefore reflective of the clinical situation of maternal chronic hypertension during pregnancy. We showed that the SHRSP exhibited a deficient uterine artery remodelling response with respect to both structure and function accompanied by a reduction in litter size relative to the WKY at gestational day (GD) 18. A previous intervention study using nifedipine in the SHRSP achieved successful blood pressure reduction from 6 weeks of age and throughout pregnancy; however uterine artery remodelling and litter size at GD18 was not improved. We concluded that the abnormal uterine artery remodelling present in the SHRSP was independent of chronic hypertension. From these findings, we hypothesised that the SHRSP could be a novel model of spontaneously deficient uterine artery remodelling in response to pregnancy which was underpinned by other as yet unidentified cardiovascular risk factors. In Chapter 1 of this thesis, I have characterised the maternal, placental and fetal phenotype in pregnant (GD18) SHRSP and WKY. The pregnant SHRSP exhibit features of left ventricular hypertrophy in response to pregnancy and altered expression of maternal plasma biomarkers which have been previously associated with hypertension in human pregnancy. I developed a protocol for accurate dissection of the rat uteroplacental unit using qPCR probes specific for each layer. This allowed me to make an accurate and specific statement about gene expression in the SHRSP GD18 placenta; where oxidative stress related gene markers were increased in the vascular compartments. The majority of SHRSP placenta presented at GD18 with a blackened ring which encircled the tissue. Further investigation of the placenta using western blot for caspase 3 cleavage determined that this was likely due to increased cell death in the SHRSP placenta. The SHRSP also presented with a loss of one particular placental cell type at GD18: the glycogen cells. These cells could have been the target of cell death in the SHRSP placenta or were utilised early in pregnancy as a source of energy due to the deficient uterine artery blood supply. Blastocyst implantation was not altered but resorption rate was increased between SHRSP and WKY; indicating that the reduction in litter size in the SHRSP was primarily due to late (>GD14) pregnancy loss. Fetal growth was not restricted in SHRSP which led to the conclusion that SHRSP sacrifice part of their litter to deliver a smaller number of healthier pups. Activation of the immune system is a common pathway that has been implicated in the development of both hypertension and adverse pregnancy outcome. In Chapter 2, I proposed that this may be a mechanism of interest in SHRSP pregnancy and measured the pro-inflammatory cytokine, TNFα, as a marker of inflammation in pregnant SHRSP and WKY and in the placentas from these animals. TNFα was up-regulated in maternal plasma and urine from the GD18 SHRSP. In addition, TNFα release was increased from the GD18 SHRSP placenta as was the expression of the pro-inflammatory TNFα receptor 1 (Tnfr1). In order to investigate whether this excess TNFα was detrimental to SHRSP pregnancy, a vehicle-controlled intervention study using etanercept (a monoclonal antibody which works as a TNFα antagonist) was carried out. Etanercept treatment at GD0, 6, 12 and 18 resulted in an improvement in pregnancy outcome in the SHRSP with an increased litter size and reduced resorption rate. Furthermore, there was an improved uterine artery function in GD18 SHRSP treated with etanercept which was associated with an improved uterine artery blood flow over the course of gestation. In Chapter 3, I sought to identify the source of this detrimental excess of TNFα by designing a panel for maternal leukocytes in the blood and placenta at GD18. A population of CD3- CD161+ cells, which are defined as rat natural killer (NK) cells, were increased in number in the SHRSP. Intracellular flow cytometry also identified this cell type as a source of excess TNFα in blood and placenta from pregnant SHRSP. I then went on to evaluate the effects of etanercept treatment on these CD3- CD161+ cells and showed that etanercept reduced the expression of CD161 and the cytotoxic molecule, granzyme B, in the NK cells. Thus, etanercept limits the cytotoxicity and potential damaging effect of these NK cells in the SHRSP placenta. Analysing the urinary peptidome has clinical potential to identify novel pathways involved with disease and/or to develop biomarker panels to aid and stratify diagnosis. In Chapter 4, I utilised the SHRSP as a pre-clinical model to identify novel urinary peptides associated with hypertensive pregnancy. Firstly, a characterisation study was carried out in the kidney of the WKY and SHRSP. Urine samples from WKY and SHRSP taken at pre-pregnancy, mid-pregnancy (GD12) and late pregnancy (GD18) were used in the peptidomic screen. In order to capture peptides which were markers of hypertensive pregnancy from the urinary peptidomic data, I focussed on those that were only changed in a strain dependent manner at GD12 and 18 and not pre-pregnancy. Peptide fragments from the uromodulin protein were identified from this analysis to be increased in pregnant SHRSP relative to pregnant WKY. This increase in uromodulin was validated at the SHRSP kidney level using qPCR. Uromodulin has previously been identified to be a candidate molecule involved in systemic arterial hypertension but not in hypertensive pregnancy thus is a promising target for further study. In summary, we have characterised the SHRSP as the first model of maternal chronic hypertension during pregnancy and identified that inflammation mediated by TNFα and NK cells plays a key role in the pathology. The evidence presented in this thesis establishes the SHRSP as a pre-clinical model for pregnancy research and can be continued into clinical studies in pregnant women with chronic hypertension which remains an area of unmet research need.

Pattern peristaltico uterino nell'adenomiosi isolata: studio prospettico

Obiettivi dello studio: valutare con l’ecografia transvaginale la peristalsi uterina in fase periovulatoria in donne con adenomiosi isolata, confrontandola con un gruppo di controllo e, secondariamente, valutare il grado di accordo tra gli sperimentatori nella descrizione dei pattern di contrattilità. Disegno dello studio: studio osservazione prospettico condotto presso il Policlinico S. Orsola- Malpighi di Bologna, Italia. Materiali e Metodi: sono state reclutate pazienti afferenti al Centro per valutazione ambulatoriale, suddivise sulla base dei criteri di inclusione ed esclusione nei gruppi A (adenomiosi) e B (controlli) e sono state sottoposte da un unico ecografista esperto a ecografia transvaginale con registrazione di un video della durata di 180 secondi della scansione sagittale dell’utero. La registrazione è stata rivalutata off line da due sperimentatori esperti ecografisti, non a conoscenza della storia clinica delle pazienti e in cieco l’uno rispetto all’altro, che hanno descritto il pattern contrattile. È stata stimata una numerosità campionaria di 18 pazienti per gruppo per ottenere una differenza del 20% nell’obiettivo primario con una significatività del 5% (power 80%). Risultati: di 51 pazienti reclutate nello studio, a seguito di drop out 36 sono state sottoposte alla videoregistrazione ecografica (18 per gruppo). Il pattern peristaltico nel gruppo A è risultato alterato in maniera statisticamente significativa rispetto al gruppo B con un p value= 0,02. Sono stati osservati un pattern retrogrado nel 27,8% vs 72,2%, anterogrado del 11,1% vs 16,7%, opposto 38,9% vs 5,6% e random nel 22,2% vs 5,6%, rispettivamente nel gruppo A e B. Il calcolo dell’accordo interosservatore ha portato a un κ value di 0,92. Conclusioni: l’adenomiosi isolata è associata a disperistalsi uterina, che concorrerebbe nello sviluppo dei sintomi tipici dell’adenomiosi. L’ecografia transvaginale rappresenta uno strumento accessibile e utile nella valutazione della contrattilità uterina in quanto il grado di accordo tra gli sperimentatori è ottimo.

Crescita volumetrica dei miomi uterini: analisi dei fattori di rischio

I miomi uterini sono la neoplasia uterina più comune e colpiscono fino al 30% delle donne in età fertile. Nonostante l’elevata prevalenza, pochi studi in Letteratura hanno analizzato i fattori di rischio per la crescita dei miomi uterini, mostrando spesso dei risultati contrastanti. Nel nostro studio osservazionale prospettico sono state arruolate pazienti che rispettassero i criteri di inclusione e di esclusione, con diagnosi di miomi uterini evidenziati mediante ecografia eseguita presso i nostri ambulatori, a partire da giugno 2017. A partire da gennaio 2019, sono state ricercate mensilmente le pazienti precedentemente arruolate che erano tornate presso i nostri ambulatori per esecuzione di ulteriore ecografia, a distanza di 24 ± 5 mesi, fino a raggiungere il campione designato, cioè 450 pazienti totali. È stato, quindi, valutato il tasso di crescita annuo del mioma di maggiori dimensioni ed è stato utilizzato l'approccio polinomiale frazionario multivariabile per selezionare i fattori di rischio anamnestici ed ecografici legati all’incremento volumetrico. Circa la metà dei miomi uterini analizzati ha mostrato stabilità dimensionale nel corso del follow-up (crescita ≤10%), mentre la restante metà ha mostrato una crescita > 10%. Il solo fattore di rischio associato alla crescita volumetrica dei miomi uterini è risultato essere il volume del mioma durante l’ecografia di arruolamento (P = 0.001), quindi miomi di piccole dimensioni presentano un tasso di crescita maggiore rispetto ai miomi di grandi dimensioni. Lo studio ha raccolto la più ampia casistica in Letteratura nella valutazione del naturale andamento di modifica dimensionale dei miomi uterini. Sebbene siano necessari ulteriori studi con campione più ampio, questi dati possono fornire un utile supporto per eseguire un adeguato counselling con le pazienti nella pratica clinica quotidiana.

Nutritional strategies to modulate metabolism and heat stress resilience in dairy cattle

Physiological and environmental stressors can disrupt barrier integrity at epithelial interfaces (e.g., uterine, mammary, intestinal, and lung), which are constantly exposed to pathogens that can lead to the activation of the immune system. Unresolved inflammation can result in the emergence of metabolic and infectious diseases. Maintaining cow health and performance during periods of immune activation such as in the peripartum or under heat stress represents a significant obstacle to the dairy industry. Feeding microencapsulated organic acids and pure botanicals (OAPB) has shown to improve intestinal health in monogastric species and prevent systemic inflammation via the gut-liver axis. Feeding unsaturated fatty acids (FA) such as oleic acid (OA) and very-long-chain omega-3 (VLC n-3) FA are of interest in dairy cow nutrition because of their potential to improve health, fertility, and milk production. In the first study, we evaluated the effects of heat stress (HS) conditions and dietary OAPB supplementation on gut permeability and milk production. In parallel with an improved milk performance and N metabolism, cows supplemented with OAPB also had an enhanced hepatic methyl donor status and greater inflammatory and oxidative stress status compared to the HS control group. In a second study, we evaluated the relative bioavailability of VLC n-3 in cows fed a bolus of rumen-protected (RP) fish oil (FO). In a third study, we proved the interaction between RPFO and RP choline to promote the synthesis of phosphatydilcholines. Lipid forms that support hepatic triglyceride export and can prevent steatosis in dairy cows. The last study, demonstrated that algae oil outperforms against a toxin challenge compared to FO and that feeding RPOA modulates energy partitioning relative to n-3 FA-containing oils. Overall, this thesis confirms the need and the effectiveness of different strategies that aimed to improve dairy cows’ health and performance under heat stress, inflammation or metabolic disease.