The RhoA guanine exchange factor ABR: a glucose-sensitive mediator of actin reorganization in feto-placental arterial endothelial cells altered by gestational diabetes mellitus.

In utero exposure to gestational diabetes mellitus (GDM) programs the fetus, increasing offspring risk for endothelial dysfunction and cardiovascular disease later in life. Hyperglycaemia is widely recognized as the driving force of diabetes-induced programming. We have previously shown that GDM exposure alters DNA methylation and gene expression associated with actin remodelling in primary feto-placental arterial endothelial cells (fpEC). Thus, we hypothesized that hyperglycaemic insults underlie programmed changes in fpEC morphology and actin organization by GDM. Therefore, arterial fpECs isolated after normal and GDM pregnancy, as well as normal fpECs that were exposed to hyperglycaemia in vitro, were analysed for the effect of GDM and hyperglycaemia on actin organization and network formation. Integration of gene expression and DNA methylation data identified the RhoA activator active BCR-related (ABR) as programmed by GDM and altered by in vitro hyperglycaemia. ABR silencing in GDM-exposed cells reduced RhoA activity by 34 ± 26% (P = 0.033) and restored normal fpEC phenotype. In fact, in vitro hyperglycaemia induced a similar fpEC phenotype as intrauterine exposure to GDM, i.e. round morphology and increased network formation on Matrigel by 34 ± 33% (P = 0.022) vs. 22 ± 20% for GDM (P = 0.004). Thus, we identified ABR as a novel glucose sensitive regulator of actin organization and cell shape, programmed by GDM and upregulated by hyperglycaemia. Identification of mechanisms induced by hyperglycaemia and affecting endothelial function in the long term will contribute to understanding GDM-induced programming of offspring endothelial dysfunction and cardiovascular disease. Future studies could focus on investigating the prevention or reversal of such malprogramming. KEY POINTS: In utero exposure to gestational diabetes mellitus (GDM) affects future health of the offspring, with an increased risk for endothelial dysfunction and cardiovascular disease in later life. GDM alters DNA methylation and expression of ABR in feto-placental arterial endothelial cells (fpEC), a model for endothelial cells exposed to the intrauterine environment of the fetus. GDM phenotype of fpECs is also induced by hyperglycaemia in vitro, and is characterized by altered actin organization and cell shape, which can be restored by ABR silencing. Revealing the cellular mechanisms induced by GDM and hyperglycaemia is important for understanding the mechanisms of how these conditions disturb endothelial function in the offspring.

Placental structural adaptation to maternal physical activity and sedentary behavior: findings of the DALI lifestyle study.

STUDY QUESTION: Are maternal levels of moderate-to-vigorous physical activity (MVPA) and sedentary time (ST) in obese pregnant women associated with placental structural adaptations for facilitating oxygen delivery to the fetus? SUMMARY ANSWER: Higher maternal MVPA and ST are associated with a higher density of villi, a proxy measure of placental surface area for oxygen delivery to the fetus, without further added placental vessels. WHAT IS KNOWN ALREADY: Physical activity during pregnancy intermittently reduces uterine blood flow, potentially limiting placental and fetal oxygen supply. The placenta can mount several adaptive responses, including enlargement of the surface area of villi and/or feto-placental vessels to accommodate fetal needs. Early research on the morphology and growth of the placenta with exercise interventions has shown inconsistencies and is lacking, particularly in non-lean pregnant women. STUDY DESIGN, SIZE, DURATION: This study is a secondary longitudinal analysis of the vitamin D and lifestyle intervention for gestational diabetes prevention (DALI) randomized controlled trial. The prospective study was conducted between 2012 and 2015 in nine European countries at 11 different sites. In this analysis, 92 pregnant women with a BMI ≥ 29 kg/m2 were combined into one cohort. PARTICIPANTS/MATERIALS, SETTING, METHODS: MVPA and percentage of time spent sedentary (% ST) were measured with accelerometers during gestation. Placental sections were immunostained for endothelial cell-specific CD34. Artificial intelligence (AI)-based stereology assessed villous density, number, and cross-sectional area of vessels on whole-slide images and in selected regions comprising peripheral villi only, where the majority of vascular adaptations occur. Expression of pro- and anti-angiogenic factors was quantified using molecular counting analysis. MAIN RESULTS AND THE ROLE OF CHANCE: In multivariable regression, higher levels of maternal MVPA (min/day) were associated with a higher density of villi in both whole-slide images (beta 0.12; 95% CI 0.05, 0.2) and selected regions (0.17; CI 0.07, 0.26). Unexpectedly, ST was also positively associated with density of villi (0.23; CI 0.04, 0.43). MVPA and ST were not associated with vessel count/mm2 villous area, vessel area, or pro- and anti-angiogenic factor mRNA expression. All estimates and statistical significance of the sensitivity analyses excluding smokers, women who developed gestational diabetes or pre-eclampsia and/or pregnancy-induced hypertension were similar in the main analysis. LIMITATIONS, REASONS FOR CAUTION: The placenta is a complex organ undergoing dynamic changes. While various adjustments were made to account for different maternal contributing factors, in addition to the outcome measures, various other factors could impact oxygen delivery to the fetus. WIDER IMPLICATIONS OF THE FINDINGS: For the first time, we evaluated the association between placental structures quantified using an AI-based approach with objectively measured physical activity and ST at multiple time points in pregnant women with obesity. The observed adaptations contribute to the advancement of our understanding of the hemodynamics and adaptations of the placental unit in response to MVPA and ST. However, our results might not be generalizable to lean pregnant women. STUDY FUNDING/COMPETING INTEREST(S): The DALI project has received funding from the European Community's 7th Framework Program (FP7/2007-2013) under grant agreement no. 242187. The funders had no role in study design, collection of data, analyses, writing of the article, or the decision to submit it for publication. The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: ISRCTN70595832.

Toxoplasma gondii spinal granuloma in a cat.

Case summary: A 7-year-old female spayed domestic shorthair cat was presented with a history of progressive pelvic limb ataxia. A CT scan of the thoracic spine identified an extradural, left lateralised and compressive soft tissue mass at the level of T9. A decompressive hemilaminectomy at the level of T8-T10 and tissue sampling of the mass were performed. Histopathological examination revealed a parasite granuloma caused by Toxoplasma gondii infection. Postoperatively, the cat improved consistently, but mild left pelvic limb lameness remained. At 5 weeks, a recheck CT scan showed a small, enhancing soft tissue lesion in the left epidural space at T9, causing a mild left lateral compression of the spinal cord. After 9 months, the cat acutely deteriorated neurologically and was euthanased without postmortem examination. Relevance and novel information: To our knowledge, this is the first report of a T gondii spinal granuloma in a cat. Such a granuloma should be considered as a differential in cats with evidence of an extradural soft tissue mass.

Physical Activity and Sedentary Time in Pregnancy: An Exploratory Study on Oxidative Stress Markers in the Placenta of Women with Obesity.

Regular moderate-to-vigorous physical activity (MVPA) and reduced sedentary time (ST) improve maternal glucose metabolism in pregnancy. More MVPA and less ST outside pregnancy increase antioxidant capacity, hence, are beneficial in preventing oxidative stress. The placenta is the first line of defense for the fetus from an adverse maternal environment, including oxidative stress. However, effects of MVPA and ST on oxidative stress markers in the placenta are unknown. The purpose of this study was to assess the association of MVPA and ST in pregnancy with oxidative stress markers in placentas of overweight/obese women (BMI ≥ 29 kg/m2). MVPA and ST were objectively measured with accelerometers at <20 weeks, 24−27 and 35−37 weeks of gestation. Using linear Bayesian multilevel models, the associations of MVPA and ST (mean and changes) with mRNA expression of a panel of 11 oxidative stress related markers were assessed in 96 women. MVPA was negatively correlated with HSP70 mRNA expression in a sex-independent manner and with GCLM expression only in placentas of female fetuses. ST was positively associated with HO-1 mRNA expression in placentas of male neonates. None of the other markers were associated with MVPA or ST. We speculate that increasing MVPA and reducing ST attenuates the oxidative stress state in placentas of obese pregnant women.

Fetal sex and maternal fasting glucose affect neonatal cord blood-derived endothelial progenitor cells.

BACKGROUND: Maternal cardiovascular risk factors (CVRF) in pregnancy, i.e., obesity and hyperglycemia, transmit to the fetus and affect placental and fetal endothelial function. Moreover, a sex dimorphism in endothelial function and susceptibility towards CVRF exists already in utero. Endothelial colony-forming cells (ECFC) are circulating endothelial progenitors highly present in neonatal cord blood and sensitive to CVRF. This study investigated whether fetal sex or subtle maternal metabolic changes within healthy range alter fetal ECFC outgrowth. METHODS: Outgrowth of ECFC from cord blood of male (n = 31) and female (n = 26) neonates was analyzed after healthy pregnancies and related to fetal sex and maternal metabolic parameters. RESULTS: Male ECFC grew out earlier (-20.57% days; p = 0.031) than female. Although all women were non-diabetic, higher levels of fasting plasma glucose (FPG) at midpregnancy increased the time required for colony outgrowth (OR: 1.019; p = 0.030), which, after stratifying for fetal sex, was significant only in the males. Gestational weight gain and BMI did not affect outgrowth. Colony number was unchanged by all parameters. CONCLUSIONS: Fetal sex and maternal FPG within normal range alter ECFC function in utero. A role of ECFC in postnatal angiogenesis and vasculogenesis has been suggested, which may be affected by altered outgrowth dynamics. IMPACT: This study is the first to report that a sexual dimorphism exists in ECFC function, as cells of female progeny require a longer period of time until colony outgrowth than ECFC of male progeny. Our data show that ECFC function is highly sensitive and affected by maternal glucose levels even in a normal, non-diabetic range. Our data raise the question of whether maternal plasma glucose in pregnancy should be considered to play a critical role even in the non-diabetic setting.

IMAGING DIAGNOSIS-CONCENTRIC PERIRADICULAR LIPOMA CAUSING LUMBAR NERVE ROOT COMPRESSION IN A DOG.

An 11-year-old, male neutered Jack Russell Terrier was presented with a nerve root signature of the right pelvic limb. Magnetic resonance imaging revealed a well demarcated, ovoid, extramedullary mass at the level of the L7 vertebral body. This showed, compared to normal spinal cord, hyperintense signal on T1- and T2-weighted images, which was suppressed on gradient echo short tau inversion recovery (GE-STIR) images. Additionally, the mass was characterized by a fat density on computed tomography images. Histopathology of the surgically excised mass was consistent with concentric periradicular lipoma, which has not been described in domestic animals yet.

Temporal lobe epilepsy in a cat with a pyriform lobe oligodendroglioma and hippocampal necrosis.

A 14-year-old male domestic shorthair cat presented with an acute onset of aggressive behaviour, fear and hypersalivation. Neurological examination revealed bilateral mydriasis and left-sided facial twitching and hemiparesis. Magnetic resonance imaging (MRI) showed moderate bilateral symmetrical T2-hyperintensity along the entire hippocampus and bilateral asymmetric T2-hyperintensity in the pyriform lobes. Marked bilateral contrast enhancement of the hippocampus was evident on post-contrast T1-weighted images. The partial complex seizures were refractory to medical treatment and the cat was euthanased 4 days after admission. The clinical and MRI findings were consistent with feline hippocampal necrosis (FHN). On histopathology, neuronal necrosis and astrocytosis were present in the hippocampi and pyriform lobes. In addition, an oligodendroglioma was detected in the right pyriform lobe. Contrary to previous reports of FHN in which no underlying cause could be identified, we believe that in this case the seizure focus arose from a neoplastic lesion within the right pyriform lobe. This unique case report represents the so-called 'dual pathology' of temporal lobe epilepsy in humans, in which an extrahippocampal lesion within the temporal lobe results in hippocampal sclerosis.

Magnetic resonance imaging features of canine incomplete humeral condyle ossification.

Incomplete ossification of the humeral condyle (IOHC) is characterized by an intracondylar fissure located where the intercondylar physis is present in growing dogs. Its radiologic and computed tomographic features have been described but the magnetic resonance (MR) features have not been characterized. Our purpose was to further describe the range of MRappearances of IOHC, to assess the diagnostic capability of MRrelative to radiology, and to determine whether MRis able to identify the disease before a fissure forms. Thirty-eight elbow MRscans and radiographs, when available, were reviewed and divided into three groups. In Group 1 (affected elbows, n = 22), there was an intracondylar defect on MRwith variable appearance; the defect was not visible radiographically in 32% of the elbows. The main difference between Group 2 (nonaffected elbows, n = 6) and Group 3 (contralaterals to IOHC or to condylar fracture, without fissure, n = 10) was the appearance of the humeral condyle in short tau inversion recovery (STIR) sequences: all elbows in Group 2 had a homogeneous humeral condyle, whereas all but one in Group 3 were heterogeneous. One dog in Group 3 developed a complete condylar fissure 7 months after the first examination, when no evidence of an intracondylar defect had been detected. The MRappearance of IOHC is variable and a heterogeneous humeral condyle in STIR images without a clear defect may warn of the possibility for the subsequent development of a condylar fissure.

Radiographic characterization of the os penis in the cat.

The os penis in the cat has not been described radiographically, as compared with the dog. However, a small linear bony radiopacity is sometimes detected in the perineal area of male cats. We hypothesized that the feline os penis might be visible on survey radiographs of the pelvis, and we aimed to investigate the frequency of its visualization using analog and computed radiography (CR) system. One hundred radiographs of the pelvis of 99 male cats were reviewed retrospectively (50 were obtained with a CR system and 50 with an analog system). Age, breed, neutering status, and reason for presentation were recorded, as well as the visualization of the os penis. An os penis was detected in 19/50 (38%) cats with CR and in eight of 50 (16%) cats with analog radiography; this difference was statistically significant. With CR, the median age of cats with a visible os penis was significantly higher than in cats where the os penis was not seen. In one cat with a visible os penis examined with CR and analog radiography, the os penis was only visible on CR images. The penile tissues were examined histopathologically in one cat and well-differentiated bone was found but there were no pathologic findings detected in surrounding tissues. Thus, the os penis can be detected on radiographs of cats and this should not be mistaken for a pathologic finding such as urolithiasis or dystrophic mineralization.

Magnetic resonance imaging findings in 15 acromegalic cats.

Feline acromegaly is characterized by chronic excessive growth hormone secretion, most commonly caused by a functional pituitary adenoma. In this study, acromegaly was diagnosed in 15 cats on the basis of compatible clinical signs, laboratory, and magnetic resonance imaging (MRI) findings. MRI findings were reviewed retrospectively. Enlargement of the pituitary gland with suprasellar extension was present in all cats. No characteristic signal patterns were identified on T1-weighted and T2-weighted sequences. Contrast enhancement was nonuniform in all cats, as was suspected involvement of the adjacent hypothalamus. A mass effect on the cavernous sinus and third ventricle was present in 13 cats. Mild peritumoral edema was present in four cats, and moderate edema in one cat. Transtentorial herniation was present in one cat. Histopathology confirmed the presence of a pituitary adenoma in two cases. MRI is a useful modality to establish the diagnosis of acromegaly.

Sonography of the soft tissue structures of the canine tarsus.

Currently, there are no available anatomic descriptions of the soft tissue structures that are visible with ultrasound in the canine tarsus. Eight cadaver hindlimbs and 10 clinically normal dogs (15-37 kg) were examined with ultrasound to establish which structures could be visualized in normal dogs. The structures always identified included the long digital extensor tendon, the tibialis cranialis tendon, the joint space, and the three bellies of the extensor digitum brevis muscle on the cranial/dorsal aspect of the tarsus. The lateral digital extensor tendon and the peroneus longus tendon were identified as a single structure on the lateral aspect of the tarsus. The caudal/plantar approach allowed identification of the calcaneal tendon, the deep and superficial digital flexor tendons, the plantar ligament, and the lumbricales and interosseus muscles. The medial collateral ligament can be identified on the medial tarsus of larger dogs. This technique has proved useful in the diagnosis of soft tissue injuries of the canine tarsus, and clinical examples of tibialis cranialis tendinopathy, deep digital flexor tendinopathy, fibrosis surrounding the medial collateral ligament and extent of invasion of a tarsal hemangiosarcoma are described.

Ultrasonographic appearance of jejunal lymph nodes in dogs without clinical signs of gastrointestinal disease.

The jejunal lymph nodes of 57 dogs without clinical signs of gastrointestinal disease were examined to characterize their ultrasonographic appearance on B-mode and power Doppler examination, and to obtain ultrasonographic measurements. The lymph nodes were mildly hypoechoic to the mesentery in 71% of dogs and isoechoic in 29%. All dogs, 6 years of age or older had jejunal lymph nodes of uniform echogenicity. In dogs less than 6 years of age, nonuniform lymph nodes with different echopatterns were observed. Although most lymph nodes had no blood flow based on power Doppler examination, hilar blood flow was detected in 33% of dogs, which were generally less than 2 years of age. The median maximum thickness of the jejunal lymph nodes was 3.9 mm (range 1.6-8.2 mm), and their median maximum width 7.5 mm (range 2.6-14.7 mm). There was a significant correlation between larger lymph node diameter and younger age and higher body weight. We concluded that patient age should be considered when interpreting the echopattern and vascularity of jejunal lymph nodes in dogs, and that the jejunal lymph nodes of dogs without clinical signs of gastrointestinal disease may exceed the previously stated upper limit of 5-6 mm thickness.