Far-red and sensitive sensor for monitoring real time H2O2 dynamics with subcellular resolution and in multi-parametric imaging applications.

H2O2 is a key oxidant in mammalian biology and a pleiotropic signaling molecule at the physiological level, and its excessive accumulation in conjunction with decreased cellular reduction capacity is often found to be a common pathological marker. Here, we present a red fluorescent Genetically Encoded H2O2 Indicator (GEHI) allowing versatile optogenetic dissection of redox biology. Our new GEHI, oROS-HT, is a chemigenetic sensor utilizing a HaloTag and Janelia Fluor (JF) rhodamine dye as fluorescent reporters. We developed oROS-HT through a structure-guided approach aided by classic protein structures and recent protein structure prediction tools. Optimized with JF635, oROS-HT is a sensor with 635 nm excitation and 650 nm emission peaks, allowing it to retain its brightness while monitoring intracellular H2O2 dynamics. Furthermore, it enables multi-color imaging in combination with blue-green fluorescent sensors for orthogonal analytes and low auto-fluorescence interference in biological tissues. Other advantages of oROS-HT over alternative GEHIs are its fast kinetics, oxygen-independent maturation, low pH sensitivity, lack of photo-artifact, and lack of intracellular aggregation. Here, we demonstrated efficient subcellular targeting and how oROS-HT can map inter and intracellular H2O2 diffusion at subcellular resolution. Lastly, we used oROS-HT with other green fluorescence reporters to investigate the transient effect of the anti-inflammatory agent auranofin on cellular redox physiology and calcium levels via multi-parametric, dual-color imaging.

Machine learning-guided engineering of genetically encoded fluorescent calcium indicators.

Here we used machine learning to engineer genetically encoded fluorescent indicators, protein-based sensors critical for real-time monitoring of biological activity. We used machine learning to predict the outcomes of sensor mutagenesis by analyzing established libraries that link sensor sequences to functions. Using the GCaMP calcium indicator as a scaffold, we developed an ensemble of three regression models trained on experimentally derived GCaMP mutation libraries. The trained ensemble performed an in silico functional screen on 1,423 novel, uncharacterized GCaMP variants. As a result, we identified the ensemble-derived GCaMP (eGCaMP) variants, eGCaMP and eGCaMP+, which achieve both faster kinetics and larger ∆F/F0 responses upon stimulation than previously published fast variants. Furthermore, we identified a combinatorial mutation with extraordinary dynamic range, eGCaMP2+, which outperforms the tested sixth-, seventh- and eighth-generation GCaMPs. These findings demonstrate the value of machine learning as a tool to facilitate the efficient engineering of proteins for desired biophysical characteristics.

En coup de sabre morphea: an uncommon condition in Africa.

The term en coup de sabre morphea refers to a lesion of linear morphea typically located in the frontoparietal scalp and/or the paramedian forehead, often resembling a strike with a sword. In literature, en coup de sabre morphea, and en coup de sabre scleroderma are terms used interchangeably and synonymously. Due to the rarity of this condition, treatment is largely based on case report series, leaving much room for speculation in terms of drugs of choice, duration of treatment, and dosages. Although it typically leaves behind notable and often permanent skin pigmentary changes and indentation of the affected areas, this condition usually remits spontaneously, even in the absence of an active form of treatment. The disease severity and prognosis vary according to the subtype: circumscribed morphea has a generally more benign course when compared with linear scleroderma and generalized morphea.

Indirect lymphography for sentinel lymph node detection in dogs with mast cell tumors.

Objective: The objective of this report is to describe the technique and diagnostic utility of indirect lymphography (IL) using water-soluble contrast for sentinel lymph node (SLN) mapping in dogs with mast cell tumors. Animals: Fifty-three dogs with 59 mast cell tumors were included. Procedure: Medical records were retrieved for dogs with a cytological diagnosis of mast cell tumor which also had IL performed for lymph node mapping. Dogs were excluded when surgery had been performed before presentation. Images were reviewed by a Board-certified radiologist for uptake of contrast within the sentinel lymph node. Results: Lymphography studies from 34 tumors (57.6%) were diagnostic (clearly identifiable lymphatics and sentinel lymph node). Lymphography studies from 12 tumors (20.3%) were partially diagnostic (identifiable lymphatics, but sentinel lymph node not highlighted). Lymphography studies from 13 tumors (22%) were non-diagnostic. Indirect lymphography studies were interpreted as either diagnostic or partially diagnostic in 77.9% of tumors. Conclusion: The results indicate that IL is a simple, available technique to allow for identification of a sentinel lymph node in dogs with mast cell tumors. Clinical relevance: Indirect lymphography is a simple and widely accessible technique for SLN mapping in dogs with mast cell tumors, particularly for the general practice environment.

Lymphographie indirecte pour la détection des ganglions lymphatiques sentinelles chez les chiens atteints de tumeurs mastocytaires. Objectif: L'objectif de ce rapport est de décrire la technique et l'utilité diagnostique de la lymphographie indirecte (IL) utilisant un contraste soluble dans l'eau pour la cartographie des ganglions lymphatiques sentinelles (SLN) chez les chiens atteints de tumeurs mastocytaires. Animaux: Cinquante-trois chiens avec 59 tumeurs mastocytaires ont été inclus. Procédure: Les dossiers médicaux ont été récupérés pour des chiens avec un diagnostic cytologique de tumeur mastocytaire qui ont également subi une IL pour la cartographie des ganglions lymphatiques. Les chiens ont été exclus lorsque la chirurgie avait été pratiquée avant la présentation. Les images ont été examinées par un radiologue certifié (ACVR) pour la prise de contraste dans le ganglion lymphatique sentinelle. Résultats: Les études de lymphographie de 34 tumeurs (57,6 %) étaient diagnostiques (lymphatiques clairement identifiables et ganglion sentinelle). Les études de lymphographie de 12 tumeurs (20,3 %) étaient partiellement diagnostiques (lymphatiques identifiables, mais ganglion sentinelle non mis en évidence). Les études de lymphographie de 13 tumeurs (22 %) étaient non diagnostiques. Les études de lymphographie indirecte ont été interprétées comme diagnostiques ou partiellement diagnostiques dans 77,9 % des tumeurs. Conclusion: Les résultats indiquent que l'IL est une technique simple et disponible pour permettre l'identification d'un ganglion lymphatique sentinelle chez les chiens atteints de tumeurs mastocytaires. Pertinence clinique: La lymphographie indirecte est une technique simple et largement accessible pour la cartographie du SLN chez les chiens atteints de tumeurs mastocytaires, en particulier dans le milieu de la pratique générale.(Traduit par Dr Serge Messier).

Public understanding of intersex: an update on recent findings.

Surgical interventions on infants with intersex characteristics are considered justified by some on the grounds that they carry a high risk of intolerable stigma. However, public understanding of intersex and its medicalization are under-researched. We review recent qualitative and quantitative studies of the understandings of intersex and its medicalization among people who have no particular professional or public experience of intersex. First, such laypeople reason about clinical dilemmas by drawing on values in similar ways as expert healthcare professionals do. Second, laypeople can over-estimate the utility of current 'umbrella terms,' including intersex, for people with direct familial experience of intersex. Third, beliefs about good and bad effects of medical intervention are affected by framing intersex as either a medical condition or the natural basis for a social identity. Fourth, sexual identity is the best evidenced predictor of opinions about early surgical intervention and its legal limitation on human rights grounds. We argue that possible stigmatizing reactions from the public may not be a solid basis on which to justify early surgical intervention on intersex characteristics.

Evaluation of zoledronate for the treatment of canine stage III osteosarcoma: A phase II study.

BACKGROUND: Greater than 90% of dogs with appendicular osteosarcoma will develop pulmonary metastasis despite the standard of care. Available treatments have limited efficacy for stage III disease. Zoledronate, a bisphosphonate, induces apoptosis of canine osteosarcoma cells and appears to modulate the tumour microenvironment. OBJECTIVES: This prospective, single institutional phase IIa trial investigated the use of single agent zoledronate in dogs with pulmonary metastases from osteosarcoma. METHODS: Zoledronate was administered once monthly, and thoracic radiographs were used to assess response. RESULTS: Eleven dogs were enrolled. Stable disease was achieved in two of eight dogs available for response assessment. The median progression-free survival was 28 days (range: 4-93 days). The median stage III-specific survival time was 92 days. Adverse events were reported in four dogs; two dogs developed grade III or higher toxicities. Notable adverse events included conjunctivitis, fever, hypocalcaemia, and hypophosphatemia. CONCLUSIONS: Zoledronate appears to have limited efficacy as a single agent for stage III osteosarcoma and may be associated with unexpected toxicity in this population. This clinical trial was registered on the AVMA Animal Health Studies Database (AAHSD004396).

Inhibitory control and problem solving in early childhood: Exploring the burdens and benefits of high self-control.

Low inhibitory control (IC) is sometimes associated with enhanced problem-solving amongst adults, yet for young children high IC is primarily framed as inherently better than low IC. Here, we explore associations between IC and performance on a novel problem-solving task, amongst 102 English 2- and 3-year-olds (Study 1) and 84 Swedish children, seen at 18-months and 4-years (Study 2). Generativity during problem-solving was negatively associated with IC, as measured by prohibition-compliance (Study 1, both ages, Study 2 longitudinally from 18-months). High parent-reported IC was associated with poorer overall problem-solving success, and greater perseveration (Study 1, 3-year-olds only). Benefits of high parent-reported IC on persistence could be accounted for by developmental level. No concurrent association was observed between problem-solving performance and IC as measured with a Delay-of-Gratification task (Study 2, concurrent associations at 4-years). We suggest that, for young children, high IC may confer burden on insight- and analytic-aspects of problem-solving.

Transcriptomic Analysis of Canine Osteosarcoma from a Precision Medicine Perspective Reveals Limitations of Differential Gene Expression Studies.

Despite significant advances in cancer diagnosis and treatment, osteosarcoma (OSA), an aggressive primary bone tumor, has eluded attempts at improving patient survival for many decades. The difficulty in managing OSA lies in its extreme genetic complexity, drug resistance, and heterogeneity, making it improbable that a single-target treatment would be beneficial for the majority of affected individuals. Precision medicine seeks to fill this gap by addressing the intra- and inter-tumoral heterogeneity to improve patient outcome and survival. The characterization of differentially expressed genes (DEGs) unique to the tumor provides insight into the phenotype and can be useful for informing appropriate therapies as well as the development of novel treatments. Traditional DEG analysis combines patient data to derive statistically inferred genes that are dysregulated in the group; however, the results from this approach are not necessarily consistent across individual patients, thus contradicting the basis of precision medicine. Spontaneously occurring OSA in the dog shares remarkably similar clinical, histological, and molecular characteristics to the human disease and therefore serves as an excellent model. In this study, we use transcriptomic sequencing of RNA isolated from primary OSA tumor and patient-matched normal bone from seven dogs prior to chemotherapy to identify DEGs in the group. We then evaluate the universality of these changes in transcript levels across patients to identify DEGs at the individual level. These results can be useful for reframing our perspective of transcriptomic analysis from a precision medicine perspective by identifying variations in DEGs among individuals.

Adjuvant Sirolimus Does Not Improve Outcome in Pet Dogs Receiving Standard-of-Care Therapy for Appendicular Osteosarcoma: A Prospective, Randomized Trial of 324 Dogs.

PURPOSE: The mTOR pathway has been identified as a key nutrient signaling hub that participates in metastatic progression of high-grade osteosarcoma. Inhibition of mTOR signaling is biologically achievable with sirolimus, and might slow the outgrowth of distant metastases. In this study, pet dogs with appendicular osteosarcoma were leveraged as high-value biologic models for pediatric osteosarcoma, to assess mTOR inhibition as a therapeutic strategy for attenuating metastatic disease progression. PATIENTS AND METHODS: A total of 324 pet dogs diagnosed with treatment-naïve appendicular osteosarcoma were randomized into a two-arm, multicenter, parallel superiority trial whereby dogs received amputation of the affected limb, followed by adjuvant carboplatin chemotherapy ± oral sirolimus therapy. The primary outcome measure was disease-free interval (DFI), as assessed by serial physical and radiologic detection of emergent macroscopic metastases; secondary outcomes included overall 1- and 2-year survival rates, and sirolimus pharmacokinetic variables and their correlative relationship to adverse events and clinical outcomes. RESULTS: There was no significant difference in the median DFI or overall survival between the two arms of this trial; the median DFI and survival for standard-of-care (SOC; defined as amputation and carboplatin therapy) dogs was 180 days [95% confidence interval (CI), 144-237] and 282 days (95% CI, 224-383) and for SOC + sirolimus dogs, it was 204 days (95% CI, 157-217) and 280 days (95% CI, 252-332), respectively. CONCLUSIONS: In a population of pet dogs nongenomically segmented for predicted mTOR inhibition response, sequentially administered adjuvant sirolimus, although well tolerated when added to a backbone of therapy, did not extend DFI or survival in dogs with appendicular osteosarcoma.

Predictive ability of fine-needle aspirate cytology for incompletely resected mast cell tumor surgical sites.

The goal of this study was to evaluate whether fine-needle aspirate cytology of a previous surgical site was predictive of recurrence for incompletely excised mast cell tumors (MCTs). Electronic medical records were searched for dogs diagnosed with MCTs; those with histologically confirmed, incompletely resected MCTs evaluated by scar aspiration cytology within 60 days after surgery were included for analysis. Variables were compared between groups using Fisher's exact test and logistic regression. Twenty-nine cutaneous and 7 subcutaneous tumors were evaluated. Local recurrence, confirmed by either histopathology or cytology, occurred in 13.8% of cases. No significant differences were identified for any variables other than surgical site cytology status. The negative predictive value of surgical site aspirate cytology without residual mast cell tumor was 93.5%, with an overall predictive accuracy of 88.9%. For the dogs evaluated in this report, surgical site aspiration cytology was predictive of local disease control for incompletely resected MCTs.

Capacité prédictive de la cytologie d'aspiration à l'aiguille fine de sites de chirurgie de résection incomplète de mastocytomes. L'objectif de la présente étude était d'évaluer si la cytologie d'aspiration à l'aiguille fine d'un site chirurgical antérieur permettait de prédire une récurrence lors de l'excision incomplète d'un mastocytome (MCT). Les dossiers médicaux électroniques furent examinés pour trouver des chiens avec un diagnostic de MCT; ceux avec confirmation histologique d'un MCT avec résection incomplète évaluée par cytologie d'une aspiration de la cicatrice en dedans de 60 jours après la chirurgie furent inclus pour analyse. Les variables furent comparées entre les groupes en utilisant le test exact de Fisher et une régression logistique. Vingt-neuf tumeurs cutanées et sept tumeurs sous-cutanées furent évaluées. Une récurrence locale, confirmée par histopathologie ou cytologie, est survenue dans 13,8 % des cas. Aucune différence significative ne fut détectée pour les différentes variables autres que le statut de la cytologie du site chirurgical. La valeur prédictive négative de la cytologie d'une aspiration du site chirurgical sans cellule résiduelle du mastocytome était de 93,5 % avec une précision prédictive globale de 88,9 %. Pour les chiens examinés dans cette étude, la cytologie d'une aspiration du site chirurgical était prédictive d'une maîtrise locale de la maladie lors de résection incomplète d'un MCT.(Traduit par Dr Serge Messier).

Evaluation of tumor immunity after administration of conditionally replicative adenoviral vector in canine osteosarcoma patients.

Osteosarcoma is one among the most common neoplasms in dogs. Current treatments show limited efficacy and fail to prevent metastasis. Conditionally replicative adenoviruses (CRAd) replicate exclusively in targeted tumor cells and release new virus particles to infect additional cells. We proposed that OC-CAVE1 (CAV2 with the E1A promoter replaced with the osteocalcin promotor) may also enhance existing immunity against tumors by overcoming immune tolerance via exposure of new epitopes and cytokine signaling. Eleven client-owned dogs with spontaneously occurring osteosarcomas were enrolled in a pilot study. All dogs were injected with OC-CAVE1 following amputation of the affected limb or limb-sparing surgery. Dogs were monitored for viremia and viral shedding. There was minimal virus shedding in urine and feces by the 6th day and no virus was present in blood after 4 weeks. CAV-2 antibody-titers increased in all of the patients, post-CRAd injection. Immunological assays were performed to monitor 1) humoral response against tumors, 2) levels of circulatory CD11c + cells, 3) levels of regulatory T cells, and 4) cytotoxic activity of tumor specific T cells against autologous tumor cells between pre-CRAd administration and 4 weeks post-CRAd administration samples. Administration of the CRAd OC-CAVE1 resulted in alteration of some immune response parameters but did not appear to result in increased survival duration. However, 2 dogs in the study achieved survival times in excess of 1 year. Weak replication of OC-CAVE1 in metastatic cells and delay of chemotherapy following CRAd treatment may contribute to the lack of immune response and improvement in survival time of the clinical patients.

Total laryngectomy and permanent tracheostomy in six dogs.

The objective of this report is to describe the surgical technique for total laryngectomy and outcome in six dogs. Laryngeal cancer is an uncommon and challenging clinical problem. Total laryngectomy can provide local disease control but is uncommonly performed. Detailed procedural descriptions are limited and similarly limited information is available regarding patient outcome. Institutional medical records were searched for dogs treated with total laryngectomy. Six dogs were identified. The procedure resulted in postoperative quality of life similar to permanent tracheostomy alone. Surgical margin status was evaluated in five of six cases and was complete in those five. All dogs survived to discharge from the hospital. Complications were mostly related to tracheostomy occlusion or collapse which is recognized as a complication associated with permanent tracheostomy. Patient quality of life was acceptable. Local recurrence was suspected in one dog. Recurrence was not observed in the case with unknown margin status.

An experimental philosophical bioethical study of how human rights are applied to clitorectomy on infants identified as female and as intersex.

Clitorectomies performed on the genitals of infants identified as female and as intersex have been described both as similar procedures and as different procedures. The former types of surgery have been recognised more consistently as human rights abuses than the latter in recent decades. We tested social psychological explanations of why human rights are differently recognised when infants are described as 'intersex' or 'female'; 122 laypeople in the UK read one of two near-identical descriptions of clitorectomies performed on intersex or female infants and reported their agreement with 22 items about the human rights of such infants. Clitorectomies were perceived as violating human rights more by women than by men, and more so when infants were described as female than intersex. Endorsement of human rights was better predicted by several psychological variables when infants were described as female than as intersex. Less politically conservative participants, as assessed by a Right-Wing Authoritarianism measure, and participants who trusted medical authority more recognised human rights violations of female infants more than intersex infants. Results are discussed with respect to human rights efforts to protect infants from medically non-necessary genital surgery on the basis of membership in identity categories or possession of sex characteristics.

A multi-level developmental approach to exploring individual differences in Down syndrome: genes, brain, behaviour, and environment.

In this article, we focus on the causes of individual differences in Down syndrome (DS), exemplifying the multi-level, multi-method, lifespan developmental approach advocated by Karmiloff-Smith (1998, 2009, 2012, 2016). We evaluate the possibility of linking variations in infant and child development with variations in the (elevated) risk for Alzheimer's disease (AD) in adults with DS. We review the theoretical basis for this argument, considering genetics, epigenetics, brain, behaviour and environment. In studies 1 and 2, we focus on variation in language development. We utilise data from the MacArthur-Bates Communicative Development Inventories (CDI; Fenson et al., 2007), and Mullen Scales of Early Learning (MSEL) receptive and productive language subscales (Mullen, 1995) from 84 infants and children with DS (mean age 2;3, range 0;7 to 5;3). As expected, there was developmental delay in both receptive and expressive vocabulary and wide individual differences. Study 1 examined the influence of an environmental measure (socio-economic status as measured by parental occupation) on the observed variability. SES did not predict a reliable amount of the variation. Study 2 examined the predictive power of a specific genetic measure (apolipoprotein APOE genotype) which modulates risk for AD in adulthood. There was no reliable effect of APOE genotype, though weak evidence that development was faster for the genotype conferring greater AD risk (ε4 carriers), consistent with recent observations in infant attention (D'Souza, Mason et al., 2020). Study 3 considered the concerted effect of the DS genotype on early brain development. We describe new magnetic resonance imaging methods for measuring prenatal and neonatal brain structure in DS (e.g., volumes of supratentorial brain, cortex, cerebellar volume; Patkee et al., 2019). We establish the methodological viability of linking differences in early brain structure to measures of infant cognitive development, measured by the MSEL, as a potential early marker of clinical relevance. Five case studies are presented as proof of concept, but these are as yet too few to discern a pattern.

Visuo-attentional correlates of Autism Spectrum Disorder (ASD) in children with Down syndrome: A comparative study with children with idiopathic ASD.

BACKGROUND: Children with Down syndrome (DS) are at increased likelihood of Autism Spectrum Disorder (ASD) relative to the general population. To better understand the nature of this comorbidity, we examined the visuo-attentional processes associated with autistic trait expression in children with DS, focusing specifically on attentional disengagement and visual search performance. METHOD: We collected eye-tracking data from children with DS (n = 15) and children with idiopathic ASD (iASD, n = 16) matched according to chronological age. Seven children with DS had a formal clinical diagnosis of ASD (DS+ASD). RESULTS: In children with iASD, but not DS, higher autistic trait levels were associated with decreased temporal facilitation on a gap-overlap task, implying increased visuospatial orienting efficiency. In all cases, higher autistic trait levels were associated with improved visual search performance according to decreased target detection latency. On a visual search task, children with DS+ASD outperformed their peers with DS-ASD, mirroring the phenotypic advantage associated with iASD. We found no evidence of a relationship between attentional disengagement and visual search performance, providing preliminary evidence of a differentiation in terms of underlying visuo-attentional mechanism. CONCLUSION: We illustrate the value of progressing beyond insensitive behavioural measures of phenotypic description to examine, in a more fine-grained way, the attentional features associated with ASD comorbidity in children with DS.

Disentangling autism spectrum and attention-deficit/hyperactivity symptoms over development in fragile X syndrome.

Even genetic disorders associated with monogenic aetiologies are characterized by complex and variable risk for poor outcomes, highlighting the need to follow trajectories longitudinally. Here, we investigated the longitudinal relationships between attention deficit hyperactivity disorder (ADHD) and autism spectrum disorder (ASD) symptoms in a population at high risk for both: boys with fragile X syndrome. 59 boys with fragile X syndrome aged 3-10 years old at entry participated in this study, and were followed up one and two years after their first visit. As expected, we found strong relationships over three timepoints for ADHD symptoms (as measured by the parent-rated Conners scale) and ASD symptoms (as measured by the Social Communication Questionnaire [SCQ]). In addition, using structural equation modeling (SEM) we found that ADHD symptoms at time 2 predicted ASD symptoms at time 3, suggestive of a causal relationship. Importantly, these relationships hold when including chronological age at entry to the study, as well as when including severity of impairment as measured by IQ, and their effects on both ASD and ADHD symptoms do not reach significance. This result highlights the need to study outcomes longitudinally and it informs the comorbidity of the two symptom domains in FXS as well as their potential directionality, both of which have been little researched. In addition, our findings may suggest a future need to study how ADHD symptoms and their treatment impact individuals with ASD.

Pharmacokinetics of multivesicular liposomal encapsulated cytarabine when administered subcutaneously in dogs.

BACKGROUND: Prolonged cytotoxic concentrations of cytarabine (CA) are required for maximum cytotoxicity. DepoCyt is a human liposomal cytarabine (LC) product that lasts longer in plasma and CSF compared with free CA (FC). The use of LC has not been evaluated in dogs. OBJECTIVES: To perform a LC pharmacokinetic (PK) study when administered SC in dogs. ANIMALS: Five healthy female beagles. METHODS: Three-period, 3-treatment, nonblinded, randomized, and crossover design, including a pilot study. LC was administered at 50 mg/m2 SC and FC was administered at 25 and 50 mg/m2 SC and IV. Plasma CA concentrations were measured until 240, 72, and 8 hours after SC LC, SC FC, and IV FC administration, respectively. CA plasma concentrations were quantitated by ultra-high-performance liquid chromatography with mass spectrometry (MS/MS) detection and concentration-time profiles were evaluated by noncompartmental analysis. RESULTS: Subcutaneous LC administration resulted in a maximum plasma concentration of 26.3 to 59.78 ng/mL, time to reach maximum plasma concentration of 2 hours, area under the concentration-time curve to last measurable concentration of 669.3 to 1126 h × ng/mL, and plasma bioavailability (%F) of 19.6% to 31.3%. The PK profiles of FC after SC and IV administration differed when compared with LC. CONCLUSIONS AND CLINICAL IMPORTANCE: In healthy dogs, SC LC administration at 50 mg/m2 results in measurable plasma CA concentrations, is apparently safe and well tolerated, but does not result in prolonged cytotoxic plasma concentrations. Poor absorption of LC prevented establishment of a complete LC PK profile.

Circulating microRNA as biomarkers of canine mammary carcinoma in dogs.

BACKGROUND: Differentiating benign from canine malignant mammary tumors requires invasive surgical biopsy. Circulating microRNAs (miRNA) may represent promising minimally invasive cancer biomarkers in people and animals. OBJECTIVES: To evaluate the serum mRNA profile between dogs with and without mammary carcinoma, and to determine if any of these markers have prognostic significance. ANIMALS: Ten healthy client-owned female dogs (5 intact, 5 spayed) and 10 dogs with histologically confirmed mammary carcinoma were included; 9 were client-owned, whereas 1 was a research colony dog. METHODS: Retrospective study. Serum miRNA was evaluated by RNA deep-sequencing (RNAseq) and digital droplet PCR (dPCR).Expression of candidate biomarkers miR-18a, miR-19b, miR-29b, miR-34c, miR-122, miR-125a, and miR-181a was compared with clinical characteristics, including grade, metastasis, and survival. RESULTS: 452 unique serum miRNAs were detected by RNAseq. Sixty-five individual miRNAs were differentially expressed (>±1.5-fold) and statistically significant between groups. Serum miR-19b (P = .003) and miR-125a (P < .001) were significantly higher in the mammary carcinoma group by dPCR. Both had high accuracy based on receiver operator characteristic area under the curve (0.930 for miR-125a; 0.880 for miR-19b). Circulating miR-18a by RNAseq was significantly higher in mammary carcinoma dogs with histologic evidence of lymphatic invasion (P = 0.03). There was no significant association with any miRNA and survival or inflammatory status. CONCLUSIONS AND CLINICAL IMPORTANCE: Circulating miRNAs are differentially expressed in dogs with mammary carcinoma. Serum miR-19b and miR-18a represent candidate biomarkers for diagnosis and prognosis, respectively.

The foundations of mathematical development in Williams syndrome and Down syndrome.

BACKGROUND: Studies in Down syndrome (DS) and Williams syndrome (WS) have suggested that mathematical abilities are impaired. However, it is unclear which domain-general or domain-specific abilities impact on mathematical development in these developmental disorders. METHOD: The current study examined the foundations of mathematical development across participants with WS (n = 24) and DS (n = 26) compared to typically developing (TD) children (n = 26) in relation to domain-general (i.e., general intelligence and visuospatial abilities) and domain-specific abilities (non-symbolic and symbolic number abilities). RESULTS: Developmental trajectories showed that mathematical abilities were delayed in line with overall mental age in DS and WS. Whilst visuospatial abilities predicted performance for DS and TD participants, this was not the case for the WS group, instead Approximate Number Sense abilities predicted mathematical development. CONCLUSIONS: These findings suggest that those with DS and WS may benefit from different mathematical intervention programmes.

Down syndrome and parental depression: A double hit on early expressive language development.

BACKGROUND AND AIMS: Down syndrome (DS) is often characterised by intellectual disability with particular difficulties in expressive language. However, large individual differences exist in expressive language across development in DS. In the general population, one of the factors associated with variability in this domain is parental depression. We investigated whether this is also the case in young children with DS. METHODS: Thirty-eight children with DS between 8 and 48 months of age participated in this study. Their parents reported on the children's receptive and expressive vocabularies (MacArthur-Bates Communicative Development Inventory) and on parental depression. Furthermore, an experimenter-led standardized developmental assessment (Mullen Scales of Early Learning) was administered to the children to test five domains: gross motor, fine motor, visual reception, receptive language, and expressive language. RESULTS: A cross-sectional developmental trajectories analysis demonstrated that expressive language developed at a slower rate in children with DS whose parent reported depression than in those whose parent did not. No differences between groups were found in any other domain. CONCLUSION: Parental depression is associated with slower rate of expressive language development in young children with DS. These findings suggest that DS and parental depression may constitute a double hit leading to increased difficulties in the development of expressive language.