Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome.

Warmblood Fragile Foal syndrome (WFFS) is an autosomal recessive condition that affects the maturation of collagen in affected foals. Foals affected with the disease typically die or are euthanised shortly after birth. WFFS is caused by a single nucleotide change at position 2032 of the equine PLOD1 gene, causing an impairment of the wild-type enzyme. A commercial test for the causative genetic mutation is currently available from companies operating under licence from Cornell University but it has limitations. This test requires amplification of a region of the PLOD1 gene encompassing the site of interest, followed by Sanger sequencing of that region and computational analysis. We describe here the development of an alternative, real-time PCR based assay that rapidly and reliably differentiates between the wild-type and WFFS associated nucleotides without the need for sequencing, thus increasing the potential for high throughput analysis of large numbers of samples in a cost-effective manner.

Warmblood fragile foal syndrome causative single nucleotide polymorphism frequency in horses in Ireland.

BACKGROUND: Warmblood Fragile Foal Syndrome (WFFS) is an autosomal recessive disorder caused by a mutation in the procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 (PLOD1) gene. Homozygosity for the mutation results in defective collagen synthesis which clinically manifests as the birth of non viable or still born foals with abnormally fragile skin. While the mutation has been identified in non Warmblood breeds including the Thoroughbred, to date all homozygous clinically affected cases reported in the scientific literature are Warmblood foals. The objective of this study was to investigate the carrier frequency of the mutation in the Thoroughbred and sport horse populations in Ireland. METHODS: A test was developed at the UCD School of Veterinary Medicine using real-time PCR to amplify the PLOD1 gene c.2032G > A variant. A subset of the samples was also submitted to an external laboratory with a licensed commercial WFFS genetic test. RESULTS: Warmblood Fragile Foal Syndrome genotyping was performed on hair samples from 469 horses representing 6 different breeds. Six of 303 (1.98%) sport horses tested and three of 109 (2.75%) Thoroughbreds tested were heterozygous for the WFFS polymorphism (N/WFFS). The WFFS polymorphism was not identified in the Standardbred, Cob, Connemara, or other pony breeds. CONCLUSIONS: The study identified a low frequency of the WFFS causative mutation in sport horses and Thoroughbreds in Ireland, highlighting the importance of WFFS genetic testing in order to identify phenotypically normal heterozygous carriers and to prevent the birth of nonviable foals.

Impaired perception of sincerity after severe traumatic brain injury.

BACKGROUND: People with a severe traumatic brain injury (TBI) often experience problems understanding non-literal utterances such as sarcasm and lies in dyadic exchanges. This study aimed to investigate whether these problems extend to settings where speakers vary in their degree of sincerity and whether such problems are associated with deficits in social cognitive abilities (emotion perception, theory of mind, and self-reported empathy) or cognitive abilities (abstract reasoning, working memory, processing speed, attentional switching). METHODS: Thirty-one adults with severe TBI (24 males) and 25 demographically matched controls (20 males) participated. They watched video vignettes depicting four actors volunteering for additional duties. Each speaker made a limited verbal response which literally suggested a willingness to be involved, but the sincerity with which the response was made was tempered by the actor's emotional demeanour. Participants rated each speaker in the vignettes for degree of sincerity (0-100%). Standardized measures of cognitive and social cognitive function were also taken. RESULTS: Control participants had excellent agreement (α = .90) in their rankings of actors according to sincerity. TBI participants were less consistent (α = .65). Overall, they were sensitive to decreasing sincerity but generally less accurate than control participants. They were poorer at differentiating between levels of sincerity and rated insincere expressions as more sincere, although they rated sincere expressions similarly. Poorer working memory and poorer social cognition were associated with poorer sincerity/sarcasm detection in the participants with TBI, but only social cognition was uniquely associated. CONCLUSIONS: Some adults with TBI have difficulty assessing the level of sincerity of speakers. Moreover, poorer social cognition abilities are associated with this difficulty.

Early verification of myocardial ischemia with a novel biomarker of acute tissue damage: C-reactive protein fractional forms.

BACKGROUND: We evaluated the utility of an independent biomarker of early ischemic cellular damage-circulating fractional forms of C-reactive protein (fracCRP), to verify the diagnostic relevance of low Troponin I (TnI) values within the context of a workup for Acute Coronary Syndrome (ACS). METHODS: On a semi-preparative scale, the molecular characteristics of fracCRP were established by electron microscopy and Western Blot, using isolates captured from patient serum on phosphorylcholine beads and purified by size exclusion high-pressure liquid chromatography (SE-HPLC). Captured on an analytical scale, the diagnostic utility of fracCRP was evaluated in first-draw plasma specimens (total CRP not exceeding 6 mg/l) recovered from 300 cardiac emergency patients with final discharge diagnoses of ACS ruled out (N=132) or ruled in (N=168). RESULTS: At a cutoff value chosen for 97.7% test specificity, the test metric (fracCRP×TnI) identified in the first blood draw 39.9% of all emergency patients ultimately diagnosed with ACS, and 17.9% of ultimately diagnosed patients who arrived with TnI values within the normal reference range (0.01-0.04 ng/ml). CONCLUSIONS: These findings suggest that the fracCRP test metric could serve as a rule-in test for ACS in a significant proportion of low to moderate risk emergency patients.

Social perception deficits after traumatic brain injury: interaction between emotion recognition, mentalizing ability, and social communication.

Thirty-four adults with severe traumatic brain injuries (TBI) and 34 matched control participants were asked to interpret videotaped conversational exchanges. Study participants were asked to judge the speakers' emotions, the speakers' beliefs (first-order theory of mind), what the speakers intended their conversational partners to believe (second-order theory of mind), and what they meant by remarks that were sincere or literally untrue (i.e., a lie or sarcastic retort). The TBI group had marked difficulty judging most facets of social information. They could recognize speaker beliefs only when this information was explicitly provided. In general, emotion recognition and first-order theory of mind judgments were not related to the ability to understand social (conversational) inference, whereas second-order theory of mind judgments were related to that ability.

TASIT: A new clinical tool for assessing social perception after traumatic brain injury.

OBJECTIVE: To develop a clinically sensitive test of social perception for people with traumatic brain injury (TBI). DESIGN: An assessment tool comprising videotaped vignettes and response probes was developed in successive stages and tested on both normal participants and those with TBI. SUBJECTS: A total of 169 normal adults and 7 adults with severe TBI (pilot studies), 283 normal adults, and 12 people with severe TBI (main studies). MAIN OUTCOME MEASURES: "The Awareness of Social Inference Test" (TASIT) comprises videotaped vignettes of everyday social interactions and has three parts, each with alternate forms. The Emotion Evaluation Test (EET) assesses recognition of spontaneous emotional expression (happy, surprised, sad, anxious, angry, disgusted, and neutral). The Social Inference-Minimal (SI-M) test assesses comprehension of sincere versus sarcastic exchanges, whereas the Social Inference-Enriched test (SI-E) assesses lies versus sarcasm. In both SI-M and SI-E speaker demeanor (voice, facial expression) indicate the intended meaning of the exchange. In addition, the SI-E vignettes have other contextual clues that reveal the speakers' intentions. Performance on SI-E and SI-E is assessed via four standard questions per item probing for understanding of the emotions, intentions, beliefs, and meanings of the speakers and their exchanges. RESULTS: Groups taken from the pool of 283 normal adults achieved a high level of performance on all aspects of the test with some influence from both education and intelligence. The 12 people with TBI were poorer at judging emotions than were matched controls, with particular difficulties recognizing neutral items, fear, and disgust. They were as capable as matched controls when understanding sincere exchanges and lies but had difficulty with sarcasm. CONCLUSIONS: TASIT is straightforward for people with a normal range of social skills while being sensitive to social perception deficits after traumatic brain injury.