More evidence on TRIO missense mutations in the spectrin repeat domain causing severe developmental delay and recognizable facial dysmorphism with macrocephaly.

TRIO is a Dbl family guanine nucleotide exchange factor (GEF) and an important regulator of neuronal development. Most truncating and missense variants affecting the Dbl homology domain of TRIO are associated with a neurodevelopmental disorder with microcephaly (MIM617061). Recently, de novo missense variants affecting the spectrin repeat region of TRIO were associated with a novel phenotype comprising severe developmental delay and macrocephaly (MIM618825). Here, we provide more evidence on this new TRIO-associated phenotype by reporting two severely affected probands with de novo missense variants in TRIO affecting the spectrin repeat region upstream of the typically affected GEF1 domain of the protein.

Genetic and phenotypic characterization of NKX6-2-related spastic ataxia and hypomyelination.

BACKGROUND AND PURPOSE: Hypomyelinating leukodystrophies are a heterogeneous group of genetic disorders with a wide spectrum of phenotypes and a high rate of genetically unsolved cases. Bi-allelic mutations in NKX6-2 were recently linked to spastic ataxia 8 with hypomyelinating leukodystrophy. METHODS: Using a combination of homozygosity mapping, exome sequencing, and detailed clinical and neuroimaging assessment a series of new NKX6-2 mutations in a multicentre setting is described. Then, all reported NKX6-2 mutations and those identified in this study were combined and an in-depth analysis of NKX6-2-related disease spectrum was provided. RESULTS: Eleven new cases from eight families of different ethnic backgrounds carrying compound heterozygous and homozygous pathogenic variants in NKX6-2 were identified, evidencing a high NKX6-2 mutation burden in the hypomyelinating leukodystrophy disease spectrum. Our data reveal a phenotype spectrum with neonatal onset, global psychomotor delay and worse prognosis at the severe end and a childhood onset with mainly motor phenotype at the milder end. The phenotypic and neuroimaging expression in NKX6-2 is described and it is shown that phenotypes with epilepsy in the absence of overt hypomyelination and diffuse hypomyelination without seizures can occur. CONCLUSIONS: NKX6-2 mutations should be considered in patients with autosomal recessive, very early onset of nystagmus, cerebellar ataxia with hypotonia that rapidly progresses to spasticity, particularly when associated with neuroimaging signs of hypomyelination. Therefore, it is recommended that NXK6-2 should be included in hypomyelinating leukodystrophy and spastic ataxia diagnostic panels.

Branchio-otic syndrome caused by a genomic rearrangement: clinical findings and molecular cytogenetic studies in a patient with a pericentric inversion of chromosome 8.

Branchio-oto-renal (BOR) syndrome is an autosomal dominantly inherited developmental disorder, which is characterized by anomalies of the ears, the branchial arches and the kidneys. It is caused by mutations in the genes EYA1,SIX1 and SIX5. Genomic rearrangements of chromosome 8 affecting the EYA1 gene have also been described. Owing to this fact, methods for the identification of abnormal copy numbers such as multiplex ligation-dependent probe amplification (MLPA) have been introduced as routine laboratory techniques for molecular diagnostics of BOR syndrome. The advantages of these techniques are clear compared to standard cytogenetic and array approaches as well as Southern blot. MLPA detects deletions or duplications of a part or the entire gene of interest, but not balanced structural aberrations such as inversions and translocations. Consequently, disruption of a gene by a genomic rearrangement may escape detection by a molecular genetic analysis, although this gene interruption results in haploinsufficiency and, therefore, causes the disease. In a patient with clinical features of BOR syndrome, such as hearing loss, preauricular fistulas and facial dysmorphisms, but no renal anomalies, neither sequencing of the 3 genes linked to BOR syndrome nor array comparative genomic hybridization and MLPA were able to uncover a causative mutation. By routine cytogenetic analysis, we finally identified a pericentric inversion of chromosome 8 in the affected female. High-resolution multicolor banding confirmed the chromosome 8 inversion and narrowed down the karyotype to 46,XX,inv(8)(p22q13). By applying fluorescence in situ hybridization, we narrowed down both breakpoints on chromosome 8 and found the EYA1 gene in q13.3 to be directly disrupted. We conclude that standard karyotyping should not be neglected in the genetic diagnostics of BOR syndrome or other Mendelian disorders, particularly when molecular testing failed to detect any causative alteration in patients with a convincing phenotype.

Dysfunction of SHANK2 and CHRNA7 in a patient with intellectual disability and language impairment supports genetic epistasis of the two loci.

Synaptopathies constitute a group of neurological diseases including autism spectrum disorders (ASD) and intellectual disability (ID). They have been associated with mutations in genes encoding proteins important for the formation and stabilization of synapses, such as SHANK1-3. Loss-of-function mutations in the SHANK genes have been identified in individuals with ASD and ID suggesting that other factors modify the neurological phenotype. We report a boy with severe ID, behavioral anomalies, and language impairment who carries a balanced de novo triple translocation 46,XY,t(11;17;19)(q13.3;q25.1;q13.42). The 11q13.3 breakpoint was found to disrupt the SHANK2 gene. The patient also carries copy number variations at 15q13.3 and 10q22.11 encompassing ARHGAP11B and two synaptic genes. The CHRNA7 gene encoding α7-nicotinic acetylcholine receptor subunit and the GPRIN2 gene encoding G-protein-regulated inducer of neurite growth 2 were duplicated. Co-occurrence of a de novo SHANK2 mutation and a CHRNA7 duplication in two reported patients with ASD and ID as well as in the patient with t(11;17;19), severe ID and behavior problems suggests convergence of these genes on a common synaptic pathway. Our results strengthen the oligogenic inheritance model and highlight the presence of a large effect mutation and modifier genes collectively determining phenotypic expression of the synaptopathy.

Newborn piglet traits associated with survival and growth performance until weaning.

Pre-weaning piglet mortality represents significant economic losses, and approximately half of this mortality occurs within the first 3 days after birth. Factors involved in postnatal mortality can also be associated with a poor growth performance until weaning. The aim of this study was to evaluate the effect of some variables measured right after birth on piglet survival during the first week of life and growth performance until weaning. Piglets included in the analysis (n=612) were born from 3 to 5 parity sows. Piglets were monitored for blood oxygen saturation (SatO2), heart rate (HR), blood glucose concentration, rectal temperature at birth (RT0 h) and at 24h after birth (RT24 h). Genetic line, birth following or not an obstetric intervention, birth order, sex, skin color, integrity of the umbilical cord, and time elapsed from birth until first attempts to stand were also recorded. Piglets were weighed at birth (BW), and at 7, 14 and 21 days after birth in order to evaluate their postnatal development. Cumulative mortality rates were 3.3%, 5.4% and 8.7% at 3, 7 and 21 days after birth, respectively. Body temperature at birth (RT0 h) did not affect (P>0.05) the survival nor the piglet growth performance. Piglets with cyanotic skin and those that took more than 5 min to stand showed higher chance of mortality (P<0.05) compared to normal skin piglets and to piglets which stood before 1 min, respectively. Piglets with broken umbilical cord had higher odds (P<0.05) of mortality up to 3 days after birth, respectively. Higher odds (P<0.05) of mortality up to 3 or 7 days were associated with later birth order (>9), low BW (<1275g), low (24-30 mg/dl) and high (45-162 mg/dl) blood glucose concentrations, or low RT24 h (<38.1°C). Piglets with BW<1545 g, low RT24 h (<38.6°C) and female piglets had higher odds of a low weight at weaning (P<0.05). Among the factors studied, cyanotic skin, delay for standing, broken umbilical cord, high birth order, low BW, low RT24 h, and both low and high blood glucose concentrations are indicators of a lower ability of piglets to survive during the first week after birth. The growth performance until weaning is compromised in piglets with a lower BW, a lower RT24 h and if they are female pigs.

Efeito da ordem de parto e da perda de peso durante a lactação no desempenho reprodutivo subsequente de matrizes suínas / The effect of parity order and lactation weight loss on subsequent reproductive performance of sows

Investigou-se o efeito da ordem de parto (OP) e da perda de peso durante a lactação no desempenho reprodutivo de 666 matrizes suínas. Foram formadas três classes de OP (OP1, OP2 e OP3-5) e duas classes de percentual de perda de peso (≤1% e >1%) durante a lactação. Foram avaliados: taxa de parto, intervalo desmame-estro (IDE) e tamanho da leitegada subsequente. Houve interação de OP versus perda de peso na taxa de parto das fêmeas (P<0,05). As fêmeas OP1 e OP2 tiveram maior chance (P<0,05) de não parir na classe >1% de perda de peso. A maior perda de peso não foi um fator de risco para não parir na condição OP3-5 (P>0,05). Não houve interação de OP versus perda de peso (P>0,05) para IDE e total de leitões nascidos. Fêmeas OP1 apresentaram IDE mais longo e menor tamanho da leitegada no parto subsequente (P<0,05) do que fêmeas OP2 e OP3-5. As perdas corporais na lactação não influenciaram o IDE (P>0,05), mas reduziram o tamanho da leitegada subsequente (P<0,05). Conclui-se que maior perda de peso na lactação resulta em aumento de falhas reprodutivas nas fêmeas mais jovens e reduz o tamanho da leitegada subsequente em todas as OP.

The effect of parity (PO) and weight loss during lactation on the subsequent reproductive performance of 666 sows was investigated. Sows were allocated into three PO classes (PO1, PO2 and PO3-5) and into two classes of weight loss percentage (≤1% and >1%) during lactation. Farrowing rate, weaning to estrus interval (WEI) and total born at next farrow were evaluated. There was an interaction effect between PO and weight loss on the farrowing rate (P<0.05). PO1 and PO2 females had more chance (P<0.05) of failure to farrow when weight loss was higher than 1%. However, the higher weight loss was not a risk factor to low farrowing rate in PO3-5 females (P>0.05). There was no interaction between PO and weight loss (P>0.05) on WEI and subsequent total born. PO1 females showed longer WEI and lower litter size on subsequent farrowing compared to PO2 and PO3-5 females. Weight loss did not affect WEI (P>0.05), but it decreased subsequent litter size (P<0.05). High weight loss during lactation results an increase in reproductive failures in young females and decreases subsequent litter size in all parity orders.

Absorção de IgG via colostro em leitões biológicos e adotados após a uniformização da leitegada / Absorption of IgG via colostrum in biological piglets and adopted piglets after crossfostering

Nove fêmeas de quinto parto (OP5) foram imunizadas com 4mg e 2mg de albumina sérica bovina (BSA) aos 70 e 100 dias de gestação, respectivamente. A uniformização da leitegada foi realizada 4,9±1,9h após o nascimento, antes de os leitões efetuarem a primeira mamada. As leitegadas foram compostas por cinco leitões biológicos (LB) e cinco leitões adotados (LA), com pesos semelhantes ao nascimento. Foram coletadas amostras de sangue dos leitões ao nascimento e 24h após, das fêmeas ao pós-parto e de colostro de cada grupo de tetos ao parto e 24h após. As amostras de soro e colostro foram quantificadas para IgG pelo ELISA indireto. A densidade ótica de IgG anti-BSA (DOIgG-BSA) dos leitões (24h de vida) foi correlacionada com a das fêmeas. A DOIgG-BSA entre LB e LA foi semelhante, assim como entre os grupos de tetos, ao parto e 24h após. Entretanto, ocorreu redução na DOIgG-BSA do parto até 24h após. LB e LA absorveram a mesma quantidade de IgG via colostro, quando a uniformização foi realizada até 5h pós-parto, independentemente do teto em que os leitões mamaram, uma vez que esses possuem a mesma concentração de IgG.

Nine sows of fifth parity (PO5) were immunized with 4mg and 2mg of bovine serum albumin (BSA) at 70 and 100d of gestation, respectively. Cross fostering was performed 4.9±1.9h after birth, before piglets had their first suckling. Litters were composed of five biological piglets (BP) and five adopted piglets (AP), with similar weight at birth. Blood samples were collected from piglets (at birth and at 24h of life) and from females (after farrowing) and colostrum from each group of teats (at farrowing time and after 24h). Samples of serum and colostrum were quantified to IgG by indirect ELISA. Optical density of IgG anti-BSA (ODIgG-BSA) from piglets (24h of life) was correlated with dams. ODIgG-BSA was similar among BP and AP, as well as among pairs of teats (at farrowing time and after 24h). However, there was a decrease in ODIgG-BSA from farrowing up to 24h after birth. BP and AP absorbed the same amount of IgG via colostrum, when cross fostering was evaluated 5h after farrowing, regardless of the teat suckled, since these have the same concentration of IgG.