Dominant mutations in GRHL3 cause Van der Woude Syndrome and disrupt oral periderm development.

Mutations in interferon regulatory factor 6 (IRF6) account for ∼70% of cases of Van der Woude syndrome (VWS), the most common syndromic form of cleft lip and palate. In 8 of 45 VWS-affected families lacking a mutation in IRF6, we found coding mutations in grainyhead-like 3 (GRHL3). According to a zebrafish-based assay, the disease-associated GRHL3 mutations abrogated periderm development and were consistent with a dominant-negative effect, in contrast to haploinsufficiency seen in most VWS cases caused by IRF6 mutations. In mouse, all embryos lacking Grhl3 exhibited abnormal oral periderm and 17% developed a cleft palate. Analysis of the oral phenotype of double heterozygote (Irf6(+/-);Grhl3(+/-)) murine embryos failed to detect epistasis between the two genes, suggesting that they function in separate but convergent pathways during palatogenesis. Taken together, our data demonstrated that mutations in two genes, IRF6 and GRHL3, can lead to nearly identical phenotypes of orofacial cleft. They supported the hypotheses that both genes are essential for the presence of a functional oral periderm and that failure of this process contributes to VWS.

Copy number variation analysis implicates the cell polarity gene glypican 5 as a human spina bifida candidate gene.

Neural tube defects (NTDs) are common birth defects of complex etiology. Family and population-based studies have confirmed a genetic component to NTDs. However, despite more than three decades of research, the genes involved in human NTDs remain largely unknown. We tested the hypothesis that rare copy number variants (CNVs), especially de novo germline CNVs, are a significant risk factor for NTDs. We used array-based comparative genomic hybridization (aCGH) to identify rare CNVs in 128 Caucasian and 61 Hispanic patients with non-syndromic lumbar-sacral myelomeningocele. We also performed aCGH analysis on the parents of affected individuals with rare CNVs where parental DNA was available (42 sets). Among the eight de novo CNVs that we identified, three generated copy number changes of entire genes. One large heterozygous deletion removed 27 genes, including PAX3, a known spina bifida-associated gene. A second CNV altered genes (PGPD8, ZC3H6) for which little is known regarding function or expression. A third heterozygous deletion removed GPC5 and part of GPC6, genes encoding glypicans. Glypicans are proteoglycans that modulate the activity of morphogens such as Sonic Hedgehog (SHH) and bone morphogenetic proteins (BMPs), both of which have been implicated in NTDs. Additionally, glypicans function in the planar cell polarity (PCP) pathway, and several PCP genes have been associated with NTDs. Here, we show that GPC5 orthologs are expressed in the neural tube, and that inhibiting their expression in frog and fish embryos results in NTDs. These results implicate GPC5 as a gene required for normal neural tube development.

To spear or not to spear: comparison of saliva collection methods.

The eye spear, or an absorbent sponge-like material, has been proposed as a useful method of obtaining repeated saliva samples from infants and young children for cortisol determination. This brief report examines possible interference effects of different types of eye spears under conditions of relatively high and low cortisol levels, with or without the use of oral stimulant, and using two common assays. In Study 1, one type of eye spear was compared to passively collected drool using two different assays (EIA, DELFIA), across high and low concentrations of cortisol. No differences were found between methods for either assay or cortisol level, indicating that the spears are potentially a viable method of collecting saliva. Study 2 compared three other types of absorbent eye spears to passive drooling under the presence or absence of oral stimulant use. This study revealed that the degree of interference varied as a function of the specific type of eye spear that was employed; stimulant use had no effect. Taken together, the results raise important considerations to take into account when selecting collection materials and procedures in the measurement of salivary cortisol.

Demystifying reflective practice: Using the DATA model to enhance evaluators' professional activities.

Reflective practice (RP), one of six essential competency domains in evaluation identified by Stevahn, King, Ghere, and Minnema (2005), refers to thinking critically about one's evaluation practice, alone or with other people, and using critical insights to improve one's practice. Currently, evaluators have minimal guidance in navigating this essential professional competency, professed to be a necessary part of their practice. This article focuses on how RP can serve as a tool for evaluators through the use of the "DATA" integrated RP framework, developed by Peters (1991, 2009). DATA is an acronym with each letter standing for a different step in the process of reflective practice. The "D" step of the acronym focuses on (D)escribing what is or has been happening in practice. The "A" step refers to (A)nalyzing the current state of practice-why is this happening the way it is? The "T" concentrates on a practice-oriented form of (T)heorizing, which comes from analysis and serves as a basis for the resulting (A)ct. The last "A" focuses on the specifics of an action plan to change one's evaluation practice in light of the practical theory developed through theorizing. This paper describes the DATA model and introduces the application of the framework in a practice context.