Review: Facial endophenotypes in non-syndromic orofacial clefting.

Cleft lip and/or palate (CL/P) is one of the most frequent congenital malformations, with a frequency of 1 in 700 live births. Non-syndromic orofacial clefting is a multifactorial condition, with both a genetic and an environmental component. Although numerous studies have been published addressing the genetic etiology of CL/P, this factor remains incompletely understood. A promising approach to find candidate gene regions for CL/P is the investigation of endophenotypes, which are characteristics associated with a certain condition and that can be an expression of underlying susceptibility genes. This review focuses on the known facial endophenotypes in CL/P (such as distortion of the orbicularis oris muscle and facial features in non-affected relatives of patients with CL/P) and genes that could be associated with these characteristics. Possibilities for further endophenotype-related studies in the field of non-syndromic CL/P are discussed.

Principal polynomial shape analysis: A non-linear tool for statistical shape modeling.

BACKGROUND AND OBJECTIVES: The most widespread statistical modeling technique is based on Principal Component Analysis (PCA). Although this approach has several appealing features, it remains hampered by its linearity. Principal Polynomial Analysis (PPA) can capture non-linearity in a sequential algorithm, while maintaining the interesting properties of PCA. PPA is, however, computationally expensive in handling shape surface data. To this end, we propose Principal Polynomial Shape Analysis (PPSA) as an adjusted approach for non-linear shape analyzes. The aim of this study was to assess PPSA's features, its model boundaries and its general applicability. METHODS: PCA and PPSA-based shape models were investigated on one verification and three model evaluation experiments. In the verification experiment, the estimated mean of the PCA and PPSA model on a data set of synthetic lower limbs of different lengths in different poses were compared to the real mean. Further, the PCA-based and PPSA shape models were tested for three challenging cases namely for shape model creation of gait marker data, for regression analysis on aging faces and for modeling pose variation in full body scans. For the latter, additionally a Fundamental Coordinate Model (FCM) and a PPSA model on Fundamental Coordinate(FC) space was created. The performances were evaluated based on model-based accuracy, generalization, compactness and specificity. RESULTS: In the verification experiment, the scaling error reduced from 75% to below 1% when employing PPSA instead of PCA for a training set with 180° angular variation. For the model evaluation experiments, the PPSA models described the data as accurate and generalized as the PCA-based shape models. The PPSA models were slightly more compact and specific (up to 30%) than the PCA-based models. In regression, PCA and PPSA-based parameterizations explained a similar amount of variation. Lastly, for the full body scans, applying PPSA to parameterizations improved the compactness and accuracy. CONCLUSIONS: PPSA describes the non-linear relationships between principal variations in a parameterized space. Compared to standard PCA-based shape models, capturing the non-linearity reduced the nonsense information in the shape components and improved the description of the data mean.

Wear patterns in knee OA correlate with native limb geometry.

Background: To date, the amount of cartilage loss is graded by means of discrete scoring systems on artificially divided regions of interest (ROI). However, optimal statistical comparison between and within populations requires anatomically standardized cartilage thickness assessment. Providing anatomical standardization relying on non-rigid registration, we aim to compare morphotypes of a healthy control cohort and virtual reconstructed twins of end-stage knee OA subjects to assess the shape-related knee OA risk and to evaluate possible correlations between phenotype and location of cartilage loss. Methods: Out of an anonymized dataset provided by the Medacta company (Medacta International SA, Castel S. Pietro, CH), 798 end-stage knee OA cases were extracted. Cartilage wear patterns were observed by computing joint space width. The three-dimensional joint space width data was translated into a two-dimensional pixel image, which served as the input for a principal polynomial autoencoder developed for non-linear encoding of wear patterns. Virtual healthy twin reconstruction enabled the investigation of the morphology-related risk for OA requiring joint arthroplasty. Results: The polynomial autoencoder revealed 4 dominant, orthogonal components, accounting for 94% of variance in the latent feature space. This could be interpreted as medial (54.8%), bicompartmental (25.2%) and lateral (9.1%) wear. Medial wear was subdivided into anteromedial (11.3%) and posteromedial (10.4%) wear. Pre-diseased limb geometry had a positive predictive value of 0.80 in the prediction of OA incidence (r 0.58, p < 0.001). Conclusion: An innovative methodological workflow is presented to correlate cartilage wear patterns with knee joint phenotype and to assess the distinct knee OA risk based on pre-diseased lower limb morphology. Confirming previous research, both alignment and joint geometry are of importance in knee OA disease onset and progression.

Structural identification of caged vanadium doped silicon clusters.

The geometry of cationic silicon clusters doped with vanadium, Si(n)V(+) (n=12-16), is investigated by using infrared multiple photon dissociation of the corresponding rare gas complexes in combination with ab initio calculations. It is shown that the clusters are endohedral cages, and evidence is provided that Si(16)V(+) is a fluxional system with a symmetric Frank-Kasper geometry.

An exploration of adolescent facial shape changes with age via multilevel partial least squares regression.

BACKGROUND AND OBJECTIVES: Multilevel statistical models represent the existence of hierarchies or clustering within populations of subjects (or shapes in this work). This is a distinct advantage over single-level methods that do not. Multilevel partial-least squares regression (mPLSR) is used here to study facial shape changes with age during adolescence in Welsh and Finnish samples comprising males and females. METHODS: 3D facial images were obtained for Welsh and Finnish male and female subjects at multiple ages from 12 to 17 years old. 1000 3D points were defined regularly for each shape by using "meshmonk" software. A three-level model was used here, including level 1 (sex/ethnicity); level 2, all "subject" variations excluding sex, ethnicity, and age; and level 3, age. The mathematical formalism of mPLSR is given in an Appendix. RESULTS: Differences in facial shape between the ages of 12 and 17 predicted by mPLSR agree well with previous results of multilevel principal components analysis (mPCA); buccal fat is reduced with increasing age and features such as the nose, brow, and chin become larger and more distinct. Differences due to ethnicity and sex are also observed. Plausible simulated faces are predicted from the model for different ages, sexes and ethnicities. Our models provide good representations of the shape data by consideration of appropriate measures of model fit (RMSE and R2). CONCLUSIONS: Repeat measures in our dataset for the same subject at different ages can only be modelled indirectly at the lowest level of the model at discrete ages via mPCA. By contrast, mPLSR models age explicitly as a continuous covariate, which is a strong advantage of mPLSR over mPCA. These investigations demonstrate that multivariate multilevel methods such as mPLSR can be used to describe such age-related changes for dense 3D point data. mPLSR might be of much use in future for the prediction of facial shapes for missing persons at specific ages or for simulating shapes for syndromes that affect facial shape in new subject populations.

Automatic 3D dense phenotyping provides reliable and accurate shape quantification of the human mandible.

Automatic craniomaxillofacial (CMF) three dimensional (3D) dense phenotyping promises quantification of the complete CMF shape compared to the limiting use of sparse landmarks in classical phenotyping. This study assesses the accuracy and reliability of this new approach on the human mandible. Classic and automatic phenotyping techniques were applied on 30 unaltered and 20 operated human mandibles. Seven observers indicated 26 anatomical landmarks on each mandible three times. All mandibles were subjected to three rounds of automatic phenotyping using Meshmonk. The toolbox performed non-rigid surface registration of a template mandibular mesh consisting of 17,415 quasi landmarks on each target mandible and the quasi landmarks corresponding to the 26 anatomical locations of interest were identified. Repeated-measures reliability was assessed using root mean square (RMS) distances of repeated landmark indications to their centroid. Automatic phenotyping showed very low RMS distances confirming excellent repeated-measures reliability. The average Euclidean distance between manual and corresponding automatic landmarks was 1.40 mm for the unaltered and 1.76 mm for the operated sample. Centroid sizes from the automatic and manual shape configurations were highly similar with intraclass correlation coefficients (ICC) of > 0.99. Reproducibility coefficients for centroid size were < 2 mm, accounting for < 1% of the total variability of the centroid size of the mandibles in this sample. ICC's for the multivariate set of 325 interlandmark distances were all > 0.90 indicating again high similarity between shapes quantified by classic or automatic phenotyping. Combined, these findings established high accuracy and repeated-measures reliability of the automatic approach. 3D dense CMF phenotyping of the human mandible using the Meshmonk toolbox introduces a novel improvement in quantifying CMF shape.

Ischiofemoral impingement: the evolutionary cost of pelvic obstetric adaptation.

The risk for ischiofemoral impingement has been mainly related to a reduced ischiofemoral distance and morphological variance of the femur. From an evolutionary perspective, however, there are strong arguments that the condition may also be related to sexual dimorphism of the pelvis. We, therefore, investigated the impact of gender-specific differences in anatomy of the ischiofemoral space on the ischiofemoral clearance, during static and dynamic conditions. A random sampling Monte-Carlo experiment was performed to investigate ischiofemoral clearance during stance and gait in a large (n = 40 000) virtual study population, while using gender-specific kinematics. Subsequently, a validated gender-specific geometric morphometric analysis of the hip was performed and correlations between overall hip morphology (statistical shape analysis) and standard discrete measures (conventional metric approach) with the ischiofemoral distance were evaluated. The available ischiofemoral space is indeed highly sexually dimorphic and related primarily to differences in the pelvic anatomy. The mean ischiofemoral distance was 22.2 ± 4.3 mm in the females and 29.1 ± 4.1 mm in the males and this difference was statistically significant (P < 0.001). Additionally, the ischiofemoral distance was observed to be a dynamic measure, and smallest during femoral extension, and this in turn explains the clinical sign of pain in extension during long stride walking. In conclusion, the presence of a reduced ischiofemroal distance and related risk to develop a clinical syndrome of ischiofemoral impingement is strongly dominated by evolutionary effects in sexual dimorphism of the pelvis. This should be considered when female patients present with posterior thigh/buttock pain, particularly if worsened by extension. Controlled laboratory study.

Separating positional noise from neutral alignment in multicomponent statistical shape models.

Given sufficient training samples, statistical shape models can provide detailed population representations for use in anthropological and computational genetic studies, injury biomechanics, musculoskeletal disease models or implant design optimization. While the technique has become extremely popular for the description of isolated anatomical structures, it suffers from positional interference when applied to coupled or articulated input data. In the present manuscript we describe and validate a novel approach to extract positional noise from such coupled data. The technique was first validated and then implemented in a multicomponent model of the lower limb. The impact of noise on the model itself as well as on the description of sexual dimorphism was evaluated. The novelty of our methodology lies in the fact that no rigid transformations are calculated or imposed on the data by means of idealized joint definitions and by extension the models obtained from them.

Multilevel principal components analysis of three-dimensional facial growth in adolescents.

BACKGROUND AND OBJECTIVES: The study of age-related facial shape changes across different populations and sexes requires new multivariate tools to disentangle different sources of variations present in 3D facial images. Here we wish to use a multivariate technique called multilevel principal components analysis (mPCA) to study three-dimensional facial growth in adolescents. METHODS: These facial shapes were captured for Welsh and Finnish subjects (both male and female) at multiple ages from 12 to 17 years old (i.e., repeated-measures data). 1000 "dense" 3D points were defined regularly for each shape by using a deformable template via "meshmonk" software. A three-level model was used here, namely: level 1 (sex/ethnicity); level 2, all "subject" variations excluding sex, ethnicity, and age; and level 3, age. The technicalities underpinning the mPCA method are presented in Appendices. RESULTS: Eigenvalues via mPCA predicted that: level 1 (ethnicity/sex) contained 7.9% of variation; level 2 contained 71.5%; and level 3 (age) contained 20.6%. The results for the eigenvalues via mPCA followed a similar pattern to those results of single-level PCA. Results for modes of variation made sense, where effects due to ethnicity, sex, and age were reflected in modes at appropriate levels of the model. Standardised scores at level 1 via mPCA showed much stronger differentiation between sex and ethnicity groups than results of single-level PCA. Results for standardised scores from both single-level PCA and mPCA at level 3 indicated that females had different average "trajectories" with respect to these scores than males, which suggests that facial shape matures in different ways for males and females. No strong evidence of differences in growth patterns between Finnish and Welsh subjects was observed. CONCLUSIONS: mPCA results agree with existing research relating to the general process of facial changes in adolescents with respect to age quoted in the literature. They support previous evidence that suggests that males demonstrate larger changes and for a longer period of time compared to females, especially in the lower third of the face. These calculations are therefore an excellent initial test that multivariate multilevel methods such as mPCA can be used to describe such age-related changes for "dense" 3D point data.

The influence of sex, age and body mass index on facial soft tissue depths.

Facial soft tissue depth charts are used in the majority of forensic facial approximation methods. In the past, based on the multitude of available soft tissue depth charts, a number of hypotheses were advanced concerning the impact of sex, BMI and age on the depth of tissues. In this study, for the first time, a multivariate analysis was performed on a large-scale study on Caucasian adults to determine the "real" impact of these attributes. The calculation of a robust multiple linear regression of soft tissue thickness versus BMI, age and sex for each landmark separately, allowed us to study the impact from a statistical as well as practical point of view. Former findings were re-evaluated. Additionally, the results confirm the dominant role of BMI in the alterations of facial soft tissue thickness. However, excluding age and sex from the equation should be considered with care and can certainly not be applied to all landmarks. Finally, the regression equation allows increase in the specificity of tissue depths used in real cases by offering practitioners the possibility of calculating individual tissue depths.

Facial soft tissue thicknesses in craniofacial identification: Data collection protocols and associated measurement errors.

Facial soft tissue thicknesses (FSTT) form a key component of craniofacial identification methods, but as for any data, embedded measurement errors are highly pertinent. These in part dictate the effective resolution of the measurements. As herein reviewed, measurement methods are highly varied in FSTT studies and associated measurement errors have generally not been paid much attention. Less than half (44%) of 95 FSTT studies comment on measurement error and not all of these provide specific quantification. Where informative error measurement protocols are employed (5% of studies), the mean error magnitudes range from 3% to 45% rTEM and are typically in the order of 10-20%. These values demonstrate that FSTT measurement errors are similar in size to (and likely larger than) the magnitudes of many biological effects being chased. As a result, the attribution of small millimeter or submillimeter differences in FSTT to biological variables must be undertaken with caution, especially where they have not been repeated across different studies/samples. To improve the integrity of FSTT studies and the reporting of FSTT measurement errors, we propose the following standard: (1) calculate the technical error of measurement (TEM or rTEM) in any FSTT research work; (2) assess the error embedded in the full data collection procedure; and (3) conduct validation testing of FSTT means proposed for point estimation prior to publication to ensure newly calculated FSTT means provide improvements. In order to facilitate the latter, a freely available R tool TDValidator that uses the C-Table data for validation testing is provided.

Statistical Shape Modeling of Skeletal Anatomy for Sex Discrimination: Their Training Size, Sexual Dimorphism, and Asymmetry.

Purpose: Statistical shape modeling provides a powerful tool for describing and analyzing human anatomy. By linearly combining the variance of the shape of a population of a given anatomical entity, statistical shape models (SSMs) identify its main modes of variation and may approximate the total variance of that population to a selected threshold, while reducing its dimensionality. Even though SSMs have been used for over two decades, they lack in characterization of their goodness of prediction, in particular when defining whether these models are actually representative for a given population. Methods: The current paper presents, to the authors' knowledge, the most extent lower limb anatomy shape model considering the pelvis, femur, patella, tibia, fibula, talus, and calcaneum to date. The present study includes the segmented training shapes (n = 542) obtained from 271 lower limb CT scans. The different models were evaluated in terms of accuracy, compactness, generalizability as well as specificity. Results: The size of training samples needed in each model so that it can be considered population covering was estimated to approximate around 200 samples, based on the generalizability properties of the different models. Simultaneously differences in gender and patterns in left-right asymmetry were identified and characterized. Size was found to be the most pronounced sexual discriminator whereas intra-individual variations in asymmetry were most pronounced at the insertion site of muscles. Conclusion: For models aimed at population covering descriptive studies, the number of training samples required should amount a sizeable 200 samples. The geometric morphometric method for sex discrimination scored excellent, however, it did not largely outperformed traditional methods based on discrete measures.

Contractile effects of 5-hydroxytryptamine (5-HT) in the equine jejunum circular muscle: functional and immunohistochemical identification of a 5-HT1A-like receptor.

REASONS FOR PERFORMING STUDY: Prokinetic drugs used to treat gastrointestinal ileus in man have equivocal results in horses. In man, prokinetic drugs have 5-hydroxytryptamine4(5-HT4) receptors as their target, but little is known about the 5-HT-receptor subtypes in the equine small intestine. OBJECTIVE: Functional and immunohistochemical identification of the serotonin receptor subtype(s) responsible for the 5-HT induced contractile response in the equine circular jejunum. METHODS: Isometric organ-bath recordings were carried out to assess spontaneous and drug-evoked contractile activity of equine circular jejunum. Histological investigations by immunofluorescence analyses were performed to check for presence and localisation of this functionally identified 5-HT receptor subtype. RESULTS: Tonic contractions were induced by 5-HT in horse jejunal circular muscle. Tetrodotoxin, atropine and NG-nitro L-arginine did not modify this response. A set of 5-HT receptor subtype selective antagonists excluded interaction with 5-HT1B, 1D, 2A, 3, 4 and 7 receptors. The selective 5-HT1A receptor antagonists WAY 100635 and NAN 190 caused a clear rightward shift of the concentration-response curve to 5-HT. The contractile effect of 5-CT, that can interact with 5-HT1A, 1B, 1D, 5 and 7 receptors was also antagonised by WAY 100635, identifying the targeted 5-HT receptor as a 5-HT1A-like receptor. Immunohistology performed with rabbit polyclonal anti-5-HT1A receptor antibodies confirmed the presence of muscular 5-HT1A receptors in the muscularis mucosae, and both longitudinal and circular smooth muscle layers of the equine jejunum. CONCLUSIONS: Contractile responses in equine jejunal circular smooth muscle induced by 5-HT involves 5-HT1A-like receptors.

Determination of the source of increased serotonin (5-HT) concentrations in blood and peritoneal fluid of colic horses with compromised bowel.

REASONS FOR PERFORMING STUDY: Increased plasma (5-HT) concentrations are reported in horses predisposed to develop laminitis and after i.v. infusion of endotoxins. In the equine jejunum contractile 5-HT1A-like receptors show tachyphylaxia upon prolonged activation with 5-HT. Therefore, increased systemic 5-HT release in colic horses could play a possible role in the pathophysiology of ileus. OBJECTIVE: To investigate possible increased systemic release of 5-HT in colic horses with compromised bowel and to identify the source of 5-HT overload. METHODS: Concentrations of 5-HT were determined in plasma and peritoneal fluid (PF) of healthy horses (n = 10), strangulating small intestinal colic horses (n = 18), nonsurgical colic horses (n = 10) and cryptorchid stallions (n = 6). It was attempted to identify the source of 5-HT overload by comparing the blood and PF 5-HT concentrations within horses and by assessing the in vivo platelet activation through determination of the beta-thromboglobulin (beta-TG)/platelet factor 4 (PF4) ratio. RESULTS: All horses in the strangulating small intestinal colic group had plasma (P = 0.006) and PF (P = 0.01) 5-HT concentrations above those found in the control group. Plasma beta-TG/PF4 ratio in these horses exceeded 2 in all cases, indicating in vivo platelet activation. Concentrations of 5-HT in PF of colic horses with compromised bowel were significantly lower than the corresponding plasma concentrations (P = 0.005). POTENTIAL RELEVANCE: In horses with compromised bowel, significant amounts of 5-HT can be released into the systemic circulation, through massive release of platelet-stored 5-HT. 5-HT is a very potent proinflammatory, vasoconstrictive and immunomodulatory agent. In view of the rapid and prolonged tachyphylaxia, shown for the jejunal 5-HT1A-like receptors, this increased systemic 5-HT release could play a role in the pathophysiology of ileus in horses.

Facial morphology and growth following surgery for congenital midline cervical cleft patients.

Congenital midline cervical cleft (CMCC) is a rare condition that consists of a cutaneous midline neck lesion with a sinus extending inferiorly towards the sternum. A fibrous band that extends superiorly to the mandible is a consistent feature of the condition. Restriction of growth of the mandible, possibly due to incomplete removal of the band, is the most significant long-term problem. It remains unclear whether early removal of the fibrous band might allow catch-up growth of the mandible. This study utilized non-invasive three-dimensional photographs to objectively evaluate the facial growth of six CMCC patients. The growth of these CMCC patients was compared to the average growth of age- and sex-matched controls from a database of three-dimensional facial photographs of clinically normal subjects. After surgical removal of the fibrous cord, CMCC patients experience growth in the chin at the same rate as in the normal population; no evidence was found for catch-up growth. As a result, individuals with CMCC are likely to require further surgical intervention to correct the residual retrognathia on completion of facial growth. Early excision of the lesion including aggressive resection of the fibrous band is still recommended, as this should optimize the early growth of the mandible in infancy.

A Comprehensive Craniofacial Study of 22q11.2 Deletion Syndrome.

The 22q11.2 deletion syndrome (22q11.2DS) is one of the most frequent microdeletion syndromes and presents with a highly variable phenotype. In most affected individuals, specific but subtle facial features can be seen. In this observational study, we aim to investigate the craniofacial and dental features of 20 children with a confirmed diagnosis of 22q11.2DS by analyzing 3-dimensional (3D) facial surface scans, 2-dimensional (2D) clinical photographs, panoramic and cephalometric radiographs, and dental casts. The 3D facial scans were compared to scans of a healthy control group and analyzed using a spatially dense geometric morphometric approach. Cephalometric radiographs were digitally traced, and measurements were compared to existing standards. Occlusal and dental features were studied on dental casts and panoramic radiographs. Interestingly, a general trend of facial hypoplasia in the lower part of the face could be evidenced with the 3D facial analysis in children with 22q11.2DS compared to controls. Cephalometric analysis confirmed a dorsal position of the mandible to the maxilla in 2D and showed an enlarged cranial base angle. Measurements for occlusion did not differ significantly from standards. Despite individual variability, we observed a retruded lower part of the face as a common feature, and we also found a significantly higher prevalence of tooth agenesis in our cohort of 20 children with 22q11.2DS (20%). Furthermore, 3D facial surface scanning proved to be an important noninvasive, diagnostic tool to investigate external features and the underlying skeletal pattern.

Large-scale in-vivo Caucasian facial soft tissue thickness database for craniofacial reconstruction.

A large-scale study of facial soft tissue depths of Caucasian adults was conducted. Over a 2-years period, 967 Caucasian subjects of both sexes, varying age and varying body mass index (BMI) were studied. A user-friendly and mobile ultrasound-based system was used to measure, in about 20min per subject, the soft tissue thickness at 52 facial landmarks including most of the landmarks used in previous studies. This system was previously validated on repeatability and accuracy [S. De Greef, P. Claes, W. Mollemans, M. Loubele, D. Vandermeulen, P. Suetens, G. Willems, Semi-automated ultrasound facial soft tissue depth registration: method and validation. J. Forensic Sci. 50 (2005)]. The data of 510 women and 457 men were analyzed in order to update facial soft tissue depth charts of the contemporary Caucasian adult. Tables with the average thickness values for each landmark as well as the standard deviation and range, tabulated according to gender, age and BMI are reported. In addition, for each landmark and for both sexes separately, a multiple linear regression of thickness versus age and BMI is calculated. The lateral asymmetry of the face was analysed on an initial subset of 588 subjects showing negligible differences and thus warranting the unilateral measurements of the remaining subjects. The new dataset was statistically compared to three datasets for the Caucasian adults: the traditional datasets of Rhine and Moore [J.S. Rhine, C.E. Moore, Tables of facial tissue thickness of American Caucasoids in forensic anthropology. Maxwell Museum Technical series 1 (1984)] and Helmer [R. Helmer, Schädelidentifizierung durch elektronische bildmischung, Kriminalistik Verlag GmbH, Heidelberg, 1984] together with the most recent in vivo study by Manhein et al. [M.H. Manhein, G.A. Listi, R.E. Barsley, R. Musselman, N.E. Barrow, D.H. Ubelbaker, In vivo facial tissue depth measurements for children and adults. J. Forensic Sci. 45 (2000) 48-60]. The large-scale database presented in this paper offers a denser sampling of the facial soft tissue depths of a more representative subset of the actual Caucasian population over the different age and body posture subcategories. This database can be used as an updated chart for manual and computer-based craniofacial approximation and allows more refined analyses of the possible factors affecting facial soft tissue depth.

Facial Characteristics and Olfactory Dysfunction: Two Endophenotypes Related to Nonsyndromic Cleft Lip and/or Palate.

Evidence exists for the presence of a specific facial phenotype in nonaffected first-degree relatives of persons with CL/P. An increased risk for olfactory dysfunction has also been reported in CL/P-relatives. These phenotypic features can probably be explained via the presence of CL/P-related susceptibility genes. We aimed at confirming the occurrence of these endophenotypic traits in first-degree CL/P-relatives, and we investigated the link between the facial phenotype and the smell capacity in this group. We studied the facial morphology of 88 nonaffected first-degree relatives of patients with CL/P and 33 control subjects without family history of facial clefting by 3D surface imaging and a spatially dense analysis of the images. Smell testing was performed in 30 relatives and compared with 23 control subjects. Nonaffected relatives showed midface retrusion, hypertelorism, and olfactory dysfunction, compared to controls. In addition, we show for the first time that olfactory dysfunction in relatives is correlated to a smaller upper nasal region. This might be explained by a smaller central olfactory system. The different facial morphology in the relatives with olfactory impairment as compared to the total group may be an illustration of the contribution of different genetic backgrounds to the occurrence of CL/P via different biological pathways.

Synthesis of delta 3-1-methylene-1-carbacephems.

The total synthesis of (+/-)-1-methylene-2,2- dimethyl-7-amino-1-carbacephem-4-carboxylic acid (1) is described. The reaction scheme was essentially that described by Christensen et al. for the synthesis of (+/-)-1-carbacephems. In vitro antibacterial activities of the 7-phenoxyacetyl and 7-D-alpha-phenylglycyl derivatives of 1 were compared with those of 7-(phenoxyacetamido)desacetoxycephalosporanic acid and cefalexin. Derivatives of 1 were 2-4 times less active against most of the sensitive organisms than the corresponding 7-aminodesacetoxycephalosporanic acid analogues. The activity of the 7-D-alpha-phenylglycyl derivative of 1 however was about 20 times lower than that of cefalexin when measured against Staphylococcus aureus ATCC 6538P.

Synthesis and antiviral activity of 5-heteroaryl-substituted 2'-deoxyuridines.

The synthesis of 5-heteroaryl-substituted 2'-deoxyuridines is described. The heteroaromatics were obtained from three different 5-substituted 2'-deoxyuridines. Cycloaddition reaction of nitrile oxides on the 5-ethynyl derivative 1 gave the isoxazoles 4a-e. The thiazole derivatives 14a-c were obtained from the 5-thiocarboxamide 11, while 5-pyrrol-1-yl-2'-deoxyuridine (17) could be synthesized directly from 5-amino-2'-deoxyuridine. The compounds were evaluated for antiviral activity. Selective activity against herpes simplex virus type 1 (HSV-1) and varicella zoster virus (VZV) was noted for 5-(3-bromoisoxazol-5-yl)-2'-deoxyuridine (4c). The compound was inactive against herpes simplex virus type 2, cytomegalovirus, and thymidine kinase (TK)-deficient mutants of HSV-1 and VZV, which indicates that, most likely, its antiviral activity depends on phosphorylation by the virus-specified TK.