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The recognition of hepatomeg aly and splenomegaly is made even simpler if ascites lasting arthritis relief buy indomethacin 25 mg cheap, which acts as an intra-abdominal contrast medium arthritis in fingers & toes purchase indomethacin 25mg mastercard, is associated (Figures 7 childhood arthritis definition discount 50 mg indomethacin amex. If hepatomegaly is very pronounced arthritis pain relief gel 50 mg indomethacin with amex, the prominence of the liver deforms the anterior abdominal wall, causing a dip at the tho Figure 7. This patient had serologically con racoabdominal junction, similarly to what happens in frmed hepatitis A infection in the frst trimester. At 19 weeks, cases of severe thoracic hypoplasia, although in this ultrasound demonstrated the following: (a) Axial view: evident hepatomegaly, with capsular macrocalcifcation and moderate case it is the abdomen that is enlarged rather than the ascites. The simultaneous presence of asci tes acts as a natural contrast medium, allowing 3D ultrasound (surface rendering) to visualize the enlarged liver (L), the spleen (on the left), and the bowel (arrow). The enlarge ment of the liver and/or the spleen does not itself repre sent a malformation; it is rather an acquired lesion due to environmental factors. This is virtually (a) On the axial view of the upper abdomen, it is possible to absent except for the already-mentioned myeloprolif recognize the enlarged, hyperechoic, and rather inhomogeneous liver (arrows). Beckwith–Wiedemann syndrome: look for ► hep we feel that the measurement of the two cross-sectional atomegaly + omphalocele + body hemihypertro diameters of the viscera remains easier and faster phy + macroglossia (Chapter 10); (Figure 7. Care megaly be diagnosed in a fetus, the first issue to consider needs only be taken not to mistake the left hepatic lobe, is a possible infective cause. Therefore, a careful interview which is larger in utero than in postnatal life, for the with the mother with the aim of disclosing any possible spleen. If other signs of severe fetal infection, searching for additional signs of fetal infection (cerebral such as cerebral calcification or ascites, are present, calcification hydrocephalus, ascites, and cardiomegaly then these are highly indicative of a poor prognosis. Storage diseases are usually lethal within a drome fetuses present hepatosplenomegaly, a karyo few months or years of life. In cases due to intrauterine typing should be recommended in those cases in which infections, the neonatal outcome will depend upon the all other causes have been excluded. This is directly related to the under ited hepatitis of moderate severity to very important lying cause of the hepatomegaly or splenomegaly. It and left) hepatic veins (the “efferent venous system”; enters the liver anteriorly and runs along its inferior Figure 7. During the fifth week of ges tation in normal embryonic development, the embry Differential diagnosis. Association with structural the blood from the trophoblast), and the vitelline or and chromosomal anomalies, which is fairly common, omphalo-mesenteric veins (taking the blood from the represents the most important poor prognostic sign. Development of the liver In cases without structural and chromosomal anoma plays a role in modifying the primitive vitelline and lies associated, the presence of hydrops represents the umbilical systems to their final form. In some cases, a persistent postnatal with other anomalies (especially trisomies 21 and 18 porto-systemic shunt may require the placement of an and Turner syndrome). However, the strong association with genetic and chromosomal anomalies in which abnor Obstetric management. Although knowledge of the malities of the lymphatic system are present may play a mechanisms involved in the genesis of these anomalies role in the pathogenesis of hydrops in these cases. A sonographic anechoic cyst is seen between the abdominal wall and the lower liver edge. Color, power, and pulsed Doppler examination indicate umbilical venous fow within the cystic intra abdominal mass. When the diagnosis of fetal umbilical vein varix is made after 26 weeks and the varix is isolated, the outcome is usually good. Associated abnormalities are usually involving the intra-abdominal part of the vein, reported in about 32% of cases, most commonly anom with an incidence in utero of about 1:1000. Therefore, any condition that increases all except one reported case, associated sonographic venous pressure could potentially dilate this portion of abnormalities were present [31]. Syndromal abnor malities were present in two of 91 cases reported in Ultrasound diagnosis. In both cases, associated structural a sonographic anechoic cyst is seen between the abdom abnormalities were present. Karyotyping measurement is above two standard deviations of the should be offered when additional fetal abnormalities mean for gestational age. In isolated cases, the cular structure (choledochal cyst, hepatic cyst, mesen prognosis is better: in 75% of cases, no clinical fin teric cyst, or enteric cyst duplication) can be excluded. Sonographic fnding of the umbilical vein curving to the left toward the stomach, usually connecting to the right portal vein rather than to the left portal vein. When associated with other anomalies, an underlying chromosomal abnor malities in 8% of cases has been reported. On sagittal and axial views of the abdomen: midline abdominal wall defect, with herniation of abdominal content. Round, solid mass that deforms the anterior abdominal wall, usually containing some bowel loops and/or right hepatic lobe. Definition Omphalocele is a defect in the closure of abdominal wall defects, with an incidence of 1 in 3000 the abdominal wall that also involves the cord inser live births [35]. It results from the failure of loops of bowel to the incidence of omphalocele is higher in the frst return to the abdominal cavity after physiological her trimester, about 1 in 384 at 11–14 weeks, according niation into the umbilical cord from the sixth to tenth to a recent publication [36]; this can be explained by week of development [33,34]. The herniated organs are its strong association with chromosomal abnormalities wrapped in a two-layered sac, with the two layers being and other structural anomalies related to a higher rate the peritoneum and the amnion. Two variants of omphalocele exist, according to the presence or absence of the liver in the sac. The two variants have a different embryogenesis and may have a completely different prognosis, due to the dif ferential risk of associated chromosomal anomalies. In this rare circumstance, as reported in this section, the differential diagnosis with gastroschisis may be difficult Figure 7. An omphalocele is sonographi imaging, showing the omphalocele on the three orthogonal cally represented by a bulging structure that (1) arises planes, which enables assessment of its content and extension from the anterior abdominal wall, (2) contains some (arrowheads indicate the abdominal wall defect; the arrow abdominal viscera (liver and/or bowel), and (3) pres indicates cord insertion). Therefore, if an ompha associated, and care should then be taken not to mistake locele containing only ileal loops is identified earlier than the ascites for amniotic fluid (Figure 7. In addi in a week’s time: if the herniation persists, then it is an tion, it should be noted that polyhydramnios, possibly omphalocele; if it has disappeared, then it was a physi secondary to bowel obstruction at the level of the wall ologic transient herniation of the intestine in the cord. Note the concurrent cord cyst (arrowhead), which is also often associated with trisomy 18. The inset shows the specimen with the small omphalocele and the bowel loops visible under the omphalocele membranes. Another entity that should be differentiated from iso lated omphalocele is cloacal exstrophy, in which ompha locele extends caudally and is associated with bladder exstrophy and anomalies of the external genitalia. In the case of ompha locele, the recognition of a normal bladder in the pelvis rules out this very rare possibility. This is high, from 15% to 40% (mainly trisomies 18 and 13, and, in a few cases, triploidy, trisomy 21, and Turner syndrome), in particular when the omphalocele is associated with other major structural abnormalities or with increased Figure 7. Huge omphaloceles containing the nosis and rule out physiologic herniation of intestine in the liver are less frequently associated with chromosomal cord, it is necessary to wait until the 12th completed week of anomalies. Surgical correction of the defect may be performed in a single intervention or may require a two-step procedure, according to the size of the ompha Risk of nonchromosomal syndromes. In fact, in those cases in which the whole liver most common syndromes detectable in utero that can has herniated, the limited intra-abdominal pressure is be associated with omphalocele are as follows: insufficient to allow normal growth of the abdomen. In these cases, the omphalocele + macroglossia + somatic hemihyper defect is only partially closed, and the viscera remain tophy + polycystic kidney (Chapter 10); ing outside the abdomen are wrapped up in a sterile. Pentalogy of Cantrell: look for ► midline anterior silastic bag (or spring-loaded silos). Cloacal exstrophy: look for ► omphalocele + muscular flaps may be used to increase the surface of exstrophy of the urinary bladder + anorectal mal the anterior abdominal wall. Should an omphalocele be diag mortality is generally due to the associated anomalies, nosed in a fetus, karyotyping is mandatory because of and it may be as high as 80% in the case of multi the high risk of chromosomal anomalies, especially if the ple anomalies or even 95%–100% in the case of severe defect is small and contains bowel only. On the contrary, if the echocardiography and a thorough anomaly scan should omphalocele is isolated and the liver is not in the sac, be performed to detect possibly associated defects. Empirically, it has been yield very different results, with significantly lower sur suggested that a Cesarean section may be indicated in vival rates and much poorer quality of life [39]. Definition this is a rare form of abdominal wall Pentalogy of Cantrell is also known as “thoracoab defect that consists of five associated anomalies: a dominal ectopia cordis” and is in part a misnomer, midline anterior ventral wall defect, a diaphragmatic given that the heart is always outside of the thorax in defect, a cleft distal sternum, a defect of the apical the latter case. The presence of a pericardial effusion implies a communication between the pericardial sac and the exomphalos.

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Referral to rheumatoid arthritis diet to loss weight generic indomethacin 50 mg free shipping a mental health professional should be made promptly to rheumatoid arthritis hives order generic indomethacin canada prevent the significant long-term morbidity and even mortality that can result from ongoing substance abuse rheumatoid arthritis upper back pain order indomethacin 75mg amex. Tobacco arthritis in hips of dogs purchase indomethacin 75mg mastercard, alcohol, and other drugs: the role of the pediatrician in prevention, identification, and management of substance abuse. You arrive to find a limp neonate with a heart rate of 40 beats/min and no respiratory effort. As you prepare to begin resuscitation, you note microcephaly, cutis aplasia on the scalp, and a cleft lip and palate. Decisions surrounding delivery room resuscitation of neonates at 23 weeks’ gestation presents a challenge to health care providers and families. Existing guidelines for delivery room resuscitation of extremely low gestation neonates remain limited and often quickly become outdated because advances in medical management lead to improved survival. A consistent recommendation is that the wishes of the parents about resuscitation should be respected when a neonate is born between 23 and 24 weeks of gestation. This is because of the high rate of mortality and high risk of diminished quality of life in surviving neonates born at this gestational age. The ethical principles of autonomy, beneficence, nonmaleficence, and justice are tightly woven into care decisions at the edge of viability. Viability has been defined as the point at which there is a reasonable chance of survival with advanced medical support, with some ethicists arguing that resuscitation should be provided to all neonates who have at least a 50% chance of survival. Neonatal outcome data published in 2010 can be seen in Item C17, but these data may not reflect subsequent improvements in neonatal care or center-to-center variability. The sixth edition of the Neonatal Resuscitation Program of the American Academy of Pediatrics and the American Heart Association has outlined clinical situations in which noninitiation of resuscitation may be appropriate. These include confirmed gestation less than 23 weeks, birthweight less than 400 g, anencephaly, and a confirmed lethal genetic disorder or malformation. Ideally, ongoing discussions about care at the time of birth should be made jointly with the family before delivery. The birthweight of 470 g, gestational age of 23 2/7 weeks, and stigmata of trisomy 13 aneuploidy in the neonate in the vignette may be indications for noninitiation of resuscitation, but only after discussion with the family. Many extremely premature infants are born with bradycardia and apnea, so it would not be appropriate to assume that this indicates birth asphyxia and withhold resuscitation. If the neonate responds to resuscitation, further discussions and decisions about ongoing care may occur in the nursery with the family as active participants. The headaches involve the left side of her head, are described as throbbing, and last 2 to 3 hours. The patient does not have any vision changes or weakness associated with the headaches, and they do not worsen with coughing, sneezing, lying down, or sitting up. She does not take any medications other than occasional ibuprofen for the headaches. Her blood pressure is 102/68 mm Hg, heart rate is 92 beats/min, respiratory rate is 22 breaths/min, and her body mass index is 21. Her physical examination, including neurological examination and fundoscopy, is unremarkable. Migraines are characterized by severe head pain, nausea, and phonophobia or photophobia. In younger children the location is typically bifrontal, and in adolescents and adults the headaches are often unilateral. Although classic teaching is that morning headaches are suggestive of a brain tumor, in pediatric patient’s migraines often occur in the morning. Time of day is not one of the diagnostic criteria for pediatric migraine, possibly because this criterion does not differentiate migraine from secondary headache very well. Red flags for pediatric headache include positional headache (worsening with lying down implies increased intracranial pressure) and headache that wakes the child from sleep. In pediatric patients who have had migraine headaches for more than 6 months, with normal neurological examination findings, and without features suggestive of neurologic dysfunction, brain imaging is not needed. Family history of migraine is another reassuring factor that supports a diagnosis of migraine headaches. If there are red flags associated with headache (positional, sleep disrupting), the preferred brain imaging method is magnetic resonance imaging. This is an ideal first imaging test if a structural malformation such as Chiari I malformation is suspected. Symptomatic Chiari I malformation presents with diffuse headache that is worse with a Valsalva maneuver, such as coughing, sneezing, laughing, or defecating. If there is an urgent clinical indication, such as new encephalopathy, new focal deficit, or recent head trauma, then computed tomography of the head is the best imaging method because it is the quickest. Other imaging modalities may be indicated when specific diagnoses are being considered. A computed tomography venogram is obtained to evaluate for cerebral sinus venous thrombosis. These cases present with headaches that have migrainous features, such as severe unilateral head pain, nausea, vomiting, and phonophobia or photophobia. Treatment of cerebral sinus venous thrombosis is anticoagulation until the thrombus is resolved. Transcranial Doppler ultrasonography is used to monitor flow velocity in the middle cerebral artery in people with sickle cell disease. The headaches in moyamoya disease can be diffuse or unilateral, and are often provoked by exercise or activity or associated with hemiparesis. The girl in the vignette does not have any signs of sickle cell disease or moyamoya disease, so a transcranial Doppler ultrasound is not indicated. He has had worsening polyuria and polydipsia for 10 days since starting perphenazine 8 mg orally once every evening for the diagnosis of schizophrenia. On physical examination, his pulse is 130 beats/min and blood pressure is 108/75 mm Hg. The goal of initial fluid therapy is expansion of the intravascular and extravascular volume and restoration of normal renal perfusion. Fluid resuscitation that is not aggressive is associated with an increased risk of complications. Increased mortality is observed in patients with unreversed shock over the first 24 hours of admission who had received less than 40 mL/kg of intravenous fluids over the first 6 hours of treatment. Therefore, aggressive isotonic fluid replacement is recommended initially to restore perfusion, even if hypernatremia is present, followed by more hypotonic (0. Although mild acidosis can occur, it is typically the result of hypoperfusion (lactic acidosis). The dose should then be titrated to achieve a decrease in glucose concentration of 50 to 75 mg/dL per hour (2. When you see him for follow-up, it becomes clear that the family is not giving him the medication as prescribed. You express your concern and review the need for medication again with an explanation of the pathophysiology. The family is very concerned about adverse effects and does not believe that the medication is necessary. Two weeks later, a check for $1,000 arrives in your office from the family with a thank you card. Acceptance of gifts may imply that you are in agreement with their choice when you are not, and may cause the family to expect preferential treatment. The American Medical Association suggests that each gift be evaluated case by case. General recommendations are that gifts that are too intimate, too expensive, or above the means of the family are not appropriate, and acceptance of them may create the wrong impression in the patient’s or family’s minds. In particular, monetary gifts of any size for personal use should not be accepted and the patient or family redirected to donate to a fund to benefit the facility or research. Any gift that makes a physician feel uncomfortable is likely one that should not be accepted. These gifts, in particular, may interfere in good patient care by influencing the physician to change their practice so as not to antagonize the family or patient. A monetary gift for personal use to the physician by patients is not recommended in any circumstance.

The transcription factor Sox9 has essential roles in successive steps of the chondrocyte differentiation pathway and is required for expression of Sox5 and Sox6 rheumatoid arthritis lyme disease generic indomethacin 75mg on-line. Hormonal and cellular regulation of Sertoli cell anti-Mullerian hormone production in the postnatal mouse arthritis of the knee nz purchase indomethacin 75mg fast delivery. Evidence that Sry is expressed in pre-Sertoli cells and Sertoli and granulosa cells have a common precursor arthritis relief for lower back purchase indomethacin 75mg otc. Targeted mutagenesis ofthe endogenous mouse Mis gene promoter: in vivo definition of genetic pathways of vertebrate sexual development arthritis knee diagram order 25mg indomethacin overnight delivery. Tet repressor-based system for regulated gene expression in eukaryotic cells: principles and advances. Abnormal sexual development in transgenic mice chronically expressing mullerian inhibiting substance. Detection of circular and linear transcripts of Sry in pre-implantation mouse embryos: differences in requirement for reverse transcriptase. A heterozygous mutation in the desert hedgehog gene in patients with mixed gonadal dysgenesis. Differentiating between parthenogenetic and "positive development" embryos in turkeys by molecular sexing. A comparison of the properties of Sox-3 with Sry and two related genes, Sox-1 and Sox-2. Identification of residues in the estrogen receptor that confer differential sensitivity to estrogen and hydroxytamoxifen. Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. The Wilms tumor gene, Wtl, is required for Sox9 expression and maintenance of tubular architecture in the developing testis. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Sexually dimorphic expression of protease nexin-1 and vanin-1 in the developing mouse gonad prior to overt differentiation suggests a role in mammalian sexual development. A gene mapping to the sex determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes. Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: a tool for temporally regulated gene activation/inactivation in the mouse. Wnt-4 deficiency alters mouse adrenal cortex function, reducing aldosterone production. Mullerian inhibiting substance messenger ribonucleic acid expression in granulosa and Sertoli cells coincides with their mitotic activity. Steroidogenic factor 1 and Dax-1 colocalize in multiple cell lineages: potential links in endocrine development. Investigation of the fate of Sry-expressing cells using an in vivo Cre/loxP system. Premeiotic endomitosis produces diploid eggs in the natural clone loach, Misgumus anguillicaudatus (Teleostei: Cobitidae). Cell-specific knockout of steroidogenic factor 1 reveals its essential roles in gonadal function. Geographic ranges, population structure, and ages of sexual and parthenogenetic snail lineages. Balancing the bipotential gonad between alternative organ fates: A new perspective on an old problem. Fgf9 and Wnt4 act as antagonistic signals to regulate mammalian sex determination. Expression of a candidate sex-determining gene during mouse testis differentiation. Conditional alleles in mice: practical considerations for tissue-specific knockouts. Zp3-cre, a transgenic mouse line for the activation or inactivation of loxP-flanked target genes specifically in the female germ line. Steroidogenic factor 1, an orphan nuclear receptor, regulates the expression of the rat aromatase gene in gonadal tissues. The role of the mesonephros in cell differentiation and morphogenesis of the mouse fetal testis. Dax-1 (dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1) gene transcription is regulated by wnt4 in the female developing gonad. Sox9 expression during gonadal development implies a conserved role for the gene in testis differentiation in mammals and birds. A common trans-acting factor, Ad4-binding protein, to the promoters of steroidogenic P-450s. Expression of the mouse anti-mullerian hormone gene suggests a role in both male and female sexual differentiation. Mechanisms of disease: Transcription factors in sex determination-relevance to human disorders of sex development. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Protamine-Cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells. Characterization of a premeiotic germ cell specific cytoplasmic protein encoded by Stra8, a novel retinoic acid-responsive gene. R-spondinl is essential in sex determination, skin differentiation and malignancy. Sox3 expression in undifferentiated spermatogonia is required for the progression of spermatogenesis. Expression of Dmrtl in the genital ridge of mouse and chicken embryos suggests a role in vertebrate sexual development. Testicular dysgenesis does not affect expression of anti-mullerian hormone by Sertoli cells in premeiotic seminiferous tubules. A shared promoter element regulates the expression of three steroidogenic enzymes. Expression of Sox8, Sfl, Gata4, Wtl, Daxl, and Fog2 in the mouse ovarian follicle: implications for the regulation of Amh expression. Twenty pairs of sox: extent, homology, and nomenclature of the mouse and human sox transcription factor gene families. Nuclear receptor steroidogenic factor 1 regulates the mullerian inhibiting substance gene: a link to the sex determination cascade. The ins and outs of transcriptional control: nucleocytoplasmic shuttling in development and disease. Mouse Daxl expression is consistent with a role in sex determination as well as in adrenal and hypothalamus function. Mesonephric cell migration induces testis cord formation and Sertoli cell differentiation in the mammalian gonad. Minifascicular neuropathy: a new concept of the human disease caused by desert hedgehog gene mutation. Sox9 is essential for outer root sheath differentiation and the formation of the hair stem cell compartment. New insights into the regulation of mammalian sex determination and male sex differentiation. Production of anti Mullerian hormone: another homology between Sertoli and granulosa cells. Molecular cloning and gene expression of Foxl2 in the Nile tilapia, Oreochromis niloticus. Fresh staining mix: 1/50 dilution of X-gal in staining base, mix well and filter before use. Appendix D; Genotype abbreviations used to describe mouse genotype in this thesis. Scotland Poster title: Conditional strategies to study gene functions during mammalian development. Angela Tye*, Christophe Galichet, Martin Cheung, Sarah Wynn, James Briscoe, Kathy Cheah and Robin Lovell-Badge Conditional strategies to study gene function during mammalian development Angela /i v*. To study these events, which take place at the genetic, molecular and cellular levels, we need Is that allow us to regulate gene expression and thereby all downstream events in spatial and temporal manner. The hple for demonstration here is Sox9, which is known to be essential for testis determination and male development in mammals.


  • Cerebral thrombosis
  • Exophthalmos
  • OSLAM syndrome
  • Oral-facial cleft
  • Bacterial gastroenteritis
  • Stevens Johnson syndrome
  • Anorexia nervosa restricting type
  • Phenobarbital antenatal infection

Following these results rheumatoid arthritis etiology buy 75 mg indomethacin otc, the 10pM library concentration was found optimal for all the subsequent samples arthritis in neck and tmj order indomethacin 25 mg otc. The mean sequencing depth was lower when a higher number of samples per chip was used (n=32) rheumatoid arthritis leg pain indomethacin 50 mg otc, albeit sufficient for comprehensive coverage of targeted markers arthritis pain bracelets buy cheap indomethacin online. Alteration to the original chip loading protocol were assessed as recommended on the Ion Torrent community forum ioncommunity. Based on the results of this study, up to 32 barcoded samples were processed on each 316 chip. The overall discrepancy among samples analysed at low stringency variant detection settings, varied between 2 to 19 markers per sample pair corresponding to 0. In general, samples sequenced at a higher depth showed less discrepancy (three to four markers per sample pair or approximately 0. However, the percentage of inconsistent calls might not be an accurate representation, since the total number of markers can be counted in various ways, such as the total number of genotyped markers or successfully genotyped markers only. Comparison between triplicates of three samples, sequenced on two chips and analysed using low stringency parameters. Three duplicates (see Table 10) sequenced at significantly higher depth, showed less discrepancy between each other, compared to the same samples, sequenced at approximately ten fold less depth. In addition, the same duplicates while analysed according to the high stringency parameters, showed significantly higher concordance (from zero to only three markers difference) per duplicate (Table 12). On the other hand, applying the higher stringency settings, resulted in less markers detected (down by approximately 30%). The same samples as in Table 10, analysed using Variant Caller, according to high stringency parameters. Comparison between the low and high stringency settings in the Variant Caller plugin. However, prior to statistical analysis (Chapter 4) the data were filtered by a minimum coverage of x10 and genotype quality of 10 (per marker), thus reducing the potential error rate. Remarkably, an application of the high stringency analysis parameters resulted in less than 0. Therefore, for any forensic use, a high coverage (x100 or higher) followed by high stringency variant analysis parameters should be considered. For example, the Ion Torrent hardware, software, reagents and laboratory manuals in this project were updated through series of optimization as follows:  the OneTouch instrument (an essential component for template preparation) was physically replaced five times in one year and its software was therefore also updated five times. The update of one platform’s component (either software, hardware or reagents) led to a subsequent adaptation of all other parts of the Ion Torrent platform, which usually were irreversible and non-compatible with previous versions. A limited validation performed in this study tested the concordance and genotyping accuracy within the same platform. Given the relatively novel nature of the Ion Torrent platform, it was decided to perform a limited comparison between at least two independent genotyping methods. Given that these two platforms use completely different methods for genotyping, this outcome can be considered a successful quality control step. However, the results of this preliminary study provided important verification of the genotyping output obtained within the Ion Torrent platform, as detailed in this section. It should be noted that additional verification of the Ion Torrent genotyping output was obtained by performing association analyses of the pigmentation traits and ancestry (as discussed in Chapter 5). The results obtained in this study were in full concordance with previously published results. Introduction the main goal of this study was to assess the craniofacial measurements reproducibility and normal distribution, as per Aims 2 and 3 of the current project (Section 1. Prior to investing in the 3D technology, the validity of the anthropometric measurements using 2D images was tested. This study was required because the 3D scanner was not available at the beginning of this project and since it is generally easier to generate 2D photographs, which can be taken at various locations without the need to attend a scanning session in a dedicated room, as required for the 3D scanning approach. Locating anatomical landmarks on the face and obtaining anthropometrical measurements from photographs is known as photogrammetry [17]. This method has been tested in numerous anthropometrical studies [29, 36, 351-353]. This process is easier and less time consuming than the direct measurements and 3D digital image processing. On the other hand, it possesses significant disadvantages, such as the quality of photographs and thus resulting in a partial coverage of facial landmarks. Although most importantly, it is unable to reflect the actual surface distances, but can only appreciate the lateral facial distances. The lateral measurement is the linear distance between two landmarks, which does not necessarily represent the actual physical (Euclidean) distance over the skin surface. As a result, facial measurements obtained from 2D images are less accurate than direct or 3D–acquired measurements, hence providing only partial and potentially misleading information on facial morphology. Conversely, 3D scanning platforms provide a more comprehensive representation of the facial morphology. The 3D scanning systems have been extensively used in anthropometric studies as well as in medical research, which usually demands a high accuracy and precision [354-357]. The Minolta Vivid V910 3D scanner that was acquired for this study, has a reported manufacturing precision of ± 0. These observations demonstrate that this platform is more appropriate for the present study and would be expected to provide a more accurate representation of the facial morphology as compared to less accurate 2D photographs. The rationale behind this comparison was that it is easier and faster to generate the lateral, rather than the surface measurements in the Geomagic software. Materials and Methods Facial measurements of ten individuals, obtained from lateral dimensions on 2D images and lateral and surface measurements generated from 3D images were compared. The 2D photographs were taken under ambient lighting, similar to the 3D scanning settings. The photographs were printed on A4 paper and analysed for a set of 13 facial measurements. Several measurements were not performed due to image quality (as discussed in Section 4. The scanner used in this study was sensitive to changes in light conditions and was calibrated every three months (according to manufactorer’s recommendations) to produce high quality standardised 3D images. The 3D facial scans (n=10) and image processing were performed as described in the Materials and Methods Chapter 2. The quality of the 3D alignment was measured automatically by Polygon software and displayed as an ”error average” and “sigma” with values between 0 and 1 (illustrated in Figure 27). The measurements obtained from 2D and 3D images of ten (10) individuals were recorded in an Excel spreadsheet and compared manually. Results and Discussion the 2D photogrammetry provided easier image processing and used inexpensive equipment, compared to 3D scanning. The 3D scanning not surprisingly, was found to be significantly more accurate and comprehensive. However, the Geomagic software (used for 3D image processing), was not designed for craniofacial measurements and 152 | P a g e elicited significant difficulties. The need for measuring the Euclidian distance between landmarks, rather than the lateral distance required a special assistance from the manufacturer, although this support was only partial and time consuming. In addition, each of the 32 measurements were performed and recorded individually, which made the landmarking procedure tedious and time-consuming, requiring approximately 20 minutes per image. Attempts to generate a ‘macro’ function, which would automatically copy a set of specific facial landmarks to a new image using Geomagic program, failed, as each face was significantly different from the previous and the software could not perform this procedure in an efficient manner. Comparison of the measurements showed that lateral and surface measurements performed on the 3D digital images were noticeably different. The 3D surface distances were longer than the lateral, with the latter more similar to the 2D measurements. The results of the comparison between 2D and 3D measurements are summarised in Tables 14, 15 and Figures 42, 43. Since it was obvious that there was a pronounce difference and given that 3D surface measurements represent the most adequate information on facial features, it was concluded that all the measurements should be calculated using Euclidean coordinates of the craniofacial landmarks (representing the actual surface distance), which was performed using Microsoft Excel automatic spreadsheet. The 3D surface measurements were subsequently used as phenotypes for genetic association study, as detailed in Chapter 5. Results of the comparison between craniofacial measurements in 2D and 3D images, including lateral and surface distance.

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