The SHOX region and its mutations.

The short stature homeobox-containing (SHOX) gene lies in the pseudoautosomal region 1 (PAR1) that comprises 2.6 Mb of the short-arm tips of both the X and Y chromosomes. It is known that its heterozygous mutations cause Leri-Weill dyschondrosteosis (LWD) (OMIM #127300), while its homozygous mutations cause a severe form of dwarfism known as Langer mesomelic dysplasia (LMD) (OMIM #249700). The analysis of 238 LWD patients between 1998 and 2007 by multiple authors shows a prevalence of deletions (46.4%) compared to point mutations (21.2%). On the whole, deletions and point mutations account for about 67% of LWD patients. SHOX is located within a 1000 kb desert region without genes. The comparative genomic analysis of this region between genomes of different vertebrates has led to the identification of evolutionarily conserved non-coding DNA elements (CNE). Further functional studies have shown that one of these CNE downstream of the SHOX gene is necessary for the expression of SHOX; this is considered to be typical "enhancer" activity. Including the enhancer, the overall mutation of the SHOX region in LWD patients does not hold in 100% of cases. Various authors have demonstrated the existence of other CNE both downstream and upstream of SHOX regions. The resulting conclusion is that it is necessary to reanalyze all LWD/LMD patients without SHOX mutations for the presence of mutations in the 5'- and 3'-flanking SHOX regions.

Halitosis: a stomatological and psychological issue.

It can be stated that halitosis is located on the ridge connecting dentistry, oral medicine, parodontology and psychology. It represents, at the same time, the manifestation of an organic malfunctioning of the oral apparatus, and a problematic element for the individual and his/her relational life. A smelly emanation comes into conflict with the wish to attract, to please, to seduce. The attitudes towards the possibility to suffer from bad breath have two typical opposite expressions, which share the common characteristic that many are bad judges of one's own breath. The dentist, in fact, is frequently involved in the management of patients who believe they have bad breath problems, which in reality are non-existent (pseudo-halitosis), and, more often, of patients who are not aware they have an halitosis condition, and who are not inclined to accept it (denied halitosis). Generally, the most adequate and suitable option is that of communicating to the unaware patient the existence and the nature of the problem. What is said, and especially the way of saying it, may play an important role in patient's acceptance of the information without producing, or reducing to a minimum, the undesirable side effects on the patient-professional relationship, and on the personal dynamics of the patient him/herself. A useful procedure is provided by employing a pre-visit questionnaire, that may suitably ask many different questions about relevant dental and mucosal aspects, for instance, dental complaints, the frequency of toothbrushing and flossing, gum bleeding, and about psychological aspects, such as dental anxiety, and degree of satisfaction as regards one's oral condition, in order to solve or alleviate the patient's problem, avoiding unnecessary personal discomfort and, at the same time, providing competent and effective professional help. Strategies for communicating in an effective way, in order to properly face both the somatic and the psychological aspects, are proposed. A questionnaire (Halitosis Questionnaire - HQ) is also provided, to facilitate the assessment and the management of the halitosis issue.

Quantitation of fetal DNA in maternal serum during the first trimester of pregnancy by the use of a DAZ repetitive probe.

Cell-free fetal DNA in maternal plasma or serum is at present widely investigated as a source of fetal genetic material, both in studies of pregnancy-related disorders and in planning strategies for non-invasive prenatal diagnosis. Despite the number of trials already performed on the quantitation of fetal DNA, data about the amount of DNA at the beginning of pregnancy, in particular in the first trimester, remain limited. A new probe mapping on the deleted in azoospermia (DAZ) repetitive region of the Yq chromosome was designed for an early assessment of fetal DNA concentration in maternal serum. Among 57 pregnant women prospectively studied in their first trimester, fetal DNA was detected already by the 5th gestational week, with the analysis becoming reliable by the 8th week of gestation when a 100% accuracy in fetal sex determination was achieved. Moreover, in the three cases of pregnancy ending in fetal loss, the amount of fetal DNA apparently decreased before the abortion was diagnosed, whereas it consistently showed an increasing trend in normal pregnancies. Real-time PCR with the use of DAZ multilocus probe can efficiently quantitate free fetal DNA in the maternal serum at the beginning of pregnancy.

A comparative study between different techniques in non-surgical periodontal treatment.

AIM: This study was carried out to compare a conventional manual instrumentation with an ultrasonic technique (Odontoson M) in scaling and root planing periodontal treatment. The effects of subgingival irrigation with 10% iodised solution in association with Odontoson M were studied. METHODS: Sixty patients were selected: they were suffering from adult periodontitis, aged between 35 and 65 and non-smokers. They were randomly divided into 3 groups (Gs1, Gs2, Gs3). Gs1 was treated with curettage using Gracey curettes, Gs2 was treated with scaling and root planing using Odontoson M, while in Gs3 scaling and root planing with Odontoson M irrigated with a 10% iodised solution were performed. Clinical parameters (probing depth, bleeding on probing, dental mobility, loss of attachment level) were measured before treatment (T0), after 30 days (T1), after 90 days (T2) and after 120 days (T3). RESULTS: No statistical significant differences between Gs1 and Gs2 were observed. The improvement of clinical parameters in Gs3 was greater and statistical significant differences with other groups treated were recorded. CONCLUSIONS: There are no significant differences between the effectiveness of scaling and root planing with manual instruments and with Odontoson M. The 10% iodised solution used such as topic antiseptic carries out greater effects in not surgical periodontal treatments.

The importance of splinting of teeth in the therapy of periodontitis.

AIM: The deep periodontal disease often leads to dental mobility with further aggravation of the symptomatology. The aim of the paper is to verify the importance of splinting of teeth in the therapy of periodontitis on the basis of parameters commonly studied [probing depth (PD), bleeding on probing (BOP), plaque index], and the role of the occlusal trauma as primary factor or second leading factor in periodontal diseases. METHODS: Thirty patients suffering from periodontitis have been treated with the splinting of teeth, neither preceded nor followed from topical and systemic pharmacological therapy, nor from surgical or non surgical treatment of the periodontal tissues involved. During the 4 visits provided by the study, T0 (0 days), T1 (30 days), T2 (90 days) and T3 (180 days), PD, BOP and plaque index have been estimated. RESULTS: From the comparison of the T0 PD and BOP with the T3 PD and BOP, a significant improvement of these parameters is observed (P<0.05). CONCLUSION: On the basis of the clinical data obtained in this research, the authors suggest that the therapy by means of splinting improves the prognosis of teeth affected by periodontal disease; occlusal trauma and dental mobility cause the aggravation of periodontal lesions.

Sonographic biometrical range of external genitalia differentiation in the first trimester of pregnancy: analysis of 2593 cases.

OBJECTIVES: The aim of this study was to establish the accuracy of fetal gender assignment by sonography in the biometrical range of 18 to 29 mm of biparietal diameter (BPD). METHODS: Transvaginal and/or transabdominal sonography was used to detect the sagittal sign as a marker of fetal gender in 2593 fetuses with BPD between 18 and 29 mm. The results of sonographic examination were compared with the gender at birth or with karyotype obtained from amniotic fluid cells or chorionic villus sampling. RESULTS: Fetal gender assignment was feasible in 2374 of 2593 cases (91%). Of the 2188 fetuses with known fetal sex outcome, 1025 were males and 1157 were females, and 6 had genital anomalies. In fetuses without genital anomalies, an accuracy rate of 100% was achieved at a BPD of >/=24 mm. The results of the six cases with genital malformations were considered separately. CONCLUSION: Sonography is a reliable method for the study of the morphological development of the external genitalia in fetuses 'in vivo'; it is possible to assign fetal gender in 95 to 99% starting at a BPD of 20 mm and to achieve an accuracy rate of 99 to 100% from a BPD of 22 mm, but fetal sex assignment should not be undertaken below a BPD of 22 mm, and especially not in cases where fetal sexing affects pregnancy management.

Early prenatal diagnosis of recurrent 46,XY partial gonadal dysgenesis.

OBJECTIVES: We present a case of early prenatal diagnosis of recurrent 46,XY partial gonadal dysgenesis, by combining early genetic and sonographic evaluations. METHODS: The conceptus of a mother with a first child affected by 46,XY gonadal dysgenesis was sonographically evaluated at 21- and 23-mm BPD (12(+2) and 12(+6) LMP-based age) and the female genitalia were observed. Karyotype analyses was performed on amniotic fluid and it revealed a 46,XY complement without mosaicism. SRY was amplified by PCR for molecular analyses. RESULTS: We observed a discordance between female phenotype detected at 21 and 23 mm of biparietal diameter (12(+2) and 12(+6) LMP-based age) and male karyotype. In the child and the fetus, seminiferous cords were not recognisable, whereas rare Leydig cells and no germ cells could be identified. Internal and external genitalia were sexually ambiguous in the child and feminized in the fetus. CONCLUSION: This is the first case of early prenatal diagnosis of recurrent 46,XY partial gonadal dysgenesis and it points to the importance of combining early analyses of genetic sex with sonography in the management of anomalies of sexual development, with particular regard to syndromes for which the risk of recurrence is little understood.

[Clinical comparison between guided tissue regeneration and induced tissue regeneration]. / Valutazione clinica tra rigenerazione tissutale guidata e rigenerazione tissutale indotta.

AIM: The aim of the present study was to evaluate, over a period of 1 y, 3 different surgical methods for the treatment of periodontal bone defects. METHODS: Thirty-six infrabone defects, at least 4 mm in depth, in non-smokers were enrolled in the study. Of these, 12 were treated with guided tissue regeneration (GTR) using a resorbable membrane with collagen, 12 with enamel matrix derivative (EMD) gel and 12 with a modified Widman flap (MWF). The pocket depth, attachment loss and gingival recession both before commencement of therapy and after an interval of 1 y are reported. RESULTS: The reduction in pocket depth was 4 mm, 4.4 mm, and 4.5 mm, respectively, for the control group, GTR group and EMD group. The attachment gain for the respective treatments was: MWF, 2 mm; GTR, 2.8 mm; EMD, 2.9 mm. Gingival recession was: MWF, 1.8 mm; GTR, 1.5 mm; EMD, 1.1 mm. CONCLUSION: These findings show the efficacy of the 3 methods in the treatment of bone defects, but none of the 3 emerges as being statistically superior to the others 2.

[Burning mouth syndrome]. / La sindrome della bocca bruciante.

Burning Mouth Syndrome (BMS) is a frequent disease characterized by a burning or painful sensation in the tongue and/or other oral sites without clinical mucosal abnormalities or lesions. The etiopathology is unknown although local, systemic and psychological factors have been connected with BMS. As this syndrome is a multifactorial disease, the diagnostic and therapeutic approach should be multidisciplinary. In this paper a review of the literature is presented and the most recent advancement on clinical, etiologic, diagnostic and therapeutic aspects of BMS are discussed.

Non-invasive first trimester fetal gender assignment in pregnancies at risk for X-linked recessive diseases.

OBJECTIVES: Prenatal diagnosis in families affected by X-linked recessive disorders should ideally be limited to the subjects at increased risk, i.e. male fetuses, in order to avoid the risk of fetal loss due to the invasive procedure in healthy female fetuses. The aim of the study was to assess the fetal sex within the first trimester of gestation by two non-invasive approaches, using ultrasonography and a molecular analysis of fetal DNA extracted from whole maternal blood with specific markers, in order to avoid invasive sampling in female fetuses. METHODS: A total number of 18 fetuses at risk for an X-linked recessive disease were included in the present investigation. Maternal peripheral blood was analysed between 7 and 12 weeks of gestation by nested PCR for the detection of fetal DNA and the prediction of fetal gender. In addition, when the biparietal diameter (BPD) was between 21 and 23 mm, an ultrasonographic examination was carried out to assess the fetal gender. CVS was then performed in male fetuses only. RESULTS: Fetal gender was correctly assigned by ultrasonography between 21 and 23 mm of BPD in all the cases studied, whereas DNA extracted from whole maternal blood accurately predicted the gender in all the female cases (10), but failed in 4 out of 8 male fetuses, erroneously assigned as females. CONCLUSION: The present study shows that sonography is able to accurately predict the fetal gender within the first trimester of pregnancy, whereas the molecular analysis of DNA extracted from whole maternal blood is biased by false-Y-negative results in 50% of the cases.

Prenatal diagnosis of female pseudohermaphroditism associated with bilateral luteoma of pregnancy: case report.

Female pseudohermaphroditism associated with luteoma of pregnancy (LP) is a rare condition characterized by varying degrees of masculinization of a female fetus. We describe a case, diagnosed at 13 weeks gestation. Transvaginal ultrasound at 5 weeks of gestation revealed a normal intrauterine gestational sac and an enlarged maternal right ovary. Re-examination at 13 weeks showed a fetus with male external genitalia. Cytogenetic investigation on amniotic fluid revealed a normal female karyotype 46,XX. Follow-up sonography confirmed the previous assignment of male external genitalia and a second amniocentesis was negative for the SRY gene. High levels of androgens were found in the maternal blood. A diagnosis of female pseudohermaphroditism associated with bilateral LP was made. A healthy girl was born by Caesarean section with complete masculinization of external genitalia (Prader V). Histology confirmed a bilateral LP. To the best of our knowledge this represents the first case of prenatal diagnosis of female pseudohermaphroditism associated with LP and demonstrates the feasibility of diagnosis by sonography from 13 weeks gestation. This is also the first case described of Prader V masculinization associated with LP.

Differential divergence of three human pseudoautosomal genes and their mouse homologs: implications for sex chromosome evolution.

The human pseudoautosomal region 1 (PAR1) is essential for meiotic pairing and recombination, and its deletion causes male sterility. Comparative studies of human and mouse pseudoautosomal genes are valuable in charting the evolution of this interesting region, but have been limited by the paucity of genes conserved between the two species. We have cloned a novel human PAR1 gene, DHRSXY, encoding an oxidoreductase of the short-chain dehydrogenase/reductase family, and isolated a mouse ortholog Dhrsxy. We also searched for mouse homologs of recently reported PGPL and TRAMP genes that flank it within PAR1. We recovered a highly conserved mouse ortholog of PGPL by cross-hybridization, but found no mouse homolog of TRAMP. Like Csf2ra and Il3ra, both mouse homologs are autosomal; Pgpl on chromosome 5, and Dhrsxy subtelomeric on chromosome 4. TRAMP, like the human genes within or near PAR1, is probably very divergent or absent in the mouse genome. We interpret the rapid divergence and loss of pseudoautosomal genes in terms of a model of selection for the concentration of repetitive recombinogenic sequences that predispose to high recombination and translocation.

X/autosomal translocations in the Xq critical region associated with premature ovarian failure fall within and outside genes.

Premature ovarian failure curtails female reproductive life and is often linked to balanced Xq/autosomal translocations in a critical region. We mapped regions around translocations at the edges of this zone (one in Xq13.3, two in Xq26) in large-insert clones and analyzed their sequence. One Xq26 region is extensively transcribed and, in agreement with a recent independent analysis, the breakpoint interrupts a gene that encodes a widely expressed peptidase. In contrast 430 kb around the second Xq26 breakpoint has no putative or detected gene content. In 260 kb around the Xq13 translocation, the breakpoint falls among a cluster of repetitive elements at least 59 kb from the only detected gene (a rarely expressed T-box family transcription factor). We discuss our results in relation to models that ascribe premature ovarian failure to interruption of ovarian genes or to a failure of interactions involving DNA of the critical region during follicle development.

Sonographic early fetal gender assignment: a longitudinal study in pregnancies after in vitro fertilization.

OBJECTIVES: A longitudinal evaluation by sonography of external genitalia in human embryos/early fetuses with a known time from fertilization is lacking. Our aim was to assign by sonographic evaluation of external genitalia the early fetal gender in a cohort of pregnancies after in vitro fertilization. Sonographic examinations were performed in each case in three subsequent sessions over a period of time early in gestation in order to establish a temporal threshold, expressed in terms of days from fertilization, at which absolute accuracy in gender prediction is achievable. METHODS: Thirty-two fetuses were included in this prospective longitudinal study. Each was examined three times for gender assignment. The first observation was performed between 65 and 69 days from fertilization, the second between 70 and 74 days and the third between 75 and 79 days. Transvaginal and/or transabdominal sonography was used to detect the 'sagittal sign' as a marker of fetal gender. The results of ultrasound examinations were compared with gender at birth or with karyotype obtained from amniotic fluid cells or chorionic villus sampling. RESULTS: Fetal gender assignment was feasible in 29 out of 32 fetuses (90%) at the first examination and in all cases at the second and third examinations. Fetal gender prediction was correct in 76% of cases in which fetal gender was assigned (22/29) at the first examination; accuracy for males was 46% (6/13) and for females 100% (16/16). At the second and third examinations, accuracy for gender prediction achieved 100% for both genders. Concerning the temporal threshold, absolute accuracy in gender prediction was achieved at 69 days from fertilization, corresponding to 11+6 weeks based on the last menstrual period. CONCLUSION: This study provides important information about the earliest stage, expressed in terms of days from fertilization, at which it is possible to make a certain diagnosis of fetal gender by sonography.

Agenesis of the scapula in Emx2 homozygous mutants.

The shoulder and pelvic girdles represent the proximal bones of the appendicular skeleton that connect the anterior and posterior limbs to the body trunk. Although the limb is a well-known model in developmental biology, the genetic mechanisms controlling the development of the more proximal elements of the appendicular skeleton are still unknown. The knock-out of Pax1 has shown that this gene is involved in patterning the acromion, while the expression pattern candidates Hoxc6 as a gene involved in scapula development. Surprisingly, we have found that scapula and ilium do not develop in Emx2 knock-out mice. In the homozygous mutants, developmental abnormalities of the brain cortex, the most anterior structure of the primary axis of the body, are associated with important defects of the girdles, the more proximal elements of the secondary axis. These abnormalities suggest that the molecular mechanisms patterning the more proximal elements of the limb axis are different from those patterning the rest of appendicular skeleton. While Hox genes specify the different segments of the more distal part of the appendicular skeleton forming the limb, Emx2 is concerned with the more proximal elements constituting the girdles.

Morphological features and measurements in a human fetus with karyotype 69, XXX: comparison with fetuses of the same CRL, without chromosomal anomalies.

A triploid fetus (karyotype 69, XXX) with crown-rump length (CRL) 94 mm, presenting micro- and retrognathia, low-set ears and crooked feet, was cleared and double-stained with alizarin red S and alcian blue for detecting the ossification patterns in the vertebral column, ribs, ischium, limbs, and face. Longitudinal measurements of some long bones in the upper (humerus, ulna, radius) and lower (femur, tibia, fibula) limb were taken. The values of both the total length (TL) and the ossified part (OL) of each long bone, as well as the OL/TL per cent ratio were considered. Reference points were located on the mandible, i.e. condylar process (Pcl), coronoid process (Pco), gnathion (GN), gonion (GO), superior symphyseal point (SSP) for measuring linear dimensions. Since the aim of this work was to assess the influence of triploidy 69, XXX the skeletal development and growth patterns, all values obtained in the examined specimen were related with those relative to a group of fetuses, without any detectable malformation and chromosomal anomalies, with a CRL mean value of 93 mm. Results evidenced that the triploid fetus presented growth restriction and that the vertebral centra ossification and the mandibular development were much delayed with the normal ossification patterns.