Randomised clinical trial investigating self-assembling peptide P11-4 in the treatment of early caries.

OBJECTIVES: This prospective, randomised, split-mouth, clinical trial compared the efficacy of the self-assembling peptide P11-4 to fluoride varnish in the treatment of early buccal carious lesions. MATERIALS AND METHODS: Subjects presenting at least two clinically affected teeth were treated at D0 (day 0) and D90 with P11-4 (test) or fluoride varnish (control). At D180, fluoride varnish was applied on all study lesions. Standardised photographs were taken at D0, D30, D90, D180 and D360 and blindly morphometrically assessed. Hierarchical linear models (HLM) under allowance of confounders were used to compare the decrease in size between test and control groups. The visual analog scale (VAS) and Global Impression of Change Questionnaire (GICQ) were used as clinical assessments. RESULTS: Overall, 37 subjects (13-36 years) with 90 early carious lesions were included. HLM analysis showed a significant difference between test and control groups, indicating a decrease in test lesions and stabilisation of control lesions size (p = 0.001). The test lesion's mean size (SD) relative to baseline decreased to D30 = 0.936(0.127), D90 = 0.874(0.173), D180 = 0.844(0.215) and D360 = 0.862(0.352), whereas control lesions remained stable at D30 = 1.018(0.209), D90 = 1.013(0.207), D180 = 1.029(0.235) and D360 = 1.068(0.401). The effect sizes ranged from 0.47 to 0.82. CONCLUSIONS: Within the limits of this study, it was shown that the size of early carious lesions treated with P11-4 was significantly reduced; this result was superior to that of fluoride varnish treatment (DRKS00012941). CLINICAL RELEVANCE: The self-assembling peptide P11-4 is the first caries treatment approach aiming to regenerate decayed enamel. P11-4 initiates formation of de novo hydroxyapatite in the depth of early carious lesions, adding a new advanced therapy option for preventive dentistry.

Estimated depth of apatite and collagen degradation in human dentine by sequential exposure to sodium hypochlorite and EDTA: a quantitative FTIR study.

AIM: To characterize chemical degradation of the principal constituents of dentine after exposure to NaOCl and EDTA using Infrared Spectroscopy (ATR-FTIR). METHODOLOGY: Ground dentine particles, from extracted permanent human molars, were passed through sieves of 38 to 1 000 µm to provide six size ranges. Portions (250 mg) of each size range were reacted with 5 mL of 2.5% NaOCl for 2-10 min; or 17% EDTA for 5-1440 min. Powders larger than 75 µm were also sequentially exposed to NaOCl/EDTA/NaOCl each for 10 min. All experiments were repeated five times. Reacted and unreacted powders were washed and dried. Particles larger than 75 µm were then reground. FTIR spectra of unground and reground reacted particles enabled assessment of particle surface versus bulk chemistry, respectively, plus estimation of reaction depth. Changes in the ratio of the 1 640 cm-1 collagen: 1 010 cm-1 phosphate peak height or its inverse were obtained. These were used to estimate surface and bulk fraction reacted and thus depth to which collagen or phosphate was reduced following immersion in NaOCl or EDTA, respectively. The data were analysed descriptively. RESULTS: Surface collagen fraction declined by ~40% within 2 min of NaOCl exposure, and plateaued at ~60% between 6-10 min. Bulk spectra showed average depth of collagen loss at 10 min was 16 ± 13 µm. Ten minute EDTA exposure caused ~60% loss of surface phosphate. Average depth of phosphate loss was 19 ± 12 µm and 89 ± 43 µm after 10 and 1 440 min EDTA immersion, respectively. Sequential NaOCl/EDTA immersion yielded a 62 ± 28 µm thick phosphate-depleted surface. Sequential NaOCl/EDTA/NaOCl treatment resulted in approximately 85 µm of collagen loss. CONCLUSIONS: Data revealed the sequential depletion of collagen by NaOCl and apatite by EDTA in dentine, simultaneously exposing the other moieties. Alternate exposure to NaOCl and EDTA therefore enhances the depth of erosion.

Gene expression measurements in the context of epidemiological studies.

BACKGROUND: Gene expression measurements became an attractive tool to assess biological responses in epidemiological studies. However, collection of blood samples poses various technical problems. We used gene expression data from two epidemiological studies to evaluate differences between sampling methods, comparability of two methods for measuring RNA levels and stability of RNA samples over time. METHODS: For the PARSIFAL study, PBLC of 1155 children were collected using EDTA tubes in two countries. In the PASTURE study, tubes containing RNA-stabilizing solutions (PAXgene) Blood RNA Tubes; PreAnalytiX) were used to collect cord blood leucocytes of 982 children in five countries. Real-time PCR (conventional single tube assay and high-throughput low density arrays) was used to quantify expression of various innate immunity genes. In 77 PARSIFAL samples, gene expression was measured repeatedly during prolonged storage. RESULTS: In PARSIFAL (EDTA tubes) the median RNA yield after extraction significantly differed between the two centres (70 and 34 ng/microl). Collecting blood into an RNA-stabilizing solution markedly reduced differences in RNA yield in PASTURE (range of medians 91-107 ng/microl). The agreement [Spearman rank correlation (r)] between repeated measurements of gene expression decreased with increasing storage time [e.g., for CD14: r (first/second measurement) = 0.35; r (first/third measurement) = 0.03]. RNA levels measured with either the conventional method or low-density arrays were comparable (r > 0.9). CONCLUSION: Collecting blood samples into tubes containing an RNA-stabilizing solution increases RNA yield and reduces its variability. Long-term storage of samples may lead to RNA degradation, requiring special attention in longitudinal studies.

Prenatal diagnosis of an isochromosome 5p in a fetus with increased nuchal translucency thickness and pulmonary atresia with hypoplastic right heart at 14 weeks.

We report on a fetus presenting with increased nuchal translucency at 11 weeks' gestation, suggesting cystic hygroma. Chorion villous sampling was performed, and cytogenetic analysis revealed a supernumerary isochromosome 5p leading to tetrasomy 5p: 47,XX,+ i(5p)[7]/46,XX[5] after short-term culture and 47,XX,+ i(5p)[20] after long-term culture. Subsequent targeted sonographic follow-up at 12 and 14 weeks revealed further increase of the NT to 6.4 mm and the additional presence of a congenital heart defect (pulmonary atresia with intact ventricular septum). Termination of pregnancy was performed, and the heart defect was confirmed. Isochromosome 5p was found in varying proportions in all examined organs. Only a few cases of mosaic tetrasomy 5p have been reported in the literature, and recent reports on prenatally detected isochromosome 5p showed a possible relationship to increased nuchal translucency in some cases and also a possible role of confined mosaicism in others. Whereas cases with confined mosaicism did not show suspicious signs on ultrasound, true mosaicism conversely showed increased nuchal thickness as well as structural abnormalities. This is the first report on the association of a cardiac defect with this chromosome aberration.

Absent or hypoplastic thymus on ultrasound: a marker for deletion 22q11.2 in fetal cardiac defects.

OBJECTIVE: Congenital heart defects (CHD), particularly conotruncal anomalies, may be associated with deletion of chromosome 22q11.2. Thymic aplasia or hypoplasia is known to be a typical feature in this condition. We aimed to establish (i) the prevalence of del22q11.2 in fetal CHD and (ii) whether ultrasound assessment of an absent or hypoplastic fetal thymus helps in preselection of a group who are at high risk for this deletion. STUDY DESIGN: In fetuses (> 16 weeks) with CHD, karyotyping and fluorescence in situ hybridization for 22q11.2 were offered and the fetal thymus was evaluated sonographically. RESULTS: One hundred and forty-nine fetuses with CHD and normal karyotype were analyzed. Seventy-six fetuses had conotruncal anomalies. 22q11.2 deletion was present in 10 cases (6.7%), all of which had conotruncal anomalies (13.1%). Thymic hypoplasia or absence was suspected in 11 cases with conotruncal anomaly. Nine of these 11 had the deletion; two cases were false positive. One fetus with a normal-sized thymus had deletion of 22q11.2 (sensitivity 90%, specificity 98.5%, positive predictive value 81.8%, and negative predictive value 99.2%). By subtype of cardiac anomaly, there was deletion in four of six fetuses with interruption of the aortic arch, two of four with absent pulmonary valve syndrome, three of nine with truncus arteriosus and one of 11 cases of tetralogy of Fallot. Pulmonary atresia with ventricular septal defect (n = 7), right-sided aortic arch (n = 4), transposition of the great arteries (n = 14), double outlet right ventricle (n = 13) and other complex malpositions of the great vessels (n = 8) were not associated with the deletion. CONCLUSION: Thymic hypoplasia or aplasia may reliably be diagnosed during fetal echocardiography. The technique allows identification of a group at high risk for 22q11.2 deletion and is more specific and sensitive than by subtype of cardiac anomaly alone.

Prenatal diagnosis of 22q11 microdeletion in an early second-trimester fetus with conotruncal anomaly presenting with increased nuchal translucency and bilateral intracardiac echogenic foci.

We report on a 16-week fetus, in which detection of increased nuchal translucency thickness and bilateral intracardiac echogenic foci led to the prenatal diagnosis of truncus arteriosus communis, interruption of the aortic arch and aplastic thymus. Cytogenetic examination confirmed a 22q11.2 microdeletion consistent with the suspected CATCH 22 syndrome. Subsequently hydrops fetalis developed and the fetus died in utero at 18 weeks. This case report supports the hypothesis that both cardiac failure and left ventricular outflow tract obstruction may cause increased nuchal translucency thickness. The association between increased nuchal translucency thickness and CATCH 22 syndrome should be considered in diagnostic procedures. The sonographic diagnosis of both increased nuchal translucency thickness and intracardiac echogenic foci requires specialist ultrasonography and echocardiography. In particular, identification of right-sided or bilateral echogenic foci should prompt further detailed examination.

[Prenatal diagnosis of heart defects and associated chromosomal aberrations]. / Pränatale Diagnostik von Herzfehlern und assoziierte chromosomale Aberrationen.

AIM: According to epidemiological studies on newborns, the association of congenital heart defects with chromosomal anomalies varies between 4 and 12%. Prenatally this rate is probably higher, due to antenatal death occurring in fetuses with chromosomal aberrations. The aim of the study was therefore to determine the rate and the distribution of chromosomal aberrations in prenatally detected heart defects. PATIENTS AND METHOD: Within a period of 7 years fetal echocardiography was performed on 2716 fetuses at high risk for CHD. The analysis of the fetal heart was achieved by the visualization of different planes. Once a heart defect was detected, karyotyping was performed after amniocentesis, cordocentesis or chorion villous sampling, or in a few cases postnatally from cord blood. Prenatal ultrasound findings were confirmed postnatally by ultrasound examination or, in case of abortion, stillbirth or neonatal death, by autopsy. RESULTS: A total of 203 fetal heart malformations were detected and 46 of them (22%) had associated chromosomal anomalies. 60% of all cases and 80% of the study group had extracardiac anomalies. Only eight out of the 46 pregnant women (17.5%) were older than 35 years. Eight out of the 15 fetuses with trisomy 18 had a ventricular septal defect, 9/13 fetuses with trisomy 21 had an atrioventricular septal defect and all 5 fetuses with monosomy X had a left heart outflow obstruction. No typical cardiac defects were found in the remaining 13 fetuses (5 trisomy 13, 2 triploidies, 6 miscellaneous). Of the 13 live births (23 terminations of pregnancy and 10 intrauterine deaths) 6 children survived (46% and overall survival rate 13%). The following rates of associations with aneuploidies were found: atrioventricular septal defect 55%, ventricular septal defect and aortic coaction both 43%, tetralogy of Fallot and double outlet right ventricle both 36%. In comparison, fetuses with isomerism, transposition of the great arteries and pulmonary atresia or stenosis had normal chromosomes. CONCLUSION: We conclude that the rate of association of heart defects and chromosomal abnormalities is higher prenatally than in the neonatal period and is approximately 22%. After detecting a fetal cardiac malformation, karyotyping is mandatory for the further management of pregnancy. The likelihood of detection of an aneuploidy increases when some typical heart defects are detected or when an association with extracardiac anomalies is found.

Fetal trisomy 10 mosaicism: ultrasound, cytogenetic and morphologic findings in early pregnancy.

We report the ultrasound, cytogenetic and morphologic findings in a case of trisomy 10 mosaicism prenatally detected by chorionic villus sampling (CVS). CVS sampling was carried out at the 13th week of gestation because of ultrasound diagnosis of hydrops fetalis and hygroma colli. Trisomy 10 mosaicism was diagnosed in cells from the cytotrophoblast (short-term culture) and the chorionic villus core (long-term culture). Fetal mosaicism was confirmed after termination of pregnancy in umbilical cord cells, placenta and fetal skin fibroblasts.

Bilaterally cleft lip, limb defects, and haematological manifestations: Roberts syndrome versus TAR syndrome.

We report on a 13-year-old patient followed since birth. He is the only offspring of young, non-consanguineous German parents. His mother has an isolated left cleft of lip and a cleft palate. At birth, our patient presented with bilaterally cleft lip/cleft palate, phocomelia of upper limbs with normal hands, and mild symmetrical deficiencies of the long bones of the lower limbs. Haematological evaluation demonstrated a leukaemoid reaction during a urinary tract infection as well as intermittent thrombocytopenia and episodes of marked eosinophilia during the first two years of life. Intellectual development has been normal. Comparison with two similar cases from the literature suggests a non-random phenotypic overlap of Roberts syndrome (MIM 268300) and TAR syndrome (MIM 274000). Such clinical constellations may be key observations to understand the genetic relationship of Roberts syndrome and TAR syndrome in future phenotype-genotype correlations.

D-thyroxine reduces lipoprotein(a) serum concentration in dialysis patients.

Uremia raises lipoprotein(a) (Lp(a)) serum concentration and the risk of arteriosclerosis in dialysis patients. The treatment of high Lp(a) levels is not satisfactory today. The decrease of Lp(a) in hypothyroid patients on L-T4 therapy raised the question of whether dextro-thyroxine (D-thyroxine) reduces not only serum cholesterol, but also Lp(a) serum concentration. In a single-blind placebo-controlled study, the influence of D-thyroxine therapy on Lp(a) serum concentration was evaluated in 30 hemodialysis patients with elevated Lp(a) serum levels. Lp(a) was quantified in parallel by two methods, i.e., rocket immunoelectrophoresis and nephelometry, and apo(a) isoforms were determined by a sensitive immunoblotting technique. Regardless of the apo(a) isoforms, 6 mg/d D-thyroxine reduced elevated Lp(a) levels significantly by 27 +/- 13% in 20 dialysis patients (P < 0.001) compared with 10 control subjects (-9.9 +/- 8.4%). In parallel, D-thyroxine therapy significantly lowered total cholesterol (P < 0.001), LDL cholesterol (P < 0.001), and LDL cholesterol/HDL cholesterol ratio (P < 0.01); raised T4 and T3 serum levels; and suppressed thyroid-stimulating hormone secretion without causing clinical symptoms of hyperthyroidism in any of the patients. D-Thyroxine reduces elevated serum Lp(a) concentration in dialysis patients. The effect in nondialysis patients can be expected but remains to be proven.

Prenatal diagnosis of classical phenylketonuria by linked restriction fragment length polymorphism analysis.

Since the isolation of a recombinant containing a cDNA sequence for human phenylalanine hydroxylase (hPH) (Woo et al., 1983; Speer et al., 1986) prenatal diagnosis by linked restriction fragment length polymorphism (RFLPs) has become possible for families in which phenylketonuria (PKU) occurs (Lidsky et al., 1985a). We describe here the application of a Hind III three-allele RFLP in a single family, which allowed the prenatal diagnosis of an affected fetus.

[Familial balanced chromosome translocation t(12;13) in a family with several deformed stillbirths]. / Familiäre balancierte Chromosomentranslokation t(12;13) in einer Familie mit mehreren missgebildeten Totgeburten.

Reported in this paper is a family which was cytogenetically examined for several stillbirths with severe neural tube defects and other malformations. Balanced translocation between chromosomes 12 and 13 [t (12; 13) (q24; q22)] was recorded from five members of that family. Partial trisomy 13 or partial monosomy 13 is discussed as the cause of the malformations. Prenatal cytogenetic diagnosis had been undertaken in two cases.