What children say and clinicians hear: accounts relating to incisor hypomineralisation of cosmetic concern.

AIM: To explore the range of impacts relating to incisor opacities as described by children, their general dental practitioners and paediatric dentists. METHODS: Participants included 50 children, aged 7-16 years, referred to a UK hospital paediatric dentistry service for management of incisor opacities. All children were subsequently diagnosed with molar incisor hypomineralisation. Following ethical approval, data were recorded as follows: patient demographics, distance travelled, waiting times, nature of any impacts relating to incisor opacities documented in referral letters and/or in subsequent paediatric dentistry assessment records. Additionally, children completed the short form Child Oral Health Impact Profile questionnaire (COHIP-SF19) as a self-report measure of their oral health-related quality of life (OHRQoL). RESULTS: Nearly, half (48%, n = 24) of the referral letters mentioned that the child was experiencing one or more negative social and/or functional impacts. Mean COHIP score was significantly lower (indicating poorer OHRQoL) for children whose referring dentist had identified a negative impact (COHIP = 42.9) compared to those with no documented impact (COHIP = 50.5; p = 0.018, independent t test). At the hospital consultation, negative impacts were elicited by a paediatric dentist in 86% (n = 43) of cases. Again, mean COHIP score was significantly lower for children whose assessment records noted a negative impact (COHIP = 44.5) compared to those with no recorded impact (COHIP = 60.2; p = 0.001). Families travelled a mean distance of 57 km (range 3-218 km) to the hospital service, with an average waiting time of 75 days from referral. CONCLUSION: It is encouraging that dental professionals seem to be aware of the negative psychosocial impacts experienced by some children with enamel opacities, and that children feel able to describe them.

The structure and composition of deciduous enamel affected by local hypoplastic autosomal dominant amelogenesis imperfecta resulting from an ENAM mutation.

In a group of families in northern Sweden, a mutation in the ENAM gene (predicted to produce a highly truncated protein) results in the local hypoplastic form of autosomal dominant amelogenesis imperfecta. In this study, sections of deciduous teeth from members of 3 of these families were examined by scanning electron microscopy (SEM) and the enamel mineral was analysed by energy dispersive X-ray spectroscopy (EDX). The sections were also probed with antibodies raised to a conserved sequence of the enamelin protein. Selected intact teeth were first analysed by digital imaging and ascribed with an 'Enamel Defects Index' (EDI) score. SEM of tooth sections revealed disrupted prism morphology and the prisms had a glass-like appearance in some areas. These areas of dysplasia were sometimes irregular but formed regular arrays in others. Comparison of EDI scores with SEM indicated that in one tooth the surface had no measurable defects but significant defects were present in the underlying enamel microstructure. SEM immunohistochemistry with the antibody raised to a fragment of the enamelin protein produced positive, but light, labelling throughout normal enamel. In dysplastic areas, however, the labelling intensity appeared to be reduced. The results indicate that the presence of functional enamelin in the correct amounts is necessary for correct prism morphogenesis. In addition, a combination of EDI and structural analysis indicate that defects in enamel microstructure are not necessarily visible as defects on the surface of the tooth, suggesting the possibility, at least, that some instances of under-diagnosis may occur.

Safeguarding children in dentistry: 2. Do paediatric dentists neglect child dental neglect?

In this second part of a two-part report, further findings of a postal questionnaire sent in March 2005 to dentists with an interest in paediatric dentistry working in varied UK settings are presented and discussed in the context of current multi-agency good practice in safeguarding and promoting the welfare of children. Using insights gained from a survey of self-reported management of children with neglected dentitions, this paper explores whether paediatric dentists neglect child dental neglect. The authors conclude that current practice already includes much that contributes to promoting children's oral health and wellbeing. However, in a society where children continue to suffer as a result of abuse and neglect, they warn that improvements are needed in communication between dentists and other health and social care professionals if children's welfare is to be safeguarded and promoted effectively and future tragedies avoided.

Safeguarding children in dentistry: 1. Child protection training, experience and practice of dental professionals with an interest in paediatric dentistry.

Following several highly publicised inquiries into the deaths of children from abuse and neglect, there has been much recent interest in the role and responsibility of all health professionals to protect children at risk of maltreatment. The findings of a postal questionnaire, sent in March 2005 to 789 dentists and dental care professionals with an interest in paediatric dentistry working in varied settings in the UK, are presented in a two-part report and discussed in the context of current multi-agency good practice in safeguarding and promoting the welfare of children. This first part explores reported child protection training, experience and practice. There was a significant gap between recognising signs of abuse and responding effectively: 67% of respondents had suspected abuse or neglect of a child patient at some time in their career but only 29% had ever made a child protection referral. The dental profession is alerted to the need to ensure necessary appropriate action to safeguard children is always taken when child abuse or neglect are suspected.

Enamel defects in extracted and exfoliated teeth from patients with Amelogenesis Imperfecta, measured using the extended enamel defects index and image analysis.

AIMS: To enhance the phenotypic description and quantification of enamel defects from a North Sweden sample of extracted and exfoliated teeth originating from families with Amelogenesis Imperfecta by use of the extended enamel defects index (EDI) and image analysis to demonstrate the comparable reliability and value of the additional measurements. METHODS AND RESULTS: The sample comprised 109 deciduous and 7 permanent teeth from 32 individuals of 19 families with Amelogenesis Imperfecta in Northern Sweden. A special holder for individual teeth was designed and the whole sample was examined using the extended EDI and an image analysis system. In addition to the extended EDI definitions, the calibrated images were measured for tooth surface area, defect area and percentage of surface affected using image analysis techniques. The extended EDI was assessed using weighted and unweighted Kappa statistics. The reliability of imaging and measurement was determined using Fleiss' intra-class correlation coefficient (ICCC). Kappa values indicated good or excellent intra-operator repeatability and inter-operator reproducibility for the extended EDI. The Fleiss ICCC values indicated excellent repeatability for the image analysis measurements. Hypoplastic pits on the occlusal surfaces were the most frequent defect in this sample (82.6%). The occlusal surface displayed the most post-eruptive breakdown (39.13%) whilst the incisal portion of the buccal surfaces showed most diffuse opacities (53.4%). Image analysis methods demonstrated the largest mean hypoplastic pit areas were on the lingual surfaces. The largest mean post-eruptive breakdown areas were on the lingual surfaces of posterior teeth. The largest mean demarcated opacity areas were found on the labial surfaces. CONCLUSIONS: The extended EDI and the standardised image acquisition and analysis system provided additional information to conventional measurement techniques. Additional phenotypic variables were described.

Tooth dimensions in hypodontia with a known PAX9 mutation.

AIM: Congenital absence of teeth is a complex condition affecting several parameters of oral development. This is the first study to measure tooth crown dimensions using image analysis in a family with hypodontia in whom the mutation has been identified, and compare them with a control group. METHODS AND RESULTS: Study models were obtained from 10 family members from three generations affected by severe hypodontia with a missense mutation in PAX9 and 10 unaffected, unrelated controls. Using established image analysis techniques all teeth up to and including the first permanent molars were digitally imaged by two operators from the occlusal (O) and buccal (B) aspects three times and an average made for the mesio-distal (MDO and MDB) bucco-lingual (BL), area (A) and perimeter (P) measurements. Intra-class correlation coefficients (ICCC) were calculated to assess intra- and inter-operator reliability. Two-sample t-tests were then used to compare these dimensions with those of the controls. Reliability of the technique was high (mean r>0.95). The majority of tooth types throughout the dentition were significantly smaller in the family members with hypodontia than in the control group for all parameters measured. The levels of significance were very high for upper lateral incisors (p<0.0001) whilst the canines and first molars were less different. The greatest number of significant differences were found in BL and P, closely followed by MD and A measurements. CONCLUSIONS: The significantly smaller tooth crown dimensions recorded in the affected family members show that the effect of the PAX9 mutation is seen not only in the congenitally missing teeth but also in smaller crown size throughout the dentition.

The aetiology of hypodontia: the prevalence, severity and location of hypodontia within families.

AIMS: Previous studies have indicated that hypodontia has a significantly higher prevalence in the relatives of affected individuals than in the general population. This study aims to examine further the roles of genetic and environmental factors in the aetiology of hypodontia by investigating the relationship between the severity and distribution of hypodontia between family members, and any discernable effect of maternal health during pregnancy and birth weight. METHODS AND RESULTS: 117 first degree relatives of 41 index patients were examined clinically and radiographically to identify the presence, severity and location of hypodontia. Both siblings and parents of index patients had a higher prevalence of hypodontia than the general population. The number and location of missing teeth was not related to the number and location of missing teeth in parents or siblings. The expression of hypodontia within a family was not affected by maternal health during pregnancy. CONCLUSIONS: The variation found in the expression of hypodontia within families suggests that its occurrence is not solely determined by genetic factors, but epigenetic and environmental factors probably also are important. This finding is consistent with a multifactorial aetiology for this condition.

Multiple crown size variables of the upper incisors in patients with supernumerary teeth compared with controls.

AIMS: As part of ongoing studies of the aetiology of dental anomalies the aims of this study were to identify multiple components of tooth size of the upper permanent incisors in 34 patients with supernumerary teeth and to compare them with those in a control group to determine whether the presence of a supernumerary tooth has a local effect on the size of the surrounding dentition. METHODS AND RESULTS: The labial and occlusal aspects of the clinical crowns of the upper permanent central and lateral incisors on the study models of 74 subjects were digitally imaged and measured using an image analysis system and automated macro (34 patients with supernumerary in the upper incisor region: 17 males and 17 females and 40 controls: 20 males and 20 females). The macro-defined 17 variables from each view. From the labial view these were: the mesio-distal and occluso-gingival length and additional measurements along 25 and 75% of the mesio-distal line and at 25, 50 and 75% along the occluso-gingival line such that all these sub-divisions extended to the periphery of the tooth. From the occlusal view these were: the mesio-distal and labio-lingual lengths, and additional variables that sub-divided the mesio-distal again at 25 and 75% along the length and at 25, 50 and 75% along the labio-lingual dimension. Principal component analysis (PCA) was used to identify the key factors with the most random variability. Comparisons were then carried out between the supernumerary cases and control group using 2-way ANOVA. Seven factors of tooth size for the upper central incisors and eight factors for the upper lateral incisors were extracted. Most of these variables were found to be larger in the supernumerary group than in the control. Statistically significant differences were found for 5 out of 7 and 4 out of 8 for the upper central and lateral incisors respectively. CONCLUSIONS: A number of factors of tooth size were identified and found to be larger in the supernumerary group compared to the control (7 for upper central and 8 for upper lateral incisors); the majority reached the 0.05 significance level. Tooth crown size of the upper central incisor was affected more than that of the upper lateral incisor, supporting a local field effect.

A new technique for symmetry determination in tooth morphology using image analysis: application in the diagnosis of solitary maxillary median central incisor.

OBJECTIVE: To develop a new technique for determining symmetry in tooth morphology and to evaluate this in the investigation of a patient with a solitary maxillary median central incisor (SMMCI) and a control group. DESIGN: A 9-year-old Caucasian female presented with SMMCI. Clinically the tooth appeared symmetrical. Morphology measurements of the maxillary central incisor were made using an image analysis system. Symmetry was determined by outlining the tooth from labial and axial views. These images were block filled, duplicated, flipped horizontally and then superimposed on the original image. The coincident area and perimeter of the two images from both views were measured. The method was repeated for the maxillary central incisors of 20 sets of control study models for young adult patients from which reference intervals for comparison with the SMMCI case were prepared. RESULTS: From the labial view, the area and perimeter of the two images of the SMMCI tooth were 98.85% and 98.97% coincident, respectively. From the axial view the area of the two images was 96.17% coincident, while the perimeter was 99.03% coincident. In all but one comparison for coincidence the SMMCI was above the upper limit of the reference range from the control group. CONCLUSIONS: This new technique is a valid method of assessing symmetry and is a useful clinical tool in cases of SMMCI.

An in-vitro and in-vivo methodology study of alveolar bone measurement using extra-oral radiographic alignment apparatus, Image Pro-Plus software and a subtraction programme.

OBJECTIVES: (a) To validate a method for acquiring reproducible radiographic images and repeatable measurements of digital images. (b) To investigate the clinical and radiographic changes following periodontal surgery and in a periodontally healthy control group. METHODS: In-vitro study. Sixteen dried human skulls had replicate intra-oral radiographs taken using an extra-oral alignment apparatus under simulated clinical conditions, and measurements were made on the images using Image Pro-Plus analysis software. In-vivo study. Seven subjects, aged 38-63 years with chronic periodontitis, were treated using replaced flap surgery. Nine patients with healthy periodontal tissues were recruited as a control group. Clinical measurements and radiographs were taken pre-surgery and 6 months afterwards, and at the same interval for untreated healthy controls using the alignment apparatus. Radiographs were analysed using Image Pro-Plus and subtraction radiography. RESULTS: Radiographs had a high degree of reliability (ICC 0.98-0.99) for the in-vitro study and intra-operator repeatability of measurements was high (ICC 0.65-0.99) for the in-vivo study. Over this interval there were both significant clinical improvements and in bone gain within infra-bony defects on radiographs. Subtracted images showed that out of 17 surgically treated sites, 13 showed evidence of bone gain, one showed no change, two showed bone loss and one could not be interpreted due to poor alignment. Overall there were no significant clinical or radiographic changes in the control group. CONCLUSIONS: Radiographic images were highly reproducible. Measurements had a high degree of repeatability. The methods used allowed accurate quantitative and qualitative measurement of change in alveolar bone. This technique would allow accurate measurement of any changes in alveolar bone levels in patients with periodontal disease.

Tooth size in patients with supernumerary teeth and a control group measured by image analysis system.

OBJECTIVE: The aim of this study was to compare tooth size measurements between patients with supernumerary teeth and a control group. METHOD: The supernumerary group consisted of 56 subjects (21 females and 35 males) and the control group of 40 subjects (20 females and 20 males). All available permanent teeth on the dental casts were imaged and measured from both buccal and occlusal views using an image analysis system. Mesio-distal, bucco-lingual or occluso-gingival dimensions, area and perimeter were measured from each view. RESULTS: Supernumerary tooth patients tended to have larger tooth size measurements for almost all variables than controls. Significant differences (p<0.05) were found for the mesio-distal dimension of the upper and lower incisors and the bucco-lingual dimensions of the lower first premolars. CONCLUSIONS: The results are compatible with supernumerary teeth being a complex dental anomaly with a multifactorial aetiology in which both genetic and environmental factors are important. There is some evidence of a local effect with greater differences in tooth dimension adjacent to the site of the supernumeraries.

Quantification of dental plaque on lingual tooth surfaces using image analysis: reliability and validation.

AIM: The aim of this study was to increase the versatility and further validate the method reported by Smith et al. (2001) by testing the reliability of plaque measurement against two well-known dental plaque quantification methodologies using image analysis in a clinical trial. METHOD: The teeth of 40 subjects were disclosed before digital images of the labial and lingual surfaces of their upper and lower incisors were acquired. The amount of plaque present was quantified using a modification of the method described by Smith et al. (2001). The method was modified for obtaining images of the lingual surfaces by incorporating the use of orthodontic occlusal mirrors and 5-mm pieces of moistened blue articulating paper used to enable calibration. Plaque measurements were made from 320 upper and lower anterior teeth from the 40 subjects by two operators. Fliess' coefficient of reliability was used to assess intra- and inter-operator reliability and the independent sample t test was used to assess statistical significance between test and control groups after checking the data for normality. For validation, measurements were recorded using the Turesky et al. (1970) (modification of the Quigley & Hein (1962) plaque index and the Addy et al. (1983) plaque area index. The results were compared with the image analysis method using Pearson's correlation coefficient. RESULTS: The results for reliability were within Fliess' range of "excellent" for both intra-operator repeatability and inter-operator reproducibility. Pearson's correlation coefficients showed highly significant values indicating the close similarity between all three methods. CONCLUSIONS: This method for the measurement of dental plaque on lingual surfaces of anterior teeth proved reliable. The combined results from the labial and lingual surfaces of anterior teeth using image analysis produced trial conclusions comparable with the alternate plaque quantification methods used, with less clinician time and further producing a permanent database of images for future use.

Further studies of a model for the etiology of anomalies of tooth number and size in humans.

A multifactorial model based on an underlying continuous distribution of tooth size, with thresholds determining hypodontia and supernumeraries, has been proposed [1]. Our aim is to investigate this model by comparing tooth morphology of affected patients and their first degree relatives with normal controls. An image analysis system [2] was used to measure teeth on study models of controls, patients with hypodontia of varying degrees and location, first-degree relatives of hypodontia index cases, and patients with supernumerary teeth in the maxillary incisor region. Mesio-distal crown size in hypodontia patients was smaller than controls, and this difference was significant for all tooth types. There was a general pattern: the more severe the hypodontia, the smaller the size of the tooth formed. Patients with supernumerary teeth had permanent maxillary central and lateral incisors and canines that were significantly larger in mesio-distal width than controls. The maxillary central incisors also differed in taper in supernumerary patients compared to controls. Hypodontia prevalence was higher in first-degree relatives (22%) of hypodontia index cases than in the general population (4.4%), and unaffected relatives had smaller teeth than controls. Thus, there were generalized and localized effects within the dentition, and these findings are compatible with the statistical expectations of the proposed multifactorial model.

Tooth dimensions in hypodontia patients, their unaffected relatives and a control group measured by a new image analysis system.

Tooth dimensions were compared between index patients with severe hypodontia (six or more congenitally missing teeth), their relatives with a full complement of teeth, and a control group. The groups consisted of 12 index cases (seven females and five males), 21 relatives without hypodontia (13 females and eight males), and a control group of 10 males and 10 females with complete dentitions, and no family history of hypodontia. All formed teeth were imaged buccally and occlusally from study models, with a digital camera linked to a computer. The images were acquired and measured using Adobe Photoshop and Image Pro Plus, respectively. Mesiodistal, buccolingual, or occlusogingival area and perimeter measurements were determined from each image. In the index hypodontia group tooth dimensions were significantly smaller (P < 0.001, Bonferroni corrected level) for maxillary and mandibular canines, and first premolars for all dimensions from the buccal view, and for maxillary and mandibular canines and first premolars, maxillary central incisors, maxillary first molars, mandibular lateral incisors, and mandibular second premolars for all dimensions from the occlusal view. In the relatives without hypodontia compared with the control group, mesiodistal dimensions from the buccal view were significantly smaller (P < 0.001) for the central incisors and maxillary first and second premolars, and for the maxillary and mandibular first premolars for all dimensions from the occlusal view. There was a trend for the tooth dimensions of all teeth in the index group to be smaller compared with the control group. The tooth dimensions of the relatives without hypodontia also tended to be smaller than the control group, but were larger than those of the index cases.

The quantification of dental plaque using an image analysis system: reliability and validation.

AIM: Established clinical indices used to measure dental plaque have limitations. The aim of this study was to develop and evaluate a new method of quantitative plaque measurement, with increased sensitivity and reproducibility, using a purpose designed image analysis system. METHOD: After plaque disclosure, the patient was positioned in a cephalometric head holder mounted in a custom-built frame with standard lighting and camera position. A 32-bit Digital Camera grabbed the image of the patient's teeth, which was analysed using Adobe Photoshop (V5.02, Adobe Systems Ltd, Europe) and Image Pro Plus software (V4.0, Media Cybernetics, USA). Measurements were made on 120 incisors in 15 patients and reproducibility was assessed using Fliess' Coefficient of Reliability. Sensitivity was assessed by comparison of results of treatment effects from a clinical trial involving 30 patients. Analysis of covariance was used to compare the image analysis system with the Turesky modification of the Quigley and Hein plaque index and the Addy plaque area index. RESULTS: The results were within the 'Excellent Reliability' category, for inter-operator reproducibility and intra-operator repeatability. CONCLUSIONS: This new system provided accurate quantitative measurements of dental plaque and labial tooth surfaces and was shown to have increased sensitivity when compared with the Turesky modification of the Quigley and Hein plaque index and the Addy plaque area index. The stored images can be re-measured for reliability and comparability between studies.

Neuropeptide expression in the ferret trigeminal ganglion following ligation of the inferior alveolar nerve.

Previous studies have found changes in neuropeptide expression in trigeminal ganglion cells after inferior alveolar nerve (IAN) section. These changes may play a part in the persistent sensory abnormalities that can be experienced after trigeminal nerve injuries. Here, neuropeptide expression after IAN ligation was studied, as this type of injury is thought to be more likely to result in sensory disturbances. The neuropeptides investigated were substance P, calcitonin gene-related peptide, enkephalin (ENK), galanin (GAL), neuropeptide Y (NPY) and vasoactive intestinal polypeptide. In anaesthetised adult female ferrets the left IAN was sectioned and the central stump tightly ligated. Recovery was allowed for 3 days, 3 or 12 weeks before perfusion-fixation. In a second procedure, 1 week before perfusion, the IAN was exposed and an injection made central to the injury site, using a mixture of 4% Fluorogold and 4% Isolectin B4 conjugated to horseradish peroxidase, to identify cell bodies with axons in the inferior alveolar nerve and cells with unmyelinated axons within this population, respectively. Control experiments involved tracer injection alone. After harvesting the tissue, sagittal sections were taken from both the right and left ganglia and immunohistochemical staining used to reveal the presence of peptides and Isolectin B4 tracer. The results showed a significant decrease in GAL expression after injury and an increase in ENK and NPY expression. No significant differences were seen in the expression of the other peptides or in the proportion of lectin-positive cells at any time after injury. When compared with previous data, significant differences were found between peptide expression following nerve ligation and nerve section. These results reveal that the changes in neuropeptide expression in the trigeminal ganglion that follow IAN injury are dependent upon the type of injury. The extent to which changes in the central neuropeptide levels contribute to the development of sensory disorders remains to be established.

Changes in neuropeptide expression in the trigeminal ganglion following inferior alveolar nerve section in the ferret.

Changes in neuropeptide expression in afferent nerve fibres may play a role in the persistent sensory abnormalities that can be experienced following trigeminal nerve injuries. We have therefore studied changes in the expression of the neuropeptides substance P, calcitonin gene-related peptide, enkephalin, galanin, neuropeptide Y and vasoactive intestinal polypeptide in the trigeminal ganglion following peripheral nerve injury. In anaesthetised adult female ferrets, the left inferior alveolar nerve was sectioned and recovery allowed for three days, three weeks or 12 weeks prior to perfusion-fixation. During a second procedure, one week prior to perfusion, the inferior alveolar nerve was exposed and an injection made central to the injury site using a mixture of 4 % Fluorogold and 4 % isolectin B4 conjugated to horseradish peroxidase to identify cell bodies with axons in the inferior alveolar nerve and cells with unmyelinated axons within this population, respectively. Control animals received tracer injection alone. After harvesting the tissue, sagittal sections were taken from both the right and left ganglia and immunohistochemical staining was used to reveal the presence of peptides and isolectin B4-horseradish peroxidase tracer. Within the Fluorogold-labelled population, cell counts revealed a significant reduction in the proportion of substance P-containing cells at three days (P = 0.0025), three weeks (P = 0.0094) and three months (P = 0.0149) after nerve section, and a significant reduction in the proportion of calcitonin gene-related peptide-containing cells at three days (P = 0.0003) and three weeks (P = 0.007). No significant changes were seen in the expression of the other peptides, or at other time periods. A significant reduction in the number of isolectin B4-horseradish peroxidase-positive cells (with unmyelinated axons) was seen at three days (P = 0.0025), three weeks (P = 0.0074) and three months after the injury (P = 0.0133). These results demonstrate a significant reduction in the expression of some neuropeptides in the early stages after inferior alveolar nerve section. Some of the results differ markedly from those reported previously in other systems, and may be related to the specific nerve studied, species variations or differences between spinal and trigeminal nerves.

A novel (6S)-4,6-dimethyldodeca-2E,4E-dienoyl ester of phomalactone and related alpha-pyrone esters from a Phomopsis sp. with cytokine production inhibitory activity.

A series of novel 6-substituted 5,6-dihydro-5-hydroxy-alpha-pyrone esters, 1 approximately 3, isolated from fermentations of a Phomopsis sp. (Xenova culture collection no. X22502) have been identified as inhibitors of lipopolysaccharide (LPS)-induced cytokine production. These include the (6S)-4,6-dimethyldodecadien-2E,4E-dienoyl ester of phomalactone, 1, and two analogues bearing a prop-2E-enoic acid moiety at the 6-position of the alpha-pyrone ring. (6S)-4,6-Dimethyl-2E,4E-dienoic acid, 4, and a hydroxylated analogue, 5, were also isolated and characterised. The most potent cytokine production inhibitor was 1, which inhibited LPS-induced tumour necrosis factor alpha (TNFalpha) production by U937 cells and LPS-induced interleukin 1beta (IL-1beta) production by peripheral blood mononuclear cells (PBMC) with IC50 values of 80 nM and 190 nM respectively. The effect of 1 in PBMC was selective for IL-1beta relative to TNFalpha. The inhibition of IL-1beta production by 1 involved a post-translational mechanism of action at the level of IL-1beta secretion as demonstrated by the lack of an effect on cell-associated IL-1beta production. 1 showed no effect on the activity of caspase 1 in cytosolic extracts from the THP1 monocytic cell line.

Angiogenesis and the placental environment.

Rapid growth and vascularization of the human placenta are characteristic of early pregnancy and are accomplished in an unusually hypoxic environment. Stimulation of placental growth through hypoxia-induced angiogenesis may therefore be of particular importance. We have previously found that several varieties of vascular endothelial growth factor (VEGF) mRNA, including VEGF165, are present in cultured placental fibroblasts. We hypothesized that hypoxia would increase the transcription and translation of VEGF by these cells and provide one mechanism linking placental development with its environment. Placental fibroblasts were grown in aerobic or anaerobic atmospheric conditions for 72 h. By 24 h the oxygen tension of the anaerobic culture media was significantly less than that of the aerobic cultures. RNA was extracted from the cells at 24, 48 and 72 h. Following reverse transcription polymerase chain reaction (RT-PCR) stronger signals for VEGF were always found in the anaerobic cultures and this was confirmed by competitive PCR. mRNA for VEGF165 was represented most strongly but the anaerobic cultures also showed clearly mRNA for VEGF121, VEGF189 and VEGF206. The VEGF protein was also measured in the aerobic and anaerobic culture medium. By 72 h the average concentration of VEGF was significantly higher (P = 0.01) in the anaerobic culture medium. VEGF production is one mechanism through which oxygen supply may influence placental development. Examples of this may include the compensatory placental hypertrophy associated with maternal anaemia and with reproduction at high altitude.

Short report: identification of a specific pattern of vascular endothelial growth factor mRNA expression in human placenta and cultured placental fibroblasts.

Reverse transcription and the polymerase chain reaction (PCR) were used to detect vascular endothelial growth factor (VEGF) mRNA in human placental tissue and cultured placental fibroblasts obtained during the first trimester of pregnancy. The primers for VEGF corresponded to areas in exon 4 and exon 8 of the VEGF gene. After one round of PCR three products, equivalent to VEGF121, VEGF165 and VEGF189, were detected within placental tissue and cultured placental fibroblasts. A further round of PCR revealed the presence of two more products equivalent to VEGF206 and VEGF145. Thus, in addition to the production of readily secreted forms of VEGF (VEGF121 and VEGF165), the placenta produces several transcripts expected to increase the growth factor pool of the extracellular matrix.