Use of the universal pain assessment tool for evaluating pain associated with TMD in youngsters with an intellectual disability.

BACKGROUND: The Universal Pain Assessment Tool (UPAT) was used to assess the level of pain in people with limited communication skills. The UPAT enables clinicians to consult a specialized pain management team more often and lead to earlier interventions. The purpose of this study was to determine, whether the UPAT could be used as an extra tool to collect data on functional TMJ pain and to assess orofacial pain levels related to temporomandibular disorder(s) (TMD) in people with intellectual disabilities (ID). MATERIAL AND METHODS: Non-down syndrome ID Athletes were screened during the Special Olympics European games in 2014. The clinical scores of possible functional jaw pain were collected using the UPAT, to indicate pain severity on a visual scale during different jaw movements (opening, closing and lateral). RESULTS: Two hundred and four youngsters were screened by calibrated dentists. The majority (65%) of participants were male (133 male and 71 female athletes); age distribution ranged from 15 to 23 years (mean 19.25 ± 2.53). The results of the UPAT have shown the existence of functional TMJ pain in 32% (n=65) of the athletes without significant prevalence (P > 0.05) in this survey group. CONCLUSIONS: According to the results of the present study, the UPAT demonstrated that it could be a useful tool to detect the existence of functional jaw pain possibly associated with TMD and also a valid instrument to score pain intensity associated with TMD in people with ID.

CXCL4L1-fibstatin cooperation inhibits tumor angiogenesis, lymphangiogenesis and metastasis.

Anti-angiogenic and anti-lymphangiogenic drugs slow tumor progression and dissemination. However, an important difficulty is that a tumor reacts and compensates to obtain the blood supply needed for tumor growth and lymphatic vessels to escape to distant loci. Therefore, there is a growing consensus on the requirement of multiple anti-(lymph)angiogenic molecules to stop cell invasion efficiently. Here we studied the cooperation between endogenous anti-angiogenic molecules, endostatin and fibstatin, and a chemokine, the Platelet Factor-4 variant 1, CXCL4L1. Anti-angiogenic factors were co-expressed by IRES-based bicistronic vectors and their cooperation was analyzed either by local delivery following transduction of pancreatic adenocarcinoma cells with lentivectors, or by distant delivery resulting from intramuscular administration in vivo of adeno-associated virus derived vectors followed by tumor subcutaneous injection. In this study, fibstatin and CXCL4L1 cooperate to inhibit endothelial cell proliferation, migration and tubulogenesis in vitro. No synergistic effect was found for fibstatin-endostatin combination. Importantly, we demonstrated for the first time that fibstatin and CXCL4L1 not only inhibit in vivo angiogenesis, but also lymphangiogenesis and tumor spread to the lymph nodes, whereas no beneficial effect was found on tumor growth inhibition using molecule combinations compared to molecules alone. These data reveal the synergy of CXCL4L1 and fibstatin in inhibition of tumor angiogenesis, lymphangiogenesis and metastasis and highlight the potential of IRES-based vectors to develop anti-metastasis combined gene therapies.

The p53 isoform, Δ133p53α, stimulates angiogenesis and tumour progression.

The tumour suppressor p53, involved in DNA repair, cell cycle arrest and apoptosis, also inhibits blood vessel formation, that is, angiogenesis, a process strongly contributing to tumour development. The p53 gene expresses 12 different proteins (isoforms), including TAp53 (p53 (or p53α), p53ß and p53γ) and Δ133p53 isoforms (Δ133p53α, Δ133p53ß and Δ133p53γ). The Δ133p53α isoform was shown to modulate p53 transcriptional activity and is overexpressed in various human tumours. However, its role in tumour progression is still unexplored. In the present study, we examined the involvement of Δ133p53 isoforms in tumoural angiogenesis and tumour growth in the highly angiogenic human glioblastoma U87. Our data show that conditioned media from U87 cells depleted for Δ133p53 isoforms block endothelial cell migration and tubulogenesis without affecting endothelial cell proliferation in vitro. The Δ133p53 depletion in U2OS osteosarcoma cells resulted in a similar angiogenesis blockade. Furthermore, using conditioned media from U87 cells ectopically expressing each Δ133p53 isoform, we determined that Δ133p53α and Δ133p53γ but not Δ133p53ß, stimulate angiogenesis. Our in vivo data using the chicken chorio-allantoic membrane and mice xenografts establish that angiogenesis and growth of glioblastoma U87 tumours are inhibited upon depletion of Δ133p53 isoforms. By TaqMan low-density array, we show that alteration of expression ratio of Δ133p53 and TAp53 isoforms differentially regulates angiogenic gene expression with Δ133p53 isoforms inducing pro-angiogenic gene expression and repressing anti-angiogenic gene expression.

Efficient gene transfer in skeletal muscle with AAV-derived bicistronic vector using the FGF-1 IRES.

IRESs (internal ribosome entry sites) are RNA elements behaving as translational enhancers in conditions of global translation blockade. IRESs are also useful in biotechnological applications as they allow expression of several genes from a single mRNA. Up to now, most IRES-containing vectors use the IRES from encephalomyocarditis virus (EMCV), highly active in transiently transfected cells but long and not flexible in its positioning relative to the gene of interest. In contrast, several IRESs identified in cellular mRNAs are short and flexible and may therefore be advantageous in gene transfer vectors such as those derived from the adeno-associated virus (AAV), where the size of the transgene expression cassette is limited. Here, we have tested bicistronic AAV-derived vectors expressing two luciferase genes separated by the EMCV- or fibroblast growth factor 1 (FGF-1) IRES. We demonstrate that the AAV vector with the FGF-1 IRES, when administrated into the mouse muscle, leads to efficient expression of both transgenes with a stable stoechiometry, for at least 120 days. Interestingly, the bicistronic mRNA containing the FGF-1 IRES leads to transgene expression 10 times superior to that observed with EMCV, in vivo. AAV vectors featuring the FGF-1 IRES may thus be advantageous for gene therapy approaches in skeletal muscle involving coexpression of genes of interest.

In vivo adenovirus-mediated delivery of a uPA/uPAR antagonist reduces retinal neovascularization in a mouse model of retinopathy.

Diabetic retinopathy and retinopathy of prematurity are among the leading causes of vision impairment throughout the world. Both diseases are characterized by pathological angiogenesis, which severely impairs vision. Extracellular proteinases play important roles in endothelial cell migration during angiogenesis. Amino-terminal fragment (ATF) is an angiostatic molecule that targets the uPA/uPAR system and inhibits endothelial cell migration. The angiostatic effect of ATF has been demonstrated in models of cancer, but has never been assessed in pathological retinal neovascularization. Endostatin also has angiostatic effects on tumor growth and retinal neovascularization. We used an adenoviral vector carrying the murine ATF (AdATFHSA) or endostatin gene coupled to human serum albumin (HSA) (AdEndoHSA) to increase the half-life of the therapeutic protein in the circulation. We induced retinopathy by exposing 7-day-old mice to high levels of oxygen. They were intravitreally injected with the vectors. Local injection of AdATFHSA or AdEndoHSA reduced retinal neovascularization by 78.1 and 79.2%, respectively. Thus, the adenovirus-mediated delivery of ATFHSA or EndoHSA reduces retinal neovascularization in a mouse model of hypoxia-induced neovascularization.

Prominent beta-5 gene expression in the cardiovascular system and in the cartilaginous primordiae of the skeleton during mouse development.

The alpha v beta (alpha(v)beta5) heterodimer has been implicated in many biological functions, including angiogenesis. We report the beta5 gene expression pattern in embryonic and foetal mouse tissues as determined by Northern blotting and in situ hybridization. During the earliest stages, beta5 mRNA is widespread in the mesoderm. During later developmental stages, it remains mostly confined to tissues of mesodermal origin, although probable inductive effects trigger shifts of beta5 gene expression from some mesenchymatous to epithelial structures. This was observed in the teeth, skin, kidneys, and gut. Of physiological importance is the beta5 labeling in the developing cardiovascular and respiratory systems and cartilages. Furthermore, early beta5 gene expression was observed within the intra- and extraembryonic sites of hematopoiesis. This suggests a major role for beta5 in the hematopoietic and angiogenic stem cells and thus in the development of the vascular system. Later, the beta5 gene was expressed in endothelial cells of the vessels developing both by angiogenesis and vasculogenesis in the lung, heart, and kidneys. Moreover, the beta5 hybridization signal was detected in developing cartilages but not in ossified or ossifying bones. beta5-Integrin is a key integrin involved in angiogenesis, vasculogenesis, hematopoiesis, and bone formation.

FIF [fibroblast growth factor-2 (FGF-2)-interacting-factor], a nuclear putatively antiapoptotic factor, interacts specifically with FGF-2.

Numerous evidence indicates that some of the activities of fibroblast growth factor 2 (FGF-2) depend on an intracrine mode of action. Recently, we showed that three high molecular mass (HMM) nuclear forms of FGF-2 are part of a 320-kDa protein complex while the cytoplasmic AUG-initiated form is included in a 130-kDa complex. Consequently, the characterization of FGF endogenous targets has become crucial to allow the elucidation of their endogenous activities. Through the screening of GAL4-based yeast two-hybrid expression libraries, we have isolated a gene encoding a nuclear protein of 55 kDa, FIF (FGF-2-interacting-factor), which interacts specifically with FGF-2 but not with FGF-1, FGF-3, or FGF-6. In this system, FIF interacts equally well with the NH2-extended 24-kDa FGF form as with the 18-kDa form, indicating that the FIF-binding motif is located in the last 155 amino acids of FGF-2. Nevertheless, coimmunoprecipitation experiments showed an exclusive association with HMM FGF-2. The predicted protein contains a canonical leucine zipper domain and three overlapping hydrophobic heptad repeats. The region spanning these repeats is, together with a region located in the N-terminal part of the FIF protein, implicated in the binding to FGF-2. In contrast to the full-length FIF protein, several deletion constructs were able to transactivate a lac-Z reporter gene. Furthermore, the COOH-terminal part, but not the full-length FIF protein, has previously been shown to exhibit antiapoptotic properties. Thus we discuss the possibility that these activities could reflect a physiological function of FIF through its interaction with FGF-2.

Comparison of clinical profiles and treatment outcomes of an elderly and a younger temporomandibular patient group.

STATEMENT OF PROBLEM: Older temporomandibular disorder patients with more general complications and health problems may have a different clinical profile and be likely to react less favorably to conservative treatment. PURPOSE: This retrospective study compared the clinical profiles of a young (20 to 30 years) and an older (50 to 70 years) group of patients with pain and dysfunction in the temporomandibular region and to analyze treatment outcomes. METHODS: Clinical profiles and treatment outcomes were studied with a standardized protocol and the Helkimo Pain and Dysfunction Index up to 1 year after initial examination. RESULTS: Younger and older patients with temporomandibular disorder differed only in pain intensity at initial examination, but the outcome of conservation treatment was equally successful. CONCLUSION: Conservative treatment resulted in a significant alleviation of pain and dysfunction in almost 85% of patients. Both the younger and the older patient groups benefitted from this treatment protocol and therefore can be treated in the same fashion.

Proximal trisomy 13q and distal monosomy 8p in a dysmorphic and mentally retarded patient with an isodicentric chromosome 13q and a 13q/8p translocation chromosome.

A 40 year-old dysmorphic and mentally retarded female is reported with a de novo unbalanced chromosomal rearrangement (karyotype: 46,XX,der(8)t(8;13)(p23;q123),idic(13)(pter-->q123: q123-->pter) resulting in an isodicentric chromosome 13 and a double aneusomy including partial trisomy 13 (13pter-q123) and distal monosomy 8p (8pter-p23). The main clinical findings consist of developmental/mental retardation, behavioural disturbances and minor congenital defects, not consistent with the clinical pattern of either of the two aneusomies. A mechanism for the chromosome rearrangement is proposed and the absence of specific physical findings in the present patient is discussed in the light of the available literature data.

FGF-2 dimerization involvement in growth factor mediated cell proliferation but not cell differentiation.

Dimerization is a prerequisite for many growth factors in their receptor activation leading to cellular response. FGF-1 and FGF-2, members of the Fibroblast Growth Factor (FGF) family, were shown to form non-covalent dimers and oligomers in vitro. Using the two-hybrid system as an in vivo binding assay we show here that of three representative members of the FGF family, only FGF-2 is able to homodimerize. Moreover the FGF-2 isoforms could heterodimerize. Two single-point mutants (T121F and W123R), defective in their dimerization capability, were isolated through random mutagenesis and were used to study the role of FGF-2 dimerization with regard to its biological activity. Remarkably, these mutant proteins were still able to induce cell differentiation, but were strongly affected in their capacity to promote cell proliferation. This study thus highlights the uncoupling between proliferation and differentiation FGF-2 signaling pathways and the crucial role of FGF-2 dimerization in the mitogenic activity of this factor.

A new technique for the three-dimensional study of the spine in vitro and in vivo by using a motion-analysis system.

We introduce a new method with a motion-analysis system (MAS) to study the vertebral model in vitro. Compared with the currently most accurate technique, roentgen stereophotogrammetric analysis (RSA), the difference between the RSA and the MAS is 0.12 degree +/- 1.64 degrees. An accuracy with an error of 0.08 degree +/- 1.15 degrees is determined by means of an angle gauge. Although a significant difference between the MAS and the goniometer (p = 0.04) is found around the X-axis (theta; transverse plane), it is limited to < 1 degree. The MAS provides an in-depth insight into the mechanism of the three-dimensional rotation at each vertebra in vivo. The backward inclination of the apical vertebra (AV) and forward inclination of the upper-end vertebra (UEV) around the Y-axis (phi) results in a correction of the hypokyphosis shown by the Cobb angle in the sagittal plane. The clockwise rotation of the UEV in the Z-axis (psi) leads to a reduction of the Cobb angle in the frontal plane. Additionally, the MAS as an intraoperative alternative shows different results of the derotation maneuver by the Cotrel-Dubousset instrumentation (CDI) compared with the computed tomography (CT) scan. Our method gives more direct details of the derotation not influenced by patient posture, as observed in the CT scan.

Otalgia in patients with temporomandibular joint disorders.

Otalgia without organic causes is a common symptom in temporomandibular dysfunction (TMD) patients even though the etiology is controversial. Investigations of the influence of TMD treatment on otalgia are scarce. This follow-up study analyzed the clinical profile of TMD patients with otalgia and evaluated the treatment outcome. A total of 400 consecutive TMD patients (75% women) were divided in two groups: group 1 consisted of 233 patients (58%) with no complaint of ear symptoms and group 2 consisted of 167 patients (42%) with complaint of otalgia. The patients were examined with a standardized protocol and treated similarly with conservative methods. Group 2 was referred and examined by otolaryngologists. Otalgia patients (group 2) had statistically significantly higher pain scores (p 0.02). They belonged to the greater dysfunction scores (Di III) according to the Helkimo Pain and Dysfunction index (41%) vs. 24%; p < 0.001). There was a statistically significant association with pain on condyle palpation and otalgia (p < 0.01). One year after the first examination, the patients exhibited no pain or occasionally mild pain in 66% (group 1) and 74% (group 2) (p 0.35). Of the otalgia patients, 48% no longer had otalgia and 32% of the patients experienced mild or occasional otalgia. The changes in dysfunction scores after 1 year revealed significant improvement. No difference was found between group 1 and 2 in pain and dysfunction score. For the dysfunction index readings 0 and I, 77% and 73% had no or only mild symptoms (Di 0 and I). The conclusions of this study are that TMD patients with otalgia are not a separate TMD group and they responded well to conservative treatment.

Physical and genetic map of the Lactococcus lactis subsp. cremoris MG1363 chromosome: comparison with that of Lactococcus lactis subsp. lactis IL 1403 reveals a large genome inversion.

A physical and genetic map of the chromosome of the Lactococcus lactis subsp. cremoris reference strain MG1363 was established. The physical map was constructed for NotI, ApaI, and SmaI enzymes by using a strategy that combines creation of new rare restriction sites by the random-integration vector pRL1 and ordering of restriction fragments by indirect end-labeling experiments. The MG1363 chromosome appeared to be circular and 2,560 kb long. Seventy-seven chromosomal markers were located on the physical map by hybridization experiments. Integration via homologous recombination of pRC1-derived plasmids allowed a more precise location of some lactococcal genes and determination of their orientation on the chromosome. The MG1363 chromosome contains six rRNA operons; five are clustered within 15% of the chromosome and transcribed in the same direction. Comparison of the L. lactis subsp. cremoris MG1363 physical map with those of the two L. lactis subsp. lactis strains IL1403 and DL11 revealed a high degree of restriction polymorphism. At the genetic organization level, despite an overall conservation of gene organization, strain MG1363 presents a large inversion of half of the genome in the region containing the rRNA operons.

Longitudinal study of functional conditions in the masticatory system in Flemish children.

A 5-yr longitudinal study on occlusal and functional parameters has been made in 75 children aged 8-11. They were selected at random from an original group of 510 studied 5 yr earlier. The following parameters were measured: frequency of frontal open bite lateral and frontal crossbite, the attrition on molars and on front teeth, the contacts on lateral excursion, pain on condyle palpation, TMJ clicking, deviations in the opening and lateral movement, and the maximal interincisal distance. There was an increase in the percentage of deep bite and crossbite. Attrition on molars was seen in 41% and on front teeth in 13% of the children. No real pattern could be observed as a function of age. The mean maximal opening range was 43.7 mm, varying between 23 and 52 mm. Restricted opening was found in one girl. In 60% a deviation in the opening has been found. In 26% TMJ sounds were observed and 28% had pain on condyle palpation. In most of the children the symptoms were very mild to moderate. No significant correlation could be found between dysfunctional signs and occlusal parameters. The present results did not provide any conclusive answer to the tremendous increase in dysfunctional signs over this 5-yr period. The treatment need is, however, low and should not be overestimated.