One-step co-culture fermentation strategy to produce high-content fructo-oligosaccharides.

An integrated process enabling the simultaneous production and purification of fructo-oligosaccharides (FOS) was explored. A co-culture fermentation with Aspergillus ibericus (used as FOS producer strain) and Saccharomyces cerevisiae YIL162 W (for small saccharides removal) was optimized. Inoculation conditions of S. cerevisiae, fermentative broth composition, temperature and pH were optimized by experimental design. Yeast extract concentration and temperature were the most significant variables affecting FOS purity. Co-culture fermentations with simultaneously inoculation of the strains, run under 30 °C, initial pH 6.0 and 17 g L-1 yeast extract led to FOS mixtures with 97.4 ± 0.2% (w/w) purity. The fermentations conducted in bioreactor, at a 0.8 vvm aeration rate, yielded 0.70 ± 0.00 gFOS.ginitial GF-1 at 45 h fermentation, with a FOS content of 133.7 ± 0.1 g L-1. A purity of FOS up to 93.8 ± 0.7% (w/w) was achieved. The one-step fermentation proved to be efficient, economical and fast.

Painful temporomandibular disorders and central sensitization: implications for management-a pilot study.

The objective was to investigate the presence of cutaneous allodynia and hyperalgesia in the trigeminal and extra-trigeminal areas, as a surrogate for central sensitization (CS), in women with a painful temporomandibular disorder (TMD) and without other painful conditions. Painful TMDs, depression, and non-specific physical symptoms (NSPS) were classified according to the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). The amount of pain in the trigeminal and extra-trigeminal areas was determined using a visual analogue scale (0-100mm) after the application of a vibrotactile stimulus and assessment of the pressure pain threshold (PPT). Statistical tests (Fisher's, χ2, and Mann-Whitney) were performed, with a significance level of 5%. The sample comprised 45 women (mean age 37.5 years; 16 with a painful TMD) who were free of any headache, fibromyalgia, or other painful condition. Painful TMD was associated with higher pain sensitivity and lower PPT values in the trigeminal (P<0.01) and extra-trigeminal regions (P<0.01). The presence of depression contributed significantly to increased pain sensitivity. The presence of hyperalgesia and allodynia in both the trigeminal and extra-trigeminal regions among women with a painful TMD indicated the presence of CS. Changes involving the central nervous system should be considered during the evaluation and management of patients with a painful TMD.

Obesity as a risk factor for temporomandibular disorders.

We conducted a clinical cross-sectional study to evaluate the association between obesity and the presence of painful temporomandibular disorders (TMD), controlling for age, gender, presence of migraine, depression, non-specific somatic symptoms and obstructive sleep apnoea syndrome (OSAS) in an adult population. A total of 299 individuals (76·6% women) with a mean age of 36·8 ± 12·8 years were evaluated. TMD were classified using the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). Depression and non-specific somatic symptoms were scored by the Symptom Checklist-90, while pain and disability was rated by the Graded Chronic Pain Scale. Bioimpedanciometry (BIA) was used to assess obesity through total body fat percentage. Migraine was diagnosed according to the criteria of the International Classification of Headache Disorders-2 (ICHD-2). OSAS was classified according to the Berlin Questionnaire. We performed univariate and multivariate models, chi-square tests and odds ratios (OR) with 95% confidence intervals (CI). In the single regression analysis, TMD-pain was significantly associated with total body fat percentage (P = 0·01). In the multivariate analysis, migraine, age, non-specific somatic symptoms and OSAS showed to be stronger predictors of TMD-pain, and obesity did not retain in the regression model. The initial association found between obesity and TMD-pain is lost when it was corrected for gender, migraine, non-specific somatic symptoms and OSAS.

Headaches and myofascial temporomandibular disorders: overlapping entities, separate managements?

There are relevant clinical overlaps between some of the painful temporomandibular disorders (TMD) and headache conditions that may hamper the diagnostic process and treatment. A non-systematic search for studies on the relationship between TMD and headaches was carried out in the following databases: PubMed, Cochrane Library and Embase. Important pain mechanisms contributing to the close association and complex relationship between TMD and headache disorders are as follows: processes of peripheral and central sensitisation which take place in similar anatomical areas, the possible impairment of the descending modulatory pain pathways and the processes of referred pain. In addition, the clinical examination does not always provide distinguishing information to differentiate between headaches and TMD. So, considering the pathophysiology and the clinical presentation of some types of headache and myofascial TMD, such overlap can be considered not only a matter of comorbid relationship, but rather a question of disorders where the distinction lines are sometimes hard to identify. These concerns are certainly reflected in the current classification systems of both TMD and headache where the clinical consequences of diagnosis such as headache attributed to or associated with TMD are uncertain. There are several similarities in terms of therapeutic strategies used to manage myofascial TMD and headaches. Considering all these possible levels of interaction, we reinforce the recommendation for multidisciplinary approaches, by a team of oro-facial pain specialists and a neurologist (headache specialist), to attain the most precise differential diagnosis and initiate the best and most efficient treatment.

Factors associated with temporomandibular disorders pain in adolescents.

To gain a better understanding of temporomandibular disorders (TMD) pain in adolescents, it is important to study the factors associated with its presence. Therefore, the aim of this study was to investigate potential predictors for TMD pain in adolescents, thereby including a diversity of factors from the biopsychosocial model to determine the strongest predictors. The sample of this cross-sectional study consisted of 1094 adolescents. The presence of TMD pain was assessed using the RDC/TMD, Axis I. Apart from demographical characteristics, the roles of parafunctional habits, psychosocial aspects, menarche and other bodily pain complaints were evaluated. Single and multiple logistic regression models were used to identify associations between the predictor variables and TMD pain. Painful TMD had a prevalence of 25·5%. Logistic regression analyses showed that TMD pain was associated with sleep bruxism (OR = 1·8 95% CI = 1·34-2·34), awake bruxism (OR = 2·1 95% CI = 1·56-2·83), other parafunctional habits (OR = 2·2 95% CI = 1·17-4·08) and bodily pain complaints (OR = 5·0 95% CI = 3·48-7·28). Parafunctional habits and other bodily pain complaints may play an important role in the presence of TMD pain in adolescents. Of course, it remains unclear whether the observed associations between the investigated factors and the adolescent's TMD pain have a true causal linkage.

Activating cAMP/PKA signaling in skeletal muscle suppresses the ubiquitin-proteasome-dependent proteolysis: implications for sympathetic regulation.

Although we have recently demonstrated that plasma catecholamines induce antiproteolytic effects on skeletal muscle (Graça FA, Gonçalves DAP, Silveira WA, Lira EC, Chaves VE, Zanon NM, Garófalo MAR, Kettelhut IC, Navegantes LCC. Am J Physiol Endocrinol Metab. 305: E1483-E1494, 2013), the role of the muscle sympathetic innervation and, more specifically, norepinephrine (NE) in regulating the ubiquitin (Ub)-proteasome system (UPS) remains unknown. Based on previous findings that chemical sympathectomy acutely reduces UPS activity, we hypothesized that muscle NE depletion induces adrenergic supersensitivity in rat skeletal muscles. We report that surgical sympathetic denervation (SDEN), a condition in which only muscle NE from both hindlimbs is depleted, transiently reduced the overall proteolysis and the UPS activity (∼25%) in both soleus and extensor digitorum longus muscles. This antiproteolytic response was accompanied by increased activity of adenylyl cyclase (112%), levels of cyclic adenosine monophosphate (cAMP; 191%), and the serine phosphorylation of cAMP response element-binding protein (32%). In extensor digitorum longus from normal rats, NE (10(-4) M) in vitro increased the levels of cAMP (115%) and the serine phosphorylation of both cAMP response element-binding protein (2.7-fold) and forkhead box class O1 transcription factor. Similar effects were observed in C2C12 cells incubated with forskolin (10 µM). In parallel, NE significantly reduced the basal UPS (21%) activity and the mRNA levels of atrophy-related Ub-ligases. Similar responses were observed in isolated muscles exposed to 6-BNZ-cAMP (500 µM), a specific PKA activator. The phosphorylation levels of Akt were not altered by SDEN, NE, forskolin or 6-BNZ-cAMP. Our results demonstrate that SDEN induces muscle adrenergic supersensitivity for cAMP leading to the suppression of UPS, and that the suppressive effects of NE on UPS activity and expression of Ub-ligases can be mediated by the activation of cAMP/PKA signaling, with the inhibition of forkhead box class O1 transcription factor.

Insulin suppresses atrophy- and autophagy-related genes in heart tissue and cardiomyocytes through AKT/FOXO signaling.

Insulin is an important regulator of the ubiquitin-proteasome system (UPS) and of lysosomal proteolysis in cardiac muscle. However, the role of insulin in the regulation of the muscle atrophy-related Ub-ligases atrogin-1 and MuRF1 as well as in autophagy, a major adaptive response to nutritional stress, in the heart has not been characterized. We report here that acute insulin deficiency in the cardiac muscle of rats induced by streptozotocin increased the expression of atrogin-1 and MuRF1 as well as LC3 and Gabarapl1, 2 autophagy-related genes. These effects were associated with decreased phosphorylation levels of Akt and its downstream target Foxo3a; this phenomenon is a well-known effect that permits the maintenance of Foxo in the nucleus to activate protein degradation by proteasomal and autophagic processes. The administration of insulin increased Akt and Foxo3a phosphorylation and suppressed the diabetes-induced expression of Ub-ligases and autophagy-related genes. In cultured neonatal rat cardiomyocytes, nutritional stress induced by serum/glucose deprivation strongly increased the expression of Ub-ligases and autophagy-related genes; this effect was inhibited by insulin. Furthermore, the addition of insulin in vitro prevented the decrease in Akt/Foxo signaling induced by nutritional stress. These findings demonstrate that insulin suppresses atrophy- and autophagy-related genes in heart tissue and cardiomyocytes, most likely through the phosphorylation of Akt and the inactivation of Foxo3a.

Airway space changes after maxillomandibular counterclockwise rotation and mandibular advancement with TMJ Concepts® total joint prostheses: three-dimensional assessment.

This study focused on three-dimensional (3D) airway space changes and stability following simultaneous maxillomandibular counterclockwise rotation, mandibular advancement, and temporomandibular joint (TMJ) reconstruction with custom-made total joint prostheses (TMJ Concepts(®)). Cone beam computed tomography (CBCT) scans of 30 consecutive female patients with irreversibly compromised TMJs were obtained at the following intervals: T1, presurgery; T2, immediately after surgery; and T3, at least 6 months after surgery. The CBCT volumetric datasets were analysed with Dolphin Imaging(®) software to evaluate surgical and postsurgical changes to oropharyngeal airway parameters. The average changes in airway surface area (SA), volume (VOL), and minimum axial area (MAA) were, 179.50 mm(2), 6302.60 mm(3), and 92.23 mm(2), respectively, at the longest follow-up (T3-T1) (P≤0.001). Significant correlations between the amount of mandibular advancement and counterclockwise rotation of the occlusal plane and 3D airway changes were also found (P≤0.01). The results of this investigation showed a significant immediate 3D airway space increase after maxillomandibular counterclockwise rotation and mandibular advancement with TMJ Concepts total joint prostheses, which remained stable over the follow-up period.

Sleep bruxism increases the risk for painful temporomandibular disorder, depression and non-specific physical symptoms.

To explore the relationship between sleep bruxism (SB), painful temporomandibular disorders (TMD) and psychologic status in a cross-sectional study. The sample consisted of 272 individuals. The Research Diagnostic Criteria for TMD (RDC/TMD) was used to diagnose TMD; SB was diagnosed by clinical criteria proposed by The American Academy of Sleep Medicine. The sample was divided into four groups: (1) patients without painful TMD and without SB, (2) patients without painful TMD and with SB, (3) patients with painful TMD and without SB and (4) patients with painful TMD and with SB. Data were analysed by Odds Ratio test with a 95% confidence interval. Patients with SB had an increased risk for the occurrence of myofascial pain (OR = 5·93, 95% CI: 3·19-11·02) and arthralgia (2·34, 1·58-3·46). Group 3 had an increased risk for moderate/severe depression and non-specific physical symptoms (10·1, 3·67-27·79; 14·7, 5·39-39·92, respectively), and this risk increased in the presence of SB (25·0, 9·65-64·77; 35·8, 13·94-91·90, respectively). SB seems to be a risk factor for painful TMD, and this in turn is a risk factor for the occurrence of higher depression and non-specific physical symptoms levels, but a cause-effect relationship could not be established.

Confiabilidade de um formulário para diagnóstico da severidade da disfunção temporomandibular / Reliability of a questionnaire for diagnosing the severity of temporomandibular disorder

CONTEXTUALIZAÇÃO: Disfunção temporomandibular (DTM) é um termo coletivo que engloba vários problemas clínicos envolvendo a musculatura da mastigação, as articulações temporomandibulares (ATM) e suas estruturas associadas, com alta prevalência nas populações. OBJETIVOS: Sabendo-se que estudos brasileiros vêm utilizando o instrumento proposto por Da Fonseca et al. (1994) para diagnóstico da severidade desta disfunção, realizou-se este estudo com o objetivo de verificar e estimar a consistência interna e a reprodutibilidade do mesmo. MÉTODOS: O delineamento amostral adotado foi o probabilístico, e participaram 1230 indivíduos moradores da cidade de Ribeirão Preto (SP), maiores de 18 anos de idade. As entrevistas foram realizadas por um único entrevistador por meio de ligações telefônicas. Para estudo da consistência interna, calculou-se o Coeficiente de Kuder-Richardson (kr-20) e para estimar a reprodutibilidade, utilizou-se a estatística Kappa (κ). RESULTADOS: A consistência interna do formulário foi de 0,5594, apontando para uma validação abaixo do desejado. Observou-se maior contribuição das questões 1, 2, 3, 6 e 7 para o coeficiente kr-20 total e maior consistência do instrumento quando composto apenas pelas mesmas (0,7044). Observou-se reprodutibilidade "Boa" e "Ótima" para as questões. CONCLUSÕES: Frente ao exposto, sugere-se que o formulário proposto por Da Fonseca et al. (1994) seja adaptado, ficando composto apenas pelas questões 1, 2, 3, 6 e 7 da versão inicial, colaborando, assim, para aumento da confiabilidade do instrumento. Deve-se ressaltar ainda a necessidade da realização de estudos de validade para assegurar adequadas características psicométricas à nova versão do instrumento.

BACKGROUND: Temporomandibular disorder (TMD) is a collective term that encompasses many clinical problems involving the masticatory muscles, temporomandibular joints (TMJ) and associated structures and it has high prevalence among populations. OBJECTIVES: Because Brazilian studies have used the instrument proposed by Da Fonseca et al. (1994) to diagnose the severity of TMD, this study was conducted to investigate and estimate the internal consistency and reproducibility of this method. METHODS: We used a probability sampling design to select 1230 participants over the age of 18 years who were living in the city of Ribeirão Preto, SP, Brazil. The interviews were conducted by a single interviewer over the phone. The internal consistency was analyzed by calculating the Kuder-Richardson coefficient (kr-20), and kappa statistics (κ) were used to estimate the reproducibility. RESULTS: The internal consistency of the questionnaire was 0.5594, thus indicating that validation was lower than desired. Questions 1, 2, 3, 6 and 7 had greater contribution towards the total kr-20 coefficient, and the consistency of the instrument was higher when it was composed only of these questions (0.7044). "Good" and "Excellent" reproducibility was observed for these same questions. CONCLUSIONS: Based on these data, it is suggested that the questionnaire proposed by Da Fonseca et al. (1994) should be adapted to include only questions 1, 2, 3, 6 and 7 of the initial version. This would help improve the reliability of the instrument. The need for validation studies must also be emphasized to ensure that the new version of the instrument has adequate psychometric characteristics.

Maxillo-mandibular counter-clockwise rotation and mandibular advancement with TMJ Concepts total joint prostheses: part IV--soft tissue response.

The purpose of this study was to evaluate soft tissue response to maxillo-mandibular counter-clockwise rotation, with TMJ reconstruction and mandibular advancement using TMJ Concepts total joint prostheses, and maxillary osteotomies in 44 females. All patients were operated at Baylor University Medical Center, Dallas TX, USA, by one surgeon (Wolford). Eighteen patients had genioplasties with either porous block hydroxyapatite or hard tissue replacement implants (Group 2) 26 had no genioplasty (Group 1). Surgically, the maxilla moved forward and upward by counter-clockwise maxillo-mandibular rotation with greater horizontal movement in Group 2. Vertically, both groups showed diversity of maxillo-mandibular mean movement. Group 1 showed a consistent 1: 0.97 ratio of hard to soft tissue advancement at pogonion; Group 2 results were less consistent, with ratios between 1: 0.84 and 1: 1.02. Horizontal changes in upper lip morphology after maxillary advancement/impaction, VY closure, and alar base cinch sutures showed greater movement in both groups, than observed in hard tissue. Counter-clockwise rotation of the maxillo-mandibular complex using TMJ Concepts total joint prostheses resulted in similar soft tissue response as previously reported for traditional maxillo-mandibular advancement without counter-clockwise rotation of the occlusal plane. The association of chin implants, in the present sample, showed higher variability of soft tissue response.

Maxillo-mandibular counter-clockwise rotation and mandibular advancement with TMJ Concepts total joint prostheses: part II--airway changes and stability.

The purpose of this study was to evaluate the anatomical changes and stability of the oropharyngeal airway and head posture following TMJ reconstruction and mandibular advancement with TMJ Concepts custom-made total joint prostheses and maxillary osteotomies with counter-clockwise rotation of the maxillo-mandibular complex. All patients were operated at Baylor University Medical Center, Dallas TX, USA, by one surgeon (Wolford). The lateral cephalograms of 47 patients were analyzed to determine surgical and post-surgical changes of the oropharyngeal airway, hyoid bone and head posture. Surgery increased the narrowest retroglossal airway space 4.9 mm. Head posture showed flexure immediately after surgery (-5.6+/-6.7 degrees) and extension long-term post surgery (1.8+/-6.7 degrees); cervical curvature showed no significant change. Surgery increased the distances between the third cervical vertebrae and the menton 11.7+/-9.1 mm and the third cervical vertebrae and hyoid 3.2+/-3.9 mm, and remained stable. The distance from the hyoid to the mandibular plane decreased during surgery (-3.8+/-5.8 mm) and after surgery (-2.5+/-5.2 mm). Maxillo-mandibular advancement with counter-clockwise rotation and TMJ reconstruction with total joint prostheses produced immediate increase in oropharyngeal airway dimension, which was influenced by long-term changes in head posture but remained stable over the follow-up period.

Caracterizaçäo de herpesvírus bovinos tipos 1 (BHV-1) e 5 (BHV-5) com anticorpos monoclonais / Monoclonal antibody characterization of bovine herpesviruses types 1 (BHV-1) and 5 (BHV-5)

O perfil antigênico de 45 herpesvírus (44 de bovinos, sendo seis amostras de referência de BHV-1 e 15 prováveis BHV-1; três amostras de referência de BHV-5 e 20 prováveis BHV-5) e uma amostra de herpesvírus bubalino (BuHV) foi examinado com um painel de anticorpos monoclonais (Acms) produzidos contra antígenos de herpesvírus bovinos. Para os exames, foi utilizada a prova de imunoperoxidase (IPX) sobre cultivos de células infectadas, tendo os Acms como anticorpos primários. A determinaçäo dos padröes de reatividade das amostras de vírus frente aos Acms permitiu a diferenciaçäo entre os tipos 1 e 5. Todas as amostras isoladas de casos de encefalite apresentaram perfil de BHV-5. Quatro amostras de BHV-5 isoladas de áreas geograficamente distintas apresentaram perfís de reatividade diferenciados em relaçäo às demais amostras do tipo 5. Duas amostras de vírus com perfil antigênico de BHV-5 foram isoladas de sêmen de animais infectados. Estes resultados comprovam a utilidade da caracterizaçäo antigênica com este painel de Acms na tipagem de amostras de BHV-1 e BHV-5