Odontología minimamente invasiva para el tratamiento del desgaste dental / Minimally invasive dentistry for the treatment of tooth wear

El desgaste dental patológico, ocasionado por el bruxismo, es una alteración que se observa frecuentemente en la clínica odontológica. En este caso clínico fue realizado un procedimiento mínimamente invasivo, a fin de restablecer una guía anterior alterada por desgaste dental, asociado a otros tratamientos, tales como, el blanqueamiento dental y el ajuste oclusal. Este abordaje restaurador directo, asociado a las otras técnicas, posibilitó la máxima preservación de estructura dental remanente, alcanzando un excelente resultado estético y funcional, con una correcta adaptación del sistema estomatognático. (AU)

The dental tooth wear due to bruxism is an alteration that is seen frequently in the today ́s clinic. In this clinical case, a mini-mal invasive procedure was conducted aiming to restore the altered anterior guide by bruxism, associating other treatments, such as occlusal adjustment and bleaching. This direct restorative approach associated to the aforementioned treatments, made possible the maximum preservation of the tooth structure, reaching excellent functional and esthetic results, with a correct adaptation of the stomatognathic system. (AU)

Multiplex-PCR for differentiation of Mycobacterium bovis from Mycobacterium tuberculosis complex

We evaluated a multiplex-PCR to differentiate Mycobacterium bovis from M. tuberculosis Complex (MTC) by one step amplification based on simultaneous detection of pncA 169C > G change in M. bovis and the IS6110 present in MTC species. Our findings showed the proposed multiplex-PCR is a very useful tool for complementation in differentiating M. bovis from other cultured MTC species.

Use of the polymerase chain reaction to detect Mycobacterium leprae in urine

Leprosy is an infectious disease caused by Mycobacterium leprae. The polymerase chain reaction (PCR) has been applied to detect M. leprae in different clinical samples and urine seems to be attractive for this purpose. PCR was used to improve the sensitivity for diagnosing leprosy by amplifying a 151-bp PCR fragment of the M. leprae pra gene (PCR-Pra) in urine samples. Seventy-three leprosy patients (39 males and 34 females, 14 to 78 years old) were selected for leprosy diagnosis at a reference laboratory in Maringá, PR, Brazil. Of these, 36 were under anti-leprosy multidrug therapy with dapsone and rifampicin for tuberculoid (TT) and dapsone, rifampicin and clofazimine for borderline (BB) and lepromatous (LL) forms. The control group contained 50 healthy individuals without any clinical history of leprosy. DNA isolated from leprosy patients’ urine samples was successfully amplified by PCR-Pra in 46.6 percent (34/73) of the cases. The positivity of PCR-Pra for patients with the TT form was 75 percent for both patients under treatment and non-treated patients (P = 0.1306). In patients with the LL form, PCR-Pra positivity was 52 and 30 percent for patients under treatment and non-treated patients, respectively (P = 0.2386). PCR-Pra showed a statistically significant difference in detecting M. leprae between the TT and LL forms of leprosy in patients under treatment (P = 0.0033). Although the current study showed that the proposed PCR-Pra has some limitations in the detection of M. leprae, this method has the potential to be a useful tool for leprosy diagnosis mainly in TT leprosy where the AFB slit-skin smear is always negative.

Uso del aparato bucal "PMpositioner" en el tratamiento del ronquido y la apnea obstructiva del sueño / Use of PMpositioner oral appliance in the treatment of snoring and mild obstructive sleep apnea

La elección de un aparato bucal apropiado para lograr los mejores resultados en el tratamiento de la apnea obstructiva del sueño es importante. El objetivo de este estudio fue evaluar el efecto de un aparato bucal específico, el PMPositioner, para el tratamiento del ronquido y la apnea obstructiva del sueño leve, a través de polisomnografía y la Escala de Epworth del sueño, después de seis meses de uso del mencionado aparato. Se incluyeron en el estudio 17 pacientes divididos en dos grupos: un grupo de roncadores (n=7) y otro grupo (n=10) con apnea obstructiva leve. Los resultados fueron significativos para el segundo grupo, revelando una disminución en el índice de apnea-hipoapnea de 7,4 +- 5,0 a 3,0 +- 2,5 (p<0.05), aumento del sueño REM de 16,0+-4,0 a 19+-6,0 y una mejora de la somnolencia en la Escala Epworth de 12,5+-5,4 a 7,4+-2,4. Se constató una disminución en los ronquidos y los síntomas subjetivos. PMPositioner, fue efectivo en el tratamiento de los ronquidos y la apnea obstructiva leve, la reducción de la somnolencia y de otros síntomas.

The choice of an adequate oral appliance is very important in the treatment of obstructive sleep apnea. The present study evaluated the effect of PMPositioner for the treatment of snoring and mild obstructive sleep apneathrough polysomnography and Epworth Sleep Scale prior to treatment and after six months. Seventeen patients, divided into 2 groups, snoring (n=7) and 10 with mild obstructive sleep apnea were enrolled. The results were significant for obstructive sleep apnea group reveling a decrease in the apnea/hypopnea index from 7.4±5.0 to 3.0±2.5 (p<0.05), anincrease in oxigen saturation from 88.0±6.0 to 90.0±2.8 (p<0.05), an increase in REM sleep from 16.0±6.0 to 19±4.0 and a sleepiness improved in from 12.5±5.4 to 7.4±2.4. Furthermore, it was noticed an improvement in snoring and subjective symptoms. The PM Positioner is efficient in the treatment of snoring and mild obstructive sleep apnea and in reduction of sleepness and others symptoms.

HFE gene mutations and iron status of Brazilian blood donors

Mutations of the HFE and TFR2 genes have been associated with iron overload. HFE and TFR2 mutations were assessed in blood donors, and the relationship with iron status was evaluated. Subjects (N = 542) were recruited at the Hemocentro da Santa Casa de São Paulo, São Paulo, Brazil. Iron status was not influenced by HFE mutations in women and was independent of blood donation frequency. In contrast, men carrying the HFE 282CY genotype had lower total iron-binding capacity (TIBC) than HFE 282CC genotype carriers. Men who donated blood for the first time and were carriers of the HFE 282CY genotype had higher transferrin saturation values and lower TIBC concentrations than those with the homozygous wild genotype for the HFE C282Y mutation. Moreover, in this group of blood donors, carriers of HFE 63DD plus 63HD genotypes had higher serum ferritin values than those with the homozygous wild genotype for HFE H63D mutation. Multiple linear regression analysis showed that HFE 282CY leads to a 17.21 percent increase (P = 0.018) and a 83.65 percent decrease (P = 0.007) in transferrin saturation and TIBC, respectively. In addition, serum ferritin is influenced by age (3.91 percent, P = 0.001) and the HFE 63HD plus DD genotype (55.84 percent, P = 0.021). In conclusion, the HFE 282Y and 65C alleles were rare, while the HFE 63D allele was frequent in Brazilian blood donors. The HFE C282Y and H63D mutations were associated with alterations in iron status in blood donors in a gender-dependent manner.

Polimorfismos no receptor Scavenger classe B tipo I e sua relação com perfil lipídico sérico e resposta a atorvastatina em indivíduos brasileiros / Scavenger receptor class B type I polymorphisms and its relation with serum lipids profile and response to atorvastatin in Braszilian individuals

O receptor scavenger BI (SR-BI) é um componente chave do transporte reverso do colesterol. Polimosfismos no gene SCARB1 foram associados com variações no perfil lipídico e outros de risco cardiovascular. Os polimosfismos de nucleotídeo único In5C>T e Ex8C>T no SCARB1 e medidas de lípides e apolipoproteínas foram avaliadas em 79 hipercolesterolêmicos (HC) e 173 normolipidêmicos (NL) provenientes do Brasil. Pacientes HC foram tratados com atorvastatina (10mg/dia/4semanas). Os polimosfismos foram identificados por PCR-RFLP. Os indivíduos HC portadores dos genótipos In5CC+TT mostraram concentrações mais elevadas de LDL-C, apoB, e menores da relação apoAI/apoB. No grupo NL, os genótipos In5CC+TT foram associados com concentrações maiores de LDL-C. Os indivíduos HC portadores de genótipo Ex8CC tiveram uma variação menor da razão apoAI/apoB em resposta à atorvastatina (p<0,05). Nos Hc portadores do haplótipo Ex8CC+CT/In5CT+TT tiveram valores basais elevados de LDL-C e relação apoAI/apoB diminuída. Após o tratamento com atorvastatina, os indivíduos Hc portadores do haplótipo Ex8CC/In5CC tiveram uma variação menor na relação apoAI/apoB. Os genótipos In5CT+TT no SCANB1 conferem um perfil lipídico mais aterogênico. O genótipo Ex8CC e o haplótipo Ex8CC estão associados com uma resposta à atorvastatina menor da razão apoAI/apoB na nossa população.

Aplicação da tecnologia de microarrays em periodontia / The use of microarray´s technique in Periodontics

O objetivo deste estudo foi revisar a literatura sobre a utilização da tecnologia de microarrays e sua aplicação na Periodontia. A tecnologia de microarrays, ou microarranjos, tem demonstrado importante papel nas pesquisas, influenciando na determinação de novos conceitos no campo da etiopatogenia e diagnósticos moleculares e definindo padrões de expressão gênica e proteica. Abrange o estudo de processos biológicos moleculares importantes na patogênese de doenças complexas, notadamente nas áreas relacionadas a doenças metabólicas, auto- imunes, oncológicas, cardiovasculares, sendo mais recentemente aplicada nas pesquisas em Periodontia. Nesta área a aplicação dessa técnica tem auxiliado no entendimento das interações celulares nos processos de saúde e doença, e também na diferenciação de respostas a estímulos específicos, como a interação com micro-organismos, apontando diferenças entre as expressões gênicas nessas diferentes condições. Sendo assim, concluímos que a aplicação da tecnologia de microarrays em Periodontia pode sugerir um perfil gênico de susceptibilidade à doença. No entanto, há a necessidade de uma padronização da técnica para validar os resultados obtidos.

The aim of this study was to review the literature about the microarray technology and its application in periodontics. Microarray analysis has been important for researchers helping the development of new concepts in the field of etiopathogenisis and molecular diagnostic, and also defining standards for gene and protein expression. It embraces the study of important molecular process, which is part of the pathogenesis of complex diseases, such as metabolic, selfimmunes, oncologic and cardiovascular diseases. Recently, microarray analysis has also been applied in periodontal research. The use of such technique has helped understanding the cellular interactions during periodontal health and disease, and also defining different responses to specific stimuli patterns, i.e, interactions between microorganisms and host cells. Differences in gene’s expression profiles can also be seen in those conditions. In conclusion, the use of microarrays technique in periodontics might be useful to indicate gene profiles related to disease susceptibility. However, it’s necessary the standardization of this technique to validate its results.

Lipoprotein lipase PvuII polymorphism is associated with variations in serum lipid levels in non-diabetic pregnant women

The aim of the present study was to determine if there is an association between the single nucleotide polymorphisms (SNPs) of the lipoprotein lipase (LPL) and apolipoprotein E (apo E) genes and the serum lipid profile in pregnancy and puerperium. Non-diabetic women of European descent in the third semester of pregnancy (N = 120) were selected. Those with diseases or other condition that could modify their lipid profile were excluded from the study (N = 32). Serum lipids were measured by routine laboratory procedures and genomic DNA was extracted by a salting out method. LPL (PvuII and HindIII) and apo E (HhaI) SNPs were detected by the polymerase chain reaction and restriction fragment length polymorphism. Categorical and continuous variables were compared by the chi-square test and Student t-test or ANOVA, respectively. Women carrying the LPL P1P1 genotype had higher serum LDL cholesterol (N = 21; 155 ± 45 mg/dL) than women carrying the P1P2/P2P2 genotypes (N = 67; 133 ± 45 mg/dL; P = 0.032). During the puerperium period, serum levels of triglycerides and VLDL cholesterol were significantly reduced in women carrying the P1P1 (73 percent, P = 0.006) and P1P2 (51 percent, P = 0.002) genotypes but not in women carrying the P2P2 genotype (23 percent, P > 0.05). On the other hand, serum concentrations of lipids did not differ between the LPL HindIII and apo E genotypes during pregnancy and after delivery. We conclude that LPL PvuII SNP is associated with variations in serum lipids during pregnancy and the puerperal period in non-diabetic women.

Efeito da posição relativa do joelho sobre a carga mecânica interna durante o agachamento / Effect of relative knee position on internal mechanical loading while squatting

CONTEXTUALIZAÇÃO: Existe um conceito difundido entre professores de educação física, fisioterapeutas e ortopedistas de que o joelho não deve ser demasiadamente anteriorizado em relação à ponta do pé na direção ântero-posterior durante qualquer tipo de agachamento de modo a diminuir a carga mecânica sobre o joelho. No entanto, são escassas as evidências quantitativas que corroboram esse conceito. OBJETIVO: Estimar forças e torque na articulação do joelho em indivíduos saudáveis durante o exercício de agachamento livre com peso em dois modos diferentes de execução: a) joelho não ultrapassando a linha vertical que passa pelos dedos do pé; b) joelho ultrapassando essa linha vertical. MÉTODOS: Análise tridimensional com câmeras de vídeo e plataforma de força do movimento de agachamento em dez adultos jovens saudáveis. Quinze repetições em cada condição do agachamento por sujeito foram executadas sobre uma plataforma de força. As forças e torques articulares no tornozelo, joelho e quadril foram calculados pelo procedimento de dinâmica inversa. RESULTADOS: Os resultados obtidos mostram que o pico do torque no joelho é, em média, cerca de 38 ± 31 por cento e a força patelofemoral é, em média, cerca de 28 ± 27 por cento maiores na condição ultrapassando o joelho que na condição não ultrapassando o joelho. CONCLUSÕES: Esses resultados demonstram que não ultrapassar o joelho da linha do pé diminui a força de compressão patelofemoral, levando assim a uma menor solicitação mecânica nessa articulação.

BACKGROUND: There is a widespread notion among physical education teachers, physical therapists and orthopedists that, during any type of squatting, the knee should not be brought forward too much in relation to the tip of the foot, so as to reduce the mechanical loading on the knee. However, there is little quantitative evidence to corroborate this notion. OBJECTIVE: To estimate the forces and torque on the knee joint in healthy individuals during free squatting exercises using weights performed in two different ways: a) knee not going beyond a vertical line going through the toes; b) knee going beyond this vertical line. METHOD: Three-dimensional analysis using video cameras and a force platform was performed on squatting movements performed by ten healthy young adults. Fifteen repetitions of each of the two squatting conditions were performed by each subject on the force plate. The forces and joint torque at the ankle, knee and hip were calculated using an inverse dynamic procedure. RESULTS: The results obtained showed that the mean peak torque on the knee was around 38 ± 31 percent greater, and the mean patellofemoral force was around 28 ± 27 percent greater, when the knee went beyond the tip of the foot, than when it did not. CONCLUSIONS: These results demonstrate that, when the knee does not go beyond the line of the foot, the patellofemoral compression force is less, which leads to lower mechanical demand on this joint.

High baseline serum total and LDL cholesterol levels are associated with MDR1 haplotypes in Brazilian hypercholesterolemic individuals of European descent

The MDR1 gene encodes the P-glycoprotein, an efflux transporter with broad substrate specificity. P-glycoprotein has raised great interest in pharmacogenetics because it transports a variety of structurally divergent drugs, including lipid-lowering drugs. The synonymous single-nucleotide polymorphism C3435T and the nonsynonymous single-nucleotide polymorphism G2677T/A in MDR1 have been indicated as potential determinants of variability in drug disposition and efficacy. In order to evaluate the effect of G2677T/A and C3435T MDR1 polymorphisms on serum levels of lipids before and after atorvastatin administration, 69 unrelated hypercholesterolemic individuals from São Paulo city, Brazil, were selected and treated with 10 mg atorvastatin orally once daily for four weeks. MDR1 polymorphisms were analyzed by PCR-RFLP. C3435T and G2677T polymorphisms were found to be linked. The allelic frequencies for C3435T polymorphism were 0.536 and 0.464 for the 3435C and 3435T alleles, respectively, while for G2677T/A polymorphism allele frequencies were 0.580 for the 2677G allele, 0.384 for the 2677T allele and 0.036 for the 2677A allele. There was no significant relation between atorvastatin response and MDR1 polymorphisms (repeated measures ANOVA; P > 0.05). However, haplotype analysis revealed an association between T/T carriers and higher basal serum total (TC) and LDL cholesterol levels (TC: 303 ± 56, LDL-C: 216 ± 57 mg/dl, respectively) compared with non-T/T carriers (TC: 278 ± 28, LDL-C: 189 ± 24 mg/dl; repeated measures ANOVA/Tukey test; P < 0.05). These data indicate that MDR1 polymorphism may have an important contribution to the control of basal serum cholesterol levels in Brazilian hypercholesterolemic individuals of European descent.

Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis

Three recombinant antigens of Treponema pallidum Nichols strain were fused with GST, cloned and expressed in Escherichia coli, resulting in high levels of GST-rTp47 and GST-rTp17 expression, and supplementation with arginine tRNA for the AGR codon was needed to obtain GST-rTp15 overexpression. Purified fusion protein yields were 1.9, 1.7 and 5.3 mg/l of cell culture for GST-rTp47, GST-rTp17 and GST-rTp15, respectively. The identities of the antigens obtained were confirmed by automated DNA sequencing using ABI Prism 310 and peptide mapping by Finningan LC/MS. These recombinant antigens were evaluated by immuno-slot blot techniques applied to 137 serum samples from patients with a clinical and laboratory diagnosis of syphilis (61 samples), from healthy blood donors (50 samples), individuals with sexually transmitted disease other than syphilis (3 samples), and from individuals with other spirochetal diseases such as Lyme disease (20 samples) and leptospirosis (3 samples). The assay had sensitivity of 95.1 percent (95 percent CI, 86.1 to 98.7 percent) and a specificity of 94.7 percent (95 percent CI, 87.0 to 98.7 percent); a stronger reactivity was observed with fraction rTp17. The immunoreactivity results showed that fusion recombinant antigens based-immuno-slot blot techniques are suitable for use in diagnostic assays for syphilis.

Interferon-alpha receptor 1 mRNA expression in peripheral blood mononuclear cells is associated with response to interferon-alpha therapy of patients with chronic hepatitis C

Interferon (IFN)-alpha receptor mRNA expression in liver of patients with chronic hepatitis C has been shown to be a response to IFN-alpha therapy. The objective of the present study was to determine whether the expression of mRNA for subunit 1 of the IFN-alpha receptor (IFNAR1) in peripheral blood mononuclear cells (PBMC) is associated with the response to IFN-alpha in patients with chronic hepatitis C. Thirty patients with positive anti-HCV and HCV-RNA, and abnormal levels of alanine aminotransferase in serum were selected and treated with IFN-alpha2b for one year. Those with HBV or HIV infection, or using alcohol were not included. Thirteen discontinued the treatment and were not evaluated. The IFN-alpha response was monitored on the basis of alanine aminotransferase level and positivity for HCV-RNA in serum. IFNAR1-mRNA expression in PBMC was measured by reverse transcription-polymerase chain reaction before and during the first three months of therapy. The results are reported as IFNAR1-mRNA/á-actin-mRNA ratio (mean ñ SD). Before treatment, responder patients had significantly higher IFNAR1-mRNA expression in PBMC (0.67 ñ 0.15; N = 5; P < 0.05) compared to non-responders (0.35 ñ 0.17; N = 12) and controls (0.30 ñ 0.16; N = 9). Moreover, IFNAR1-mRNA levels were significantly reduced after 3 months of treatment in responders, whereas there were no differences in IFNAR1 expression in non-responders during IFN-alpha therapy. Basal IFNAR1-mRNA expression was not correlated with the serum level of alanine and aspartate aminotransferases or the presence of cirrhosis. The present results suggest that IFNAR1-mRNA expression in PBMC is associated with IFN-alpha response to hepatitis C and may be useful for monitoring therapy in patients with chronic hepatitis C.

Polymorphisms of the low-density lipoprotein receptor gene in Brazilian individuals with heterozygous familial hypercholesterolemia

Familial hypercholesterolemia (FH) is a metabolic disorder inherited as an autosomal dominant trait characterized by an increased plasma low-density lipoprotein (LDL) level. The disease is caused by several different mutations in the LDL receptor gene. Although early identification of individuals carrying the defective gene could be useful in reducing the risk of atherosclerosis and myocardial infarction, the techniques available for determining the number of the functional LDL receptor molecules are difficult to carry out and expensive. Polymorphisms associated with this gene may be used for unequivocal diagnosis of FH in several populations. The aim of our study was to evaluate the genotype distribution and relative allele frequencies of three polymorphisms of the LDL receptor gene, HincII1773 (exon 12), AvaII (exon 13) and PvuII (intron 15), in 50 unrelated Brazilian individuals with a diagnosis of heterozygous FH and in 130 normolipidemic controls. Genomic DNA was extracted from blood leukocytes by a modified salting-out method. The polymorphisms were detected by PCR-RFLP. The FH subjects showed a higher frequency of A+A+ (AvaII), H+H+ (HincII1773) and P1P1 (PvuII) homozygous genotypes when compared to the control group (P<0.05). In addition, FH probands presented a high frequency of A+ (0.58), H+ (0.61) and P1 (0.78) alleles when compared to normolipidemic individuals (0.45, 0.45 and 0.64, respectively). The strong association observed between these alleles and FH suggests that AvaII, HincII1773 and PvuII polymorphisms could be useful to monitor the inheritance of FH in Brazilian families

Expression and biological activity of two recombinant polypeptides related to subunit 1 of the interferon-alpha receptor

Abnormal production of interferon alpha (IFN-a) has been found in certain autoimmune diseases and can be also observed after prolonged therapy with IFN-a. IFN-a can contribute to the pathogenesis of allograft rejection in bone marrow transplants. Therefore, the development of IFN-a inhibitors as a soluble receptor protein may be valuable for the therapeutic control of these diseases. We have expressed two polypeptides encoding amino acids 93-260 (P1) and 261-410 (P2) of the extracellular domain of subunit 1 of the interferon-a receptor (IFNAR 1-EC) in E. coli. The activities of the recombinant polypeptides and of their respective antibodies were evaluated using antiproliferative and antiviral assays. Expression of P1 and P2 polypeptides was achieved by transformation of cloned plasmid pRSET A into E. coli BL21(DE3)pLysS and by IPTG induction. P1 and P2 were purified by serial sonication steps and by gel filtration chromatography with 8 M urea and refolded by dialysis. Under reducing SDS-PAGE conditions, the molecular weight of P1 and P2 was 22 and 17 kDa, respectively. Polyclonal anti-P1 and anti-P2 antibodies were produced in mice. P1 and P2 and their respective polyclonal antibodies were able to block the antiproliferative activity of 6.25 nM IFN-aB on Daudi cells, but did not block IFN-aB activity at higher concentrations (>6.25 nM). On the other hand, the polypeptides and their respective antibodies did not inhibit the antiviral activity of IFN-aB on Hep 2/c cells challenged with encephalomyocarditis virus.