HPV16 Tumor Associated Macrophages Suppress Antitumor T Cell Responses

Purpose: High-risk human papillomavirus (HPV) is the main etiologic factor for cervical cancer. The severity of HPV-associated cervical lesions has been correlated to the number of infiltrating macrophages. The objective of this work is to characterize the role of tumor-associated macrophages (TAM) on the immune cellular response against the tumor. Experimental Design: We used the HPV16 E6- and E7-expressing TC-1 mouse tumor model to study the effect of TAM on T-cell function in vitro, and depleted TAM, using clodronate-containing liposomes, to characterize its role in vivo. Results: TAM, characterized by the positive expression of CD45, F4/80, and CD11b, formed the major population of infiltrating tumor cells. TAM displayed high basal Arginase I activity, producing interleukin-10 (IL-10); they were resistant to iNOSll activity induction, therefore reversion to M1 phenotype, when stimulated in vitro with lipopolysaccharide/IFN gamma, indicating an M2 phentoype. In cultures of isolated TAM, TAM induced regulatory phenotype, characterized by IL-10 and Foxp3 expression, and inhibited proliferation of CD8 lymphocytes. In vivo, depletion of TAM inhibited tumor growth and stimulated the infiltration of tumors by HPV16 E7(49-57)-specific CD8 lymphocytes, whereas depletion of Gr1(+) tumor-associated cells had no effect. Conclusions: M2-like macrophages infiltrate HPV16-associated tumors causing suppression of antitumor T-cell response, thus facilitating tumor growth. Depletion or phenotype alteration of this population should be considered in immunotherapy strategies.

Prevalência da infecção por Chlamydia Trachomatis em mulheres atendidas no Programa de rastreamento do câncer de colo do útero

Genital infection with Chlamydia trachomatis is now recognized as one of the most prevalent sexually transmitted infections (STDs). Despite major advances in laboratory diagnosis techniques, primarily the character of asymptomatic chlamydial infection in both men and in women constitutes the basis for the formation of reservoirs that perpetuate transmission and acquisition of this and other STDs. The asymptomatic in women favors the rise of infection to the upper genital tract, causing injuries that can result in infertility. An examination of population screening for early detection and treatment of asymptomatic infections is the key step in combating this major public health problem. The present study aimed to evaluate the prevalence of infection by C. trachomatis in sexually active women attended the screening program for cervical cancer of the uterus in health facilities in municipalities in different regions of the State of Rio Grande do Norte, and identify factors that may contribute to the spread of this pathogen and its relationship with the lesions of the uterine cervix. It is a cross-sectional study aimed at detecting the presence of genital tract infection by C. trachomatis either in isolated form or in association with human papilloma virus (HPV) infection in asymptomatic women. Were included in this study, a total sample of 1,134 women aged 13-76, mean 34.4 years, from March 2008 to September 2012. Specimens containing exfoliated cells of the epithelium of the uterine cervix were analyzed by examining Pap cytology for the detection of possible injuries, and the polymerase chain reaction (PCR) for detection of plasmid DNA from C. trachomatis and HPV. Infection with C. trachomatis was detected with overall prevalence rate of 8.1% in the isolated form and 2.8% in co-infection with HPV. The infection was detected in 7.4% of women with normal cytology 11.5% of those with atypical cells of undetermined significance (ASC-US) and 16.7% of those with low-grade squamous intraepithelial lesion (LSIL). We observed an association between C. trachomatis and incidence of low-grade squamous intraepithelial lesion (LSIL). The genital tract infection by C. trachomatis alone was associated with education level, ethnicity and parity, revealing that women with higher education, those of non-white ethnicity and those who had three or more pregnancies were more likely to acquire infection. Levels very close to statistical significance were observed for chronological age, age at first sexual intercourse and first pregnancy. There was no association with marital status, number of sexual partners. Co-infection with C. trachomatis and HPV was detected in 2.3% of women with normal cytology, who had 5.1% in ASC-US and 10.4% in those with LSIL. No association was found between infection C. trachomatis and increased risk of HPV infection, but women with simultaneous infection by both pathogens showed greater risk for LSIL. Co-infection was more prevalent among single women, who had in the first sexual intercourse under 18 years and those who had two or more sexual partners over a lifetime

Avaliação in vitro da eficácia de produtos homeopáticos contendo momordica charantia através de bioensaios

Homeopathic medicines have been used for over two hundred years without the examination of their effects on in vivo and in vitro assays, due to the peculiarity of homeopathic preparations, the high dilution, which creates a challenge for the use of usual analytical techniques of quality control of medicine.Although there is scarcity of literature and variety of experiments, recently there have been some studies with few in vitro assays which have shown positive responses when evaluating the mechanism of action of homeopathic medicines which are able to act on a specific system.The present study aims to evaluate the efficacy of homeopathic products containing Momordica charantia through bioassays.Homeopathic products were tested by the MTT to assess cytotoxicity in RAW 264.7 (macrophage-like cells) and in tumor cells HeLa (human cervical adenocarcinoma cells), CHO K1 (Chinese hamster ovary cells), PANC-1 (human pancreas cancer cells) and PC-3 (human prostate cancer cells), dosage of inflammatory mediators NO, TNF-α and IL-6 released by RAW 264.7 cells, analysis of the death process and cell cycle changes of PC-3 by flow cytometry. The data demonstrate that homeopathic products of Momordica charantia did not show cytotoxicity to RAW 264.7, increased the production of inflammatory mediators by RAW 264.7 synergistically with LPS, showed cytotoxicity to PC-3 with change in its cell cycle inhibiting its proliferation, being the 30CH the most potent sample. Correlation studies were conducted in order to evaluate the possible in vitro applicable models to the quality control of homeopathic products with Momordica charantia. The data showed that the best applicable models in assessing the quality are the MTT to assess cytotoxicity in RAW 264.7 and PC-3 in 24 hours for Momordica charantia fruit products and dosage of NO production by RAW 264.7 with and without LPS

Estratégias de intervenção utilizadas por enfermeiros da ESF do município de Natal/RN no controle do câncer do colo de útero

Cancer of the cervix (cervical cancer) is the second most prevalent cancer among Brazilian women. The high rates of cervical cancer in Brazil justify the implementation of effective strategies to control this, which include actions to promote health, primary prevention, early detection, screening, treatment and palliative care. Despite the existence of the National Programme for Control of the CCU there was no reduction in the incidence and mortality of this disease in Brazil. The Family Health Strategy (FHS) has the potential to facilitate such control and, in this context, one should consider that nurses play a central role. The study aimed to know the general intervention strategies used by nurses FHS of Natal / RN in CCU control, and how specific: analyzing the knowledge of these nurses on the CCU, the actions developed in the ESF for the control of CCU and identify the difficulties faced by them to perform it. This is a descriptive exploratory quantitative developed through a structured interview guide with 106 nurses who have experience in controlling the CCU in FHS teams of Natal / RN. Data analysis was performed using descriptive statistic s. The results pointed to actions taken in the FHS to control the CCU, collection of cervical cancer screening, health education activities, nursing consultation, referral of suspected cases for medical monitoring and active women with abnormal test result . The actions that were not mentioned by the nurses included: forming groups of prevention and health promotion; expand coverage of exams and office hours of consultations, establishment of alternatives to end the pent-up demand in the health units, participation in treatment or rehabilitation process users with the CCU; interventions for pain management, alliances and partnerships with schools, in dustry and the use of protocols. This study can be seen that the practice nurses partially shares to the CCU in Natal / RN. The participants of this study, when asked about the CCU, specifically for signs and symptoms of disease and risk factors in general showed important gaps. Difficulties such as lack of materials for collection of Pap smear; inadequate physical space in the Health Units; pent-up demand in the service, delay in arrival of the test results; obstacles in the actions of referral and counter-referral and cultural factors make the CCU control is compromised. It is believed in this research contributed to a reflection on the importance of the role of nurses in the development of the ESF control actions CCU, pointing out the factors that affect these. It is important to involve all nurses who comprise the ESF as knowledgeable of the risk factors, signs and symptoms, and existing tools for the early detection of cervical cancer in the pursuit of quality improvement actions to promote women`s health, contributing in planning future interventions that may reduce mortality from this disease in Natal / RN.

Avaliação da coluna vertebral: relação entre gibosidade e curvas sagitais por método não-invasivo

Os objetivos do estudo foram avaliar o alinhamento, no plano sagital, da coluna de indivíduos com alterações na medida da gibosidade, comparando com um grupo sem alterações; testar a confiabilidade do instrumento utilizado e verificar se existem correlações entre as medidas da gibosidade e os valores das curvaturas vertebrais. Foram avaliados 40 jovens, divididos em grupo controle - ausência ou presença de gibosidades inferiores a 0,5 cm na curvatura torácica e 0,7 cm na lombar (n=20) e, grupo experimental - gibosidades superiores às descritas (n=20). A gibosidade e as curvaturas no plano sagital foram mensuradas com um instrumento adaptado a um nível d'água e o teste de Adams. As coletas foram realizadas em duas datas distintas, nos dois grupos. Após aplicação do teste Mann-Whitney não foi encontrada diferença entre as ocasiões de coletas e, emparelhando-se os grupos, foi encontrada diferença apenas na medida cervical. Na verificação de existência de relação entre as medidas coletadas, foi encontrada correlação linear (Spearmann) no grupo controle - curvatura torácica e gibosidade torácica; em ambos os grupos - curvaturas torácica e lombar; e no grupo experimental - gibosidade torácica e as curvaturas lombar e sacral e, curvatura sacral e curvaturas torácica e lombar. Pôde-se concluir que a medida da gibosidade tem relações com as curvaturas no plano sagital. Por ser um método confiável, simples e acessível, pode ser reproduzido sem altos custos financeiros e sem causar prejuízo à saúde do paciente.

Internal carotid artery pseudoaneurysm after tonsillectomy treated by endovascular approach - A case report

Surgery on the head and neck region may be complicated by vascular trauma, caused by direct injury on the vascular wall. Lesions of the arteries are more dangerous than the venous one. The traumatic lesion may cause laceration of the artery wall, spasm, dissection, arteriovenous fistula, occlusion or pseudoaneurysm.We present a case of a child with a giant ICA pseudoaneurysm after tonsillectomy, manifested by pulsing mass and respiratory distress, which was treated by endovascular approach, occluding the lesion and the proximal artery with Histoacryl. We reinforce that the endovascular approach is the better way to treat most of the traumatic vascular lesions.

Isolation of coccidioides brasiliensis from armadillos (Dasypus noveminctus) captured in an endemic area of paracoccidioidomycosis

Paracoccidioides brasiliensis, the causative agent of paracoccidioidomycosis (PCM), was first isolated from armadillos from the Amazonian region where the mycosis is uncommon. In the present study, we report on the high incidence of PCM infection in armadillos from a hyperendemic region of the disease. Four nine-banded armadillos (Dasypus novemcinctus) were captured in the endemic area of Botucatu, São Paulo, Brazil, killed by manual cervical dislocation and autopsied under sterile conditions. Fragments of lung, spleen, liver, and mesenteric lymph nodes were processed for histology, cultured on Mycosel agar at 37 degrees C, and homogenized for inoculation into the testis and peritoneum of hamsters. The animals were killed from week 6 to week 20 postinoculation and fragments of liver, lung, spleen, testis, and lymph nodes were cultured on brain heart infusion agar at 37 degrees C. Paracoccidioides brasiliensis was isolated from three armadillos both by direct organ culture and from the liver, spleen, lung, and mesenteric lymph nodes of hamsters. In addition, one positive armadillo presented histologically proven PCM disease in a mesenteric lymph node. The three armadillos isolates (Pb-AL, Pb-A2, and Pb-A4) presented thermodependent dimorphism, urease activity, and casein assimilation, showed amplification of the gp43 gene, and were highly virulent in intratesticularly inoculated hamsters. The isolates expressed the gp43 glycoprotein, the immunodominant antigen of the fungus, and reacted with a pool of sera from PCM patients. Taken together, the present data confirm that armadillos an a natural reservoir of P. brasiliensis and demonstrate that the animal is a sylvan host to the fungus.

Cloning, sequencing and expression analysis of the equine hepcidin gene by real-time PCR

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Periodontal tissue response to coverage of root cavities restored with resin materials: A histomorphometric study in dogs

Background: the purpose of this study was to histomorphometrically evaluate the response of periodontal tissues covering Class V resin restorations in dogs.Methods: After raising a mucoperiosteal flap, bony defects measuring 5 x 5 mm were created on the buccal aspect of the canines of five dogs followed by cavity preparations on the root surface measuring 3 x 3 x 1 mm. Before repositioning the flap to cover the bone defect, the cavities were restored with composite resin (CR) or resin-modified glass ionomer cement (RMGIC) or were left unrestored as control (C). The dogs were euthanized 90 days after surgery. Specimens comprising the tooth and periodontal tissues were removed, processed routinely, cut into longitudinal serial sections in the bucco-lingual direction, and stained with hematoxylin and eosin (H&E) or Masson's trichrome. The most central sections were selected for histomorphometric analysis.Results: Histomorphometric analysis revealed apical migration of epithelial tissue onto the restorative materials (RMGIC and CR). The C group presented significantly longer connective tissue attachment (P < 0.05) than the RMGIC and CR groups and significantly higher bone regeneration (P < 0.05) compared to the RMGIC group. Histologically, the cervical third (CT) of all groups had the most marked chronic inflammatory infiltrate.Conclusions: Within the limits of this study, it can be concluded that the restorative materials used exhibit biocompatibility; however, both materials interfered with the development of new bone and the connective tissue attachment process.

Resolution of a titanium implant fracture after a recurrent trauma

Post-traumatic complications occasionally lead to tooth loss as well as the need for future implants. However, rehabilitation with endosseous osseointegrated implants does not protect the patient from the risk of suffering a new trauma. Implant fracture and the damage of the hexagon are post-traumatic complications that guide the clinician to preparing a more intricate treatment plan. The authors present a clinical case of a recurrent trauma of maxillary implant fracture. The treatment plan was to remove the implants followed by autogenous bone grafting to correct the defect. Two titanium implants were replaced, followed by connective tissue graft after allowing complete the healing process of the bone graft to occur. In the postoperative period of 6 months, satisfactory results have been shown as regards soft and hard tissues wound healing.

Intraoral approach for surgical treatment of Eagle syndrome

Eagle syndrome is symptomatic elongation of the styloid process, or calcification of the stylomandibular ligament. Difficulty with deglutition. phonation. cervical movement. and the sensation of a foreign body in the oropharynx are all symptoms of this syndrome. Its treatment consists of partial removal of the styloid process. leaving it within the range of normality. We describe the technique, report it case, and point Out Sortie of the advantages. such as the simplicity of the procedure, reduced operating time, and lack risks of operative complications. (C) 2008 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

Effect of dentin conditioning time on nanoleakage

Objectives: To study the nanoleakage pattern in the dentin hybrid layer by using different dentin adhesives. The null hypotheses tested in this study were: 1) dentin conditioning time does not affect nanoleakage within the hybrid layer; 2) the type of dentin adhesive used does not affect nanoleakage. Methods: Standardized Class V cavities were prepared in 30 intact human molars on the buccal and lingual surfaces. The specimens were randomly assigned to 2 total-etch dentin adhesives (OptiBond SOLO Plus [OPS, Kerr] and One-Step [ONS, BISCO Inc]) and 2 self-etch dentin adhesives (Clearfil SE Bond [CSE, Kuraray] and Adper Prompt L-Pop [APL, 3M ESPE]). The specimens were etched or conditioned for 15 seconds, 30 seconds or 60 seconds. Upon restoration of the Class V cavities with the proprietary resin composite, the specimens were isolated with nail polish except for a 2.0-mm rim around the restoration, and they were immersed in 50 wt% ammoniacal silver nitrate solution (pH=9.5) for 24 hours followed by 8 hours of immersion in photo-developing solution to reduce the silver ions to metallic silver. The specimens were fixed, dehydrated and processed for FESEM and TEM. Silver penetration was measured along the cervical wall, and data were analyzed with Kruskal-Wallis non-parametric tests at a significance level of 95%. Results: There were no statistically significant differences among the experimental groups for the factor conditioning time (p>0.926). There were significant differences for the variable dentin adhesive (p<0.0001). The least amount of nanoleakage within the hybrid layer occurred with CSE, while ONS resulted in the greatest penetration of silver ions. The adhesives OPS and APL ranked in the intermediary subset. Under TEM, all adhesives resulted in some degree of nanoleakage within the hybrid layer. Both spotted/reticular and water-tree nanoleakage patterns were observed. Significance: Longer conditioning times did not increase nanoleakage within the hybrid layer. Nanoleakage varied with the type of adhesive used.

Morphometrical analysis of cleaning capacity of a hybrid instrumentation in mesial flattened root canals

The cleaning capacity of hybrid and rotary instrumentation techniques in mesial flattened canals of mandibular first molars was evaluated by morphometrical analysis in this study. Twenty human mandibular first molars were randomly assigned into two groups, according to instrumentation technique, as follows: group 1, instrumentation with ProTaper Starter Kit (Dentsply/Maillefer) rotary system; group 2, manual instrumentation using K files (Dentsply/Maillefer) by crown-down technique in middle and apical thirds, cervical preparation with Gates-Glidden #1 and #2 (Dentsply/Maillefer) burs, and to finalise the preparation, ProTaper F2 and F3 rotary files. Serial transverse cross-sections (5 mu m) of the apical third, stained with hematoxylin and eosin, were analysed at 100 x original magnification. The images were submitted to morphometrical analysis with an integration grid to determine the percentage of root canal area with debris. Statiscal analysis (t-Student, P < 0.05) showed significant difference between the techniques (P < 0.05), although neither completely cleaned the root canal.

Marginal adaptation of Class 2 adhesive restorations

Objective: Control of microleakage represents a challenge for posterior composite restorations. The technique for composite placement may reduce microleakage. The null hypothesis of this in vitro study was that centripetal incremental insertion of composite resin would result in less microleakage than that obtained with the oblique incremental technique or bulk technique. Method and Materials: Standardized Class 2 preparations were made in 60 caries-free extracted third molars and randomly assigned to 3 groups ( n = 20): ( 1) oblique incremental insertion technique ( control), ( 2) centripetal incremental insertion technique, and ( 3) bulk insertion. The teeth were restored with a total-etch adhesive and micro-hybrid composite resin. The specimens were isolated with nail varnish except for a 2-mm-wide area around the restoration and then thermocycled ( 1,000 thermal cycles, 5 degrees C/ 55 degrees C; 30-second dwell time). The specimens were immersed in an aqueous solution of 50% silver nitrate for 24 hours, followed by 8 hours of immersion in a photo-developing solution and subsequently evaluated for leakage. The microleakage scores ( 0 to 4) obtained from the occlusal and cervical walls were analyzed with median nonparametric tests ( P <.05). Results: The null hypothesis was rejected. All techniques attained statistically similar dentin microleakage scores ( P =.15). The centripetal insertion technique displayed significantly less microleakage than the oblique technique at the enamel margins ( P =.04). Conclusion: None of the techniques eliminated marginal microleakage in Class 2 preparations. However, in occlusal areas, the centripetal technique performed significantly better than the other techniques.

Intestinal anti-inflammatory activity of paepalantine, an isocoumarin isolated from the capitula of Paepalanthus bromelioides, in the trinitrobenzenesulphonic acid model of rat colitis

The preventative intestinal anti-inflammatory activity of paepalantine, an isocoumarin isolated from the capitula of Paepalarithus bromelioides, was tested in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis. This was performed in two different experimental settings, i. e. when the colonic mucosa is intact or when the mucosa is in process of recovery after an initial insult. The results obtained revealed that the paepalantine pretreatment, at doses of 5 and 10 mg/kg, significantly attenuated the colonic damage induced by TNBS in both situations, as it was evidenced both histologically and biochemically. This beneficial effect was associated with an improvement in the colonic oxidative status, since paepalantine prevented the glutathione depletion that occurred as a consequence of the colonic inflammation. In addition, the intestinal anti-inflammatory effect exerted by this isocoumarin was associated with an inhibition of colonic nitric oxide activity, which is upregulated as a consequence of the inflammatory process. In conclusion, the preventative effect exerted by paepalantine in the TNBS model of rat colitis is probably related with its antioxidant properties.