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Abstract Background De novo retrotransposition of Alu elements has been recognized as a major driver for insertion polymorphisms in human populations. In this study, we exploited Alu-anchored bisulfite PCR libraries to identify evolutionarily recent Alu element insertions, and to investigate their genetic and epigenetic variation. Results A total of 327 putatively recent Alu insertions were identified, altogether represented by 1,762 sequence reads. Nearly all such de novo retrotransposition events (316/327) were novel. Forty-seven out of forty-nine randomly selected events, corresponding to nineteen genomic loci, were sequence-verified. Alu element insertions remained hemizygous in one or more individuals in sixteen of the nineteen genomic loci. The Alu elements were found to be enriched for young Alu families with characteristic sequence features, such as the presence of a longer poly(A) tail. In addition, we documented the occurrence of a duplication of the AT-rich target site in their immediate flanking sequences, a hallmark of retrotransposition. Furthermore, we found the sequence motif (TT/AAAA) that is recognized by the ORF2P protein encoded by LINE-1 in their 5'-flanking regions, consistent with the fact that Alu retrotransposition is facilitated by LINE-1 elements. While most of these Alu elements were heavily methylated, we identified an Alu localized 1.5 kb downstream of TOMM5 that exhibited a completely unmethylated left arm. Interestingly, we observed differential methylation of its immediate 5' and 3' flanking CpG dinucleotides, in concordance with the unmethylated and methylated statuses of its internal 5' and 3' sequences, respectively. Importantly, TOMM5's CpG island and the 3 Alu repeats and 1 MIR element localized upstream of this newly inserted Alu were also found to be unmethylated. Methylation analyses of two additional genomic loci revealed no methylation differences in CpG dinucleotides flanking the Alu insertion sites in the two homologous chromosomes, irrespective of the presence or absence of the insertion. Conclusions We anticipate that the combination of methodologies utilized in this study, which included repeat-anchored bisulfite PCR sequencing and the computational analysis pipeline herein reported, will prove invaluable for the generation of genetic and epigenetic variation maps.

Background and Aim: Infectious bovine keratoconjunctivitis is the most crucial ophthalmic disease among ruminants worldwide. Moraxella is the bacteria generally associated with this disease and leads to keratitis, conjunctivitis, corneal ulcers, or blindness. Platelet-rich plasma (PRP) effects in corneal ulcers and different ocular superficial diseases in animals and humans are beneficial and enhance rapid healing and improvement, but the effects in infectious keratoconjunctivitis in ruminants are uncertain. This study aimed to examine the effect of PRP on re-epithelization, corneal tissue, clinical signs, and matrix metalloproteinase (MMP) expression in sheep with infectious keratoconjunctivitis., Materials and Methods: Eighteen sheep were divided into three groups and subjected to a disease-induction experiment. Group 1 (G1) was administered 1.0 mL PRP subconjunctivally, Group 2 (G2) was administered 1.0 mL PRP subconjunctivally and 50 μL gentamicin drops, and the control group (CG) was administered 50 μL saline solution topically every 12 h. Clinical ophthalmologic examination, fluorescein staining, and photography were carried out. Ulcerated areas were measured employing J-Image software. Five and eleven days following the procedure, half of the animals from each group were euthanized, and their corneas were evaluated by histopathology and zymography., Results: Control Group and G2 epithelialized more rapidly. The CG exhibited fewer clinical signs of ocular disease. In histopathological analysis, in G2, alterations were observed only in the epithelium. The CG and G1 exhibited alterations in the epithelium, stroma, and Descemet's membrane. In zymography, a decline in MMP-2 expression in the animals treated with PRP was detected. Matrix metalloproteinase-9 was significantly expressed in the animals treated with PRP monotherapy, whereas PRP + gentamicin and CG caused a decrease., Conclusion: Platelet-rich plasma alone did not demonstrate any beneficial effect on re-epithelialization, a decline in clinical signs, tissue alterations, and expression of metalloproteinases. Platelet-rich plasma combined with gentamicin was capable of suppressing MMPs, primarily MMP-9, but do not display positive effects in re-epithelization, reduction of clinical signs, or tissue effects. These outcomes are similar to those discovered in untreated animals, so the use of PRP in patients with infectious keratoconjunctivitis does not offer greater benefits in sheep. Additional research is required to validate the results of PRP use in natural disease presentation., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Piso, et al.)

Background and Purpose: Miller Fisher syndrome (MFS) is a subtype of Guillain-Barré syndrome characterized by the triad of ophthalmoparesis, areflexia, and ataxia. Although cases of MFS have been associated with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, no studies have synthesized the clinical characteristics of patients with this condition., Methods: In this rapid systematic review, we searched the PubMed database to identify studies on MFS associated with SARS-CoV-2 infection., Results: This review identified 11 cases, of whom 3 were hospitalized with motor and/or sensory polyneuropathy as the first sign of SARS-CoV-2 infection. SARS-CoV-2 RNA was not detected in analyses of cerebrospinal fluid, suggesting a mechanism of immune-mediated injury rather than direct viral neurotropism. However, antiganglioside antibodies were found in only two of the nine patients tested. It is possible that target antigens other than gangliosides are involved in MFS associated with SARS-CoV-2 infection., Conclusions: The present patients exhibited clinical improvement after being treated with intravenous immunoglobulin. Although rare, patients with SARS-CoV-2 infection may present neurological symptoms suggestive of MFS. Early recognition of the MFS clinical triad is essential for the timely initiation of treatment., Competing Interests: The authors have no potential conflicts of interest to disclose., (Copyright © 2021 Korean Neurological Association.)

Objective: To determine the course of serological tests in subjects with chronic Trypanosoma cruzi infection treated with anti-trypanosomal drugs., Methods: A systematic review and meta-analysis was conducted using individual participant data. Survival analysis and the Cox proportional hazards regression model with random effects to adjust for covariates were applied. The protocol was registered in the PROSPERO database (http://www.crd.york.ac.uk/PROSPERO; CRD42012002162)., Results: A total of 27 studies (1296 subjects) conducted in eight countries were included. The risk of bias was low for all domains in 17 studies (63.0%). Nine hundred and thirteen subjects were assessed (149 seroreversion events, 83.7% censored data) for enzyme-linked immunosorbent assay (ELISA), 670 subjects (134 events, 80.0% censored) for indirect immunofluorescence assay (IIF), and 548 subjects (99 events, 82.0% censored) for indirect hemagglutination assay (IHA). A higher probability of seroreversion was observed within a shorter time span in subjects aged 1-19 years compared to adults. The chance of seroreversion also varied according to the country where the infection might have been acquired. For instance, the pooled adjusted hazard ratio between children/adolescents and adults for the IIF test was 1.54 (95% confidence interval 0.64-3.71) for certain countries of South America (Argentina, Bolivia, Chile, and Paraguay) and 9.37 (95% confidence interval 3.44-25.50) for Brazil., Conclusions: The disappearance of anti-T. cruzi antibodies was demonstrated along the course of follow-up. An interaction between age at treatment and country setting was found., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Retirement research is now expanding beyond the post-World War II baby boomers' retirement attitudes and plans to include the nature of their workforce exit and how successfully they adjust to their new life. These elements are collectively known as the process of retirement. However, there is insufficient research in developing countries to inform the management of their ageing populations regarding this process. This review aims to facilitate national and cross-cultural research in developing and non-English speaking countries by reviewing the existing measures of the retirement process published in English and Portuguese. The review identified 28 existing measures assessing retirement attitudes, planning, decision making, adjustment and satisfaction with retirement. Information on each scale's item structure, internal reliability, grammatical structure and evidence of translations to other languages is presented. Of the 28 measures, 20 assessed retirement attitudes, plans and decision-making, 5 assessed adjustment to retirement and only two assessed retirement satisfaction. Only eight of the 28 scales had been translated into languages other than English. There is scope to translate measures of retirement attitudes and planning into other languages. However there is a paucity of translated measures of retirement decision-making and adjustment, and measures of retirement satisfaction in general. Within the limitations of this review, researchers are provided with the background to decide between translating existing measures or developing of more culturally appropriate assessment tools for addressing their research questions.

Introduction: Injuries and/or dysfunctions in the somatosensory system can lead to neuropathic pain. Transient receptor potential vanilloid sub‑type 1 (TRPV1) play an important role in the development of allodynia and hyperalgesia following injury and the ensuing inflammatory conditions. Resiniferatoxin (RTX) is an ultrapotent synthetic TRPV1 agonist and many different administration routes are available for different mechanisms and different effects. RTX is used intraperitonially as a model of neuropathic pain or epidurally and topically to produce prolonged analgesic effects. However, the use of RTX is controversial because its neurotoxicity and margin of safety have not been addressed adequately. The present study evaluates the effect of intrathecal RTX on the induction and allodynia behavior of animals submitted to neuropathic pain by chronic constriction injury (CCI)., Methods: 160 Swiss mice were randomly distributed into two groups: intrathecal pre‑treatment group (PRE) aiming the effect in induction of allodynia and late intrathecal treatment group (POST) to evaluate the antiallodynic effect of the RTX on mechanical nociceptive threshold evaluated by the Von Frey hair filaments. Additionally, we evaluated the expression of TRPV1 in dorsal root ganglia (DRG) by western blotting after PRE‑ and POST‑treatment with RTX., Results: Our results showed that the CCI mice developed prolonged mechanical allodynia‑like behavior in ipsilateral paw after surgery up to 24 hours. The PRE‑ and POST‑treatment groups presented significant antiallodynic effects in ipsilateral paw for 24 hours. Only the POST‑treatment group showed a significant reduction of expression of the TRPV1 receptor after CCI., Conclusion: The presented data demonstrated that both PRE‑ and POST‑treatment with RTX given intrathecally produced potent antiallodynic activities in CCI mice and that POST‑treatment can reduce TRPV1 expression in DRG, suggesting that POST‑treatment RTX can revert central sensitization and its associated allodynia.

Neuropathic pain can be defined as pain initiated or caused by a primary lesion or dysfunction in the central or peripheral nervous system. The low level laser therapy (LLLT) has gained great prominence as a treatment in this type of pain; however, the application parameters are still controversial in the literature. This study aimed to review the literature on the use of LLLT in neuropathic pain with the goal of establishing a "therapeutic window" for the effective use of this treatment. We analyzed 14 articles, 10 in experimental animals and 4 in humans. The results are presented in three tables, the first being for comparison of the studies' application parameters, the second showing the average and median parameters experimental studies and third showing the clinical studies embodiment. The experimental studies revealed better results for LLLT and infrared laser powers above 70mW. Clinical studies are inconclusive as to the application parameters, due to the discrepancy; however all demonstrate the effectiveness of LLLT. According to the data presented, it was concluded that LLLT has positive effects on the control of analgesia for neuropathic pain, but further studies with high scientific rigor are needed in order to define treatment protocols that optimize the action LLLT in neuropathic pain., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Introduction: Exercise performed with blood flow restriction simultaneously enhances the acute responses to both myogenic and mitochondrial pathways with roles in training adaptation. We investigated isoform-specific gene expression of the peroxisome proliferator-activated receptor gamma coactivator 1 and selected target genes and proteins regulating skeletal muscle training adaptation., Methods: Nine healthy, untrained males participated in a randomized, counterbalanced, crossover design in which each subject completed a bout of low-intensity endurance exercise performed with blood flow restriction (15 min cycling at 40% of V˙O2peak, BFR-EE), endurance exercise (30 min cycling at 70% of V˙O2peak, EE), or resistance exercise (4 × 10 repetitions of leg press at 70% of one-repetition maximum) separated by at least 1 wk of recovery. A single resting muscle biopsy (vastus lateralis) was obtained 2 wk before the first exercise trial (rest) and 3 h after each bout., Results: Total PGC-1α mRNA abundance, along with all four isoforms, increased above rest with EE only (P < 0.05) being higher than BFR-EE (P < 0.05). PGC-1α1, 2, and 4 were higher after EE compared with resistance exercise (P < 0.05). EE also increased vascular endothelial growth factor, Hif-1α, and MuRF-1 mRNA abundance above rest (P < 0.05), whereas COXIV mRNA expression increased with EE compared with BFR-EE (P < 0.05)., Conclusion: The attenuated expression of all four PGC-1α isoforms when EE is performed with blood flow restriction suggests this type of exercise provides an insufficient stimulus to activate the signaling pathways governing mitochondrial and angiogenesis responses observed with moderate- to high-intensity EE.

Many changes in the socioeconomic scenario led to the emergence of different models of career guidance, among which the protean career stands out. This model works with the prospect of a career that is self-directed and aligned with personal values, with important propositions for both professionals and students entering the work market. In the Brazilian scenario, however, there is a lack of appropriate measures to evaluate protean aspects among college students without work experience. Thus, the present study aimed at adapting and validating the attitudes towards the Protean Career Scale to this population. The sample consisted of 902 students aging from 18 to 30 years old (M = 22.52; SD = 6.53) attending 34 different undergraduate courses. Exploratory and confirmatory analysis attested the two-dimensional nature of the scale structure. The reliability indexes were satisfactory: over .65. The correlation between the protean models and factors such as personality, values, and locus of control provided adequate evidence of the measure's predictive validity (p < .05).

Unlabelled: Objetctive: Study was to translate and culturally adapt the modified Rowe score for overhead athletes., Methods: The translation and cultural adaptation process initially involved the stages of translation, synthesis, back-translation, and revision by the Translation Group. It was than created the pre-final version of the questionnaire, being the areas "function" and "pain" applied to 20 athletes that perform overhead movements and that suffered SLAP lesions in the dominant shoulder and the areas "active compression test and anterior apprehension test" and "motion" were applied to 15 health professionals., Results: During the translation process there were made little modifications in the questionnaire in order to adapt it to Brazilian culture, without changing the semantics and the idiomatic concept originally described., Conclusion: The questionnaire was easily understood by the subjects of the study, being possible to obtain the Brazilian version of the modified Rowe score for overhead athletes that underwent surgical treatment of the SLAP lesion.

Background: The epidermal growth factor receptor (EGFR) and the extracellular matrix metalloproteinase inducer (EMMPRIN) have been identified as oncologically important targets. This study aimed to evaluate the immunoexpression of EGFR and EMMPRIN in a series of cases of head and neck squamous cell carcinoma (HNSCC)., Methods: Forty-five cases of HNSCC were selected for this study and evaluated with anti-EGFR and anti-EMMPRIN antibodies. The percentage of positive cells was determined assessing to the following categories: score 1 (staining in 0-50% of cells), score 2 (staining in 51-75% of cells), and score 3 (staining in >75% of cells). Immunostaining intensity was graded according to the following parameters: score 1 (absent/weak expression) and score 2 (strong expression)., Results: For EGFR, a predominance of high median scores was observed in cases of both histological grades of malignancy and in different clinical stages (p>0.05). For EMMPRIN, a statistically significant difference was observed between the histological grades of malignancy (p=0.030). Regarding the immunostaining intensity of EMMPRIN, it was observed a predominance of score 1 in cases with stages I/II, whereas most cases with stages III/IV presented score 2 (p=0.032). Considering the anatomical location, most cases of buccal floor presented higher median score of EMMPRIN in comparison with the other sites (p=0.015)., Conclusions: These findings suggest that both proteins are potential targets for cancer therapy and EMMPRIN can be used as a prognostic marker of a more aggressive biological behavior in patients with HNSCC., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Aim: To evaluate and compare the immunoexpression of tryptase in samples of periapical granulomas (PGs) and radicular cysts (RCs) correlating it with the type of lesion, localization, intensity of the inflammatory infiltrate and thickness of the cystic epithelial lining, in order to gain insight into the phlogistic role of these cells in the lesions studied., Methodology: Twenty-five PGs and twenty-five RCs obtained from human teeth without endodontic treatment were submitted to morphological and immunohistochemical analysis using anti-tryptase antibody. Mast cells were identified and counted in three regions: intra-epithelial, central/superficial and deep portions. The data were analysed using the Mann-Whitney U-test (P < 0.05)., Results: In comparison with RCs, PGs exhibited higher immunoexpression of tryptase-positive mast cells located in both central/superficial and deep regions (P < 0.001 and P < 0.001, respectively). When considering the total number of mast cells and disregarding the location, the number of tryptase-positive mast cells increased gradually from RCs to PGs (P < 0.001). Lesions with inflammatory infiltrate grade III had greater number of tryptase-positive mast cells located in both central/superficial and deep regions than lesions with inflammatory infiltrates grade II (P = 0.045 and P = 0.025). When the location was ignored, the lesions with inflammatory infiltrate grade III also exhibited higher immunostaining of tryptase-positive mast cells (P = 0.01)., Conclusions: Tryptase-positive mast cells were present in chronic periapical lesions in a larger number in periapical granulomas than in radicular cysts, in both central/superficial and deep regions., (© 2014 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Canine mesenchymal cells (MSCs) derived from Wharton's jelly were co-cultured, then supplemented or not supplemented with platelet rich plasma (PRP) and demineralized bone matrix (DBM) to verify osteogenic differentiation. Osteoblastic differentiation followed by mineralized bone matrix production was found to be significantly higher (p < 0.05) when MSCs were associated with PRP/DBM in culture after 14-21-days of induction. Osteopontin and osteocalcin gene expression were significantly superior (p < 0.05) under the same culture conditions after 21 days of observation. In conclusion, addition of PRP to DBM co-cultured with MSCs successfully induced osteogenesis in vitro.

Scientific development and increasing technological progress in the field ofdentistry have allowed the use of many resources in theoral rehabilitationplanning. In complex cases, in order to achieve projected final results,many procedures should be undertakenpriorto implant surgery. Prosthetic rehabilitation planningand bone graft procedures canbetter restorethe functional, aesthetic, social and emotional needs of the patient. This clinical case report shows the oral conditions of a woman who underwent several surgical proceduresfor maxillary reconstruction with autogenous bone grafts from the iliac crest. In this case, a bone volume guide was made to evaluate the bone volume requirement for correct labial support for metalloceramic fixed prosthesis. After graft integration, dental implantsurgery was performed. After the osseointegration period, the prostheses was fabricated and installed on the implants, thus concluding the oral rehabilitation. In conclusion, the bone guide was a fundamental tool for final aesthetic outcome predictability.

Actinomycosis is a relatively rare infection caused by saprophytic bacteria of the oral cavity and gastrointestinal tract that can become pathogenic. The chronic hyperglycemia of diabetes mellitus induces events that promote structural changes in various tissues and are associated with problems in wound healing. This infection remains largely unknown to most clinicians because of its different presentations, and palatal involvement is extremely rare. This report describes the case of a 46-year-old woman who was diagnosed with actinomycosis involving the hard palate. The main clinical, histopathologic, and therapeutic characteristics and differential diagnosis of actinomycosis are reviewed. To date, 3 cases of actinomycosis involving the hard palate have been reported., (Copyright © 2014 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

Objective: To assess quantitative real-time polymerase chain reaction (q-PCR) for the sputum smear diagnosis of pulmonary tuberculosis (PTB) in patients living with HIV/AIDS with a clinical suspicion of PTB., Method: This is a prospective study to assess the accuracy of a diagnostic test, conducted on 140 sputum specimens from 140 patients living with HIV/AIDS with a clinical suspicion of PTB, attended at two referral hospitals for people living with HIV/AIDS in the city of Recife, Pernambuco, Brazil. A Löwenstein-Jensen medium culture and 7H9 broth were used as gold standard., Results: Of the 140 sputum samples, 47 (33.6%) were positive with the gold standard. q-PCR was positive in 42 (30%) of the 140 patients. Only one (0.71%) did not correspond to the culture. The sensitivity, specificity and accuracy of the q-PCR were 87.2%, 98.9% and 95% respectively. In 39 (93%) of the 42 q-PCR positive cases, the CT (threshold cycle) was equal to or less than 37., Conclusion: q-PCR performed on sputum smears from patients living with HIV/AIDS demonstrated satisfactory sensitivity, specificity and accuracy, and may therefore be recommended as a method for diagnosing PTB.

The study describes the main chest radiographic changes in people living with HIV/AIDS and pulmonary tuberculosis, confirmed by sputum culture. This was a descriptive study involving a total of 42 sputum tests from 42 people living with HIV/AIDS and a clinical suspicion of pulmonary tuberculosis. All patients attended two referral hospitals in Recife-PE, Brazil, between August 2009 and January 2012. The most common isolated radiological change was parenchymal consolidation, encountered in six (14.3%) patients, followed by patterns of interstitial infiltrate, diffuse micronodular (miliary), and an association between interstitial infiltrate and parenchymal consolidation, each being encountered in five (11.9%) patients. No statistically significant difference was observed between the radiological findings and CD4 T-cell counts, p = 0.680.

Objective: : The purpose of this study was to clinically evaluate patients who underwent Arthroscopic Rotator Cuff Repair (RC) using the Modified Manson-Allen technique., Methods: : We evaluated 79 patients who underwent shoulder arthroscopy. The lesions were repaired using the modified Mason-Allen suture between 2003 and 2009, divided by Cofield classification and clinically evaluated by the scoring system of the University of Los Angeles (UCLA) in the pre- and postoperative periods., Results: : The evaluation of lesion sizes showed 7 small lesions (<1cm), 55 average lesions (1-3cm) and 17 large lesions (3-5cm), and in this last group there were 5 reruptures and the patients were reoperated by the same technique. Comparing the pre (14.1) and postoperative (32.6) values by UCLA system there was a significant improvement of score (142.3%), regardless of lesion size. The modified Mason-Allen suture provided satisfactory clinical results, regardless of lesion size, similar to those found in literature. The rerupture rate was high in large lesions. New suture techniques have been developed with the aim of reducing the incidence of rerupture., Conclusion: : The modified Mason-Allen suture technique provided clinical improvement, regardless of lesion size. Level of Evidence IV, Cases Series .

Objective: To translate and culturally adapt the Rowe score for use in Brazil., Methods: The translation and cross-cultural adaptation process initially involved the steps of translation, synthesis, back-translation and revision by the Translation Group. The pre-final version of the questionnaire was then created. The Stability and Function fields were applied to 20 patients with anterior shoulder luxation, and the Mobility field was applied to 20 health professionals., Results: It was found that some of the patients had difficulty understanding some of the expressions of the questionnaire, so these were replaced with terms that were easier to understand. All health professionals understood the translation of the Mobility field. The altered questionnaire was then reapplied to another 20 patients, and this time it was understood by all the assessed subjects., Conclusion: After a careful process of translation and cultural adaptation, a definitive version of the Rowe questionnaire was obtained in Brazilian Portuguese. Level of Evidence II, Development of diagnostic criteria on consecutive patients.

Molluscum contagiosum (MC) is a contagious disease caused by a virus of the poxvirus family. In children, the disease commonly manifests as a variable number of discrete umbilicated papules on the face and trunk. In healthy and immunosuppressed adults, the disease appears on or near the genital organs and is often sexually transmitted. MC involving the intraoral mucosa has been documented but is rare. We report a case of MC involving the oral mucosa exclusively and discuss the main clinical, histopathologic, and therapeutic characteristics, comparing the findings with cases of this rare oral presentation described in the literature., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Coccidiosis is a serious obstacle to sheep production, which is becoming a limiting factor, especially with regard to lamb production. However, there are few studies on this parasite in the State of Rio Grande do Norte. The aim of this study was to evaluate the action of decoquinate, added to mineral salt, for controlling Eimeria infection in lambs, and to identify which species are infecting sheep in the eastern region of the state. This study was carried out from August 2009 to January 2010, and used 76 animals. These were divided into two treatment groups: one with common mineral salt, and the other with mineral salt enriched with 6% micronized decoquinate. Fecal samples and body weight measurements were taken every 14 days for parasitological diagnosis, weight gain follow-up and quantitative analysis. The study showed that there was a significant difference in OPG only at the 7th collection, but no significant difference in weight gain. The Eimeria species found were E. ahsata. E. crandallis. E. granulosa. E. intrincata. E. ovina. E. faurei. E. ovinoidalis. E. pallida and E. parva. It was concluded that addition of decoquinate to mineral salt gave rise to lower oocyst elimination, thus favoring eimeriosis control in sheep.

PspA is one of the most well studied pneumococcal proteins and a promising candidate for a future protein-based anti-pneumococcal vaccine. Nevertheless, its structural and serological variability suggests the inclusion of more than one PspA molecule in order to broaden protection. Since different PspAs exhibit variable levels of cross-reactivity, the selection of the protein combination with the highest coverage potential is an essential step for PspA-based vaccine development. This work investigated the level of cross-reactivity within family 1 PspAs, and established a complement based antibody mediated opsonophagocytic assay for measuring the level of cross-protection. Among a panel of ten family 1 PspA molecules, two of them, one belonging to clade 1 and another from clade 2, induced antibodies capable of enhancing complement deposition and mediating the phagocytic killing by mouse peritoneal macrophages of all pneumococci bearing PspA family 1 strains tested, regardless of their serotype. Therefore, we suggest the inclusion of either one in a PspA-based vaccine, as a representative of family 1. Furthermore, our results suggest that opsonophagocytosis by mouse peritoneal cells can be an efficient means of evaluating the induction of protective immune responses in mice across a large number of strains., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Purpose: To detect the occurrence and expression of the suppressor gene p53 and of the oncogene c-Myc in eyelid tumors of dogs using the PCR, RT-PCR, PCR-ELISA and RT-PCR-ELISA techniques. These genes have not been described in dog eyelid tumors before., Methods: Nine samples of eyelid or third eyelid epithelial tumors were obtained from the archives of the Department of Veterinary Pathology. Tumor diagnosis was confirmed by evaluation of hematoxylin-eosin stained sections, and immunohistochemistry for cytokeratin AE1/AE3 and vimentin V9. A canine mammary tumor was used for positive control. Agarose gel electrophoresis, PCR-ELISA and RT-PCR-ELISA were used to detect p53 and c-Myc genes., Results: The occurrence of p53 was detected in most of the eyelid tumors and third eyelid tumors studied (88.8%, n = 8) and was expressed in 75% of the positive samples, as indicated by ELISA. The c-Myc gene was found in 77.7% (n = 7) of the samples and was expressed in eight samples., Conclusions: Eyelid and third eyelid tumors of dogs express both the p53 and the c-Myc genes as shown by PCR and RT-PCR. However, PCR ELISA and RT-PCR ELISA were more efficient in assessing occurrence and expression of these genes because they identified amplified products that were not detected by agarose gel electrophoresis.

Objective: To compare the costs and benefits of pneumococcal conjugate vaccination compared with no vaccination from the perspectives of the health care system and society., Methods: Using data from established sources, we estimated the incidence and mortality due to invasive pneumococcal disease, pneumonia, and acute otitis media (AOM) for a hypothetical birth cohort of children from birth to 5 years., Results: A universal pneumococcal conjugate vaccination program was estimated capable of annually avoiding 1 047 cases of invasive disease, 58 226 cases of pneumonia, and 209 862 cases of AOM. When herd immunity effects were considered, the program prevented 1.3 million cases of pneumococcal disease and over 7 000 pneumococcal deaths. At a vaccination cost of R$ 51.12 (US$ 26.35) per dose, vaccination would cost annually R$ 4 289 (US$ 2,211) per disability-adjusted life years averted. This does not take into account herd immunity effects., Conclusions: At the current vaccine price, conjugate vaccination could be a cost-effective investment compared to other options to control childhood diseases. Further analysis is required to determine whether vaccination at the current price is affordable to Brazil.

Pneumococcal surface protein A (PspA) has been considered a potential candidate for human vaccines because of its serotype-independent protective immunity. Nasopharyngeal (NP) pneumococcal colonization is highly prevalent in infants and precedes the invasive disease. Thus, prevention of NP colonization may reduce the burden of pneumococcal disease in children. Scarce information focusing on PspA from pneumococcal carriage in humans is available. We examined the genetic diversity of PspA from NP isolates obtained during an ongoing pneumococcal surveillance study with children. PspA families and clades of 183 community-acquired Streptococcus pneumoniae NP isolates from healthy children (n = 97) and children with respiratory tract infections (n = 48), pneumonia (n = 33), or meningitis (n = 5) were investigated. Overall, 79.8% (n = 146) of the pneumococcal isolates were classified as PspA family 1 (35.5%) and family 2 (44.3%), whereas 20.2% of the isolates could not be typed. The distribution of PspA families and clades did not differ significantly according to the clinical status of the children. A dendrogram comparing the genetic relationship between the amino acid sequences of the clade-defining region of PspA from NP strains together with 24 invasive reference strains (GenBank) closely reproduced the profile of the families and clades previously reported for pneumococcal invasive strains. These findings strengthen the idea that the use of PspA as a vaccine antigen may protect children against carriage as well as invasive pneumococcal disease.

One of the candidate proteins for a mucosal vaccine antigen against Streptococcus pneumoniae is PsaA (pneumococcal surface antigen A). Vaccines targeting mucosal immunity may raise concerns as to possible alterations in the normal microbiota, especially in the case of PsaA, which was shown to have homologs with elevated sequence identity in other viridans group streptococci. In this work, we demonstrate that intranasal immunization with a cholera toxin B subunit-PsaA fusion protein is able to protect mice against colonization with S. pneumoniae but does not significantly alter the natural oral or nasopharyngeal microbiota of mice.

Objective: To identify evidence of the impact of Haemophilus influenzae type b (Hib) conjugate vaccine on the epidemiology of invasive Hib disease., Sources of Data: This review was based on a search of MEDLINE, LILACS, technical reports, national and international guidelines (publications from 1991 to 2005). The keywords Haemophilus influenzae type b, immunization, impact, and effectiveness, alone or in combination, were used to retrieve the articles. Studies published before 1991 and cited in the references of the studies reviewed were analyzed for useful information., Summary of the Findings: Introduction of the Hib conjugate vaccine produced great decline in the incidence of invasive Hib disease in childhood in countries where this vaccine was introduced into the routine immunization schedule. Nevertheless, the resurgence of invasive Hib disease in some regions has challenged several researchers to identify the reasons for this epidemiological pattern, as well as the measures to be implemented in order to avoid such a phenomenon., Conclusions: The use of Hib conjugate vaccine on a population scale has been greatly effective; nonetheless, changes in the vaccination scheme seem to be necessary to keep invasive Hib disease under control.

The aim of this study was to describe the frequency of antimicrobial-resistance and serotypes of nasopharyngeal pneumococcal isolates from adolescents. Clinical data and nasopharyngeal specimens for culture were collected from 1,013 adolescents as a part of a population-based study. A total of 83 isolates of Streptococcus pneumoniae were identified (8.2%). Seventy-four of the 83 isolates were serotyped. The median age of the 83 adolescents colonized by pneumococci was 14 years (mean 14 +/- 2.2 yrs); 55.4% were males. Intermediate resistance to penicillin was detected in 7.2% (6/83). No strain showed high resistance to penicillin. All isolates were susceptible to clindamycin, chloramphenicol, rifampin, and vancomycin; 37.3%, 18.1%, and 4.8% were resistant to trimethoprim-sulfamethoxazole, tetracycline, and erythromycin, respectively. The most frequent serotypes (5-10% of strains each) were 6B, 6A, 23F, and 18C among 28 serotypes/serogroups identified; 18.9% of the strains were nontypeable (NT). Intermediate resistance to penicillin was detected in serotypes 6B, 14, and NT. The rate of resistance to penicillin of nasopharyngeal isolates is low considering data from other studies about invasive strains recovered from children in Brazil. Serotype patterns are similar, except for type 14, which was unusually infrequent.

Nasopharyngeal carriage of Streptococcus pneumoniae is a key factor in the development of invasive disease and the spread of resistant strains within the community. A single nasopharyngeal swab was obtained from 648 unvaccinated children aged <5 years, either healthy or with acute respiratory tract infection or meningitis, during the winters of 2000 and 2001. The overall pneumococcal carriage rate was 35.8% (95% CI 32.1-39.6). The pneumococcal serotypes found most frequently in the nasopharynx were 14, 6B, 6A, 19F, 10A, 23F and 18C, which included five of the seven serotypes in the currently licensed seven-valent conjugate vaccine (PCV7); serotypes 4 and 9V were less common. Serotypes 1 and 5 were isolated rarely from the nasopharynx. A comparison of 222 nasopharyngeal isolates with 125 invasive isolates, matched for age and time to the carrier isolates, showed a similar prevalence of penicillin non-susceptible pneumococci (PNSp) (19.8% and 19.2%, respectively). PNSp serotypes were similar (6B, 14, 19F, 19 A, 23B and 23F) for carriage and invasive disease isolates. The coverage of PCV7 for carriage isolates (52.2%) and invasive isolates (62.4%) did not differ significantly (p 0.06); similarly, there was no significant difference in PCV7 coverage for carriage isolates (34.5%) and invasive isolates (28.2%) of PNSp. These data suggest that PCV7 has the potential to reduce pneumococcal carriage and the number of carriers of PNSp belonging to vaccine serotypes.

We present a case of invasive pneumococcal infection in a healthy 10-month-old infant from whom Streptococcus pneumoniae serotype 23F was isolated from the blood and serotype 23B was isolated from the cerebrospinal fluid. Both serotypes were penicillin nonsusceptible. Pulsed-field gel electrophoresis analysis demonstrated that the two serotypes had distinct DNA patterns, indicating that infection did not occur as a result of capsular transformation but as a result of a mixed infection with two distinct pneumococcal serotypes.

Background: The conjugate Haemophilus influenzae type b (Hib) vaccine has been highly efficacious in reducing type b H. influenzae meningitis. However, information is limited about its impact on childhood pneumonia after vaccine introduction into the routine programme. This study evaluated the effectiveness of Hib conjugate vaccine in the reduction of community-acquired pneumonia among infants in Central Brazil., Methods: A matched case-control study was built into an ongoing prospective population-based surveillance of pneumonia, enrolling 1293 participants between May 2000 and August 2001. Cases (n = 431) were children

To assess the magnitude of discrepant results obtained by routine Haemophilus influenzae serotyping, 258 isolates, collected by the epidemiological surveillance system in Brazil from individuals with invasive diseases or carriage, were evaluated by two slide agglutination (SlAg) methods: SlAg method 1, by which strains were initially screened with a serotype b-specific antiserum, and SlAg method 2, by which strains were tested against all serotype-specific antisera in parallel. Investigators comparing results of the two SlAg methods with those obtained by capsule type-specific PCR were blinded to the method used. The serotype prevalence rates found by the three methods were significantly different, involving discrepancies mainly between serotype b and noncapsulated (NC) isolates. For invasive isolates (n = 131), the overall agreement rate between SlAg method 1 or 2 and PCR was 68.0 or 88.3%, respectively, whereas for colonizing isolates (n = 127) the corresponding rate was 46.5 or 94.2%, respectively. SlAg method 2 improved the ascertainment of serotypes over that obtained with SlAg method 1, demonstrating good correlation with PCR. Use of the polyvalent antiserum as a screening reagent for SlAg for invasive and colonizing isolates showed poor discriminatory power, with a sensitivity of 65.8% and a specificity of 91.7%. We stress the importance of using a well-standardized SlAg methodology and suggest that reference laboratories should utilize PCR routinely to confirm SlAg results and to check all nonspecific SlAg reactions and apparent NC isolates by SlAg in order to provide reliable data on the prevalence of H. influenzae serotypes in the H. influenzae type b vaccine era.

Acute bacterial meningitis is an important cause of morbidity and mortality among children less than five years old, Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis being the most important agents of bacterial meningitis in developing countries. The development of the conjugate vaccines in the beginning of the 90's, especially type b H. influenzae (Hib), and more recently the heptavalent pneumococcal and the serogroup C meningococcal vaccines, have contributed directly to changes in the epidemiological profile of these invasive diseases (direct effect) and of their carriage status (indirect effect). We review the impact of the Hib conjugate vaccine in Latin American countries, where this vaccine has been implemented, and the potential of pneumococcal and meningococcal conjugate vaccines for the reduction of meningitis worldwide. We also address constraints for the development and delivery of these vaccines and review new candidate state-of-the-art vaccines. The greatest challenge, undoubtedly, is to implement these vaccines worldwide, especially in the developing regions.

The molecular epidemiology of Streptococcus pneumoniae isolates from carriage and cerebrospinal fluid (CSF) concurrently recovered from the same individual has not yet been reported. By using pulsed-field gel electrophoresis, we demonstrated the genetic linkage among strains from CSF and nasopharynges of two children with pneumococcal meningitis.

The distribution of pneumococcal serotypes in Brazil was analyzed by age group and clinical diagnosis, using data obtained during 20 years of national surveillance. Serotypes 1 and 5 remained among the main serotypes in all age groups, increasing in frequency with age. Serotype 14 was prevalent among children, whereas serotypes 3 and 4 were most prevalent among the adult population. The potential impact of the 7- and 9-valent conjugate vaccines on children up to age 5 years with severe pneumococcal diseases was 58.2% and 73%, respectively; the highest coverage of the 7-valent vaccine for pneumonia was achieved for children aged 7 months to 2 years (70%), whereas, for meningitis, it was observed for children aged 7 months to 5 years (58.6%). The use of conjugate vaccine may be of potential benefit by reducing the childhood sequelae and mortality of pneumococcal infection in Brazil.

Haemophilus influenzae is a relevant cause of morbidity and mortality among children under 5 years of age in the developing world. In Latin America, H. influenzae type b (Hib) conjugate vaccine and surveillance of H. influenzae antimicrobial susceptibility have been implemented in recent years. We have undertaken a systematic review and a pooled analysis on H. influenzae antimicrobial resistance, including reports of 15 Latin America countries over a 10-year period (1990-2000). We have found that 450 (21.4%) of 2,100 invasive isolates were beta-lactamase producers compared to 145 (14.5%) of 998 isolates of noninvasive isolates (p < 0.05). Ampicillin resistance was detected among 783 (21.9%) of 3,577 invasive isolates compared to 111 (17.2%) of 646 noninvasive strains (p < 0.05). In contrast, 568 (41.9%) of 1,355 noninvasive strains were trimethoprim-sulfamethoxazole (TMP-SMX) resistance against 241 (26.9%) of 897 invasive ones (p < 0.05). Therefore, TMP-SMX resistance was more common in nonsterile fluids than in sterile fluids. Over time, rates of beta-lactamase-producing strains were stable in Brazil and Mexico, whereas rates of TMP-SMX resistance were increasing in Brazil. It is predictable that following the Hib immunization, Latin America countries will be faced with increased nontypeable H. influenzae infection. Although standing by the nontypeable H. influenzae vaccine, in this novel epidemiological scenario of post-Hib vaccination in Latin America settings there is a need to improve H. influenzae resistance monitoring to guide clinicians to choose efficacious antimicrobial therapy.

This report describes clinical and pathological findings in 2 flocks in Brazil where blindness and deaths in sheep occurred after closantel overdosage. Depression, weakness, and blindness affected 37 animals and 17 died in 2 flocks of 190 animals. Two animals submitted for ophthalmic examination showed no inflammation in the anterior segment of both eyes; posterior segment evaluation by indirect ophthalmoscopy suggested retinal degeneration. One postmortem evaluation local spongy vacuolization was in several regions of the brain and the optical nerves had severe axonal degeneration.

As part of a major epidemiologic study on Chagas' disease, we compared the prevalence of electrocardiographic (ECG) abnormalities among 141 school children 7-12 years of age and seropositive for Trypanosoma cruzi, and 282 age-, sex-, and school-matched seronegative children in an endemic area in Brazil. The prevalence of ECG abnormalities was 11.3% among seropositive children and 3.5% among seronegative children (odds ratio = 3.5, 95% confidence interval [CI] = 1.5-8.4). The prevalence rate of ECG alterations was 10.7% for seropositive males versus 8.9% for seropositive females. Complete right bundle branch block (CRBBB), which is highly suggestive of Chagas' disease cardiopathy, was diagnosed in nine (6.4%) seropositive children and in only one (0.3%) seronegative child (odds ratio = 18.5, 95% CI = 2.3-146.5, attributable fraction = 58.3%). Five incident new cases of CRBBB were diagnosed after a 36-month follow-up of seropositive children who were enrolled in an independent clinical field trial. No case of frequent and/or multifocal ventricular premature beats was found in the cohort of children. The surprisingly high frequency of early ECG abnormalities, which indicates a rapid evolution from infection to disease, suggests the existence of endemic areas with a particular accelerated disease progression that was not described before. Under such conditions, a public health chemotherapy program focusing on the treatment of young seropositive children would be recommended.

The anti-phenolic glycolipid-I (PGL-I) assay as currently applied for leprosy is conceived as an early marker of asymptomatic infection, early disease diagnosis and cure monitoring. Its use as a prognostic marker of reaction is still a matter of controversy. We conducted a case-control study to investigate whether IgM and IgG anti-PGL-I antibodies could discriminate patients at increased risk of developing reactions. Eligible cases were untreated leprosy patients at the onset of type 1 and type 2 reactions recruited from among 600 concurrent, newly detected, untreated leprosy patients attending an outpatient clinic in central Brazil. For the patients with reaction, approximately the same number of leprosy cases without reaction matched as to bacterial index (BI), age and gender were randomly selected. Individuals without clinical leprosy were evaluated as healthy controls. Sera from type 1 reaction (N = 43) and type 2 reaction (N = 26) patients were tested by an ELISA using PGL-I synthetic disaccharide-BSA antigen and 1:300 sera dilution (cut-off point > or = 0.2 OD). Antibody profiles were evaluated by exploratory data analysis and reverse cumulative distribution curves. The IgG anti-PGL-I response did not have a defined pattern, being detected only at low levels. Our results indicate that leprosy patients, independently of their reactional status, produce high levels of IgM anti-PGL-I, demonstrating a strong correlation between the magnitude of antibody response and the BI. Patients with a higher BI were at least 3.4 times more prone to produce an antibody response compared to healthy controls.

Background: Benznidazole, a nitroimidazole derivative, has been recommended for the treatment of acute and congenital Trypanosoma cruzi infection (Chagas' disease). We have examined the safety and efficacy of this drug in the treatment of the early chronic phase of T cruzi infection., Methods: Between 1991 and 1995, we carried out a randomised, double-blind, placebo-controlled trial in a rural area of Brazil with endemic Chagas' disease. 82% of 2434 schoolchildren (aged 7-12 years) identified in a census were screened for antibodies to T cruzi by indirect immunofluorescence, indirect haemagglutination, and ELISA. 130 were positive in all tests and were randomly assigned benznidazole (7.5 mg/kg daily for 60 days by mouth) or placebo. The primary endpoint for efficacy was the disappearance of specific antibodies (negative seroconversion) by the end of 3-year follow-up. The secondary endpoint was the reduction of antibody titres on repeated serological tests. One child moved away from the area just after randomisation and was excluded from the analyses. Insecticidal measures were taken throughout the trial to reduce the risk of reinfection., Findings: Minor side-effects requiring no specific medication were recorded in a small proportion of individuals. On a chemiluminescent ELISA with purified trypomastigote glycoconjugate, serum from all participants was positive at the beginning of the trial. At the end of follow-up, 37 (58%) of the 64 benznidazole-treated participants and 3 (5%) of those who received placebo were negative for T cruzi antibodies. The efficacy of benznidazole treatment estimated by intention to treat was 55.8% (95% CI 40.8-67.0). At the end of follow-up, children who received benznidazole had five-fold lower geometric mean titres by indirect immunofluorescence than placebo-treated children (196[147-256] vs 1068[809-1408], p < 0.00001)., Interpretation: The trial showed that a 60-day course of benznidazole treatment of early chronic T cruzi infection was safe and 55.8% effective in producing negative seroconversion of specific antibodies. The results are very encouraging and justify the recommendation of treatment for seropositive children as public health policy.

An active entomologic survey was conducted by a team of trained health workers in a rural area endemic for Chagas' disease in central Brazil. They used pyrethrum as a flushing agent and 4,232 houses were inspected for triatomine bugs both inside and in the immediate environs. Houses with Triatoma infestans or evidence of an established colony were identified and defined as infested houses (cases). The building and environmental characteristics of 161 randomly selected infested houses were compared with 161 matched, noninfested houses (controls) that were the shortest distance from the infested house. Domestic and peridomestic potential risk factors associated with house infestation by Triatoma infestans were assessed by logistic regression analysis. Incomplete house construction (odds ratio [OR] = 2.5, 95% confidence interval [CI] = 1.5-4.1) was confirmed as a risk factor related to the presence or evidence of Triatoma infestans in the dwellings. The study also disclosed a statistically significant association between the presence of rats (OR = 1.6, 95% CI = 1.1-2.6) and indoor crop storage (OR = 2.3, 95% CI = 1.1-5.2) and house infestation. Further experimental field studies using tagged rodents should be conducted to assess their epidemiologic role in the domestic chain of Trypanosoma cruzi transmission.

The distribution of T. cruzi infection overlaps regions with high prevalence of child malnutrition. We examined the possible association between T. cruzi infection and chronic malnutrition. In a cross-sectional survey conducted among 1900 7-12 year-old schoolchildren in 60 village schools in central Brazil, anthropometric measurements (NCHS) taken from 153 children with at least two positive serological tests for antibodies against T. cruzi (IIF, ELISA, IHA) were compared to two age and sex seronegative matched classmates. Information on children's medical history and socio-economic status (SES) were collected from parents of the participants. Seropositive children had a 2.4-fold risk (95% CI 1.4-4.0) of being stunted (z-score < 2.0 of height-for-age) when compared to uninfected children even after adjusting for confounding variables. Being underweight (z-score < -2.0 of weight-for-age) was also statistically associated with seropositivity to T. cruzi (OR = 2.8, 95% CI 1.4-5.6). No statistical evidence of multiplicative interaction between nutritional status and SES was detected. Further studies on nutrition and metabolism are required to look into a possible physiopathological mechanism for this association.