Gamification and Oral Health in Children and Adolescents: Scoping Review.

BACKGROUND: Oral health is a determinant of overall well-being and quality of life. Individual behaviors, such as oral hygiene and dietary habits, play a central role in oral health. Motivation is a crucial factor in promoting behavior change, and gamification offers a means to boost health-related knowledge and encourage positive health behaviors. OBJECTIVE: This study aims to evaluate the impact of gamification and its mechanisms on oral health care of children and adolescents. METHODS: A systematic search covered multiple databases: PubMed/MEDLINE, PsycINFO, the Cochrane Library, ScienceDirect, and LILACS. Gray literature, conference proceedings, and WHOQOL internet resources were considered. Studies from January 2013 to December 2022 were included, except for PubMed/MEDLINE, which was searched until January 2023. A total of 15 studies were selected following PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. The eligibility criteria were peer-reviewed, full-text, and empirical research related to gamification in oral health care, reports of impact, and oral health care outcomes. The exclusion criteria encompassed duplicate articles; unavailable full texts; nonoriginal articles; and non-digital game-related, non-oral health-related, and protocol studies. Selected studies were scrutinized for gamification mechanisms and outcomes. Two main questions were raised: "Does gamification in oral health care impact oral health?" and "Does oral health care gamification enhance health promotion and literacy?" The PICO (Patient, Intervention, Comparison, Outcome) framework guided the scoping review. RESULTS: Initially, 617 records were obtained from 5 databases and gray literature sources. After applying exclusion criteria, 15 records were selected. Sample size in the selected studies ranged from 34 to 190 children and adolescents. A substantial portion (11/15, 73%) of the studies discussed oral self-care apps supported by evidence-based oral health. The most clearly defined data in the apps were "brushing time" (11/11, 100%) and "daily amount brushing" (10/11, 91%). Most studies (11/15, 73%) mentioned oral health care behavior change techniques and included "prompt intention formation" (11/26, 42%), "providing instructions" (11/26, 42%), "providing information on the behavior-health link" (10/26, 38%), "providing information on consequences" (9/26, 35%), "modeling or demonstrating behavior" (9/26, 35%), "providing feedback on performance" (8/26, 31%), and "providing contingent rewards" (8/26, 31%). Furthermore, 80% (12/15) of the studies identified game design elements incorporating gamification features in oral hygiene applications. The most prevalent gamification features were "ideological incentives" (10/12, 83%) and "goals" (9/16, 56%), which were found in user-specific and challenge categories, respectively. CONCLUSIONS: Gamification in oral health care shows potential as an innovative approach to promote positive health behaviors. Most studies reported evidence-based oral health and incorporated oral health care behavior change techniques.

Patient reported outcomes in oncology: changing perspectives-a systematic review.

In public health context, oncology is associated with severe negative impact on patients and on their relatives' quality of life. Over the last decades, survival has remained at 50% worldwide for some tumor locations. Patient reported outcomes (PROs) assessment and, the corresponding use in clinical practice, help establishing patient individualized profiling involving caregivers. The purpose of this systematic review was to examine critical success factors for PROs assessment in daily clinical oncology practice. Additionally, we investigated how PROs collection can change oncology perspectives for patients and caregivers. According to PRISMA guidelines, 83 studies were included in this systematic review, whether related with implementation in daily clinical practice or associated with its use in oncology. PROs assessment gathers multi-professional teams, biomedical and clinical expertise, patients, families and caregivers. Institutional involvement, first line for caregiver's adherence, team continuous formation, encompassing training and support, design of clear workflows, continuous monitoring, and data analysis are crucial for implementation. PROs measures are decisive in oncology. Several items were improved, including caregiver-patient-physician communication, patient risk groups identification, unmet problems and needs detection, disease course and treatment tracking, prognostic markers, cost-effectiveness measurement and comfort/support provision for both patients and caregivers. Routine assessment and implementation of PROs in clinical practice are a major challenge and a paradigm transformation for future.

Molecular identification of hookworm species infecting free-roaming and owned dogs from an urban area in inner São Paulo State, Brazil.

Dogs are the most popular pet animals worldwide, but on the other hand, they are main hosts of pathogens potentially transmissible to humans. The aim of this study was to assess the occurrence of intestinal parasites in free- roaming and owned dogs in an urban area in southeastern Brazil and to identify the hookworm species infecting them. Faecal samples (80 from free-roaming and 53 from owned dogs) were examined for intestinal parasites using concentration methods. DNA extracted from hookworm microscopy-positive samples were tested by PCR targeting the ITS1-5.8S-ITS2 region and the amplicons retrieved were sequenced. Intestinal parasites were detected in 43.60% (58/133) of the dogs and hookworm infection was found at the highest prevalence rate (38.30%), followed by Toxocara canis (10.50%), Trichuris vulpis (2.25%), Giardia spp. (0.75%) and Cystoisospora spp. (0.75%). Out of the 51 samples positive for hookworm eggs, 26 (50.90%) were successfully amplified and sequenced. Single infections with Ancylostoma caninum and Ancylostoma braziliense were recorded in 18 (69.20%) and two (7.70%) isolates, respectively, and mixed infections were found in the remaining six samples (23.10%). Both species were found infecting free-roaming and owned animals, but A. caninum was more common. These findings highlight the public health relevance of dogs as reservoirs of zoonotic parasites, with emphasis on hookworm species commonly implicated in cutaneous larva migrans (CLM) in poor and deprived areas.

Sarcoidosis Presenting as Cranial Multinevritis.

A 29-year-old woman presented to the emergency department with red eyes and hypoacusia since the previous weeks associated with a sudden right facial palsy. Ophthalmologic examination revealed bilateral uveitis and lumbar puncture was compatible with aseptic meningitis. Thoracic computed tomography scan showed pulmonary nodules and mediastinal and hilar lymphadenopathy. Bronchoalveolar lavage revealed a CD4/CD8 ratio of 8, and lymph node biopsy confirmed non-caseating granulomas. The diagnosis of sarcoidosis with ocular, pulmonary, and neurologic involvement was made. Due to sarcoidosis severity at presentation, the patient was started on systemic steroids and methotrexate with improvement. Sarcoidosis involvement of the central nervous system is a rare condition (5-10%) but can be the first manifestation of the disease. Its presentation can be heterogeneous; therefore, symptoms are frequently devalued by patients and physicians, leading to a late diagnosis and permanent damage. Clinicians' awareness is crucial to early diagnosis and treatment.

Left Lung and Pulmonary Artery Hypoplasia: A Rare Case of Hemoptysis.

Pulmonary hypoplasia or agenesis is a rare congenital disorder that results in lung underdevelopment. This disease is usually found in children but rarely encountered in adults. We describe the case of an 84-year-old woman diagnosed with a unilateral pulmonary hypoplasia presenting simultaneously with left pulmonary artery hypoplasia. Due to this condition, the patient had a lifelong history of pulmonary infections that resulted in several bronchiectases in the affected lung. Moreover, the pulmonary artery hypoplasia led to the development of pulmonary hypertension and collateral circulation causing hemoptysis, giving rise to the patient attending the emergency department. Although we were able to medically manage the hemoptysis, it can be fatal and require surgical intervention. Hence, an early diagnosis is essential so that appropriate follow-up and prompt prevention and treatment of complications, such as pulmonary infections, hemoptysis and pulmonary hypertension, are achieved. LEARNING POINTS: Although rare and typically associated with childhood, lung and pulmonary artery hypoplasia can in exceptional cases be present in adult life.The diagnosis of lung and pulmonary artery hypoplasia is challenging in adults as they can easily be mistaken for more common diseases.Symptoms and complications may be life-threatening, including dyspnea, hemoptysis, recurrent pulmonary infections and pulmonary hypertension.An early diagnosis is extremely important because it allows a close follow-up with consequent timely detection and treatment of serious complications, especially pulmonary hypertension.

Variable immunodeficiency study: Evaluation of two European cohorts within a variety of clinical phenotypes.

INTRODUCTION: Given the wide heterogeneity of common variable immunodeficiency (CVID), several groups have proposed clinical and immunological classifications to better define follow-up and prognostic algorithms. The present study aims to validate recent clinical and laboratory algorithms, based on different combinations of CVID biomarkers, to provide more personalized treatment and follow-up strategies. METHODS: We analysed clinical and immunological features of 80 patients with suspected or diagnosed CVID, in two reference centres of Portugal and Spain. Clinical manifestations were categorized into clinical phenotyping proposed by Chapel et al. [1] that included cytopenia; polyclonal lymphocytic infiltration; unexplained enteropathy; and no disease-related complications. RESULTS: 76% of patients in our cohort entered one of the four categories of clinical phenotyping, without overlap (cytopenia; polyclonal lymphocytic infiltration; unexplained enteropathy; and no disease-related complications). The most prominent phenotype was "cytopenia" (40%) followed by "polyclonal lymphocytic infiltration" (19%). The remaining 24% patients of our cohort had overlap of 2 clinical phenotypes (cytopenia and unexplained enteropathy mainly). A delay of CVID diagnosis in more than 6 years presented 3.7-fold higher risk of developing lymphoproliferation and/or malignancy (p < 0.05), and was associated with increased CD8+CD45RO + T-lymphocytes (p < 0.05). An association between decreased switched-memory B cells with lymphoproliferation and malignancy was observed (p < 0.03 and p < 0.05, respectively). CD4 + T-lymphocytopenia correlated with autoimmune phenotype, with 30% prevalence (p < 0.05). HLA-DR7 expression was related to CVID onset in early life in our patients (13 vs 25 years), and DQ2.5 or DQ2.2 with unexplained enteropathy (p < 0.05). CONCLUSIONS: The phenotypic and genetic study is crucial for an adequate clinical orientation of CVID patients. In these two independent cohorts of patients, classification based in clinical and laboratory algorithms, provides more personalized treatment and follow-up strategies.

Head and Neck Cancer: Improving Patient-Reported Outcome Measures for Clinical Practice.

OPINION STATEMENT: Head and neck cancer includes a wide range of tumors that occur in several areas of the upper aerodigestive tract. Most head and neck cancer patients report treatment-related late effects (both physical and psycho-social). High-quality and patient-centered care in head and neck cancer depend on the understanding of the continuum patient's experience-the disease pathway. Healthcare has been improved by involving patients more actively in the disease process, and a few reports support that patient-reported outcomes-built around the patient's experience-given in a timely manner to oncologists are extremely valuable in oncology clinical care. Implementation and clinical use of patient-reported outcomes requires some procedures involving head and neck cancer patients, clinicians, researchers, and institutional leaders The unified and integrated vision is still absent and some current concerns are being discussed to optimize benefits of patient-reported outcomes use in clinical practice. The inclusion of all first-line caregivers, team formation and training, continuous monitoring improvement, and analysis are critical success factors to consider. Our team developed a broader and inclusive understanding of patient-reported outcomes. Patient-reported outcome (Health-Related Quality of Life) assessment is implemented as a systematic and routine process in Head and Neck Unit. Head and neck cancer patients consider the questionnaire administration as part of the clinical approach. We are currently working in a program (PROimp) using mathematical models to identify common head and neck cancer patterns and building prognostic predictive models, to predict future outcomes, to appraise risk/benefit of treatments (standard or new), and to estimate patient's risk of future disease development. It is our aim to better comprehend the singular and unexpected perceptions to really provide directed and personalized cancer care defining the patient pathway. The future holds promising for PROs that are ascending as a nuclear outcome in head and neck oncology.

Forensic Medicine and the Military Population: International Dental Records and Personal Identification Concerns.

INTRODUCTION: The first goal of this research was to perceive the global commitment towards the organization and archiving of dental records and to compare it with each country's security risk rating. The second one was to study dental records in a sample of the Portuguese military population, using the available national dental records. MATERIAL AND METHODS: An e-mail was sent to representative dentistry associations in several countries, requesting some information concerning the professionals' awareness of this issue. After obtaining permission from the Ethics Committee, the information was collected through the Forensic Dental Symbols® system into the Dental Encoder®, as an extension of a Spanish study, and a generic codification was used (unrestored, restored, missing and crowned teeth). RESULTS: The most common dental record retention period is ten years after treatment. Observing the samples' dental records (595 files), we found a total of 19 040 analyzed teeth, with the following frequencies: unrestored (89.6%), restored (7.0%), missing (2.2%) and crowned (1.1%). DISCUSSION: There is a wide range of guidelines on how long dentists should keep dental records. Especially for the military population, dental records must include detailed information concerning each tooth situation, in order to support the process of human identification. CONCLUSION: This article reinforces the need for mandatory quality dental records in all countries, which must be efficiently stored and easily accessible in case dental identification is necessary. For the military population, these requirements are especially important, due to the added risks to which this group is subject.

Introdução: Esta investigação teve o intuito de procurar conhecer o comprometimento global relativamente à organização e arquivo dos registos dentários e compará-lo com o risco de segurança de cada país. Por outro lado, procurou-se estudar os processos clínicos de uma amostra da população militar Portuguesa, utilizando-se para o efeito os registos dentários. Material e Métodos: Foi enviado um e-mail para associações dentárias e solicitada informação sobre o tempo de guarda dos registos dentários. Após autorização prévia da Comissão de Ética, a informação foi recolhida através do sistema Forensic Dental Symbols® para Dental Encoder®, como uma extensão de uma investigação realizada em Espanha e utilizada a codificação genérica (dentes sãos, com restaurações, ausentes e coroas). Resultados: Globalmente, dez anos após o último tratamento, foi o procedimento mais comum relativamente ao tempo de guarda dos documentos. Após observação dos registos dentários da amostra (595 militares) verificou-se um total de 19 040 dentes analisados, com as seguintes frequências: dentes sãos (89,6%), com restauração (7,0%), ausentes (2,2%) e coroas (1,1%). Discussão: Existe grande variedade de orientações sobre quanto tempo têm que ser guardados pelos profissionais de saúde os seus registos. Nos registos dentários da população militar deve-se incluir informação detalhada de cada dente, de maneira a suportar o processo de identificação humana. Conclusão: Este artigo reforça a necessidade de registos dentários de qualidade em todos os países, com manutenção eficiente para a identificação humana. Na população militar torna-se especialmente importante, devido ao facto de este ser um grupo sujeito a riscos acrescidos.

Proteolytic activity of excretory/secretory products of Cochliomyia hominivorax larvae (Diptera: Calliphoridae) / Atividade proteolítica de produtos de excreção/secreção de Cochliomyia hominivorax (Diptera: Calliphoridae)

The protein profiles and proteolytic activity of the excretory secretory products (E/SP) of the first (L1), second (L2) and third (L3) larval stages of Cochliomyia hominivorax were studied in the laboratory. Analysis on the E/SP protein profile was carried out using polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). The E/SP of each larval stage (L1, L2 and L3) treated with protease inhibitors, containing 30µg, 40µg and 50µg of protein, was applied to the 10% polyacrylamide gel. The proteolytic activity of the crude E/SP was analyzed in gels copolymerized with gelatin and by colorimetric assays using azocasein as a substrate, with the characterization of the proteases using synthetic inhibitors. Different protein profiles were observed for the larval instars, with L1 presenting the most complex profile. Nevertheless, various protein bands were observed that were common to all the larval instars. The E/SP of all the instars showed proteolytic activity on gelatin, evidenced by proteolysis zones, predominantly with apparently higher molecular masses in L1, while for L2 and L3 the proteolysis zones could also be observed in regions with lower masses. Tests with protease inhibitors using gelatin as substrate showed that the E/SP of larvae were mainly composed of serine proteases. Additionally, inhibition was observed in L2 E/SP treated previously with EDTA, an inhibitor of metalloproteases. The assays with azocasein revealed a gradual increase of proteolytic activity on this substrate with larval development progress, with the strongest inhibitions being observed after treatments with 3,4-dichloroisocoumarin (DCI) for E/SP of L1, L2 and L3. These results suggest that C. hominivorax larvae produce different proteases, a fact that can be related to the parasite's vital processes for survival, such as penetration into the host's tissues and nutrition during the larval stage.(AU)

Os perfis protéicos e a atividade proteolítica dos produtos de excreção/secreção (PE/S) das larvas de primeiro (L1), segundo (L2) e terceiro (L3) estágios de Cochliomyia hominivorax foram estudados em laboratório. Os perfis protéicos foram obtidos por eletroforese em géis de poliacrilamida (SDS-PAGE). Os PE/S de cada fase larval (L1, L2 e L3), tratados com inibidores de proteases, contendo 30µg, 40µg e 50µg de proteína, foram aplicados em géis de poliacrilamida a 10%. A atividade proteolítica dos PE/S na sua forma nativa, foi analisada em géis co-polimerizados com gelatina e por testes colorimétricos usando a azocaseína como substrato, com a caracterização das proteases feita por meio de inibidores sintéticos. Diferentes perfis protéicos foram observados para os instares larvais, com L1 apresentando o perfil mais complexo. Apesar disso, foram observadas várias bandas protéicas comuns a todos os estágios larvais. Os PE/S de todos os instares mostraram atividade proteolítica sobre a gelatina, evidenciada por zonas de proteólise, com predominância de massas moleculares aparentes mais altas em L1, enquanto que para L2 e L3 as zonas de proteólise puderam ser observadas também em regiões de menores massas. Os testes com inibidores de proteases usando a gelatina como substrato mostraram que os PE/S de L1, L2 e L3 eram compostos principalmente de serina-proteases. Adicionalmente, inibição foi observada nos PE/S de L2 tratada previamente com EDTA, um inibidor de metalo-proteases. Os ensaios com a zocaseína revelaram um aumento gradual da atividade proteolítica sobre este substrato com o progresso do desenvolvimento larval, com a mais forte inibição sendo observada após o tratamento com 3,4 dicloroisocumarina (DCI) para os PE/S de L1, L2 e L3. Estes resultados sugerem que as larvas de C. hominivorax produzem diferentes proteases, fato que pode estar relacionado a processos vitais para a sobrevivência do parasita, tais como a penetração nos tecidos dos hospedeiros e nutrição durante os estágios larvais.(AU)

Molecular characterization of intestinal protozoa in two poor communities in the State of São Paulo, Brazil.

BACKGROUND: Several species of protozoa cause acute or chronic gastroenteritis in humans, worldwide. The burden of disease is particularly high among children living in developing areas of the world, where transmission is favored by lower hygienic standards and scarce availability of safe water. However, asymptomatic infection and polyparasitism are also commonly observed in poor settings. Here, we investigated the prevalence of intestinal protozoa in two small fishing villages, Porto Said (PS) and Santa Maria da Serra (SM), situated along the river Tietê in the State of São Paolo, Brazil. The villages lack basic public infrastructure and services, such as roads, public water supply, electricity and public health services. METHODS: Multiple fecal samples were collected from 88 individuals in PS and from 38 individuals in SM, who were asymptomatic at the time of sampling and had no recent history of diarrheal disease. To gain insights into potential transmission routes, 49 dog fecal samples (38 from PS and 11 from SM) and 28 river water samples were also collected. All samples were tested by microscopy and PCR was used to genotype Giardia duodenalis, Blastocystis sp., Dientamoeba fragilis and Cryptosporidium spp. RESULTS: By molecular methods, the most common human parasite was Blastocystis sp. (prevalence, 45% in PS and 71% in SM), followed by D. fragilis (13.6% in PS, and 18.4% in SM) and G. duodenalis (18.2% in PS and 7.9% in SM); Cryptosporidium spp. were not detected. Sequence analysis revealed large genetic variation among Blastocystis samples, with subtypes (STs) 1 and 3 being predominant, and with the notable absence of ST4. Among G. duodenalis samples, assemblages A and B were detected in humans, whereas assemblages A, C and D were found in dogs. Finally, all D. fragilis samples from humans were genotype 1. A single dog was found infected with Cryptosporidium canis. River water samples were negative for the investigated parasites. CONCLUSIONS: This study showed a high carriage of intestinal parasites in asymptomatic individuals from two poor Brazilian villages, and highlighted a large genetic variability of Blastocystis spp. and G. duodenalis.

EFFECT OF Bifidobacterium animalis ON MICE INFECTED WITH Strongyloides venezuelensis / Efeito da administração de Bifidobacterium animalis sobre a infecção por Stongyloides venezuelensis em camundongos

The administration of viable Bifidobacterium animalis was tested to induce resistance against Strongyloides venezuelensis infection in mice. Effects on parasite burden, worm length, egg output, and intestinal mucosal histology were evaluated. The oral administration of B. animalis, strain 04450B, starting 14 days before the inoculation of nematode larvae significantly decreased the worm burden and egg output. In probiotic treated animals, the percent reduction of adult worms in the intestine was of 33% and the reduction of egg production was of 21%, compared with those of the control group. The duodenum villous height and villous/crypt ratio were significantly higher in probiotic-treated mice, indicating that this group could be experiencing less intestinal damage. The present findings revealed that the administration of B. animalis for the amelioration of host response to nematode infections is biologically plausible and could have some potential for impacting public health. Meanwhile, further study is needed to delineate the nature and identity of the factor(s) involved in these beneficial effects.

Os efeitos da administração de Bifidobacterium animalis viáveis sobre a infecção por Strongyloides venezuelensis foram avaliados em camundongos experimentalmente infectados. Os parâmetros analisados incluíram a carga parasitária, o comprimento dos vermes, a quantidade de ovos eliminados e a histologia da mucosa intestinal. A administração oral da cepa 04450B de B. animalis, iniciada 14 dias antes da inoculação de larvas do nematódeo, foi acompanhada de uma redução significativa do número de vermes que se estabeleceu no intestino e do número de ovos eliminados nas fezes. Nos animais tratados com o probiótico, o percentual de redução de vermes adultos no intestino foi de 33% e da produção de ovos foi de 21%, em comparação com os do grupo controle. O comprimento das vilosidades do duodeno e a relação vilus/cripta foram significativamente maiores nos animais tratados, indicando que nestes animais as lesões intestinais foram mais leves. Os resultados do presente trabalho revelaram que a administração de B. animalis com o propósito de modular a resposta do hospedeiro contra infecções por nematódeos é uma possibilidade biologicamente plausível com impacto potencial em saúde pública. No entanto, são ainda necessários mais estudos para esclarecer os mecanismos de ação destes microrganismos e identificar os fatores envolvidos na produção dos efeitos benéficos.

Effect of Bifidobacterium animalis on mice infected with Strongyloides venezuelensis.

The administration of viable Bifidobacterium animalis was tested to induce resistance against Strongyloides venezuelensis infection in mice. Effects on parasite burden, worm length, egg output, and intestinal mucosal histology were evaluated. The oral administration of B. animalis, strain 04450B, starting 14 days before the inoculation of nematode larvae significantly decreased the worm burden and egg output. In probiotic treated animals, the percent reduction of adult worms in the intestine was of 33% and the reduction of egg production was of 21%, compared with those of the control group. The duodenum villous height and villous/crypt ratio were significantly higher in probiotic-treated mice, indicating that this group could be experiencing less intestinal damage. The present findings revealed that the administration of B. animalis for the amelioration of host response to nematode infections is biologically plausible and could have some potential for impacting public health. Meanwhile, further study is needed to delineate the nature and identity of the factor(s) involved in these beneficial effects.

Quantitative study of Babesia bovis infection in beef cattle from São Paulo state, Brazil.

The qPCR technique with SYBR Green was used to estimate the prevalence and level of Babesia bovis infection in beef cattle raised in areas endemic for babesiosis in Brazil, where the animals were continuously exposed to ticks (Rhipicephalus microplus). This is the first report in which qPCR was used to quantify and compare B. bovis DNA in blood of different cattle breeds. Blood samples were collected from 150 animals (75 cows and 75 calves) of the Angus and Nelore breeds and the first generation of an Angus and Nelore cross (AxN). Blood samples from the jugular vein were used for DNA extraction and determination of packed cell volume (PCV), while samples from peripheral veins were used for microscopic parasite detection. Although no piroplasms of B. bovis were found in blood smears, DNA amplification using qPCR revealed that all of the 150 animals, except two calves and one cow, were positive. The number of copies of B. bovis DNA was higher (p<0.05) in the Angus than in the Nelore and AxN animals, for both calves and cows, but no significant difference was found between the Nelore and AxN groups. These results suggest that a heterotic effect was present, since the results from the crossbred animals significantly deviated from the mean of the two parental groups, while closely approaching that of the Nelore group. In the Nelore and AxN groups, calves showed higher infection levels than cows (p<0.05), while for the Angus group the difference was found to be non-significant. Within each animal age group, the breed groups with higher infection levels were those with lower PCV values. However, within each breed group, no significant correlations were found between the number of DNA copies and PCV according to animal age. The qPCR method applied here allowed the observation that although there are no differences in the prevalence of infection among breed groups, Nelore and AxN cattle are able to maintain infection by B. bovis at lower levels than the Angus cattle.

In vitro antigiardial activity of the cysteine protease inhibitor E-64.

The quest for new antiparasitic alternatives has led researchers to base their studies on insights into biology, host-parasite interactions and pathogenesis. In this context, proteases and their inhibitors are focused, respectively, as druggable targets and new therapy alternatives. Herein, we proposed to evaluate the in vitro effect of the cysteine protease inhibitor E-64 on Giardia trophozoites growth, adherence and viability. Trophozoites (105) were exposed to E-64 at different final concentrations, for 24, 48 and 72 h at 37 °C. In the growth and adherence assays, the number of trophozoites was estimated microscopically in a haemocytometer, whereas cell viability was evaluated by a dye-reduction assay using MTT. The E-64 inhibitor showed effect on growth, adherence and viability of trophozoites, however, its better performance was detected in the 100 µM-treated cultures. Although metronidazole was more effective, the E-64 was shown to be able to inhibit growth, adherence and viability rates by ≥ 50%. These results reveal that E-64 can interfere in some crucial processes to the parasite survival and they open perspectives for future investigations in order to confirm the real antigiardial potential of the protease inhibitors.

Molecular typing of Giardia duodenalis isolates from nonhuman primates housed IN a Brazilian zoo.

Giardia infections in captive nonhuman primates (NHP) housed at a Brazilian zoo were investigated in order to address their zoonotic potential. Fresh fecal samples were collected from the floors of 22 enclosures where 47 primates of 18 different species were housed. The diagnosis of intestinal parasites after concentration by sedimentation and flotation methods revealed the following parasites and their frequencies: Giardia (18%); Entamoeba spp. (18%); Endolimax nana (4.5%); Iodamoeba spp. (4.5%); Oxyurid (4.5%) and Strongylid (4.5%). Genomic DNA extracted from all samples was processed by PCR methods in order to amplify fragments of gdh and tpi genes of Giardia. Amplicons were obtained from samples of Ateles belzebuth, Alouatta caraya, Alouatta fusca and Alouatta seniculus. Clear sequences were only obtained for the isolates from Ateles belzebuth (BA1), Alouatta fusca (BA2) and Alouatta caraya (BA3). According to the phenetic analyses of these sequences, all were classified as assemblage A. For the tpi gene, all three isolates were grouped into sub-assemblage AII (BA1, BA2 and BA3) whereas for the gdh gene, only BA3 was sub-assemblage AII, and the BA1 and BA2 were sub-assemblage AI. Considering the zoonotic potential of the assemblage A, and that the animals of the present study show no clinical signs of infection, the data obtained here stresses that regular coproparasitological surveys are necessary to implement preventive measures and safeguard the health of the captive animals, of their caretakers and of people visiting the zoological gardens.

In vitro ANTIGIARDIAL ACTIVITY OF THE CYSTEINE PROTEASE INHIBITOR E-64 / Atividade in vitro do inibidor de cisteína-proteases E-64 sobre trofozoítos de Giardia

The quest for new antiparasitic alternatives has led researchers to base their studies on insights into biology, host-parasite interactions and pathogenesis. In this context, proteases and their inhibitors are focused, respectively, as druggable targets and new therapy alternatives. Herein, we proposed to evaluate the in vitro effect of the cysteine protease inhibitor E-64 on Giardia trophozoites growth, adherence and viability. Trophozoites (105) were exposed to E-64 at different final concentrations, for 24, 48 and 72 h at 37 °C. In the growth and adherence assays, the number of trophozoites was estimated microscopically in a haemocytometer, whereas cell viability was evaluated by a dye-reduction assay using MTT. The E-64 inhibitor showed effect on growth, adherence and viability of trophozoites, however, its better performance was detected in the 100 µM-treated cultures. Although metronidazole was more effective, the E-64 was shown to be able to inhibit growth, adherence and viability rates by ≥ 50%. These results reveal that E-64 can interfere in some crucial processes to the parasite survival and they open perspectives for future investigations in order to confirm the real antigiardial potential of the protease inhibitors.

As cisteína-proteases estão entre os alvos mais promissores para o desenvolvimento de novos agentes terapêuticos, visto que participam de eventos fundamentais do ciclo de vida de muitos microorganismos, inclusive Giardia. Como a atividade das proteases pode ser controlada por inibidores específicos, essas substâncias têm sido avaliadas quanto ao potencial antiparasitário. Diante disso, o presente estudo teve por objetivo avaliar o efeito in vitro do inibidor de cisteína-proteases E-64 sobre o crescimento, a aderência e a viabilidade de trofozoítos de cepa de Giardia isolada em Botucatu. Nos ensaios de crescimento e aderência, o número de trofozoítos foi estimado microscopicamente em hemocitômetro, enquanto que a viabilidade celular foi avaliada pelo método do MTT. No presente estudo, embora o metronidazol tenha se apresentado bastante efetivo, o E-64 mostrou ser capaz de inibir o crescimento, a aderência e a viabilidade em taxas superiores a 50%, especialmente nos cultivos expostos à concentração de 100 µM. A despeito de preliminares, esses resultados demonstram que o inibidor E-64 pode interferir em processos primordiais para a sobrevivência do parasita, além do que, abrem novas perspectivas para investigações futuras a fim de se avaliar o real potencial giardicida dos inibidores de proteases.

MOLECULAR TYPING OF Giardia duodenalis ISOLATES FROM NONHUMAN PRIMATES HOUSED IN A BRAZILIAN ZOO / Genotipagem de isolados de Giardia duodenalis de primatas não humanos mantidos em zoológico do Brasil

Giardia infections in captive nonhuman primates (NHP) housed at a Brazilian zoo were investigated in order to address their zoonotic potential. Fresh fecal samples were collected from the floors of 22 enclosures where 47 primates of 18 different species were housed. The diagnosis of intestinal parasites after concentration by sedimentation and flotation methods revealed the following parasites and their frequencies: Giardia (18%); Entamoeba spp. (18%); Endolimax nana (4.5%); Iodamoeba spp. (4.5%); Oxyurid (4.5%) and Strongylid (4.5%). Genomic DNA extracted from all samples was processed by PCR methods in order to amplify fragments of gdh and tpi genes of Giardia. Amplicons were obtained from samples of Ateles belzebuth, Alouatta caraya, Alouatta fusca and Alouatta seniculus. Clear sequences were only obtained for the isolates from Ateles belzebuth (BA1), Alouatta fusca (BA2) and Alouatta caraya (BA3). According to the phenetic analyses of these sequences, all were classified as assemblage A. For the tpi gene, all three isolates were grouped into sub-assemblage AII (BA1, BA2 and BA3) whereas for the gdh gene, only BA3 was sub-assemblage AII, and the BA1 and BA2 were sub-assemblage AI. Considering the zoonotic potential of the assemblage A, and that the animals of the present study show no clinical signs of infection, the data obtained here stresses that regular coproparasitological surveys are necessary to implement preventive measures and safeguard the health of the captive animals, of their caretakers and of people visiting the zoological gardens.

A pesquisa de infecções por Giardia e a caracterização genotípica deste protozoário foi realizada em primatas não humanos (PNH) mantidos em Zoológico a fim de avaliar o seu potencial zoonótico. As amostras dos animais consistiram de fezes colhidas do piso de 22 baias onde eram mantidos 47 primatas de 18 diferentes espécies. Exames coproparasitológicos foram realizados pelos métodos de concentração por sedimentação e centrífugo-flutuação e revelaram a presença dos seguintes parasitas e suas respectivas frequências: Giardia (18%); Entamoeba spp. (18%); Endolimax nana (4.5%); Iodamoeba spp. (4.5%); oxiurídeos (4.5%) e estrongilídeos (4.5%). O DNA extraído de todas as amostras fecais foi submetido à técnica de PCR para a amplificação dos genes gdh e tpi de Giardia, porém, só foram obtidos amplicons das quatro amostras positivas provenientes de Ateles belzebuth, Alouatta caraya, Alouatta fusca and Alouatta seniculus. O seqüenciamento dos fragmentos amplificados foi possível apenas para as amostras oriundas de Ateles belzebuth (BA1), Alouatta fusca (BA2) e Alouatta caraya (BA3), cuja análise fenética de ambos os genes revelou pertencerem ao genótipo A. As análises das sequências de tpi revelaram que todas as amostras pertencem ao subgenótipo AII. No que se refere ao gene gdh as análises revelaram uma amostra pertencente ao subgenótipo AII (BA3) e duas ao subgenótipo A1 (BA1 e BA2). Considerando o potencial zoonótico do genótipo A e o fato de que os animais não apresentavam sintomas de infecção, os dados do presente trabalho salientam a importância de se realizar, periodicamente, exames coproparasitológicos dos animais de zoológico, para implementação de medidas preventivas para resguardar a saúde dos animais em cativeiro, a de seus tratadores e dos visitantes de parques zoológicos.

[Head and neck cancer: health related quality of life assessment considering clinical and epidemiological perspectives]. / Oncologia de Cabeça e Pescoço: enquadramento epidemiológico e clínico na avaliação da Qualidade de Vida Relacionada com a Saúde.

The importance of oncology diseases as a cause of morbidity and mortality is increasing worldwide, and their social impact is being recognized due to economic and social costs involved in prevention, treatment and rehabilitation. Head and neck cancer is one of the six most prevalent neoplasms worldwide, with an estimated 900,000 new cases diagnosed annually. Regardless of tumor site, deterioration of basic functions affecting head and neck areas are perceived and affect patients' lives. Appropriate cancer registration may provide a better analysis of health-related quality of life outcomes. In this study, 380 head and neck cancer patients were evaluated. The study showed that women have lower overall Quality of Life results. It also emphasizes the importance of early diagnosis, which often relates to stages with better prognosis and better Quality of Life outcomes. The study concluded that tumor location has an impact on Quality of Life self-perception. Values of Health Related Quality of Life should be analyzed along with socio-demographic and clinical variables in order to better understand the epidemiology, pathogenesis, and prevention of Head and Neck Cancer.

Oncologia de Cabeça e Pescoço: enquadramento epidemiológico e clínico na avaliação da Qualidade de Vida Relacionada com a Saúde / Head and Neck Cancer: Health Related Quality of Life Assessment considering clinical and epidemiological perspectives

A importância das doenças oncológicas como causa de morbilidade e mortalidade está em crescimento, sendo reconhecido o seu impacto social e peso global pelos custos económicos e sociais envolvidos na sua prevenção, tratamento e reabilitação. As patologias oncológicas de cabeça e pescoço representam um dos seis tumores malignos mais prevalentes em todo o mundo, com um valor estimado de 900.000 novos casos diagnosticados anualmente em escala mundial. Estes doentes oncológicos apresentam deterioração de funções básicas que, quando percepcionadas, têm impacto negativo na sua Qualidade de Vida. Um registo oncológico adequado permite uma análise mais rigorosa dos resultados obtidos na avaliação da Qualidade de Vida Relacionada com a Saúde. Este estudo incluiu 380 doentes oncológicos de cabeça e pescoço e demonstra que as mulheres apresentam resultados de Qualidade de Vida globalmente inferiores. Salienta-se ainda a importância do diagnóstico precoce em oncologia, que se relaciona frequentemente com melhores scores e conclui-se que a localização do tumor tem impacto sobre a autopercepção de Qualidade de Vida. Os valores de Qualidade de Vida Relacionada com a Saúde devem ser interpretados à luz das variáveis sociodemográficas e clínicas, para melhor se avaliar a Patologia Oncológica de Cabeça e Pescoço numa perspectiva epidemiológica no sentido de melhor compreender o processo saúde - doença.

The importance of oncology diseases as a cause of morbidity and mortality is increasing worldwide, and their social impact is being recognized due to economic and social costs involved in prevention, treatment and rehabilitation. Head and neck cancer is one of the six most prevalent neoplasms worldwide, with an estimated 900,000 new cases diagnosed annually. Regardless of tumor site, deterioration of basic functions affecting head and neck areas are perceived and affect patients' lives. Appropriate cancer registration may provide a better analysis of health-related quality of life outcomes. In this study, 380 head and neck cancer patients were evaluated. The study showed that women have lower overall Quality of Life results. It also emphasizes the importance of early diagnosis, which often relates to stages with better prognosis and better Quality of Life outcomes. The study concluded that tumor location has an impact on Quality of Life self-perception. Values of Health Related Quality of Life should be analyzed along with socio-demographic and clinical variables in order to better understand the epidemiology, pathogenesis, and prevention of Head and Neck Cancer.