Mestrado em Doenças Infecciosas

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Abortamento humano : detecção molecular de AAV e de HPV em decídua e vilosidade coriônica
(Universidade Federal do Espírito Santo, 2007-11-14) Pereira, Christiane Curi; Vargas, Paulo Roberto Merçon de; Spano, Liliana Cruz; Miranda, Angelica Espinosa Barbosa; Oliveira, Ledy do Horto dos Santos
Pregnancy failure is a common event often of unknown cause. Some viruses are suggested as cause of abortion and, among them, adeno-associated virus (AAV), that has never been implicated as a cause of disease. AAV (AAV1-11 types), belonging to Parvoviridae family, requires helper virus function for replication, as from human papillomavirus (HPV), that belongs to Papillomaviridae family, also suggested as being an etiologic agent of abortion. Thus, this study aimed to detect nucleic acid of AAV and HPV in decidua and chorionic villi (CV) from human abortion occurred up to 22nd gestational week. A total of 118 fragments (66 decidua and 52 CV) were obtained from tissues from 81 abortion cases (68 non-intentional and 13 intentional ones). Viral DNA was extracted by DNAzolTM, TRIzolTM and/or QIAampTM DNA mini Kit methodologies and b-globina gene was amplified by PCR as a control reaction. AAV2/5 and HPV genome were amplified (nested-PCR and PCR, respectively) using primer pairs Pan1/Pan3 and Nest1/Nest2 and MY09/11, respectively. HPV positive cases were submitted to PCR-typing for the most common types (6, 11, 16, 18, 33). In situ hybridization (ISH) was developed in paraffin embedded tissues from AAV positive cases, using digoxigenin labeled probe. Frequency of AAV and HPV were observed in 28% (23/81) and 10% (8/81) of the cases, respectively. Only type AAV2 was detected. AAV2 was present in 18 and in 7 decidua and CV fragments, respectively, while HPV, in 4 and 5 fragments, respectively. AAV2 occurred in 32% (22/68) and in 8% (1/13) of non-intentional and intentional abortions, respectively, and HPV, in 10% (7/68) and 8% (1/13), respectively. Only one HPV were typed, corresponding to HPV11. ISH showed AAV DNA in 3 cases: in decidua, CV or chorionic plate and extravillous trophoblast. Co-infection rate between AAV and HPV was 26%, and with CMV (previously studied), 9%. Significant evidence of AAV infection in abortion tissues was observed in the present study, however, in lower frequency than those found in literature. Only AAV2 type revealed in the cases, instead of AAV5, suggests that it is the most frequent in population and/or shows tissue tropism. Infected cells with AAV2, observed by ISH in decidua and in extravillous trophoblast, suggest that cellular invasiveness of infected cells could be compromised and that gestational loss may occur. Frequency of HPV in CVs is in accordance to literature. HPV was found in similar frequency in the two abortion groups. AAV presence found in cases without co-infection with helper virus, could represent latency, autonomous replication or co-infection with other helper virus. These results do not allow inference to a causal association between AAV and abortion, albeit mostly detected in non-intentional abortion.
Alterações craniofaciais e tomográficas em pessoas atingidas pela hanseníase atendidas no Programa de Unidade Básica de Saúde e no Hospital Colônia Pedro Fortes, Cariacica (ES), Brasil
(Universidade Federal do Espírito Santo, 2017-04-10) Serafim, Rachel Azevedo; Deps, Patrícia Duarte; Falqueto, Aloísio; Floriano, Marcos Cesar
Introduction: Leprosy is an infectious disease caused by Mycobacterium leprae, and, more recently, Mycobacterium lepromatosis, a slow evolution, manifested mainly by dermatoneurological signs and symptoms. In addition to affecting the skin and peripheral nerves, it can also compromise the nasal tissues, eyes, testicles and bones. Skeletal facial and extremity deformities are historical markers and part of the stigma of this disease. Objective: To study craniofacial changes in leprosy. Methods: A case series study was carried out. Fifty-five patients were evaluated, 26 ex-leprosy from the Pedro Fontes Hospital and 29 leprosy patients from the UBS of Jardim América, Cariacica/ES, from September 2015 to December 2016. They were submitted to a Questionnaire with epidemiological data, otolaryngological evaluation and tomographic examination, when necessary. We attempted to correlate some variables with nasal / facial changes clinically and radiologically such as: gender, age, delay time in the diagnosis of leprosy, classification of leprosy, group of participants, leprosy face characteristics. Results: Of the main results found, ten (18.2%) patients presented features of the leprosy face, nine (16.4%) had nasal tip projection smaller than normal, 12 (21.8%) had no nasal tip support, nine (16.4%) had no nasal dorsal support, and the columella was reduced in 14 (25.5%). Regarding the tomographic findings of the 38 patients evaluated, the nasal bones were altered in nine (23.6%), there was resorption of the anterior (partial or total) nasal spine in 21 (55.2%) and septal perforation In five (13.2%), and 16 (42.1%) had lower and middle atrophy of the turbinates. In addition, between the toothed teeth (with three or more upper incisors) only one (2.6%) presented partial resorption with root exposition of the alveolar maxilla process, about upper toothless 27 (71%) presented resorption (partial or even palatal process), and 28 (73.7%) had lost all four upper incisors.
Análise da concordância de um teste rápido treponêmico com um teste VDRL com vistas ao seu emprego na triagem reversa da sífilis em gestantes durante o pré-natal no Brasil
(Universidade Federal do Espírito Santo, 2013-09-13) Degaut, Andressa Bolzan; Dietze, Reynaldo; Barros, Geisa Baptista; Miranda, Angelica Espinosa Barbosa
Introduction: The present study used information from the Sentinel Childbirth Study for 2010/2011 (Estudo Sentinela Parturientes) under the Ministry of Health as source of data. Objectives: To analyze the concordance of a rapid treponemal test with a VDRL test and with laboratory routine flow sheet testing used for syphilis diagnosis in Brazil. To assess how often the ministerial protocol recommendations for syphilis diagnosis during the mother’s prenatal and delivery periods have been complied. Methods: serum samples from 2.426 women ready to give birth were selected out of a total of 38.393, who met the criteria for blood sampling: positive results for syphilis diagnosed by means of rapid test at the time of admission in a hospital or the absence of a syphilis screening during prenatal period. These samples were sent to the study reference laboratory where the VRDL test would be carried out. In case of a positive finding, the test TPHA – study flow chart, would be carried out. The Kappa test was used to analyze the concordance between rapid and VDRL tests. The frequency of agreement between the final results of the flow chart and the rapid test performed at the maternity hospital was also calculated. The descriptive analysis of the study results also includes socio-demographic data from the women ready to give birth and information on prenatal medical consultations. Results: The rapid treponemal test presented a concordance of 83.4% (Kappa) when compared to the non-treponemal VDRL test. The rapid test was positive to 95.6% of the 298 expectant mothers who had a syphilis positive laboratory result according to the procedures used (VDRL and reagent TPHA). Of the 303 expectant mothers with a positive result for syphilis, 23.3% were between 30-39 years old; 38% were between the 5th and the 8th grade (incomplete) of middle school or its equivalent; and 51.7% were brown-skinned individuals. Most of the expectant mothers (91.7%) had prenatal tests performed; 74.3% (225) had the test 1 for VDRL, of which 41.3 had a non-reactive result. Of the 38.3% (116) expectant mothers who got the test 2 for VDRL, 36.2% had a non-reactive result. Conclusion: The rapid treponemal test presented an outstanding concordance with the VDRL test, and considering the growth in its use during the prenatal period, it can be considered as a parameter to indicate the immediate beginning of treatment for syphilis in expectant mothers. Due to the high frequency of agreement (95.6%) with laboratory results (VDRL e positive TPHA), the rapid test can improve diagnosis and treatment to expectant mothers during prenatal medical consultations, with no need to wait for the laboratory result. Considering women with a positive result at the delivery moment, we can conclude that expectant mothers are still acquiring syphilis during pregnancy. We can also conclude that the compliance of the ministerial protocols for both tests during prenatal period and at the delivery moment should be broadened. In this context, the rapid test can perform a crucial role in promoting early diagnosis for expectant mothers and provide the due treatment, aiming to reduce congenital syphilis and to break the syphilis transmission chain.
Análise de fatores geográficos e climáticos relacionados à distribuição de Phlebotominae (Diptera: Psychodidae) em área de transmissão de leishmaniose tegumentar americana no Estado do Espírito Santo, Brasil
(Universidade Federal do Espírito Santo, 2014-08-07) Farias, Priscila Camargo Granadeiro; Fux, Blima; Falqueto, Aloísio; Shimabukuro, Paloma Helena Fernandes; Cerutti Júnior, Crispim
American cutaneous leishmaniasis (ACL) affects the skin and mucous membranes caused by dermotropic parasites of the genus Leishmania Ross, 1903. The parasites are transmitted through the bite of small insects of the family Psychodidae, the sandflies. The municipality of Cariacica, Espírito Santo, Brazil, has been in the last five years (2009-2013) among the five municipalities which had the largest numbers of reported cases in the, according to the State Department of Health (SESA-ES, 2014). The locality of Roda DÁgua has shown high epidemiological importance, due to the large concentration of cases contributing to the municipality notifications. Evaluating the registered cases from the region, from the medical reference service, Unidade de Medicina Tropical da Universidade Federal do Espírito Santo, it was observed that they occurred up to 550 m of altitude, in an area between 20-718 m above sea level. The most likely hypothesis is that the phenomenon is related to the vector distribution, since humans and animals are be present in all altitudes. From February 2002 to January 2004 sand flies were simultaneously captured in monthly collections in Roda D’Água , at three altitude levels: level 1 - up to 250 m, level 2 - between 250 and 500 m and level 3 - above 500 m. In each level, collection of sand flies was carried out in two environments: sylvatic and peridomicile. Collections were made using modified Shannon traps and by active search with Castro pooter. We evaluated flight activity of the species as the rainfall (rainy and dry seasons) and the seasonality. Only species known to be anthropophilic and with epidemiological importance were statistically analyzed. Abundance, richness, diversity, equitability, and dminance ecological indices were estimated. A total of 13,233 sand flies from 23 species were collected. The most abundant species was Nyssomyia intermedia (61.12%), followed by Pintomyia fischeri (18.20%) and Migonemyia migonei (8.68%), all species are anthropophilic. Pintomyia monticola represented 1.67% of the total specimens collected and is also antropophilic. The other species comprised 10.10% of the total specimens collected. The altitude affected the distribution of four species, and Ny. intermedia and Pi. fischeri were more abundant at level 2, Mg. migonei most abundant at level 1 and Pi. monticola at level 3. Nyssomyia intermedia and Mg. migonei were statistically more abundant in peridomestic areas and Pi. monticola in sylvatic environment. Pintomyia fischeri occurrence was not statistically significant according to environment, but it was the only one affected by the rain and showed seasonality, most commonly found in dry periods and during the winter. Nyssomyia intermedia seems to be the most important vector of ACL in Roda D’Água and Mg. migonei presents probably acting as asecondary vector. Pintomyia fischeri does not seem to be involved in disease transmission to humans in the locallity, despite having already been incriminated in others regions. According to its distribuition, probably Pi. monticola does not play a role in ACL transmission in Roda D’Água.
Análise de um procedimento simplificado de coleta de escarro para avaliação da atividade bactericida precoce de fármacos contra o Mycobacterium tuberculosis
(Universidade Federal do Espírito Santo, 2011-04-26) Nascimento, Cristina Paula do; Palaci, Moisés; Conde, Marcus Barreto; Dietze, Reynaldo
In clinical trials studies on new drugs against tuberculosis, the main methodology used is the evaluation of the early bactericidal activity (EBA), which quantifies the decrease of CFU of Mycobacterium tuberculosis in sputum samples during the first 2 days of treatment, through 12 to 16 hours of nocturnal sputum collections. However, the sputum collection for a period of 12 to 16 hours has several shortcomings. In this context, we proposed to develop and evaluate a simple, less time-consuming procedure for sputum collection that does not require hospitalization and presents the same efficiency as a 12-hour collection procedure. We performed quantitative cultures of sputum to measure the bacterial load after three different procedures: 5-hour and 12-hour sputum collections. So this study was divided into two stages. In the first step, the bacillary loads observed in sputum were compared to those obtained after a 5-hour (6,17 log10CFU/mL) and 12 hour (6,23 log10CFU/mL) collection, and there was no statistically significant difference (p =0,27). For the second stage, the patients underwent 4 procedures for sputum collection on consecutive days - two 5-hour collections and two 12-hour collections, in order to analyze the bacterial load variation intra and inter-patients. When the intra-patient variation in the bacillary load was compared between the procedures lasting 5 hours (0,037 Log10CFU/mL) and 12 hours (- 0,022 Log10CFU/mL), no statistically significant difference was observed (p = 0,56). Moreover, the morning collection has reduced the inter-patient variation and improved the accuracy for estimation of the average change in bacterial load from a collection of one day to the other (standard error for 5-hour =0,019 and standard error for 12-hour =0,028). These results show that a 5-hour morning sputum collection is as representative as that provided by a 12-hour overnight collection. In conclusion, the simplified procedure of sputum collection for a period of 5 hours proposed by this study could be used in clinical assessment of EBA as an alternative to 12-hour collections, with no impairment to the EBA evaluation of the study drug.
Análise do potencial preditivo de marcadores de exaustão celular como indicadores de sucesso da resposta terapêutica antituberculose em pacientes com tuberculose pulmonar ativa
(Universidade Federal do Espírito Santo, 2016-07-01) Portela, Bruna Sousa de Mendonça; Gomes, Daniel Cláudio de Oliveira; Santos, Eduardo Caio Torres dos; Rodrigues, Rodrigo Ribeiro
Tuberculosis (TB) infection caused by Mycobacterium tuberculosis (Mtb) continues to represent a serious risk to public health. Current treatment routines require long period of care and also are toxic, which often lead to low adhesion and increase of multidrugresistant strains. Infectious processes that result in antigen persistence or even chronic inflammatory syndromes affect the effector function of antigen-specific T cells reducing its ability to proliferate and to produce inflammatory mediators and develop the cytotoxic functions. The phenomenon described as cell exhaustion, which is characterized by a high expression of PD-1, TIM-3, CTLA-4 and KLRG-1 receptors has been described as the main mechanism involved in this immune deactivation. In addition, the severity of infections caused by pathogens such as HIV, HBV and HCV are correlated with increased expression of these markers, which suggest their association in the pathogenesis and also indicate a potential target as predictive marker of immunologic/therapeutic efficacy. In this work we aimed to evaluate the expression of the cell exhaustion receptors PD-1 and TIM-3 as a potential target for therapeutic efficacy during chemotherapy against active pulmonary Tuberculosis. We showed higher frequencies of PD-1 and TIM-3 in patients with TB and the consequent loss the expression after treatment initiation. Moreover, we observed an significant increase of IFN-ɣ and decreased IL-10 production, which directly reflected on the microbicidal capacity observed in the whole blook killing assay. Additionally a positive correlation between TIM3+ and PD1+TIM3+ expression in CD8 T cell and bacillary load was observed indicating a possible association of this receptor to clinical evolution. Our results extend the knowledge for development of new strategies in predictive diagnosis and therapeutic efficacy for TB, which may be exploited to elimination of tuberculosis.
Análise do tratamento da infecção latente pelo Mycobacterium tuberculosis na faixa etária de 0 a 15 anos nos programas de referências para tuberculose nos municípios da Grande Vitória/ES-Brasil
(Universidade Federal do Espírito Santo, 2012-06-29) Gomes, Sandra Mary Jardim; Maciel, Ethel Leonor Noia; Hadad, David Jamil; Miranda, Angelica Espinosa Barbosa; Alves, Rosana
Controling of Tuberculosis (TB) one of the measures used in Brazil is the treatment of latent infection by Mycobacterium tuberculosis (ILTB), which is made in the TB Control Program (PCT). The evolution of the infection to disease during the life is estimated at 5-10% related to immunocompetent adult, however, in child is estimated 43%.This research was an observational and analytical study, which aimed to analyze the epidemiological profile and factors related to renunciation of treatment for ILTB. Individuals were analyzed until 15 years old who were undergoing treatment for ILTB, in the PCT of the priority municipalities of Vitória, Vila Velha, Serra and Cariacica, which are the sites that used the notification form of treatment for ILTB. The form was used as a tool for data collection. The research period was between July 2009 and August 2010. 144 individuals were found in treatment for ILTB, and 140 (97,22%), the majority of nomination was TB contacts, 46 (31.94%) abandoned the treatment, and 98 (68.06%) completed the treatment. Most individuals were contacts of TB, and when was analyzed the situation of the bacilloscopy (BAAR) of the index case of contact, the majority of both the group that left, and the group that completed treatment, 97,62% e 87,91% (p=0,06), respectively, were positive. Regarding the type of contact, the home was the most frequent, and also between the two groups, both those abandoned and those who completed, totaled 81,82% e 84,21% (p=0,72), respectively, followed by others, who were uncles, grandparents , who lived nearby. Children and teenagers were evaluated mostly had scar of BCG (Bacillus of Calmette and Guérin) present, and had mostly a normal chest X-ray. Teenagers (≥ 10 years old), were checked if they had more chances to leave the treatment of ILTB, comparing with less than 10 years old, because the same need caregivers help for use of medication, however, the majority of both groups, concluded the treatment, teenagers 75% and children 64,13% (p=0.17). This study reinforces the need to control the contacts of patients, identifying priority children and teenagers and developing strategies for treatment adherence. The study can also stimulate the improvement of notification s system since, despite the orientation of MOH, its implementation has not yet been standardized
Análise dos óbitos por AIDS no Brasil e sua relação com o uso de terapia antirretroviral no período de 2009 a 2013
(Universidade Federal do Espírito Santo, 2016-04-06) Freitas, Marcelo Araújo de; Pascom, Ana Roberta Pati; Miranda, Angélica Espinosa Barbosa; Mesquita, Fábio Caldas de; Cerutti Junior, Crispim
INTRODUCTION: Despite the introduction of universal access to antiretroviral therapy (ART) in Brazil 20 years ago, around 12,000 people still die of AIDSrelated causes every year. The aim of this study was to describe antiretroviral treatment of people living with HIV (PLHIV) who died of AIDS-related causes between 2009 and 2013. METHODS: We conducted a cross-sectional, populationbased study to establish the distribution of AIDS-related deaths among PLHIV between 2009 and 2013 in Brazil according to antiretroviral therapy status. Data were generated from a linkage process between Mortality Information System (SIM) and National ART dispensing database (SICLOM). Trends were modeled using linear regression analysis. RESULTS: A total of 61,425 AIDS-related deaths were registered in Brazil between 2009 and 2013. Median age at death was 41 years (IQR: 33-49) and 65.7% (40,337) of deaths were among men; 47.2% (29,004) of PLHIV who died during the study period never started treatment, 7.0% (4,274) had discontinued treatment, 15.9% (9,775) were on ART for six months or less and 29.9% (18,372) were on ART for more than six months. Only 1.3% of the PLHIV were on third-line ARV regimens at the time of death. CONCLUSIONS: Almost all of those who died of AIDS-related causes in Brazil did not get the full benefit of antiretroviral therapy. Even in a context of universal access to antiretroviral treatment for many years, robust health policies targeting gaps in HIV continuum of care are crucial to further reduce the impact of AIDS-related mortality in Brazil.
Análise espacial de coinfecção TB/HIV em microrregiões do Brasil de 2007 a 2011
(Universidade Federal do Espírito Santo, 2017-05-16) Loriato, Karina Covre; Miranda, Angélica Espinosa Barbosa; Cerutti Junior, Crispim; Zandonade, Eliana
Introduction: Human immunodeficiency virus (HIV) infection is an important risk factor for tuberculosis (TB). Knowledge of the spatial distribution and dependence of tuberculosis and HIV/AIDS cases in Brazil will allow the visualization of its geographical distribution and the frequency of occurrence of coinfection in space/time. Objectives: To characterize the demographic and clinical aspects of the cases of TB/HIV/AIDS coinfection and to analyze the spatial dependence of epidemiological data on these cases in the micro-regions of Brazil with the use of geoprocessing tools. Methods: The study comprises 558 micro regions of the Brazilian territory. It is an ecological study in which the SINAN tuberculosis notification database was used. Demographic and clinical epidemiological profile was performed. For the spatial analysis, the Moran index, local empirical bayesian estimator (LEbayes), the global empirical Bayesian estimator (GEbayes) and the Poisson model were used. Results: The final sample of the study was 33,773 notifications of new cases of tuberculosis in HIV/AIDS patients. In the profile, 23,621 individuals (69.94%) were male; 15,882 (54.46%) were declared black or brown; 19,216 (56.9%) were aged 15 to 39 years; and 10,484 (41.09%) had between one and three years of study. Regarding clinical aspects, 24,654 (84.71%) had suspected X-ray of TB and sputum smear microscopy was positive in 12,194 (51.11%) collected samples. The most common form of tuberculosis was pulmonary tuberculosis, in 21,100 (62.5%). Cure was reported in 17,288 (53.93%) cases, abandonment occurred in 4,575 cases (14.27%) and 5,340 (16.65%) died. The Moran index showed a weak and significant spatial correlation, being the index value 0.265481 and p-value of 0.01. Conclusions: The results showed that TB/HIV/AIDS coinfection occurs mainly in men aged 15-39, browns and blacks with up to six years of study, and does present an weak overall pattern of dependence in space.
Análise fenotípica e funcional de células dendríticas e monócitos, em pacientes com tuberculose pulmonar infectados por helmintos intestinais
(Universidade Federal do Espírito Santo, 2009-04-28) Silva, Flávia Dias Coelho da; Rodrigues, Rodrigo Ribeiro; Pereira, Fausto Edmundo Lima; Carvalho, Andréa Teixeira de
The Tuberculosis (TB) remains a global threat to humanity, which no effective vaccine available until now. Epidemiological studies have shown that Mycobacterium tuberculosis (MTB) and helminths are co-endemic in many regions in the world, and that co-infections with these pathogens are frequent. The intestinal helminths infection may be a risk factor to the development of active pulmonary TB. Through phenotypic markings and dosage of cytokines and chemokines, we aimed to show that the worms are capable to deviate the Th-1 response TB protective to the Th-2 response, characteristic in intestinal helminthiasis. Blood culture for cytokines and chemokines dosage was done by the technique of marked microspheres for reading on flow cytometry, in conjunction with immunophenotyping of dendritic cells (DCs) and monocytes / macrophages through extra and intracellular markers by cytometry were used to study some aspects of innate immunity in TB patients, TB and intestinal helminths patients (TB + HELM), patients with helminths, and healthy controls. Were analyzed: a) The number and phenotype of DCs and monocytes/macrophages in peripheral blood, and b) The production of cytokines (IL-10, IL-12, TNF- , IL-6 and IL-1 ) and of some chemokines (CCL2, CCL5, CXCL8, CXCL9 and CXCL10) in the culture supernatant of whole blood after stimulation with MTB antigens. The analysis of circulating dendritic cells and macrophages, and evaluation of cytokines and chemokines in patients with TB + HELM, should us conclude that: a) The majority of DCs were immature cell phenotype. Among the cells with mature cell phenotype, there were cells mDC DC-SIGN + and CD40 + CD123 + pDC, phenotypes consistent with mature DCs to induce Th-2 response; b) Most of the activated macrophages was assessed and expressed DC-SIGN (phenotype compatible with M2 macrophages); c) The production of cytokines was significantly greater for IL-10, TNF-and IL-6, while IL-1had a tendency to increase. However, the levels of IL-12 remained similar to control group; d) significant reduction of the production of CCL-2, a trend to lower production of CXCL-8 and no significant increase of CXCL-10 were observed, whereas the chemokines CXCL CCL-5 and -9 were not detected. Although the number of cases does not permit a conclusive statement, the observations allow the possibility that infection with intestinal helminths in TB patients may be interfering with the Th-1 response to antigens of MTB
Análise histopatológica e econográfica post mortem das tireóides de pacientes infectados pelo vírus da imunodeficiência humana
(Universidade Federal do Espírito Santo, 2011-08-22) Fraga, Alana Rocha Puppim; Pereira, Fausto Edmundo Lima; Musso, Carlos; Costalonga, Everlayny Fiorot; Miranda, Angelica Espinosa Barbosa
The thyroid follicular cells have a close interaction with the immune system and are often injured by several factors determining autoimmune inflammatory response. Immunosuppression caused by Human Immunodeficiency Virus (HIV) infection provides changes in diverse organic systems, including the endocrine system. To assess the consequences of immunosuppression on the thyroid inflammation response, were examined 63 autopsy thyroid glands from patients infected with HIV, of whom 46 were matched by age and gender with patients without HIV infection. In addition to the medical records, representative thyroid gland fragments were analyzed under optic microscope. Also immunohistochemistry for identification of antigens p24, CD4, CD8 and CD20 were performed. Volume measures and glandular ecogenicity were taken correlating with microscopic findings in order to detect pathologic processes. There was no statistical difference in ecogenicity and volume measurements between the groups. The antigen p24 was not identified in any glands. Sixty seven per cent of thyroids sampled, either HIV group or control group, showed inflammatory infiltrate. There was a significant decrease in CD4+ cells in inflammatory infiltrate of HIV group. Microorganisms in thyroid tissue were observed in 19,04% of HIV group and and 3% of control group. CD8+ intraepithelial and interfollicular cells were identified in all glands. CD20+ cells were identified with no significant difference between groups. The inflammatory response may be related, in both groups, of the occurrence of sepsis and thyrotoxic drugs. The reduction of CD4+ cells contributes to changes in the thyroid inflammatory response. The CD8+ intraepithelial cells detected in all glands with little predominance in HIV group may indicate changes in CD4+/CD8+ cells as observed in other organic systems. The measurement of thyroid volume and ecogenicity were not able to identify the specific occurrence of pathological processes between the groups.
Análise histopatológica e imuno-histoquímica da erupção papular prurítica associada ao vírus da imunodeficiência humana
(Universidade Federal do Espírito Santo, 2012-06-22) Tozzi, Brunela Bastos; Deps, Patrícia Duarte; Oliveira, Norma Suely; Miranda, Angélica Espinosa Barbosa
Introduction: Papular pruritic eruption (PPE) is characterized by the presence of pruritic erythematous papules as chronic disease that affects oftenly seropositive patients. The lesions are seen in different stages, preferably in the trunk and extremities, and end with post inflammatory hyperpigmentation and / or scars resulting from scratching. Objectives: To evaluate the hitopathological and immunohistochemical features of the PPE. Methods: The study enrolled HIV-positive patients with clinical diagnosis of PPE carried out at University Hospital Cassiano Antonio Moraes of the Federal University of Espirito Santo in Vitória, ES. Patients were dermatological examinated and skin lesions photographed. Skin biopsies were obtained to evaluate the histopathological and immunohistochemical features. Results: Thirty-nine patients with EPP were biopsied, 17 were women and 22 men, mean age was 40.2 years ranging from 25 to 63 years. Among the histopathological biopsies, 6 patterns were found and these often: 1) dermal perivascular inflammatory infiltrate, 2) inflammatory infiltrate affecting the follicular unit; 3) interstitial inflammatory infiltrate, 4) inflammatory infiltrate in the subcutaneous tissue, 5) inflammatory infiltrate affecting at least one nerve, 6) signal of the scratching in the epidermis. About immunohistochemistry features, Langerhans cells are found in normal amounts in the epidermis and relatively increased in dermal inflammatory infiltrate. There are predominantly CD8 + T lymphocyte and macrophages in the inflammatory infiltrate compared with CD4 + T lymphocytes. No microorganisms were found using the Ziehl-Neelsen staining and Grocott. Conclusions: It was identified six patterns present in biopsy, in order to obtain a set of histopathologic and immunohistochemical to describe and characterize the PPE histopathological diagnosis. The dermal perivascular inflammatory infiltrate is essential for diagnosis, and other tissue alterations are not mandatory that may be present together or separately, at different stages of the skin lesions.
Análise proteômica de isolados de Leishmania (Leishmania) chagasi sensíveis e resistentes à miltefosina
(Universidade Federal do Espírito Santo, 2011-04-05) Trindade, Juliana Brambilla Carnielli; Figueiredo, Suely Gomes de; Lemos, Elenice Moreira; Pereira, Fausto Edmundo Lima; Escobar, Patrícia Cuervo
Visceral leishmaniasis is a systemic disease that is fatal if untreated and is caused by the Leishmania donovani complex, which include the Leishmania (L.) chagasi. Visceral leishmaniasis treatment relies on a few chemotherapeutic drugs including Sb(V), amphotericin B and miltefosine. Miltefosine was recently approved as the first oral drug active against visceral leishmaniasis in India. Miltefosine resistance mechanisms are being elucidated in laboratory Leishmania spp. isolates but are less clear in clinical isolates. In this study, we used comparative two-dimensional gel electrophoresis and mass spectrometry methodologies to highlight and identify proteins that are differentially expressed between miltefosine-sensitive (S: S1 and S2) and resistant (R: R1 and R2) L. (L.) chagasi isolates from kala-azar patients included in clinical trial in Brazil, in order to assess the effectiveness of this drug. We describe here a high-resolution proteome for L. (L.) chagasi promastigotes comprising an average of 459 spots, which corresponds to 5,7% of gene products predicted for Leishmania spp. Following comparison of the whole proteome profiles between sensitive and resistant L. (L.) chagasi clinical field strains, 80 differentially expressed spots were detected. Eighteen spots were found to be specific of a sensitive group and only one of a resistant group, while 48 spots changed in intensity between these groups. The others spots were present in a unique isolates (7 in S1, 3 in S2 and 2 in R1) or in three isolates simultaneously (1 in S1, S2 e R1). MALDI/TOF-TOF mass spectrometry allowed the identification of 49 spots (61,3%) corresponding to 32 distinct protein and 7 hypothetical proteins. Among the proteins identified, the comparative proteomics screen highlighted two proteins, peroxidoxin (overexpressed in R group) and calpain-like cisteína peptidase (exclusively detected in S), differentially expressed, suggesting that programmed cell death is reduced in resistant parasite. These data suggest that these proteins may be related to resistance phenotype to miltefosine.
Aspectos epidemiológicos e clínicos da paracoccidioidomicose no Estado do Espírito Santo
(Universidade Federal do Espírito Santo, 2012-08-31) Peçanha, Paulo Mendes; Falqueto, Aloísio; Araujo, Mariceli Lamas; Miranda, Angelica Espinosa Barbosa; Wanke, Bodo
In a serie of cases of paracoccidioidomycosis we retrospectively reviewed the records of 546 patients treated at the Hospital Universitario Cassiano Antonio Moraes (HUCAM), Federal University of Espírito Santo - UFES, from 1978 to 2012. The patients ages ranged from 7 to 83 years, with 512 males and 34 females in a ratio of 15:1. Most of the patients( 81,34%) were from the state of Espírito Santo (ES), 18,66% came from other states, mainly from rural areas and 77,6% of them were farmers. The map of geographical distribution in ES showed higher concentration of cases in counties in the western border, along Minas Gerais state.Looking at the clinical features 60 (10.99%) patients presented the acute / subacute form of the disease, 485 (88.83%) the chronic form and one patient the subclinical form. The organs most often affected were the lungs, oropharyngeal mucosa, lymph nodes, skin and larynx. Several other organs were also affected less frequently. The diagnosis was established by histopathology in 252 (46.15%), direct examination in 168 (30.77%), direct examination and histopathology in 111 (20.33%) and serology in 15 (2.75%) patients. The infectious diseases more frequently associated were tuberculosis, AIDS, leishmaniasis and intestinal parasites. Patients were treated with sulfonamides, or azole in mild to moderate forms and amphotericin B in severe forms. The length of the treatment was analysed in 366 patients. Only 146 (40%) completed the minimum of 18 months needed when taking sulphonamides. The most common sequelae were residual pulmonary lesions and scars on skin and mucous membranes. The number of cases in this serie highlights the state of Espírito Santo as important endemic area of paracoccidioidomycosis in Brazil and the results follow the clinical and epidemiological patterns demonstrated in other series.
Avaliação custo-efetividade da utilização de um teste rápido como método alternativo no fluxograma para a detecção de anticorpos anti-HIV
(Universidade Federal do Espírito Santo, 2008-03-27) Borges, Cristina Betim; Rodrigues, Rodrigo Ribeiro; Valli, Luiz Carlos Pedrosa; Dietze, Reynaldo
Even with the advances that have allowed a better understanding of pathogenesis, epidemiology and prevention of infection, approximately 33.2 million people worldwide are suffering from HIV / AIDS. The HIV / AIDS is considered a pandemic, causing a major demographic impact due to the devastating infection among young adults. According to the Organization of the United Nations fighting AIDS (UNAIDS), despite a reduction in the overall number of new infections, this index is still very high, and the sexually transmission still the more frequent form of HIV infection. In order to interrupt transmission efficiently an accurate and fast diagnosis is pivotal. The great majority of diagnostic methods for HIV are based in the detection of antibodies specific to different proteins of the virus or the detection of its genetic material. Therefore delay in obtaining accurate results affects negatively prevention and treatment strategies. In the present study, we examined the effectiveness and cost of a rapid test (Rapid Check) as an alternative method in the serological diagnosis workflow for HIV infection. Three hundred and eight serum samples sent to LACEN-ES between October 2006 and May 2007 were analyzed. Both the Rapid Check and the prototype Tri-line depicted a high degree of concordance (98.97%) compared to other tests used for diagnosis of HIV infection at the LACEN-ES. In parallel, from 16 samples with results considered as "inconclusive" by the normal workflow due to absence or low reactivity of anti-gp41, only 3 out of these 16 were still considered inconclusive after using the Rapid Check. In addition to its efficiency, when cost were considered, rapid tests had the lowest cost/test, and required the shortest time to provide a final result (less than 15 minutes) not requiring additional equipment, accessories or specific infrastructure. Therefore, we believe that rapid tests with performance equal to or greater than the Rapid Check could be used as an alternative method at the Immunofluorescence, thus promoting a reduction in the cost and time of release of results.
Avaliação da acurácia de métodos fenotípicos propostos por manuais de referência na classificação de Klebsiella pneumoniae carreadora do gene blaKPC
(Universidade Federal do Espírito Santo, 2017-04-12) Pegoretti, Jaqueline; Schuenck, Ricardo Pinto; Ferreira, Ana Paula Nunes; Spano, Liliana Cruz; Pereira, Polyana Silva
Hospital infections caused by multidrug resistant bacteria constitute a serious public health problem worldwide associated with treatment failure and higher mortality rates. Among Enterobacteriaceae family, KPC-producing K. pneumoniae is an important pathogen responsible for these infections and have received the most attention because they have high potential to spread. Accurate detection of KPC producers is essential for infection control measures and antibiotic therapy. However, this is a major issue in microbiology laboratories, because the current clinical breakpoints used by reference guidelines are not capable to detect all carbapenemase producers and none confirmatory tests have 100% sensibility and specificity. The aim of the study was to evaluate methods and interpretative criteria proposed in Clinical and Laboratory Standards Institute (CLSI) and Brazilian Committee on Antimicrobial Susceptibility Testing (BrCAST) manuals to detect carbapenemase production in 47 K. pneumoniae clinical samples carrying blaKPC gene. The carbapenems susceptibility testing and the screening criteria to select samples to perform confirmatory tests (Modified Hodge Test and Inhibitor-based method) were performed through disk diffusion method and minimum inhibitory concentration (MIC) determination and was interpreted according clinical breakpoints proposed by both guidelines. All clinical samples were classified as MDR (non-susceptibility to at least three antimicrobial classes). The results support that carbapenems clinical breakpoints plus criteria of screening to select strains proposed by BrCAST presented better performance to correctly select K. pneumoniae possessing blaKPC strains to confirmatory tests. It is important to use all three carbapenems to ensure higher accuracy in susceptible testing. Furthermore, was observed that disk diffusion method applying BrCAST criteria was the only capable to correctly select all strains to confirmatory tests. Both confirmatory tests were able to identify carbapenemase production in the same 40 strains. Seven strains showed negative results on confirmatory tests in despite of the presence of blaKPC gene.
Avaliação da atividade antifúngica de extratos vegetais e antissépticos bucais em Candida albicans
(Universidade Federal do Espírito Santo, 2012-08-10) Balliana, Roberta Carvalho Sthel; Jamal, Cláudia Masrouah; Araújo, Mariceli Lamas de; Velloso, Tânia Regina Grão; Nunes, Ana Paula Ferreira
Yeasts are opportunistic fungi responsible for most fungal infections in humans and Candida albicans is the most often species associated with these infections. Oral candidiasis is distinguished by high impact and by the frequent use of antifungal drugs, which can lead to the occurrence of resistance and treatment failure for this infection. There is a constant need for research about new agents for the prophylaxis and effective treatment of oral candidiasis. The main objective of this study was to evaluate the potential antifungal effect of extracts of plants Gossypium hirsutum L., Arctium lappa, Equisetum sp. Cecropia pachystachya Trécul. and Pogostemon heyneanus against not adhered cells and biofilm of Candida albicans and evaluation of the effect of the mouthwash Cepacol ® Ice Cool, Cool Mint Listerine ®, Periogard ® and Plax ® Whitening against not adhered cells and biofilms of Candida albicans. The methods used were based on inhibition of cell growth by turbidimetry and for determination of the Minimum Inhibitory Concentration (MIC) of clinical isolates of C. albicans. The results obtained in tests with non-adherent cells were promising with extracts of Cecropia pachystachya Trécul., presenting MIC values between 7.81 and 3.91 mg / mL. The antiseptic Periogard Cepacol ® had the lowest MIC values, between 0.20 and 0.10%; Plax Whitening ® ranged between 1.56% and 0.39% and Listerine ® showed values between 6.25% and 0,39%. Statistical differences (Kruskal-Wallis test, p <0.0001) revealed that the mouthwashes Cepacol ® Cool Ice and Periogard ® showed the best results. Regarding the inhibition of biofilm formation, the mouthwashes MICs were similar, ranging between 50% and 0.20%, with no statistical differences between among them (Kruskal-Wallis test, p = 0.6915).
Avaliação da concordância entre os testes tuberculínico e o QuantiFERON®-TB Gold in tube no diagnóstico da infecção latente pelo Mycobacterium tuberculosis em crianças e adolescentes
(Universidade Federal do Espírito Santo, 2015-07-06) Boni, Patrícia Marques Rodrigues; Miranda, Angelica Espinosa Barbosa; Dietze, Reynaldo; Palaci, Moisés; Dalcolmo, Margareth Maria Pretti
Background: One of the main characteristics of M. tuberculosis (Mtb) is its capacity to produce latent infection. The World Health Organization (WHO) estimates that 2 billion people are infected by this bacillus, only 10% of whom develop active disease. Until recently the tuberculin skin test (TST) was the only test available for the diagnosis of latent infection. However, because of its potential limitations an effort was made to develop a more accurate method to diagnose latent tuberculosis. The interferon-gamma release assays (IGRA’s) were the result. These tests measure the interferon-gamma production by T-lymphocytes in response to a challenge of three synthetic antigens, specific for Mtb (ESAT-6, CFP-10 e TB7.7). The objective of our study was to evaluate the performance of the TST and the QuantiFERON®-TB Gold assay-in-tube (QFT-GIT), for the diagnostic of latent Mtb infection in children and adolescents identified as household contacts of smear positive pulmonary Mtb index cases. Methods: This was a comparative study conducted in the Metropolitan area of Vitória, Espírito Santo, Brazil from March 2008 through October 2013. Clinical, demographic and epidemiological data were collect from all participants. We defined latency as a positive TST (induration ≥ 10mm) or a positive QFT-GIT test (Nil ≤8.0 IU/ml, TB Antigen minus Nil ≥0.35 IU/ml and ≥25% of Nil value). Both tests were performed in each volunteer between eight and ten weeks after the identification of the respective index cases. Concordance, as defined by kappa testing, was used to compare the results of the two methods of diagnosing latency. The factors associated with positivity of both tests in bivariate analysis (p ≤0.30) were included in the multivariate model (logistic regression), and the odds ratios (OR) and 95%CI were calculated. Results: 291 subjects were enrolled in the study. The global concordance between TST and QFT-GIT was high (87,6%) - k 0.75 [CI 95%: 0.63- 0.86]. However in children age ≤ 5 years the concordance was even higher (93.65%) - κ 0.87 [CI 95%: 0,63-1,12]. The presence of BCG scar was not statistically associated with tests positivity. There was a positive correlation between the size of TST induration and the probability of a positive QFT-GIT. Conclusion: There was no advantage in the use of QFT-GIT in relation to the TST. We recommend the twomonth timeframe after the identification of the index case to perform the tests in the household contacts.
Avaliação da eficácia da claritromicina e da amicacina no tratamento da infecção experimental de camundongos BALB/c pelo Mycobacterium massiliense
(Universidade Federal do Espírito Santo, 2012-08-21) Colombo, Débora Corona; Palaci, Moisés; Zanini, Marcos Santos; Pereira, Fausto Edmundo Lima
Nosocomial infections caused by rapidly growing mycobacteria (RGM) have increased significantly in recent years in developed and developing countries. In Brazil, the occurrence of outbreaks due to RGM was reported since last decade. In most cases, a single strain of M. massiliense, named BRA100, was the causative agent. M. massiliense, as well as other species of the group RGM, is resistant to a broad-spectrum of antimicrobial agents, which limits the therapeutic options for the treatment of patients. Due to the lack of clinical trials to establish appropriate therapeutic regimens, the current guidelines for the treatment of infections caused by these microorganisms are based on in vitro antimicrobial activities and clinical case reports. However, there are variations in the correlation between the in vitro susceptibility tests results and the therapeutic response observed, which creates a controversy about their use to support the pharmacotherapy of infections by RGM. In this context, we conducted a study to evaluate the activities of clarithromycin and amikacin against M. massiliense in BALB/c mice. The Minimal Inhibitory Concentration (MICs) of clarithromycin and amikacin for the M. massiliense isolate obtained from a nosocomial outbreak that occurred in 2007, were determined by the broth microdilution assay using cation-adjusted Mueller–Hinton. For in vivo tests, BALB/c mice were infected intravenously with 1 X 107 to 2 x 107 CFU/mL of M. massiliense. Treatments with amikacin or clarithromycin lactobionate were initiated one day post-infection. The mice received daily by subcutaneous injection, 0.1 ml of clarithromycin lactobionate (50mg/kg/12h) or 0.1 ml of amikacin (100mg/kg/day) for 14 days. The mice were sacrificed 24 to 48h after administration of the last dose. Spleen and liver were aseptically removed for determination of weight and number of CFU (log10) per organ. Our results demonstrated in vitro activities of clarithromycin and amikacin against the M. massiliense isolate. Treatment of BALB/c mice with clarithromycin prevented the development of hepatosplenomegaly, as well as the formation of granulomatous lesions for 15 days of infection with M. massiliense. Nevertheless, the clarithromycin and even amikacin were not effective, inducing no reduction of the bacterial population of M. massiliense in the organs. The in vitro susceptibility for bacteria isolated from organs of the mice demonstrated that there was no development of inducible resistance to clarithromycin and amikacin during 12 the treatment. In conclusion, there was no correlation between the activities in vitro and in vivo of clarithromycin and amikacin against M. massiliense.
Avaliação da reação em cadeia da polimerase para identificação do complexo Mycobacterium tuberculosis em cultura de escarro no meio BACTEC 12B
(Universidade Federal do Espírito Santo, 2006-09-27) Gouveia, Ana Cláudia Carvalho; Vinhas, Solange Alves; Palaci, Moisés; Lemos, Elenice Moreira; Hadad, David Jamil
Use of the most rapid and reliable laboratory tests for Mycobacterium tuberculosis detection and identification is important for tuberculosis (TB) control. Diagnostic techniques based on molecular biology methods are able to dramatically reduce the time of detection as well as increase the sensitivity for detecting the bacilli. A polymerase chain reaction DNA amplification method for direct detection of Mycobacterium tuberculosis Complex (PCR IS6110) in BACTEC 12B broth cultures was evaluated. A total of 107 sputum samples were processed by standard methods, and then inoculated into Ogawa slants and BACTEC 12B vials. The PCR assay was used on BACTEC 12B broth cultures with a growth index (GI) of equal to or greater than 30. Molecular results were compared to those obtained by phenotypic identification methods. Of the 107 broth cultures evaluated, 90 were all culture and PCR positive for M. tuberculosis. Except for one, all cultures (n = 8) which grew mycobacteria other than M. tuberculosis were PCR negative. In two cultures which grew both mycobacteria and other organisms than acid-fast bacilli a phenotypic identification was not possible, and both of them were PCR negative. The remaining seven cultures that did not contain mycobacteria were PCR negative. Of particular interest were all 44 cultures positive from smear-negative sputum specimens and three contaminated BACTEC 12B broth cultures yielding mycobacterial growth which a M. tuberculosis Complex, which were successfully identified by PCR, resulting in a mean time to identify M. tuberculosis of 5,5 and 16 days before phenotypic identification, respectively. In light of an overall sensitivity and specificity of 100% and 87,5%, respectively, coupled with the ability to identify the bacilli days or weeks before other methods can be applied, we conclude that PCR might prove to be a rapid alternative for identification of M. tuberculosis Complex in culture, even in the context of paucibacillary patients.

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