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OBJECTIVE: Mohs micrographic surgery is a specialized surgical procedure used to treat skin cancer. The purpose of this study was to better understand the profile of the patients who underwent the procedure and to determine how histology might be related to complications and the number of stages required for complete removal. METHODS: The records of patients who underwent Mohs micrographic surgery from October 2008 to November 2013 at the Dermatology Division of the Hospital of the Campinas University were assessed. The variables included were gender, age, anatomical location, histology, number of stages required and complications. RESULTS: Contingency tables were used to compare the number of stages with the histological diagnosis. The analysis showed that patients with superficial basal cell carcinoma were 9.03 times more likely to require more than one stage. A comparison between complications and histological diagnosis showed that patients with superficial basal cell carcinoma were 6.5 times more likely to experience complications. CONCLUSION: Although superficial basal cell carcinoma is typically thought to represent a less-aggressive variant of these tumors, its propensity for demonstrating “skip areas” and clinically indistinct borders make it a challenge to treat. Its particular nature may result in the higher number of surgery stages required, which may, as a consequence, result in more complications, including recurrence. Recurrence likely occurs due to the inadequate excision of the tumors despite their clear margins. Further research on this subtype of basal cell carcinoma is needed to optimize treatments and decrease morbidity.

FUNDAMENTO: A psoríase é uma dermatose inflamatória crônica caracterizada por lesões eritemato-descamativas que atingem extensas áreas da pele, comprometendo a qualidade de vida dos pacientes por interferir na sua vida pessoal, no relacionamento social e nas atividades diárias. O tratamento permite um bom controle, mas a impressão do paciente, quase sempre, é pouco valorizada. OBJETIVOS: Avaliar a qualidade de vida de pacientes psoriáticos, verificar se há correlação desta com melhoras clínicas e se há equivalência entre dois questionários de qualidade de vida. MÉTODOS: Foram incluídos no estudo pacientes maiores de 18 anos que estavam no início ou troca de tratamento no ambulatório de psoríase do Hospital de Clínicas da Unicamp. Eles responderam dois questionários de qualidade de vida (Psoriasis Disability Index e Dermatological Life Quality Index), até completar 180 atendimentos, para avaliar se haveria correlação entre os questionários. Receberam também um índice clínico (Índice de Área e de Severidade da Psoríase) no tempo inicial da pesquisa e nos retornos subsequentes. RESULTADOS: A pesquisa foi realizada com 138 pacientes (76 homens, idade média de 50 anos). Observou-se correlação entre os questionários aplicados e o índice clínico, mostrando que alterações clínicas se refletem na qualidade de vida, e que há equivalência entre os questionários. CONCLUSÃO: Constatou-se que a maioria dos pacientes atendidos no ambulatório tem apresentado melhora clínica e da qualidade de vida e que há equivalência entre os questionários de qualidade de vida.

Summary Background The strongest genetic marker for psoriasis is Cw*06. Polymorphisms in the tumor necrosis factor (TNF)-alpha promoter region, especially replacement of guanine with adenine in positions -238 and -308 are related to higher TNF-alpha production and higher risk for psoriasis in Caucasoid populations, not found in Asians. We performed a case-control study of 69 patients with psoriasis type I and 70 controls, characterized clinical progression along 10-years of follow-up in mild or severe disease and determined HLA class I, II, and TNF single nucleotide polymorphisms (SNPs) -238 and -308 polymorphisms to demonstrate whether these polymorphisms may be genetic risk for susceptibility to psoriasis or severity of the disease in Brazilians. Methods Polymorphisms were identified using PCR/SSP. Alleles, genotypes, and haplotypes frequencies were compared using Fisher’s test. Results More severe disease was found in male patients. It may be suggested that alleles B*37, Cw*06, Cw*12, and DRB1*07 were associated with severe disease course, while B*57 with mild disease. No statistical difference was found between the patients and controls regarding polymorphisms frequencies in TNF SNPs. This study pointed to a higher TNF-238 G/G genotype frequency (OR: 3.21; CI: 1.06–9.71; P = 0.04) in the group with severe disease. Conclusions Polymorphisms in the TNF-alpha SNPs do not seem to be a more important genetic risk factor for psoriasis than the already known Cw*06 in Brazilian patients, but these markers may be related to clinical manifestations.

This article was migrated. The article was not marked as recommended. The electronic resources available for educational use have rapidly developed in recent years, thus increasing the use of new assessment and evaluative pedagogical models. Pre- and post-course testing using a virtual learning environment (VLE) can be useful for analyzing student learning, as well as for evaluating the course itself. This study aimed to identify and evaluate the use by the students of a VLE assessment model as part of a dermatology course at the School of Medical Sciences (SMS) - UNICAMP and to evaluate the course itself.The research sample included 145 students who were enrolled in the medical degree program in 2015 and 2016. The data was obtained using the Modular Object-Oriented Dynamic Learning Environment (MOODLE) platform, an identification tool containing 10 questions called "Knowing You" and a bank of 80 multiple-choice questions with a similar level of difficulty that had been previously analyzed by residents and dermatology professors. Evaluations were conducted before the beginning of the module (pre-test), shortly after the dermatological practicum (post-test) and at the end of the module (final test). Among the students invited to participate in the study, 81.38% responded to the "Knowing You" questionnaire, and the majority of students displayed good or reasonable knowledge of technology and preferred to study alone at night. Only 2.6% of the students used the VLE. During the assessment step, 50.34% of the participants completed all three tests. The post-practicum test scores were higher than those of the final-module test, which in turn were higher than the pre-module test scores. The analysis was adjusted for the groups in regard to the completion time of the partial post-practicum test. Varying time did not influence the test results among different classes (p=0.7456) or among other groups (p=0.9073).The method enabled the creation of tests with a similar degree of difficulty as no significant difference was observed between the groups and students. We observed that few of the students in the medical course at SMS - UNICAMP used the VLE and that the system of assessment was useful for documenting the module learning.The results also induced changes in the learning model used in the discipline of dermatology, with the goal of better knowledge retention by the end of the module. Therefore, it may be beneficial for students, professors, educational institutions, and development agencies to re-examine their use of VLEs in dermatology education. Questions submitted by dermatology professors from other well-recognized medical schools can be added to the question pool. Thus, this method can also allow for an external evaluation of the discipline or the school.

Some Bartonella species are able to invade red blood cells (RBC) and may cause persistent infection in the susceptible host. Use of transmission electron microscopy (TEM) demonstrates, inside erythrocytes, the typical triple-walled agents. However, when examining ultrathin sections of blood cells, the authors have, on several occasions, detected intraerythrocytic abnormalities that mimic but are not typical of Bartonella spp. Small endovesicles, pseudoinclusions, cavities, and irregular hemoglobin granules distribution, resulting in regions of increased or decreased electron density, may be observed in the erythrocytes and platelets, which may be confused with bartonellas. So far, detailed ultrastructural findings of Bartonella spp. in blood cells have not yet been described. Aiming to improve TEM interpretation of blood cells changes, in routine examination of blood sections of patients with suspected bartonellosis, the authors studied the morphological findings they have observed, and present their putative nature, according to information in the literature.

The authors present the case of a young man with aplastic anemia who went into shock and died after several red blood cell unit transfusions. Immunohematological studies did not show any abnormality and blood cultures from patients and blood bags were negative. The ultrastructural findings, allied with current scientific knowledge, permitted the diagnosis of Bartonella sp. infection. In face of this diagnosis, two possibilities should be considered: the first one is that the patient was already infected by the bacteria before the last RBC unit transfusion. The pathogen could be involved in aplastic anemia etiology and in the failure to recover hemoglobin levels, in spite of the transfusions. The second possibility is that the RBC unit was contaminated with a Bartonella sp., which would have led to a state of shock, causing the death of the patient.

A criocirurgia com nitrogênio líquido é recurso cirúrgico utilizado para o tratamento de várias lesões cutâneas, benignas, pré-malignas e malignas. Promove destruição dos tecidos acometidos por congelamento e alterações da resposta imunológica. Os autores enfocam a utilização da criocirurgia no tratamento das dermatoses infecciosas, principalmente as verrugas virais, leishmaniose e ccromoblastomicose. Discutem os resultados como tratamento primário ou coadjuvante. O artigo também apresenta os cuidados com os instrumentos no tratamento das doenças infecciosas. Cryosurgery using liquid nitrogen is a surgical resource to treat several types of benign, pre-malignant and malignant skin lesions. It promotes the destruction of tissues and changes the immune response. The authors identify several uses of cryosurgery in treating skin disorders, particularly viral warts, chromoblastomycosis and leishmaniasis. The results of primary and adjuvant treatments are also discussed. The article focuses on precautions when using surgical instruments during treatment of infectious diseases.

Background: Keratoacanthoma (KA) is easily confused with squamous cell carcinoma (SCC) on a clinical or a histopathologic basis. However, KA undergoes spontaneous regression, whereas SCC does not. Objective: Our objective was to study the histopathologic features associated with clinical regression in KA-like lesions to support the therapeutic option. Methods: Forty-three biopsies of KA-like lesions were taken at patient admission. One month later, surgical excision was performed in 18 growing lesions. Regressing lesions were left untreated. Classic histopathologic features and diagnosis were blindly recorded in both biopsies and surgical specimens. Results: On a clinical and a histologic basis, 32 lesions were assessed as KA and 11 as SCC. Features that indicated malignancy were observed in both groups, but the probability of SCC was 31 times higher in tumors with five or more of such features. Several of the histologically atypical lesions were found to regress. Conclusion: SCCs and KAs have more pathologic similarities than differences, especially in the proliferative phase. The combination of the most useful features did not allow the nosologic diagnosis in difficult cases but helped. Differential diagnosis was easier to determine after the 1-month follow up. Complete surgical excision should be indicated in nonregressing and growing lesions. Antecedents: II est facile de confondre le kerato-acanthome (KA) avec le carcinome squameux (CS) sur une base clinique ou histopathologique. Toutefois, les KA subissent des regressions spontanees, ce qui n'est pas le cas des CS. Objectif: Etudier les caracteristiques histopathologiques associees a la regression clinique des lesions ressemblant au KA afin de trouver la meilleure option therapeutique. Methodes: Quarante-trois biopsies de lesions ressemblant au KA ont ete pratiquees sur admission des patients. Un mois plus tard, une excision chirurgicale a ete pratiquee sur 18 lesions en etat de croissance. Les lesions en regression n'ont pas ete traitees. Les caracteristiques histopathologiques classiques et le diagnostic des biopsies et des excisions ont ete enregistres en insu. Resultats: Les evaluations cliniques et histologiques ont decele 32 lesions comme KA et 11 comme CS. Les caracteristiques d'une tumeur maligne ont ete observes dans les deux groupes mais la probabilite que ce soit un CS etaient 31 fois plus elevees dans les tumeurs presentant cinq caracteristiques ou plus. Plusieurs des lesions atypiques du point histologique ont regresse. Conclusion: Les CS et les KA ont davantage de similitudes que de differences au niveau pathologique, surtout dans la phase de proliferation. La combinaison des caracteristiques les plus importantes n'a pas permis un diagnostic nosologiques dans les cas complexes, mais y a contribue. Le diagnostic differentiel etait plus facile a etablir apres un mois de suivi. L'excision complete doit etre indiquee dans le cas de lesions non regressives ou en pleine croissance.

Bartonella henselae, a facultative intracellular bacterium, has been known as the agent of cat scratch disease, bacillary angiomatosis, peliosis hepatis, endocarditis, and bacteremic syndrome in humans. Bartonella species can cause intraerythrocytic infections and have been isolated from the bloodstream of patients by several methods. It was demonstrated that B. bacilliformis and B. quintana infect human endothelial cells and human erythrocytes and B. henselae infects erythrocytes of cats. The aim of this study was to investigate through transmission electron microscopy whether B. henselae infects mature human erythrocytes. One red blood cell (RBC) unit received an experimentally standard strain of B. henselae. Blood aliquots were collected from the infected unit immediately after inoculation, at 30 min and 1, 5, 10, and 72 h for ultrastructural evaluation. B. henselae was seen adhering to human erythrocytes 10 h after inoculation and inside the erythrocyte after 72 h. This study demonstrates that B. henselae adheres to and invades mature human erythrocytes. The results favor the possibility that erythrocytes can serve as a primary target in Bartonella spp. infections. From this observation, further studies are warranted to prevent Bartonella spp. transfusional transmission.

A frequência de dermatoses é alta, apresentando grande impacto na qualidade de vida dos pacientes. Contudo, há poucas políticas públicas nesta área de saúde, e o tempo destinado ao ensino dermatológico na graduação médica é restrito. Objetivos Definir as hipóteses diagnósticas e as condutas mais frequentes adotadas nas consultas realizadas em uma Unidade Básica de Saúde (UBS). Métodos Foram utilizados cadernos de registro de atendimento da UBS para análise de variantes como idade, sexo, diagnóstico, conduta e encaminhamento dos pacientes. Os dados foram analisados estatisticamente pelo programa SPSS 15.0. Resultados As hipóteses diagnósticas mais frequentes foram: eczemas, tumores benignos, transtornos pigmentares, onicopatias, infecções fúngicas, bacterianas e virais, lesões acneiformes foliculares e eritematodescamativas. Conclusão As hipóteses diagnósticas mais frequentes nas consultas de um médico generalista diferem daquelas feitas pelo especialista. Os resultados deste estudo deverão promover uma discussão sobre o ensino da Dermatologia nos cursos de Medicina. The frequency of skin disorders is high and they have a considerable impact on patients’ quality of life. However, there are very few public policies for this area of public health and the time devoted to studying Dermatology at medical school is limited. Objectives to define the most common diagnostic hypotheses and procedures in consultations performed at a primary health care unit. Methods medical notebooks entries were used in order to analyze variables such as age, gender, diagnosis, patient conduct and referral. Statistical analysis was performed using the program SPSS 15.0. Results the most frequent diagnostic hypotheses were: eczema, benign tumors, pigment disorders, onychopathy, fungal infections, bacterial infections, viral infections, acneiform and erythemato-squamous conditions. Conclusion the most frequent hypotheses diagnosed by a general practitioner differ greatly from those diagnosed by specialists. The results of this study should promote further discussions on dermatology teaching in Medical courses.

Psoriasis is a chronic inflammatory dermatosis characterized by erythematous, scaly lesions covering extensive areas of the skin and negatively affecting patients' quality of life by interfering in their personal life, social relationships and routine activities. Treatment offers reasonable control of the condition; however, little importance is generally given to the patient's impression.To evaluate the quality of life of patients with psoriasis, to verify whether there is a correlation between quality of life and clinical improvement and to compare two quality of life questionnaires and evaluate their equivalence.Patients over 18 years of age attending the psoriasis outpatient clinic at the Teaching Hospital of the University of Campinas (UNICAMP), who were either initiating treatment or changing to a new form of treatment, were included in the study. Patients answered two quality of life questionnaires (the Psoriasis Disability Index and the Dermatological Life Quality Index) up to a total of 180 consultations to evaluate whether there is a correlation between the questionnaires. Patients were also rated using a clinical index (Psoriasis Area and Severity Index) at baseline and at follow-up visits.The study involved 138 patients (76 men; mean age 50 years). A correlation was found between the questionnaires and the clinical index, showing that clinical changes are reflected in quality of life. The two questionnaires were found to be equivalent.It was found that both clinical condition and quality of life improved in the majority of patients receiving care at this outpatient clinic and that the quality of life questionnaires are equivalent.

The connexins are a family of proteins whose major function is as part of the gap junctions of cell-to-cell channels. They are expressed in several tissues including brain, skin, and cochlea. Mutations in connexin genes play a major role in non-syndromic sensorineural deafness, but have been also described in individuals with variable dermatological features. In recent years, many genes responsible for hereditary skin diseases have been discovered. These genes may encode different proteins that participate in the terminal differentiation of the epidermis. Therefore alteration or absence of these proteins causes a keratinization disorder. It has been demonstrated that distinct germline mutations within six connexin (Cx) genes GJB2 (Cx26), GJB6 (Cx30), GJB3 (Cx31), GJA1 (Cx43), GJB4 (Cx30.3), and GJB5 (Cx31.1), may cause sensorineural hearing loss and various skin disease phenotypes. The crucial functional importance of each of these connexins in the mentioned ectodermic tissues is reflected by the finding that genetic defects in their genes produce a wide spectrum of genetic disorders comprising sensorineural hearing loss, disorders of cornification of the skin, hair, and nails, and keratitis. Here, we report on different mutations in the connexin genes in individuals with or without hearing loss and different skin disorders illustrating the clinical and genetic heterogeneity of the condition.

Orientador: Maria Helena Stangler Kraemer Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas Resumo: Psoríase é uma dermatose inflamatória crônica, determinada por desregulação do sistema imune e associada a várias comorbidades. O marcador genético mais associado à psoríase em todas as populações é o HLA-CW06. Polimorfismos na região promotora do TNF-a, especialmente a troca de uma guanina por uma adenina nas posições -238 e -308 estão relacionados à alta produção de TNF-a e risco aumentado para psoríase nas populações caucasóides e não em asiáticos. Com o objetivo de determinar se polimorfismos destes genes podem ser fatores de risco para susceptibilidade ou gravidade da doença em pacientes brasileiros, foi realizado um estudo caso-controle com 69 pacientes com psoríase de início até 40 anos, com acompanhamento por dez anos para caracterização de sua evolução clínica em doença leve (grupo I) e grave (grupo II), e 70 indivíduos sadios. Foi feita a identificação dos alelos HLA classe I e II e SNPs da região promotora do TNF-a -238 e -308. Coletaram-se 10 mL de sangue periférico dos indivíduos e se realizou a extração do DNA através do método salting out. O DNA foi amplificado pela reação em cadeia da polimerase (PCR), com primers sequência-específica. As freqüências alélicas e gênicas foram estimadas por contagem direta e a comparação entre as frequências dos grupos foi efetuada por Teste de Fisher (programa GraphPad InStat 3.05). Dois métodos computacionais foram usados para determinar os haplótipos dos indivíduo: (1) o algoritmo ELB implementado pelo software Arlequin 3.1 e (2) um método de base coalescente implementado pelo software PHASE v2, e as freqüências de cada haplótipo foram comparadas por Teste de Fisher. No grupo II, observou-se maior associação com fatores desencadeantes como estresse, início na adolescência e predominância do sexo masculino. Pode-se sugerir que os alelos HLA-B*37, -Cw*06, -Cw*12 e -DRB1*07 foram associados ao curso mais grave da doença, enquanto - B*57 à doença mais leve. O aielo DRBT04 teve tendência a associação negativa. Ao se comparar o grupo I com o grupo II, o alelo HLA-B*37 pode ser interpretado como fator de mau prognóstico. Não houve diferença estatística entre polimorfismos da região promotora do TNF-a entre pacientes e controles. Este estudo apontou uma alta frequência do genótipo TNF-a -238 G/G {OFt 3,21; Cl:1,06-9,71; p=0,04), assim como do alelo -238 G, no grupo com doença mais grave e, ao contrário, o genótipo -238 G/A com frequência maior no grupo de boa evolução. O haplótipo -238A-308G mostrou frequência reduzida conferindo um efeito protetor. Estes dados não correspondem ao reportado para as populações caucasianas, considerando que a população brasileira é miscigenada. Polimorfismos dos SNPs do TNF-a não parecem ser um fator de susceptibilidade genética mais importante do que o já conhecido HLA-Cw*06 em pacientes brasileiros, mas podem ter relação com as manifestações e evolução da doença. Abstract: Psoriasis is an erythematous, scaly inflammatory dermatosis with a complex immunologic basis. The strongest genetic marker for psoriasis is HLA-Cw*06. Polymorphisms in the TNF-a promoter region, especially replacement of guanine with adenine in positions -238 and -308 are related to higher TNF-a production and higher risk for psoriasis in Caucasoid populations, not found in Asians. We did a case-control study of 69 patients with psoriasis type I and 70 controls, characterized clinical progression along 10-years of follow-up in mild or severe disease and determined HLA class I, II and TNF-a SNPs -238 and -308 polymorphisms to demonstrate whether these polymorphisms may be genetic risk for susceptibility to psoriasis or severity of the disease in Brazilians. Peripheral blood (10 ml) was collected. Genomic DNA from both psoriasis patients and controls was isolated using a salting out procedure. Polymorphisms were identified by PCR/SSP. Alleles and genotypes frequencies were compared by Fisher's test (GraphPad InStat 3.05 software). Two computational methods were used to determine the haplotypes of each subject, without taking into account any prior information: (1) the ELB algorithm implemented by the ARLEQUIN 3.1 software and (2) a coalescence based method as implemented by the PHASE v2 software. The haplotype frequencies were compared between group pairs by Fisher's test. Severe disease was found more frequently in male patients, associated with environmental factors and onset at adolescence. It may be suggested that alleles HLA- B*37, -Cw*06, -Cw*12 and -DRB1*07 were associated with severe disease course, while -B"57 with mild disease. No statistical difference was found between the patients and controls regarding polymorphisms frequencies in TNF SNPs. This study pointed to a higher TNF-238 G/G genotype frequency (OR 3,21; Cl:1,06-8,71; p=0,04) in the group with severe disease and -238A-308G haplotype was found in reduced frequencies revealing a protective effect. These data do not correspond to those reported for the Caucasian population, considering that Brazilian population is admixed, and this is the first consideration about TNF-a SNPs in psoriasis in this population. Polymorphisms in the TNF-a SNPs do not seem to be a more important genetic risk factor for psoriasis than the already known Cw*06 in Brazilian patients, but these markers may be related to clinical manifestations. Doutorado Clínica Médica Doutor em Clínica Médica

Teaching of Dermatology is little valorized in medical courses, and sexually transmitted diseases are generally presented to the students in a non-systematized way. In a study about the teaching of Dermatology, only ten out of 83 medical doctors taking part in the project correctly answered a question about syphilis serology. More research regarding the teaching of the sexually transmitted diseases is needed in our country. Moreover, dermatologists should take the opportunity brought about by the current discussion on the re-elaboration of medical course curricula to retake the responsibility on teaching about these diseases.

Differential diagnosis between keratoacanthoma (KA) and squamous cell carcinoma (SCC) is difficult due to their similarities. The mechanisms that drive their distinct biological behavior are poorly understood. To investigate whether the assessment of microvessel density (MVD) could be helpful in KA and SCC differential diagnosis and to gain insight into the pathogenesis of KA-like neoplasms, we compared the density of CD105- and CD34-stained vessels in KAs and SCCs and their relation to the expression of the p53 oncoprotein and proliferation marker Ki67. This is an observational retrospective cohort study. Forty lesions with clinical appearance of KAs (29 KAs and 11 SCCs) entered the study. A biopsy was taken from each lesion at presentation and the natural clinical course was monitored for at least 1 month. Growing or minimally regressing lesions were submitted to complete surgical excision. The diagnoses were established on combined clinical, histological, and follow-up evaluations. The MVD and p53 or Ki67 expression in neoplastic cells were assessed through morphometry. The MVD did not show discriminating power between KAs and SCCs. The Ki67 proliferation rate was significantly higher in SCCs. Although neoangiogenesis (CD105-MVD) in KAs was associated with cell proliferation, in SCCs it was not. There was significant correlation between p53 expression and neoplasia size in SCCs but not in KAs. From our results, we may conclude that KA and SCC have similarities, as CD105- and CD34-MVD. However, the low Ki67 proliferation index and the positive correlation between Ki-67 index and neovascularization in KA suggest a dependence in neovascularization to grow in KA, pointing to involvement of distinct pathogenesis.

We analyzed 100 control individuals and 60 patients with psoriasis vulgaris from the population of Campinas, Brazil. Typification of class II HLA alleles (HLA-DRB1-5 and -DQB1) was carried out through the DNA/PCR/SSP at medium and high resolution. DNA was extracted through a salting-out procedure: 13 DRB1 alleles, 3 DRB3 alleles, 1 DRB4 allele, 2 DRB5 alleles, and 5 DQB1 alleles were identified at a medium resolution using the PCR/SSP, and 45 DRB1 alleles were identified at a high resolution in analyzed patients. Results showed associations with psoriasis vulgaris: positive associations HLA-DRB3*02 (p0.05, chi(2) = 5.10, RR = 2.14); HLA-DRB1*0102 alleles (p0.05, RR = 5.44). Negative associations were found for HLA-DRB4*01 (chi(2) = 3.23, RR = 0.55) and HLA-DRB1*1302 alleles (p0.05, RR = 0.23). The haplotypes revealed positive association for HLA-DRB1*0102/DQB1*05 (p0.05, RR = 5.44) and HLA-DRB1*0701/DQB1*03 alleles (p0.02, RR = 9.00). These findings suggest a possible association of the DRB1 allele with the group of patients showing an early onset of the illness, as well as an association with haplotypes HLA-DRB1*0102/DQB1*05 and HLA-DRB1*0701/DQB1*03.

Introduction: The genus Bartonella comprises a unique group of emerging, Gram-negative, facultative intracellular bacteria, which can cause Carrión’s disease, trench fever, cat scratch disease and bacillary angiomatosis. Man is a reservoir host of Bartonella species. Ten to 15% of the populations in areas that are hyperendemic for Carrión’s disease are asymptomatic carriers of B. bacilliformis. In addition, positive serologic tests for B. henselae were found in 2 to 6% of Americans, 48% of Swiss, 19% of Germans, and 13.7% of Brazilians. Blood donors can be asymptomatic carrier of Bartonella spp. and the risk for transmission by transfusion should be considered. The aim of this study was to demonstrate that B. henselae remains viable in red blood cell (RBC) units at the end of the storage period. Methods: Two RBC units from healthy blood donors were collected in CPDA1 and each one split into two portions via a sterile connecting device. One bag from each split unit received experimentally B. henselae (Adolpho Lutz Institute, Sao Paulo, Brazil) and the second one served as a control. A brain and heart infusion (BHI) suspension of B. henselae colonies (Houston 1, American Type Culture Collection, Rockville, MD, ATCC 49882T) was used to obtain equivalence with tube 10 of McFarland scale, which determined an initial suspension with approximately 3×109 colony-forming units (CFU)/mL. This bacterial suspension was inoculated (0,33mL of the initial suspension) in one split RBC unit using a Sampling Site Coupler. Thus approximately 5,5×106 CFU of B. henselae were obtained per mL of RBC concentrate. All split units were stored (4 ±2°C) for 35 days. Aliquots were collected weekly using, Sample Site Coupler for, culture in dish with chocolate agar (incubated at 37°C in environment with 5% CO2), samples for blood culture bottles Bact/Alert (BioMérieux, Inc, USA) and Karnovisky medium for future evaluation by transmission electron microscopy. Results: All dishes with chocolate agar culture medium from infected units (seeded on days D0, D7, D14, D21, D28 and D35) presented growth of exuberant and mucoid colonies, after the fourth day of incubation. The electron microscopy from these samples showed typical Gram-negative cell wall in the interior of red blood cells. Samples from infected units presented negative blood culture bottles. All cultures from control units had negative results. Discussion: Bartonella ssp. is fastidious bacteria. Incubation for a short period of time and in standard medium does not allow for B. henselae growth as observed in the bottle culture of this study. Cultivation in blood-enriched media and incubation in environment with high CO2 levels of infected smears resulted positive. In conclusion, this study has demonstrated that B. henselae remain viable in RBC units during the storage period, at 4±2°C. These data reinforce the possibility of infection by transfusion of blood units collected from asymptomatic blood donors. Receptors of blood transfusion many times are or may become immunocompromissed, with risk of developing severe forms of bartonelloses.