Avaliação da qualidade de vida, grau de incapacidade e do desenho da figura humana em pacientes com neuropatias na hanseníase / Quality of life, physical disability, and the human figure drawing assessment of patients with neuropathies in leprosy

Na hanseníase, a presença de sintomas dermatoneurológicos com potencial evolução para incapacidades físicas pode comprometer a qualidade de vida (QV) e a imagem corporal do paciente. Objetivo: Avaliar as possíveis associações entre a QV, o Grau de Incapacidade (GI) e o Desenho da Figura Humana (DFH) em indivíduos com neuropatia hansênica. Método: Este estudo consiste em um estudo descritivo, com abordagem quanti-qualitativa. Foram utilizados quatro instrumentos de avaliação: Questionário sociodemográfico, NeuroQol (Neuropathy ­ Specific Quality of Life Questionnaire), DFH e Formulário de avaliação do GI. Foram incluídos pacientes com GI 1 ou 2 nos pés e idade igual ou superior a 18 anos. Resultados: Foram avaliados 100 indivíduos. Entre aqueles com GI 2, houve uma tendência à omissão do nariz (p=0,050) e DFH no tamanho pequeno (p=0,047). Houve associação entre o DFH e o domínio QV Sintomas difuso sensitivo-motores (p=0,035), sugerindo que a omissão dos pés no DFH pode representar perda da QV. Conclusão: Indivíduos com neuropatia hansênica apresentam QV boa à moderada. A omissão de segmentos do corpo pode indicar conflitos e sentimentos de insegurança. Há indícios de perda de autonomia quando o paciente omite ou corta os pés no DFH

In Leprosy, the presence of dermatoneurological symptoms with potential evolution to physical disabilities may influence the quality of life (QoL) and the patient's body image. Objective: To evaluate possible associations between the QoL, degree of disability (DoD) and human figure drawing (HFD) in individuals with leprosy neuropathy. Method: This is a descriptive study with a quanti-qualitative approach. Four evaluation instruments were used: a sociodemographic questionnaire, quality of life in neurological disorder questionnaire (Neuro-QoL), HFD and DoD evaluation form. Patients with DoD grades 1 or 2 of the feet and age equal to or greater than 18 were included in this study. Results: One hundred individuals were evaluated. Individuals with DoD grade 2 tended to omit the nose (p=0.050) and the HFD was small (p=0.047). An association was identified between the HFD and the QoL with dominance of diffuse sensory-motor symptoms (p=0.035) suggesting that omission of the feet in the HFD may represent a loss of QoL. Conclusion: Individuals with leprosy neuropathy have a good to moderate QoL but the omission of body segments may indicate conflicts and feelings of uncertainty. There is evidence of loss of autonomy when the subject omits or cuts off the feet in the HFD

Desempenho funcional de pacientes com deformidades visíveis na hanseníase / Functional performance of patients with visible deformities in leprosy

Esse estudo tem por objetivo avaliar a capacidade funcional da mão das pessoas atingidas pela hanseníase na realização das atividades básicas e instrumentais da vida diária. A amostra foi constituída por 50 pacientes, maiores de 18 anos, que estavam sendo acompanhados no ambulatório do Instituto Lauro de Souza Lima. Para avaliação das características sociodemográficas e clínicas foi elaborado um questionário próprio. As dificuldades manuais para realizar atividades nas áreas de vestuário, alimentação, higiene pessoal, cuidados com a casa, escrita e outros foram avaliados por meio da Avaliação Funcional das Mãos em Hanseníase. Os resultados demonstram que em todas as atividades houve algum grau de dificuldade, porém, a maioria dos pacientes as realiza com independência. As atividades consideradas mais difíceis de serem executadas pelos pacientes com garra ulnar foram: pegar objetos pequenos em superfície plana, abrir/fechar fecho de corrente e cortar unhas. Na população de pacientes com garra ulnar/mediano foram: pegar objetos pequenos em superfície plana, abotoar/desabotoar, dar laço/ amarrar cadarço e abrir/fechar fecho de corrente. As atividades são realizadas com algum grau de dificuldade, porém, com independência pela maioria dos indivíduos que tem ou tiveram hanseníase e apresentam deformidades visíveis. Muitas dessas dificuldades podem ser minimizadas por meio de transferências tendinosas ou pela indicação, confecção e uso de tecnologia assistiva(AU).

This study aims to assess the functional capacity of the hand of people affected by leprosy in carrying out basic and instrumental activities of daily living. The sample consisted of 50 patients, aged over 18, who were being followed up at the outpatient clinic of the Instituto Lauro de Souza Lima. A questionnaire was developed to assess sociodemographic and clinical characteristics. Manual difficulties in carrying out activities in the areas of clothing, food, personal hygiene, home care, writing and others were assessed through the Functional Hand Assessment in Leprosy. The results show that in all activities there was some degree of difficulty, however, most patients perform them independently. The activities considered most difficult to be performed by patients with an ulnar claw were: picking up small objects on a flat surface, opening / closing the chain clasp and cutting nails. In the population of patients with an ulnar / median claw were: picking up small objects on a flat surface, buttoning / unbuttoning, looping / tying shoelaces and opening / closing chain fastening. The activities are performed with some degree of difficulty, however, independently by the majority of individuals who have or have had leprosy and have visible deformities. Many of these difficulties can be minimized by means of tendon transfers or by the indication, preparation and use of assistive technology(AU).

Mirror visual feedback: elaboração de protocolo para aplicação em pacientes com hanseníase / Mirror visual feedback: elaboration of protocol for application in patients with leprosy

Os primeiros estudos acerca da técnica Mirror visual feedback, técnica essa introduzida por Ramachandran e conhecida como terapia do espelho, foram em pacientes com amputação dos membros superiores e inferiores, na busca por respostas sobre a origem da dor no membro fantasma. Após o êxito em sua aplicação relativa às dores fantasmas começou a ser também testada em outras patologias. Uma doença que desperta curiosidade para métodos alternativos de tratamento é a hanseníase, que compromete pele e nervos periféricos, podendo causar hipersensibilidade do nervo, edema, déficit motor, sensitivo e dor intensa interferindo na funcionalidade e estabilidade emocional, social e produtiva. O objetivo deste trabalho foi elaborar um protocolo terapêutico com foco na futura aplicação do Mirror Visual Feedback em pacientes atingidos pela hanseníase que apresentam dor neuropática e dor fantasma, através do levantamento de conhecimentos acerca da técnica na literatura nacional e internacional.

Functioning and disability in leprosy

This study has been carried out to investigate the frequency of musculoskeletal symptoms in people affected by leprosy and correlate this to their functional capacity to perform daily living and working activities and to their degree of disability. This cross-sectional study was performed on 100 clients who answered a questionnaire on personal and clinical data, the Nordic Musculoskeletal (QNSO), and the shoulder, arm and hand dysfunction (DASH) questionnaires. The mean age was 53.63 years and there was a predominance of men (67%), low education (53%), and multibacillary disease (84%). Physical impairment affected 95% of respondents and 92% indicated pain and/or paresthesia in the arms that interfered in functional capacity.Respondents with musculoskeletal symptoms have more difficulty to perform daily living and working activities compared to those without musculoskeletal symptoms (p-value<0.05). The presence of physical disabilities was not associated with difficulties to perform daily living (p-value=0.29) and labor activities (p-value=0.87). The majority of patients experienced pain and/or paresthesia of the arms, and this was associated with difficulties to perform daily living and working activities. However, the presence of physical disability does not seem to cause major impediments or limitations to perform these activities.

Altitudinal population structure and microevolution of the malaria vector Anopheles cruzii (Diptera: Culicidae).

BACKGROUND: In Brazil, the autochthonous transmission of extra-Amazonian malaria occurs mainly in areas of the southeastern coastal Atlantic Forest, where Anopheles cruzii is the primary vector. In these locations, the population density of the mosquito varies with altitude (5-263 m above sea level), prompting us to hypothesise that gene flow is also unevenly distributed. Describing the micro-geographical and temporal biological variability of this species may be a key to understanding the dispersion of malaria in the region. We explored the homogeneity of the An. cruzii population across its altitudinal range of distribution using wing shape and mtDNA gene analysis. We also assessed the stability of wing geometry over time. METHODS: Larvae were sampled from lowland (5-20 m) and hilltop (81-263 m) areas in a primary Atlantic Forest region, in the municipality of Cananéia (State of São Paulo, Brazil). The right wings of males and females were analysed by standard geometric morphometrics. Eighteen landmarks were digitised for each individual and a discriminant analysis was used to compare samples from the hilltop and lowland. A 400-bp DNA fragment of the mitochondrial cytochrome oxidase gene subunit I (CO-I) was PCR-amplified and sequenced. RESULTS: Wing shapes were distinct between lowland and hilltop population samples. Results of cross-validated tests based on Mahalanobis distances showed that the individuals from both micro-environments were correctly reclassified in a range of 54-96%. The wings of hilltop individuals were larger. The CO-I gene was highly polymorphic (haplotypic diversity = 0.98) and altitudinally structured (Фst = 0.085 and Jaccard = 0.033). We found 60 different haplotypes but only two were shared by the lowland and hilltop populations. Wing shape changed over the brief study period (2009-2013). CONCLUSIONS: Wing geometry and CO-I gene analysis indicated that An. cruzii is vertically structured. Wing shape varied rapidly, but altitude structure was maintained. Future investigations should identify the biotic/abiotic causes of these patterns and their implications in the local epidemiology of malaria.

The contribution of HLA molecules to Dupuytren's contracture in a Southeast Brazilian population.

PURPOSE: The aim of the present study was to investigate the HLA phenotype in Dupuytren's contracture (DC) patients in order to verify the correlation of these alleles with risk factors for development of DC in the Brazilian population. METHODS: This was a case-controlled study of 25 DC patients and 443 healthy individuals with no history of HLA-associated diseases. HLA class I and class II typing was performed using the polymerase chain reaction sequence-specific primer method. RESULTS: The HLAB*18 phenotype was observed in 32% of the patients and 10.5% of controls. However, P values did not remain significant after correction. DISCUSSION: Although we observed an increased tendency of DC patients to possess the HLA-B*18 allele, the results were not statistically significant after correction. This allele was higher in patients of Italian and/or Spanish ethnicity, localities with frequencies higher than 18.0% and 14.0% respectively. Further investigation with a larger cohort of DC patients is required to confirm the potential role of HLA in this disease.

Associação entre sintomas depressivos, trabalho e grau de incapacidade na hanseníase / Relationship between depression, work, and grade of impairment in leprosy

A depressão é o transtorno psiquiátrico mais comum na hanseníase e com alto índice de sintomas depressivos. Objetivo: Verificar a frequência dos sintomas depressivos e sua relação com o grau de incapacidade (GI) da OMS e variáveis sociodemográficas. Método: Aplicou-se um questionário, contendo aspectos sociodemográficos, clínicos e o GI. Foi aplicada a escala original do BDI para identificar a frequência dos sintomas depressivos (21 itens) e a subescala cognitiva chamada BDI-Short Form - BDI-SF (1-13 itens), recomendada para avaliar sintomas depressivos em indivíduos com diagnóstico de alguma patologia. Foi utilizada análise estatística descritiva, com distribuição de frequência para a caracterização da casuística e para o cruzamento das variáveis, foi utilizado oTeste Chi-square-corrected (Yates), considerando resultados significantes valor - p < 0,05. Resultados: Foram avaliados 130 pacientes que tem ou tiveram hanseníase. A idade média dos pacientes foi de 49,64 (SD 14,04).Houve predomínio do sexo masculino (64,6%), dos que vivem com familiares (87,7%), com ensino fundamental incompleto (66,2%), união civil estável (61,6%), não trabalham (75,4%) e recebem aposentadoria ou auxíliosaúde (63,9%). Em relação aos aspectos clínicos, 94,5% são multibacilares, 74,6% concluíram a poliquimioterapia e a maioria apresenta perda da sensibilidade protetora e/ou deformidades (31,5% grau 1 e 37% grau 2). Dentre os casos avaliados 43,1% apresentou sintomas depressivos de intensidade moderada a grave. Não houve correlação significativa entre BDI-SF e GI (valor - p = 0,950), mas, ?não trabalhar? associou-se com sintomas depressivos (BDI-SF) (valor - p = < 0,05). Preocupação somática foi o sintoma mais frequente (80,7%), seguido de dificuldade no trabalho (78,5%), irritabilidade (68,5%), fadiga (67,7%), auto-acusação (62,3%) e choro fácil (60%). Conclusão: Conclui-se que sintomas depressivos moderados e graves acometeram 43,1% dos casos avaliados,independentemente de ter ou não deficiências físicas (GI 1 e 2). As pessoas que não trabalhavam foram mais acometidas por sintomas depressivos em comparação aos que exerciam alguma atividade profissional.

Depression is the most common psychiatric disorder in leprosy and with high depressive symptoms. Objective: The aim of this study was to determine the prevalence and frequency of depressive symptoms andtheir relationship to WHO impairment grading (IG), and sociodemographic variables. Method: We applied a survey containing sociodemographic, clinical aspects and IG. The original scale of the BDI was applied to identifythe frequency of depressive symptoms (21 items), as well as the cognitive subscale BDI-Short Form-BDI-SF (items 1-13), recommended to assess depressive symptoms in individuals with defined pathologies. Descriptivestatistical analysis was used, with the frequency distribution to characterize the sample, and to the intersection of the variables, the Chi-square Test-corrected (Yates) was applied, considering significant results p - value < 0.05. Results: 130 patients who have or have had leprosy were evaluated. The mean age of patients was 49.64 (SD = 14.04). There was a predominance of males (64.6%), those living with family (87.7%), with incomplete primary education (66.2%), stable civil union (61.6%), which did not work (75.4%) and receive retirement or health aid (63.9%). Regarding clinical aspects, 94.5% were multibacillary, 74.6% had multidrug therapy and most have loss of protective sensation and/or deformities (31.5% IG1; 37% IG2). Among the cases evaluated, 43.1% had depressive symptoms of moderate to severe intensity. There was no significant correlation between BDI-SF and IG (p - value = 0.950). However, the individuals without an occupation or job (?which did not work?) were associated with depressive symptoms (BDI-FS; p - value < 0.05). Somatic apprehension was the most frequent symptom (80.7%), followed by difficulty with work (78.5%), irritability (68.5%), fatigue (67.7%), self-blame (62.3%)and tearfulness (60%). Conclusion: It is concluded that moderate and severe depressive symptoms assaulted 43.1% of the cases evaluated, regardless of whether or not physical disabilities (IG1 and 2). Individuals who did not work were the most affected by depressive symptoms compared to those who had some sort of occupation.

Contribuicao das moleculas de antigeno de histocompatibilidade leucocitaria (HLA) para a contratura de Dupuytren em uma populacao do Sudeste do Brasil / The contribution of HLA molecules to Dupuytren's contracture in a Southeast Brazilian population

Objetivo: O objetivo deste estudo foi investigar o fenótipo do HLA em pacientes com contratura de Dupuytren (CD) para verificar a correlação desses alelos com os fatores de risco para o desenvolvimento da CD na população brasileira. Métodos: Este foi um estudo de caso-controle de 25 pacientes com CD e 443 indivíduos saudáveis sem histórico de doenças associadas ao HLA. As tipagens classe I e classe II do HLA foram feitas utilizando o método iniciador de sequências específicas da reação em cadeia da polimerase. Resultados: O fenótipo HLA-B*18 foi observado em 32% dos pacientes e 10,5% do grupo controle. Contudo, os valores de p não permaneceram significativos após correção. Discussão: Apesar de termos observado um aumento na tendência de os pacientes com CD terem o alelo HLA-B*18, os resultados não foram estatisticamente significativos após correção. Esse alelo foi maior em pacientes de etnia italiana e/ou espanhola, locais com frequências superiores a 18% e 14%, respectivamente. São necessárias investigações adicionais com uma coorte maior de pacientes com CD para confirmar o possível papel do HLA nessa doença. .

Purpose: The aim of the present study was to investigate the HLA phenotype in Dupuytren's contracture (DC) patients in order to verify the correlation of these alleles with risk factors for development of DC in the Brazilian population. Methods: This was a case-controlled study of 25 DC patients and 443 healthy individuals with no history of HLA-associated diseases. HLA class I and class II typing was performed using the polymerase chain reaction sequence-specific primer method. Results: The HLAB*18 phenotype was observed in 32% of the patients and 10.5% of controls. However, P values did not remain significant after correction. Discussion: Although we observed an increased tendency of DC patients to possess the HLA-B*18 allele, the results were not statistically significant after correction. This allele was higher in patients of Italian and/or Spanish ethnicity, localities with frequencies higher than 18.0% and 14.0% respectively. Further investigation with a larger cohort of DC patients is required to confirm the potential role of HLA in this disease. .

Morphometrical diagnosis of the malaria vectors Anopheles cruzii, An. homunculus and An. bellator.

BACKGROUND: Anopheles (Kerteszia) cruzii is a primary vector of Plasmodium parasites in Brazil's Atlantic Forest. Adult females of An. cruzii and An. homunculus, which is a secondary malaria vector, are morphologically similar and difficult to distinguish when using external morphological characteristics only. These two species may occur syntopically with An. bellator, which is also a potential vector of Plasmodium species and is morphologically similar to An. cruzii and An. homunculus. Identification of these species based on female specimens is often jeopardised by polymorphisms, overlapping morphological characteristics and damage caused to specimens during collection. Wing geometric morphometrics has been used to distinguish several insect species; however, this economical and powerful tool has not been applied to Kerteszia species. Our objective was to assess wing geometry to distinguish An. cruzii, An. homunculus and An. bellator. METHODS: Specimens were collected in an area in the Serra do Mar hotspot biodiversity corridor of the Atlantic Forest biome (Cananeia municipality, State of Sao Paulo, Brazil). The right wings of females of An. cruzii (n= 40), An. homunculus (n= 50) and An. bellator (n= 27) were photographed. For each individual, 18 wing landmarks were subjected to standard geometric morphometrics. Discriminant analysis of Procrustean coordinates was performed to quantify wing shape variation. RESULTS: Individuals clustered into three distinct groups according to species with a slight overlap between representatives of An. cruzii and An. homunculus. The Mahalanobis distance between An. cruzii and An. homunculus was consistently lower (3.50) than that between An. cruzii and An. bellator (4.58) or An. homunculus and An. bellator (4.32). Pairwise cross-validated reclassification showed that geometric morphometrics is an effective analytical method to distinguish between An. bellator, An. cruzii and An. homunculus with a reliability rate varying between 78-88%. Shape analysis revealed that the wings of An. homunculus are narrower than those of An. cruzii and that An. bellator is different from both of the congeneric species. CONCLUSION: It is possible to distinguish among the vectors An. cruzii, An. homunculus and An. bellator based on female wing characteristics.

Mosquito (Diptera: Culicidae) assemblages associated with Nidularium and Vriesea bromeliads in Serra do Mar, Atlantic Forest, Brazil.

BACKGROUND: The most substantial and best preserved area of Atlantic Forest is within the biogeographical sub-region of Serra do Mar. The topographic complexity of the region creates a diverse array of microclimates, which can affect species distribution and diversity inside the forest. Given that Atlantic Forest includes highly heterogeneous environments, a diverse and medically important Culicidae assemblage, and possible species co-occurrence, we evaluated mosquito assemblages from bromeliad phytotelmata in Serra do Mar (southeastern Brazil). METHODS: Larvae and pupae were collected monthly from Nidularium and Vriesea bromeliads between July 2008 and June 2009. Collection sites were divided into landscape categories (lowland, hillslope and hilltop) based on elevation and slope. Correlations between bromeliad mosquito assemblage and environmental variables were assessed using multivariate redundancy analysis. Differences in species diversity between bromeliads within each category of elevation were explored using the Renyi diversity index. Univariate binary logistic regression analyses were used to assess species co-occurrence. RESULTS: A total of 2,024 mosquitoes belonging to 22 species were collected. Landscape categories (pseudo-F value = 1.89, p = 0.04), bromeliad water volume (pseudo-F = 2.99, p = 0.03) and bromeliad fullness (Pseudo-F = 4.47, p < 0.01) influenced mosquito assemblage structure. Renyi diversity index show that lowland possesses the highest diversity indices. The presence of An. homunculus was associated with Cx. ocellatus and the presence of An. cruzii was associated with Cx. neglectus, Cx. inimitabilis fuscatus and Cx. worontzowi. Anopheles cruzii and An. homunculus were taken from the same bromeliad, however, the co-occurrence between those two species was not statistically significant. CONCLUSIONS: One of the main findings of our study was that differences in species among mosquito assemblages were influenced by landscape characteristics. The bromeliad factor that influenced mosquito abundance and assemblage structure was fullness. The findings of the current study raise important questions about the role of An. homunculus in the transmission of Plasmodium in Serra do Mar, southeastern Atlantic Forest.

Culicídeos associados às bromélias na Mata Atlântica do município de Cananéia, São Paulo, Brasil / Bromeliad associated mosquitoes from Atlantic Forest, in Cananéia, São Paulo, Brasil

A Mata Atlântica não possui fisionomia uniforme, pois seu relevo determina diferentes condições ambientais que definem a composição de sua fauna e flora. Em uma mesma região, a diversidade de espécies pode variar de acordo com a heterogeneidade dos habitats. Objetivo: Caracterizar a fauna de Culicidae em fitotelmatas de bromélias (Bromeliaceae), no município de Cananéia, São Paulo, Brasil. Métodos: Larvas e pupas de culicídeos foram obtidas, mensalmente, do conteúdo aquático de bromélias de três ambientes: planície, encosta e morro. As coletas ocorreram entre julho de 2008 e junho de 2009. Para estimar a variedade de espécies foram utilizados os índices de Margalef e de Menhinick. O índice de Jaccard e o de Sorensen indicaram a semelhança entre bromélias de dois ambientes. Para avaliar a dominância, empregou-se o índice de Simpson e de Berger-Parker. A equidade na distribuição das espécies foi avaliada pelo índice de Pielou e a constância em cada ambiente pelo índice c. Os testes estatísticos empregados foram: análise de variância (ANOVA) com o teste de Fisher; teste Kruskal-Wallis e valor p; teste de associação qui-quadrado e regressão linear simples por meio do coeficiente de correlação de Pearson. Resultados: O teste Kruskal-Wallis mostrou diferenças significantes para o volume hídrico das bromélias de cada ambiente (p=0,002, IC= 95 por cento) mas não para o valor do pH (p= 0,775). Foram identificados 2024 mosquitos, 786 (38,83 por cento) na planície, 606 (29,94 por cento) na encosta e 632 (31,23 por cento) no morro, dentre eles: Culex (Microculex) (62,25 por cento), Culex ocellatus (21,20 por cento), Anopheles (Kerteszia) (15,17 por cento), Wyemoyia (Phoniomyia) (1,33 por cento) e Runchomyia (Runchomyia) (0,05 por cento). O índice de riqueza de Margalef foi maior na planície (d1=2,55) e menor no morro (d1=1,86), e o de Sorensen indicou encosta e morro como os ambientes mais similares (QS=0,79) e planície e morro como os menos similares (QS=0,65)...

Atlantic Forest is not uniform and its relief determines different environmental conditions that define the composition of its fauna and flora. Thus, in the same region, species diversity may vary according to habitat heterogeneity. Objective: To assess mosquito (Culicidae) fauna in bromeliads phytotelmate (Bromeliaceae), in Cananéia, São Paulo, Brazil. Methods: The larvae and pupae of mosquitoes were collected monthly in the water content of bromeliads from three regions: the lowland, hillside and hilltop. Sampling occurred between July 2008 and June 2009. To estimate richness, Margalef and Menhinick indices were used. Jaccard and Sorensen indices indicated the similarity between bromeliads from two regions. To assess dominance, we employed Simpson and Berger-Parker índices. Equity in the distribution of species was assessed by Pielou index and c index evaluated species constancy in each region. Statistical analyses applied were: ANOVA with Fisher's exact test, Kruskal-Wallis test and p value; Chi-square association test and linear regression using Pearson correlation coefficient. Results: Kruskal-Wallis test showed significant differences in water volume for bromeliads from each region (p=0,002, CI=95 per cent) but not for the pH value (p=0,775). In all, 2024 mosquitoes were identified, 786 (38,83 per cent) in the lowland, 606 (29,94 per cent) on the hillside and 632 (31,23 per cent) on the hilltop, among them: Culex (Microculex) (62,25 per cent), Culex ocellatus (21,20 per cent), Anopheles (Kerteszia) (15,17 per cent), Wyemoyia (Phoniomyia) (1,33 per cent) and Runchomyia (Runchomyia) (0,05 per cent). Margalef richness index was higher in the lowland (d1=2,55) and lower on the hilltop (d1=1,86). According to the Sorensen similarity index, species from the hillside are more likely to be found on the hilltop (QS=0,79) than in the lowland (QS=0,73). The greatest differences occurred between the hilltop and the lowland (QS=0,65). The 2 test showed...