Intraoral primary syphilis mimicking lymphoproliferative disorder.

Syphilis can mimic, clinically and microscopically, many other diseases. By microscopy, typically syphilis presents with plasma cell infiltration, admixed with lymphocytes and macrophages, in lichenoid and/or perivascular/perineural distribution pattern. When exuberant, this inflammatory infiltrate can mimic a lymphoproliferative disorder (LPD), notably plasma cell neoplasia or lymphoma. To date, about 12 cases of secondary syphilis, all but one in extraoral location, suggesting initially a LPD, have been published. Here, to our knowledge, we report an unusual case of intraoral primary syphilis initially suggesting LPD, notably lymphoid hyperplasia (pseudolymphoma); however, mucosa-associated lymphoid tissue (MALT) lymphoma and follicular lymphoma could not be disregarded. Polyclonality of plasma cells on immunohistochemistry, in strict clinical correlation, was essential to arrive at the correct diagnosis.

Round Bodies Detected by Treponema pallidum Immunohistochemical Stain in Two Cases of Cutaneous Syphilitic Gummata.

ABSTRACT: Tertiary syphilis may present a diagnostic challenge due to negative nontreponemal serologies in up to 30% of cases and frequent lack of identifiable spirochetes on histopathology or other direct detection tests. We report 2 cases of round bodies staining with Treponema pallidum immunohistochemistry by light microscopy in biopsies from cutaneous syphilitic gummata. In 1 case, the finding was validated 3 times by 2 independent laboratories; in the other case, T. pallidum was detected by polymerase chain reaction in the biopsy sample. Spirochete round bodies have previously been reported in the setting of electron microscopy and fluorography, but to the best of our knowledge, have not been reported by light microscopy in a routine skin biopsy. Although the clinical implications are unclear, this may represent a helpful new paradigm for the diagnosis of tertiary syphilis.

The outer membrane protein Tp92 of Treponema pallidum delays human neutrophil apoptosis via the ERK, PI3K/Akt, and NF-κB pathways.

Syphilis is a persistent sexually transmitted disease caused by infiltration of the elusive pathogen Treponema pallidum. Despite the prevalence of human polymorphonuclear neutrophils (hPMNs) within cutaneous lesions, which are characteristic of incipient syphilis, their role in T. pallidum infection remains unclear. Tp92 is the only T. pallidum helical outer membrane protein that exhibits structural features similar to those of outer membrane proteins in other gram-negative bacteria. However, the functional mechanism of this protein in immune cells remains unclear. Neutrophils are short-lived cells that undergo innate apoptosis in response to external stimuli that typically influence this process. In this study, we determined that Tp92 impedes the activation of procaspase-3 via the ERK MAPK, PI3K/Akt, and NF-κB signaling pathways, consequently suppressing caspase-3 activity within hPMNs, and thereby preventing hPMNs apoptosis. Furthermore, Tp92 could also modulate hPMNs apoptosis by enhancing the expression of the anti-apoptotic protein Mcl-1, stimulating IL-8 secretion, and preserving the mitochondrial membrane potential. These findings provide valuable insights into the molecular mechanisms underlying T. pallidum infection and suggest potential therapeutic targets for syphilis treatment.

Diagnostic utility of fine-needle aspiration cytology in syphilitic lymphadenitis: A case report.

Syphilis is a sexually transmitted infection that causes serious health problems without treatment. Detection of syphilis is necessary to stop the spread of the infection. Cytological screeners and pathologists may experience fine-needle aspiration cytology (FNAC) for syphilitic lymphadenitis; however, its characteristic cytological features are rarely reported. We present FNAC cytological features of syphilitic lymphadenitis in a case of a 21-year-old man. He presented with a sore throat and painful neck lymphadenopathy. His swollen and painful neck persisted even with antibiotic treatment. Necrotizing lymphadenitis and lymphoma were clinically suspected. FNAC was taken from the swollen lymph node. Cytologic findings of the specimen showed various inflammatory cells with small-sized vessels arranged in a branching/arborizing fashion. The vessels were surrounded by inflammatory cells, including plasma cells, neutrophils, and macrophages. Perivascular plasma cell cuffing was focally seen along with inconspicuous granulomas. Neutrophils appeared to involve the vascular wall. The cytological findings suggested syphilitic lymphadenitis, and clinical findings and serological tests confirmed primary syphilis with concomitant human immunodeficiency virus infection. Branching/arborizing vessels associated with many plasma cells, vascular involvement of neutrophils, and granulomas may suggest syphilitic infection if the specimen is obtained via FNAC. (189 words).

Primary syphilis of the oropharynx mimicking a neoplasm.

Syphilis is surging in Japan and worldwide among both homosexual and heterosexual individuals. Diagnosis can be challenging because syphilis is "the great imitator" and clinical manifestations are highly variable. Oral manifestations of syphilis are usually seen in the second stage and the primary syphilis in the oral cavity is rare. We describe a heterosexual man with isolated tonsillar lesions initially misdiagnosed as pharyngeal lymphoma and subsequently diagnosed as primary syphilis. Clinicians should be aware that isolated oropharyngeal involvement can occur in the primary syphilis and can mimic a neoplasm.

Primary syphilis with a tongue ulcer mimicking tongue cancer: a case report.

The main symptom in primary syphilis is a small, painless, sore or ulcer called a chancre on the penis, vagina, or around the anus, although chancres can sometimes appear in the mouth or on the lips, fingers, or buttocks. We present the case of a man in his early 60 s with a chief complaint of a painful tongue ulcer. An ulcerated, indurated, and hemorrhagic lesion (23 × 14 mm) was found on the ventral tongue surface, near the oral floor. Palpation identified several swollen, mobile, elastic cervical lymph nodes, with no tenderness. We initially diagnosed tongue cancer; however, during a subsequent detailed examination for a malignant tumor, including biopsy and obtaining additional history, his disease was finally identified as primary syphilis with multiple swollen cervical lymph nodes. Oral amoxicillin and probenecid were started, and after 14 days, there was partial reduction in the size of the submandibular lymph nodes and the ulcer on the left tongue margin. The number of patients with syphilis in Japan increased by eight times from 2012 to 2018. We suggest that dentists consider primary syphilis as a differential diagnosis for oral refractory ulcer with induration and obtain a detailed patient history.

Erythematous scaly plaques and nodules on scalp.

Cutaneous manifestations of syphilis are varied and may present with non-specific features. We describe a 45-year-old man who presented with erythematous scaly plaques and nodules on his scalp. In previously reported cases, there were only descriptions of nodules as well as tumors. However, in our case, the patient presented with plaques and nodules on his scalp that quickly resolved with treatment for syphilis. It is important to recognize and treat syphilis at an early stage.

Sífilis mimetizando lúpus eritematoso sistêmico: um relato de caso / Syphilis mimicking systemic lupus erythematosus: a case report / Sífilis que simula lupus eritematoso sistémico: informe de un caso

O Lúpus Eritematoso Sistêmico (LES) é uma patologia crônica, de origem autoimune e inflamatória. As diversas manifestações clínicas existentes em pacientes acometidos pelo LES, sejam elas sistêmicas ou órgãos-alvo, possibilitam variados diagnósticos diferenciais. Dentre as manifestações clínicas que possibilitam estes diagnósticos está o acometimento cutâneo, com vasta variabilidade de apresentação. Da mesma forma, a sífilis também possui apresentação cutânea, tornando possível o diferencial de diagnóstico com outras patologias, inclusive o próprio LES. O presente estudo tem como objetivo relatar um caso de sífilis mimetizando lúpus eritematoso sistêmico, descrever o quadro clínico apresentado pelo paciente, bem como as ferramentas utilizadas para diagnóstico, e a posterior abordagem terapêutica. O caso relatado refere-se a um paciente de 29 anos, do sexo masculino, procedente de Campos Novos (SC), que apresentou um quadro clínico e laboratorial de lúpus-like induzido por uma infecção aguda de sífilis. A resolução completa de critérios inflamatórios de LES ocorreu após tratamento correto da doença infecciosa, com total melhora clínica e sorológica.

Systemic lupus erythematosus (SLE) is a chronic autoimmune inflammatory disease. The various clinical manifestations in SLE patients, both systemic and in target organs, allow for various differential diagnoses. Among the clinical manifestations that aid in diagnosis are the cutaneous injuries, which have a wide range of presentations. Syphilis also has cutaneous manifestations, which aid in the differential diagnosis from other pathologies, including SLE. The present study aims to report a case of syphilis mimicking SLE, describe the clinical condition presented by the patient, the tools used for diagnosis, and the therapeutic approach. The case reported refers to a 29- year-old male patient from Campos Novos (SC), who showed a clinical and laboratory lupus-like condition induced by an acute syphilis infection. The full resolution of SLE inflammatory criteria occurred following appropriate treatment for the infectious disease, with complete clinical and serological improvement.

El lupus eritematoso sistémico (LES) es una enfermedad inflamatoria autoinmune crónica. Las diversas manifestaciones clínicas de los pacientes con LES, tanto sistémicas como en órganos diana, permiten realizar varios diagnósticos diferenciales. Entre las manifestaciones clínicas que ayudan al diagnóstico se encuentran las lesiones cutáneas, que tienen una amplia gama de presentaciones. La sífilis también tiene manifestaciones cutáneas, que ayudan al diagnóstico diferencial con otras patologías, incluido el LES. El presente estudio tiene como objetivo comunicar un caso de sífilis que simula un LES, describir el cuadro clínico presentado por la paciente, las herramientas utilizadas para el diagnóstico y el abordaje terapéutico. El caso relatado se refiere a un paciente masculino de 29 años, natural de Campos Novos (SC), que presentó un cuadro clínico y de laboratorio semejante al lupus, inducido por una infección aguda por sífilis. La resolución completa de los criterios inflamatorios del LES ocurrió tras el tratamiento adecuado de la enfermedad infecciosa, con mejoría clínica y serológica completa.

Tendência temporal e distribuição espacial da sífilis gestacional e congênita no Brasil, período de 2008-2018 / Temporal trend and spatial distribution of gestational and congenital syphilis in Brazil, period 2008-2018

Introdução: A sífilis tornou-se um problema de saúde pública em várias regiões no mundo. Objetivo: Analisar a tendência temporal e distribuição espacial da sífilis gestacional e congênita no Brasil, no período de 2008 a 2018. Método: Trata-se de um estudo de diferentes métodos. 1) Estudo de série temporal da taxa de detecção da sífilis gestacional e correlação entre variáveis socioeconômicas e de serviços de saúde. Os dados foram extraídos de bancos de dados nacionais de acesso público. Utilizou-se o software Joinpoint Regression e o coeficiente de correlação de Pearson. 2) Estudo ecológico de análise espacial da taxa de detecção da sífilis gestacional e correlação espacial entre variáveis socioeconômicas e de serviços de saúde. Extraídos dados secundários, agregados em 482 Regiões Imediatas de Articulação Urbana. Utilizou-se o software GeoDa. 3) Estudo ecológico de tendência temporal e distribuição espacial da taxa de incidência da sífilis congênita e correlação espacial entre variáveis socioeconômicas e de serviços de saúde. Extraídos dados secundários. 4) Protocolo para uma revisão de escopo com o objetivo de identificar e mapear o uso da Análise Espacial como ferramenta em pesquisas sobre sífilis na área da saúde. Baseou-se no manual do Joanna Briggs Institute e no guia PRISMA-ScR. Resultados: A região Sul apresentou a maior tendência; enquanto a Centro-Oeste, menor. Detectada correlação com o Índice de Desenvolvimento Humano Municipal, taxa de analfabetismo, percentual de cobertura da atenção primária à saúde e proporção de médicos, enfermeiros e unidades básicas de saúde por habitante. A taxa de detecção de sífilis gestacional distribuiu-se desigualmente e mostrou correlação espacial com o Índice de Desenvolvimento Humano Municipal, percentual de cobertura da atenção básica e proporção de médicos por habitantes. A taxa de incidência de sífilis congênita mostrou tendência ascendente e distribuíção desigual. Apresentou correlação espacial com o percentual de indivíduos com abastecimento de água e saneamento inadequados e percentual de nascidos vivos com 1 a 3 consultas de pré-natal. O protocolo para a revisão de escopo explicitou a questão de pesquisa, as bases de dados para buscas, critérios de inclusão e exclusão, planilha de extração de dados e o tipo de análise dos dados. Considerações finais: Investimentos em políticas de saúde e sociais são necessárias para mitigar as vulnerabilidades sociais e fortalecer a atenção primária à saúde para o controle da sífilis (AU).

Introduction: Syphilis has become a public health problem in several regions of the world. Objective: To analyze the temporal trend and spatial distribution of gestational and congenital syphilis in Brazil, from 2008 to 2018. Method: It is a study of different methods. 1) Time series study of the detection rate of gestational syphilis and correlation between socioeconomic and health service variables. Data were extracted from publicly accessible national databases. The Joinpoint Regression software and Pearson's correlation coefficient were used. 2) Ecological study of spatial analysis of the detection rate of gestational syphilis and spatial correlation between socioeconomic variables and health services. Extracted secondary data, aggregated in 482 Immediate Urban Articulation Regions. GeoDa software was used. 3) Ecological study of temporal trends and spatial distribution of the incidence rate of congenital syphilis and spatial correlation between socioeconomic variables and health services. Secondary data extracted. 4) Protocol for a scoping review with the objective of proposing a protocol to identify and map the use of Spatial Analysis as a tool in research on syphilis in the health area. It was based on the Joanna Briggs Institute manual and guided by PRISMA-ScR. Results: The South region showed the greatest trend; while the Midwest, smaller. Correlation detected with the Municipal Human Development Index, illiteracy rate, percentage of primary health care coverage and proportion of doctors, nurses and basic health units per inhabitant. The detection rate of gestational syphilis was unevenly distributed and showed a spatial correlation with the Municipal Human Development Index, percentage of primary care coverage and proportion of physicians per population. The incidence rate of congenital syphilis showed an upward trend and uneven distribution. It showed a spatial correlation with the percentage of individuals with inadequate water supply and sanitation and the percentage of live births with 1 to 3 prenatal consultations. The protocol for the scope review explained the research question, the databases for searches, inclusion and exclusion criteria, data extraction worksheet and the type of data analysis. Final considerations: Investments in health and social policies are necessary to mitigate social vulnerabilities and strengthen primary health care for syphilis control (AU).

Syphilitic Folliculitis: A Case Report With Demonstration of Spirochetes Showing Follicular Epitheliotropism.

ABSTRACT: We report a case of a 59-year-old man presenting with a widespread follicular-based papular rash with a several-month history of myalgias, lymphadenopathy, fatigue, and weight loss who was diagnosed with acute syphilitic folliculitis by tissue biopsy analysis with immunohistochemical demonstration of spirochetes in hair follicle epithelium. Serologic analysis also showed evidence of Treponema sp. infection. Owing to the rising number of syphilis cases in the last decade, it is important to recognize classic cutaneous findings of syphilis in addition to unusual presentations such as syphilitic folliculitis.

FDG PET/CT in a Patient With Syphilis Suspected of Having Penile Cancer.

ABSTRACT: A 58-year-old man with large penile wound and enlarged regional lymph node was suspected of having disseminated penile cancer. FDG PET/CT for primary staging showed high FDG uptake on penis and in several enlarged lymph nodes. However, biopsies revealed no signs of malignancy, but ulceration, inflammation, fibrosis, and spirochetes. Furthermore, Wassermann test was positive. The patient was then treated for syphilis. To our knowledge, this is the first report on FDG PET/CT in a patient suspected of having penile cancer that turned out to be syphilis. Thus, syphilis can be added to the list of benign pitfalls in FDG PET/CT.

Comparison of five-year survival rates among patients with oral squamous cell carcinoma with and without association with syphilis: a retrospective case-control study.

BACKGROUND: Syphilis is an infectious disease that is at least discussed to be premalignant. This potential, combined with its general pathological impact, raises the question if syphilis increases mortality in oral cancer patients. The aim of the study was to assess if the five-year survival rates among patients suffering from oral squamous cell carcinoma (OSCC) with (cohort I) and without association with syphilis (cohort II) differ. METHODS: Retrospective clinical data of patients diagnosed with OSCC (International Classification of Diseases [ICD]-10 codes C01-06) within the past 20 years from the access date September 25, 2021 were retrieved from the TriNetX network (TriNetX, Cambridge, Massachusetts, USA) to gain initial cohort 0. Subjects also diagnosed with syphilis (ICD-10 codes A51-53) were assigned to cohort I. Cohort II was comprised of the remaining individuals of cohort 0 by creating a group with the same number of patients as cohort I, and by matching for age and gender. Subsequently, Kaplan-Meier analysis and Cox proportional hazards regression were performed, and risk, odds and hazard ratios were calculated. RESULTS: Of a total of 73,736 patients in cohort 0, 199 individuals were each assigned to cohort I and II. During the five-year period after tumor diagnosis, 39 and 30 patients in cohort I and II died. The five-year survival probabilities did not significantly differ between the cohorts (I vs. II = 74.19% vs. 75.01%; p = 0.52; Log-Rank test), nor the risk of dying (I vs. II = 19.6% vs. 15.08%; risk difference = 4.52%; p = 0.23). The calculated risk, odds and hazard ratios were 1.3 (95% confidence interval [CI] = 0.84; 2.00), 1.37 (95% CI = 0.81; 2.31) and 1.17 (95% CI = 0.73; 1.88), respectively. CONCLUSIONS: The obtained results indicate that the survival rate of individuals with OSCC might not be negatively influenced if syphilis is present/associated. However, the results need to be interpreted cautiously due to limitations related to the retrospective approach, especially as data on the tumor staging were not accessible. TRIAL REGISTRATION: Due to the retrospective nature of the study, no registration was necessary.

Hepatic Secondary Syphilis Can Cause a Variety of Histologic Patterns and May Be Negative for Treponeme Immunohistochemistry.

The rate of syphilis in the United States has been increasing steadily in the past decade, but it remains an uncommon diagnosis in tissue biopsies. Most of the pathology literature on hepatic syphilis consists of older series or case reports. This study aimed to systematically characterize the histologic spectrum of hepatic syphilis in a contemporary cohort. Clinicopathologic features of 14 hepatic syphilis cases between 2012 and 2018 were analyzed to characterize the broad spectrum of histologic changes. Thirteen patients were men (age range: 19 to 59 y); 6 had known human immunodeficiency virus, 7 were men known to have sex with men, and no patient had known prior syphilis. Hepatic syphilis was the primary clinical suspicion in only 1 patient. Common symptoms included jaundice, rash, and abdominal pain. Thirteen had elevated transaminases, and 12 had elevated alkaline phosphatase. Pathologic changes were grouped into 5 histologic patterns: biliary-pattern injury (n=5), acute hepatitis (n=4), autoimmune hepatitis-like (n=1), fibroinflammatory mass-forming lesion (n=2), and no particular pattern (n=2). Nearly all showed portal and lobular lymphocytes and plasma cells; 12 had prominent histiocytes/Kupffer cells, 9 had ductular reaction, and 7 had duct inflammation. Occasional focal findings included dropout (n=7), phlebitis (n=7), and loose granulomata (n=5). Treponeme immunohistochemistry was positive in 10 and negative in 4, though treatment was given before biopsy in 3 of those 4. Thirteen patients had rapid plasma reagin testing either before or after biopsy, with 1:64 or higher titer. All patients who received treatment recovered. Hepatic syphilis is rare but likely underrecognized. It exhibits a variety of histologic appearances and therefore should be considered in several hepatic differential diagnoses, especially in men who have sex with men. Kupffer cells, granulomata, and phlebitis may suggest the diagnosis regardless of predominant histologic pattern. Negative treponeme immunohistochemical staining does not exclude the diagnosis, including in untreated patients.

Case report of gastric syphilis in Korea: Clinical features, pathology, management, and prognosis.

RATIONALE: Syphilis is a contagious infectious disease caused by Treponema pallidum. Gastric involvement of syphilis is rare and has nonspecific gastrointestinal symptoms and endoscopic findings. To date, 16 cases have been reported in Korea. Here, we report 2 additional cases of gastric syphilis in men in their 30 second. PATIENTS CONCERNS: Two 35- and 33-year-old men presented with epigastric pain. DIAGNOSIS: The serum venereal disease research laboratory and fluorescent treponemal antibody absorption tests were positive. Esophagogastroduodenoscopy showed multiple variable-sized flat elevated lesions and geographic ulcers with whitish exudates in the antrum and body. Warthin-Starry silver staining of endoscopic biopsy specimens confirmed gastric syphilis. INTERVENTIONS: The patients were treated with an intramuscular injection of 2.4 million units of benzathine penicillin once a week for 3 weeks. OUTCOMES: Clinical symptoms and gastric lesions were completely resolved. LESSONS: First, gastric syphilis, despite its rarity and nonspecific symptoms and endoscopic findings, should be considered in a rare extracutaneous presentation of syphilis. Second, a high index of clinical suspicion and an accurate diagnosis based on a combination of clinical, radiological, endoscopic, serologic, and histopathologic findings provide an opportunity to identify and treat patients with gastric syphilis.

Analysis of 23 Years of Cornea Donor Data From an Eye Bank in Turkey.

OBJECTIVES: We evaluated 23 years of data for cornea donors at the Istanbul Faculty of Medicine Lions Eye Bank. MATERIALS AND METHODS: Annual statistics of corneal donors between 1996 and 2019 were reviewed retrospectively. Records for 2008 and previous years were compared with records for 2009 and years thereafter,to assess donor demographics and reasons for discard of corneas. RESULTS: A total of 3849 corneas were obtained from 2018 donors during a 23-year period. Of these, 26 donors (11.2%) were registered, whereas 1792 (88.8%) did not register any decision for donation. There were 210 (5.46%) corneas discarded for positive serology and 291 (7.56%) for unsuitable tissue morphology, and 3348 (86.98%) corneas were determined to be suitable for transplant. For the cause of death in 2009 and subsequent years, the incidence of trauma was lower (P = .001) compared with the years previous to 2009, whereas incidence of cardiac pathology (P = .014) was higher. The number of donors older than 50 years was higher for 2009 and years thereafter, compared with data from earlier years (P = .024). The rate of discarded corneas for unsuitable tissue morphology (P = .004) and total discarded corneas (P = .027) decreased in 2009 and years thereafter. The rate of discarded corneas for HIV was higher after for 2009 and years thereafter (P = .047), whereas the rate of discarded corneas for syphilis was significantly higher for 2015 and years thereafter (P = .001). CONCLUSIONS: This study evaluated the most extensive eye bank data in Turkey and found that advances in lamellar surgery techniques and improvements in specular microscopic facilities have expanded the pool for cornealtransplants to include tissues from older donors. Moreover, the incidence rates for HIVpositive and syphilis-positive serology tests in discarded corneas have increased over time.

Unusual clinical manifestation and challenging serological interpretation of syphilis: insights from a case report.

BACKGROUND: The clinical manifestations of recent syphilis can be variable, with typical and atypical patterns. Several conditions may cause atypical clinical aspects, including human immunodeficiency virus (HIV) co-infection. Besides the clinical features, co-infections may completely alter syphilis serological tests, causing interpretative difficulties and diagnostic delays. Aim of the work is to describe the difficulties encountered during the diagnostic evaluation of atypical skin manifestations and of the serology for syphilis of an HIV-infected patient who had contracted it several times. CASE PRESENTATION: In 2020, a 52-year old HIV-positive bisexual male patient was admitted to our department with a 4-month history of moderately itchy cutaneous lesions localized at his neck, trunk and arms. In 2013, the patient presented with a classic syphilitic roseola of the trunk and a secondary syphilis was diagnosed, with increased levels of rapid plasma reagin (RPR), Treponema pallidum hemagglutination assay (TPHA), anti-Treponema pallidum IgM and IgG Index. A second episode occurred in 2018, as a primary syphilis with multiple ulcerative lesions of the penis, and increased levels of RPR, IgG and IgM. In 2019, a further episode of secondary syphilis was treated with Doxycycline. In 2020, erythematous and papular lesions with vesicular components and urticarial erythema multiforme (EM)-like lesions were present at the neck, trunk and arms. Serological tests and Nucleic Acid Amplification Test (NAAT) for Treponema Pallidum were performed, as well as a cutaneous biopsy with histological and immunohistochemical evaluation of one lesion. NAAT was negative for T. pallidum. Serological test results were discordant with a new syphilis infection, showing only increased levels of RPR and anti-Treponema IgG. The cutaneous biopsy revealed a non specific histological pattern, while the immunohistochemical evaluation with anti-spirochetal antibodies was mandatory for the diagnosis of recent syphilis, showing clusters of rod-shaped elements, some of which with spiral form, focally present at the epidermis and adnexal structures. CONCLUSIONS: Nowadays, syphilis may present with atypical clinical and serological features. Physicians should be aware of these possible alterations and consider syphilis even in case of uncommon clinical aspect and unclear serological tests. Cutaneous biopsy and immunohistochemical exam may be mandatory for the diagnosis.