Spirulina Platensis Attenuates Rebound Dyspeptic Symptoms After Proton Pump Inhibitors' Discontinuation: Phase 2 Placebo-controlled Trial.

Context: Rebound acid hypersecretion after cessation of proton pump inhibitors (PPIs) can provoke dyspeptic symptoms. The search for alternatives to minimize the dyspeptic rebound symptoms after PPI discontinuation is warranted. Spirulina platensis, a dietary supplement made from blue-green algae, might be an alternative. Objective: The study intended to assess whether Spirulina platensis, through its anti-inflammatory and analgesic properties, can minimize rebound symptoms after PPI withdrawal. Design: The research team performed a randomized, phase 2, double-blinded, placebo-controlled clinical trial. Setting: The study took place at São Vicente de Paulo Hospital (trial registry number NCT04988347) in Passo Fundo, Brazil. Participants: Participants were 45 Brazilian patients in the clinical practice of two of the research team's member between November 2010 and February 2012, who were using PPIs regularly. Interventions: Participants underwent clinical and endoscopic evaluations after a 28-day run-in phase of 40 mg/day of pantoprazole. In the absence of a large hiatal hernia, peptic ulcer, or severe reflux esophagitis, participants stopped using PPIs, and the research team randomly assigned them to receive either 1.6g/day of spirulina or of a placebo for two months, followed by clinical and endoscopic reevaluations. Outcome measures: Using an intention-to-treat analysis, the primary outcomes postintervention were dyspepsia and typical reflux symptoms, either the appearance or maintenance of symptoms of >50% from baseline. Results: The median time of continuous PPI use was 32 months. The research team excluded two participants due to large hiatal hernias. Among the remaining 43 participants, 18 received spirulina (42%), and 25 used a placebo (58%). Two months later, 12 participants who had received spirulina (67%) and 18 who had received the placebo (72%) completed the study (P = .968). Rebound dyspepsia occurred in 10 out of 18 patients treated with spirulina (55.56%) and in 22 out of 25 patients treated with placebo (88%), with relative risk=0.63, CI95% (0.41-0.98), and P = .039. Reflux symptoms postintervention occurred in 72% and 76%, with the relative risk=0.95, CI95% (0.66-1.36), and P > .05, respectively. No significant side effects occurred in either group. The findings from endoscopy and gastric histology didn't differ between groups. Conclusions: A two-month course of Spirulina platensis was able to attenuate rebound dyspepsia but not reflux symptoms after PPI discontinuation. Considering its good safety profile, spirulina might be useful to relieve dyspeptic symptoms after PPI discontinuation.

Diagnostic criteria and case definitions for abdominal angiostrongyliasis: a systematic review from the Brazilian experience.

Although rare, Angiostrongylus costaricensis infection may be a more prevalent etiology of inflammatory bowel disease than ulcerative colitis and Chron's disease in endemic areas in Central and South America. The present study reviewed the occurrence of A. costaricensis in Brazil, its clinical presentation and pathology; and proposed diagnostic criteria and case definitions for abdominal angiostrongyliasis (AA). Southern and southeastern Brazilian regions are the main endemic areas, and AA affects both genders and all age groups. A review of all 23 published reports of 51 Brazilian patients highlighted the following features that were subsequently classified as minor diagnostic criteria: abdominal pain, palpable mass in the right lower abdominal quadrant, history of exposure, ileocecal tumor, and intestinal perforation with wall thickening. Proposed major criteria include right lower quadrant abdominal pain, blood eosinophilia, positive serology (antibody detection), intense eosinophilic infiltration that involves all strata of the intestinal wall, eosinophilic granulomatous reaction, and eosinophilic vasculitis. In addition to the definitions of suspected and possible cases according to increasing strength of evidence of this infection, demonstration of worms/eggs/larvae in tissues or Angiostrongylus DNA in tissues or serum are required for a confirmed diagnosis. The application of the proposed criteria and definitions may improve patient management, epidemiologic surveillance, and identification of new endemic areas.

Molecular diagnosis of abdominal angiostrongyliasis by PCR using serum samples.

Abdominal angiostrongyliasis (AA) is a zoonotic disease caused by the nematode Angiostrongylus costaricensis, which is endemic in southern Brazil. Humans become infected by ingesting third-stage (L3) larvae and are considered accidental hosts since neither eggs nor first-stage (L1) larvae are found in feces. The definitive diagnosis can be made by histopathologic examination of surgical specimens or intestinal biopsies. The present study assessed the use of PCR to carry out the molecular detection of AA from serum samples. A total of 62 human serum samples were divided into three groups: (i) 28 serum samples from human patients with presumptive histopathological diagnosis of AA; (ii) 23 serum samples from individuals with unknown serology for AA; (iii) 11 serum samples from patients that suffered from different parasitosis were included. The serum samples were initially tested by in-house indirect ELISA and then by PCR. A total of 14 samples were positive by ELISA, and 6 were positive by PCR. Six samples that were negative by ELISA were positive by PCR. Amplicons were sequenced, and Angiostrongylus DNA was confirmed. We conclude that PCR amplification can be used to confirm Angiostrongylus DNA in serum, which is especially important in cases where antibody levels are too low to be detected. It may also serve as a useful target for survey studies.

Abdominal angiostrongyliasis can be diagnosed with a immunochromatographic rapid test with recombinant galactin from Angiostrongylus cantonensis.

Angiostrongylus costaricensis is the causative agent of abdominal angiostrongyliasis, a zoonotic infection that may produce severe eosinophilic enterocolitis or hepatitis in humans. Parasites are usually not released in stools and serology has an important role in diagnosis. Since cross-reactivity is demonstrated between A. costaricensis and another metastrongylid worm, A. cantonensis, we tested heterologous recombinant galectin as a probe in an immunochromatographic rapid diagnostic test (ICT-RDT) for detection of anti-A. costaricensis antibodies. Almost all (11/12) positive control sera from A. costaricensis infected patients were positive at ICT RDT. These are preliminary indications that r-galectin ICT-RDT is useful for diagnosing A. costaricensis infection.

A practical guide for the diagnosis of abdominal angiostrongyliasis caused by the nematode Angiostrongylus costaricensis.

Abdominal angiostrongyliasis (AA) is a severe parasitic infection caused by the nematode Angiostrongylus costaricensis. This disease is characterized by abdominal pain, a strong inflammatory eosinophilic response in the blood and tissues, and eventually intestinal perforation. Diagnosis of AA is challenging since there are no commercially available serological kits for A. costaricensis, and thus, histopathological analysis remains the gold standard. Herein we provide a decision flowchart for clinicians to improve the diagnosis of AA based on a patient's clinical manifestations, laboratory findings, macroscopic observations of the gut lesions, as well as characteristic microscopic alterations in biopsies. A brief discussion of the available polymerase chain reaction and in-house serological methods is also presented. The aim of this mini-review is to improve the diagnosis of AA, which should lead to prompt detection of cases and better estimates of the epidemiology and geographical distribution of A. costaricensis.

Artificial intelligence as a tool for diagnosis in digital pathology whole slide images: A systematic review.

Digital pathology had a recent growth, stimulated by the implementation of digital whole slide images (WSIs) in clinical practice, and the pathology field faces shortage of pathologists in the last few years. This scenario created fronts of research applying artificial intelligence (AI) to help pathologists. One of them is the automated diagnosis, helping in the clinical decision support, increasing efficiency and quality of diagnosis. However, the complexity nature of the WSIs requires special treatments to create a reliable AI model for diagnosis. Therefore, we systematically reviewed the literature to analyze and discuss all the methods and results in AI in digital pathology performed in WSIs on H&E stain, investigating the capacity of AI as a diagnostic support tool for the pathologist in the routine real-world scenario. This review analyzes 26 studies, reporting in detail all the best methods to apply AI as a diagnostic tool, as well as the main limitations, and suggests new ideas to improve the AI field in digital pathology as a whole. We hope that this study could lead to a better use of AI as a diagnostic tool in pathology, helping future researchers in the development of new studies and projects.

Elimination of Angiostrongylus costaricensis larvae in feces from experimentally infected Swiss mice: circadian rhythm and correlation with survival.

Angiostrongylus costaricensis is a nematode which harbors mesentery arteries of rodents. In these animals, a circadian rhythm of elimination of first-stage larvae (L1) and a relation between the amount of L1 in feces and survival are unknown. We assessed fecal elimination of A. costaricensis L1 from experimentally infected Swiss mice and tried to correlate L1 elimination with survival. Thirteen Swiss mice were infected by gavage with ten A. costaricensis L3 larvae obtained from Phyllocaulis slugs. Feces were weighed at 7 A.M: . and 7 P.M: . starting from the 24th day post-infection until animal death. Feces sediment was examined in microscope for L1 counting. The mice were dead after a period ranging 19-61 days post-infection. Compared to diurnal samples, both feces' weight (2.3 ± 0.7 vs. 1.8 ± 0.5 g; P < 0.0001) and L1 total count [median 1,950 vs. 1,250; P = 0.015] were higher in feces eliminated at night. No difference was observed between diurnal and nocturnal elimination when counting L1 by gram of feces (725 vs. 650 L1/g; P = 0.821). A significant correlation was observed between survival and total number of L1 in feces (r = 0.84; P = 0.0007). This study suggests that mice experimentally infected with A. costaricensis eliminate more L1 at night due to higher fecal volume at this period. The correlation between number of L1 in feces and survival suggests a phenomenon of tolerance to A. costaricensis infection in mice with longer survival.

Outcomes in mice with abdominal angiostrongyliasis treated with enoxaparin.

Abdominal angiostrongyliasis (AA) is caused by the nematode Angiostrongylus costaricensis. Parasite-associated thrombosis of mesenteric vessels may lead to intestinal infarction, which might be prevented with anti-thrombotic agents. This study assessed the effect of enoxaparin on survival and pathological findings in Swiss mice with AA. In this experiment, 24 mice were infected with A. costaricensis (10 L3 per animal) followed by treatment with subcutaneous enoxaparin (40 mg/kg/day) or water (sham), starting from 15 days post-infection (dpi) and continued until animal death. Animals were monitored until death or sacrifice at the 50th dpi. Ten mice (42%) were dead after 36 ± 8 dpi. Of these, five (50%) were treated with enoxaparin. Animals treated with enoxaparin and sham did not differ in terms of weight loss (median, 1.3 vs. 4.2 g; P = 0.303) and macroscopical findings. Microscopically, no difference was found in regard to vascular granuloma (median grade, 2 vs. 3; P = 0.293) and presence of either vasculitis (75% vs. 100%; P = 0.217), mesenteric thrombosis (33% vs. 50%; P = 0.680), or bowel necrosis (25% vs. 50%; P = 0.400). Mice dead before the 50th dpi showed more pneumonia (90% vs. 21%; P = 0.002), bowel infarction (40% vs. 0%; P = 0.02), and purulent peritonitis (60% vs. 7%; P = 0.008) compared to survivors. Prophylactic enoxaparin in mice did not prevent tissue damage and mortality related with AA. The lower prevalence of mesenteric thrombosis and bowel infarction regardless of treatment were notorious. Frequent septic complications suggest the need of studies addressing the effect of antibiotics in AA.

Abdominal angiostrongyliasis in the Americas: fifty years since the discovery of a new metastrongylid species, Angiostrongylus costaricensis.

Angiostrongylus costaricensis is a zoonotic parasitic nematode described for the first time in 1971 by Pedro Morera and Rodolfo Céspedes in Costa Rica. This parasite causes an infection known as abdominal angiostrongyliasis, affecting mainly school-aged children and young adults. Infection with A. costaricensis has been associated with a myriad of rodent and mollusk species in the Americas and the Caribbean, as its natural hosts and reservoirs. In this commemorative review, we highlight the extensive research collected through a 50-year journey, which includes ecological, pathological, and molecular studies on A. costaricensis and its implicated disease. We also identify major knowledge gaps in its evolutionary history, the ecological role of imported and invasive mollusk species, and immune response. We propose that the advent of -omics analyses will allow us to gather novel information regarding A. costaricensis biology and infection dynamics, as well as to promote the design of much-needed sensitive and specific diagnostic tools.

Apical root canal cleaning after preparation with endodontic instruments: a randomized trial in vivo analysis.

OBJECTIVES: This study aimed to evaluate vital pulp tissue removal from different endodontic instrumentation systems from root canal apical third in vivo. MATERIALS AND METHODS: Thirty mandibular molars were selected and randomly divided into 2 test groups and one control group. Inclusion criteria were a positive response to cold sensibility test, curvature angle between 10 and 20 degrees, and curvature radius lower than 10 mm. Root canals prepared with Hero 642 system (size 45/0.02) (n = 10) and Reciproc R40 (size 40/0.06) (n = 10) and control (n = 10) without instrumentation. Canals were irrigated only with saline solution during root canal preparation. The apical third was evaluated considering the touched/untouched perimeter and area to evaluate the efficacy of root canal wall debridement. Statistical analysis used t-test for comparisons. RESULTS: Untouched root canal at cross-section perimeter, the Hero 642 system showed 41.44% ± 5.62% and Reciproc R40 58.67% ± 12.39% without contact with instruments. Regarding the untouched area, Hero 642 system showed 22.78% ± 6.42% and Reciproc R40 34.35% ± 8.52%. Neither instrument achieved complete cross-sectional root canal debridement. Hero 642 system rotary taper 0.02 instruments achieved significant greater wall contact perimeter and area compared to reciprocate the Reciproc R40 taper 0.06 instrument. CONCLUSIONS: Hero 642 achieved higher wall contact perimeter and area but, regardless of instrument size and taper, vital pulp during in vivo instrumentation is not entirely removed.

First case report of nematode parasitic myelopathy in a wild feline in Brazil.

Parasitic myelopathy caused by Gurltia paralysans in domestic cats is a disease commonly reported in several South American countries. The adult parasite is lodged in the meningeal veins and spinal cord, often causing clinical manifestations of vascular proliferation, thrombophlebitis, and medullary compression. Wild felines are believed to be the definitive hosts of this parasite. The infection occurs through the ingestion of paratenic hosts, but the life cycle of G. paralysans is not yet clearly understood. In this paper, we discuss a case of parasitic myelopathy in a margay (Leopardus wiedii) that died during post-surgical care. Necropsy revealed focal hemorrhages in the thoracolumbar spinal cord. A microscopic examination revealed adult nematodes and eggs inside the veins of subarachnoid space in spinal cord, suggesting G. paralysans infection. This is first description of parasitic myelopathy in a margay in Brazil.

Effects of ZnSO4-induced peripheral anosmia on zebrafish behavior and physiology.

Olfaction plays a key role in modulating behavioral and physiological responses of various animal species, including fishes. Olfactory deficits can be induced in fish experimentally, and utilized to examine the role of olfaction in their normal and pathological behaviors. Here, we examine whether experimental anosmia, evoked by ZnSO4 in adult zebrafish can be associated with behavioral and/or physiological responses. We show that experimental ZnSO4-induced anosmia caused acute, but not prolonged, anxiogenic-like effects on zebrafish behavior tested in the novel tank test. The procedure also elevated whole-body cortisol levels in zebrafish. Moreover, ZnSO4 treatment, but not sham, produced damage to olfactory epithelium, inducing overt basal cell vacuolization and intercellular edema. The loss of olfaction, assessed by the fish food preference behavior in the aquatic Y-maze, was present 1h, but not 24h, after the treatment. Collectively, this suggests that transient experimental anosmia by ZnSO4 modulates zebrafish behavior and olfaction, which can be used to evoke and assess their stress-related anxiety-like states.

Mucinous cystic neoplasm of the liver

A 55-year-old woman was investigated for occasional epigastric pain and weight loss. T2-weighted abdominal magnetic resonance imaging and magnetic resonance cholangiography revealed a multilocular cyst with multiple septa and a solid component in the liver, measuring 6.1 × 4.8 × 6.5 cm. Given the patient's symptoms and malignant potential, a laparoscopic segmentectomy with partial resection of segments IV B and V was performed to completely remove the cystic lesion, associated with cholecystectomy. Histopathology demonstrated a cyst lined by columnar mucinous epithelium. Therefore, the diagnosis was mucinous cystic neoplasm of the liver. This article presents a case report and literature review of this entity.

Umbilical cord constriction as a cause of intrauterine fetal death / Constrição de cordão umbilical como causa de morte fetal intrauterina

ABSTRACT The umbilical cord constriction (UCC) is an uncommon condition and an important etiology for stillborn fetuses. The main goal of this study was to verify the UCC occurrence as the cause of intrauterine fetal death, the associated etiology and its pathological characteristics. Therefore, a descriptive retrospective cross-sectional study was developed using the database from a Pathology Institute, in Brazil, from 1995 to 2017. The results presented a total of 1,359 embryo/fetus deaths - 69 (5.07%) due to UCC, 60.9% males and 39.1% females. The average age of pregnant women was 27.5 years ± 7.2 years of standard deviation (SD). The majority of deaths occurred during the second trimester (76.5%), followed by the first (14.7%) and third (8.8%) trimesters, respectively. One constriction alone was found in 87% of cases, 11% had two constrictions and only 1% had three or more. The presence of congenital malformations was detected in 20.2% of necropsies, the identification of chronic fetal distress was described in 71% of the technical reports and 17% of the cases had obstructive vasculopathy characteristics in microscopy analysis. Regarding the anatomopathological characteristics between the male and female sexes, no significant difference was found (p > 0.05) correlating gestational age, weight or congenital malformations. UCC was a cause of fetal death found in 5% of the cases, and it was linked to congenital malformations, fetal distress and obliterative vasculopathy.

RESUMEN La constricción del cordón umbilical (CCU) es una condición infrecuente que no está bien descrita en la literatura, a pesar de ser una importante etiología observada en mortinatos. El objetivo de este estudio fue verificar la ocurrencia de CCU como causa de muerte fetal y las características patológicas asociadas. Para ello, se llevó a cabo un estudio retrospectivo transversal con informaciones de la base de datos de un Instituto de Patología, en Brasil, de 1995 hasta 2017. Los resultados fueron: 1.359 muertes de embriones/fetos, de las cuales 69 (5,07%) se debieron a CCU; 60,9% eran del sexo masculino y 39,1%, del femenino. El promedio de edad de las mujeres embarazadas fue de 27,5 anos; desviación estándar de ± 7,2 anos. La mayoría de las muertes ocurrió en el segundo trimestre (76,5%), seguido del primero (14,7%) y del tercero (8,8%). Entre el total de casos de CCU observados, 87% tuvieron sólo una constricción; 11%, dos; y 1%, tres o más. La presencia de malformaciones congénitas fue encontrada en 20,2% de las necropsias; la identificación de sufrimiento fetal crónico fue reportada en 71% de los informes; y características de vasculopatía obstructiva en el análisis de microscopía, en 17% de los casos. Con respecto a las características anatomopatológicas entre los sexos masculino y femenino, no se encontró ninguna diferencia significativa (p > 0,05) correlacionando edad gestacional de los fetos/embriones, peso o malformaciones congénitas. La CCU es una condición rara de muerte fetal asociada a bajo peso fetal y restricción de crecimiento. Nuestra investigación sugiere una posible relación entre la CCU y la vasculopatía obliterativa.

RESUMO A constrição de cordão umbilical (CCU) é uma condição infrequente, não bem descrita na literatura, apesar de ser uma importante etiologia observada em fetos natimortos. O objetivo deste estudo foi verificar a ocorrência de CCU como causa de morte fetal e as características etiopatológicas associadas. Para tanto, um estudo retrospectivo transversal foi realizado a partir das informações do banco de dados do Instituto de Patologia de Passo Fundo, durante os anos de 1995 a 2017. Os resultados foram: 1.359 mortes de embriões/fetos, das quais 69 (5,07%) foram devidas a CCU; 60,9% eram do sexo masculino e 39,1%, do feminino. A média de idade das gestantes foi de 27,5 anos; desvio padrão (DP) de ± 7,2 anos. A maioria das mortes ocorreu no segundo trimestre (76,5%), seguido pelo primeiro (14,7%) e terceiro (8,8%). Entre o total de CCU observadas, 87% tiveram ocorrência de apenas uma constrição; 11%, de duas; e 1%, de três ou mais. A presença de malformações congênitas foi encontrada em 20,2% das necropsias; a identificação de sofrimento fetal crônico foi relatada em 71% dos laudos; e características de vasculopatia obstrutiva na análise da microscopia, em 17% dos casos. Em relação às características anatomopatológicas entre os sexos masculino e feminino, nenhuma diferença significativa (p > 0,05) foi encontrada correlacionando idade gestacional dos fetos/embriões, peso ou malformações congênitas. A CCU é uma condição incomum de morte fetal associada a baixo peso fetal e restrição de crescimento. Nossa pesquisa sugere uma possível relação entre a CCU e a vasculopatia obliterativa.

Longitudinal clinical and serological survey of abdominal angiostrongyliasis in Guaporé, southern Brazil, from 1995 to 1999.

Abdominal angiostrongyliasis is a zoonotic infection caused by Angiostrongylus costaricensis, a nematode with an intra-vascular location in the mesentery. Our objective was to address several aspects of the natural history of this parasitosis, in a longitudinal clinical and seroepidemiological study. A total of 179 individuals living in a rural area with active transmission in southern Brazil were followed for five years (1995-1999) resulting in yearly prevalence of 28.2%, 4.2%, 10%, 20.2% and 2.8% and incidences of 0%, 5.9%, 8% and 1.5%, respectively. Both men and woman were affected with higher frequencies at age 30-49 years. In 32 individuals serum samples were collected at all time points and IgG antibody reactivity detected by ELISA was variable and usually persisting not longer than one year. Some individual antibody patterns were suggestive of re-infection. There was no association with occurrence of abdominal pain or of other enteroparasites and there was no individual with a confirmed (histopathologic) diagnosis. Mollusks were found with infective third-stage larvae in some houses with an overall prevalence of 16% and a low parasitic burden. In conclusion, abdominal angiostrongyliasis in southern Brazil may be a frequent infection with low morbidity and a gradually decreasing serological reactivity.

First case report of nematode parasitic myelopathy in a wild feline in Brazil / Primeiro relato de caso de mielopatia parasitária causada por nematódeo em um felídeo selvagem no Brasil

Abstract Parasitic myelopathy caused by Gurltia paralysans in domestic cats is a disease commonly reported in several South American countries. The adult parasite is lodged in the meningeal veins and spinal cord, often causing clinical manifestations of vascular proliferation, thrombophlebitis, and medullary compression. Wild felines are believed to be the definitive hosts of this parasite. The infection occurs through the ingestion of paratenic hosts, but the life cycle of G. paralysans is not yet clearly understood. In this paper, we discuss a case of parasitic myelopathy in a margay (Leopardus wiedii) that died during post-surgical care. Necropsy revealed focal hemorrhages in the thoracolumbar spinal cord. A microscopic examination revealed adult nematodes and eggs inside the veins of subarachnoid space in spinal cord, suggesting G. paralysans infection. This is first description of parasitic myelopathy in a margay in Brazil.

Resumo Mielopatia parasitária causada por Gurltia paralysans em gatos domésticos é uma doença comumente relatada em vários países da América do Sul. O parasita adulto aloja-se nas veias das meninges e da medula espinhal, muitas vezes causando proliferação vascular, tromboflebite e compressão medular, que se manifestam como sinais clínicos. Acredita-se que os felídeos selvagens sejam hospedeiros definitivos deste parasita e que a infecção ocorre por ingestão de hospedeiros paratênicos, entretanto seu ciclo de vida completo é desconhecido. Aqui, apresentamos um caso de um gato-maracajá (Leopardus wiedii) que morreu durante a assistência pós-cirúrgica. Na necropsia, foram observadas hemorragias focais na medula espinhal toracolombar. A microscopia revelou presença de nematódeos adultos e ovos, localizados dentro das veias do espaço subaracnoide da medula espinhal, o que sugeriu a infecção por G. paralysans. Esta é a primeira descrição de mielopatia parasitária em um gato-maracajá no Brasil.

Abdominal angiostrongyliasis can be diagnosed with a immunochromatographic rapid test with recombinant galactin from Angiostrongylus cantonensis

Angiostrongylus costaricensis is the causative agent of abdominal angiostrongyliasis, a zoonotic infection that may produce severe eosinophilic enterocolitis or hepatitis in humans. Parasites are usually not released in stools and serology has an important role in diagnosis. Since cross-reactivity is demonstrated between A. costaricensis and another metastrongylid worm, A. cantonensis, we tested heterologous recombinant galectin as a probe in an immunochromatographic rapid diagnostic test (ICT-RDT) for detection of anti-A. costaricensis antibodies. Almost all (11/12) positive control sera from A. costaricensis infected patients were positive at ICT RDT. These are preliminary indications that r-galectin ICT-RDT is useful for diagnosing A. costaricensis infection.

Dogs may be a reservoir host for Angiostrongylus costaricensis.

Angiostrongylus costaricensis is a parasitic nematode of wild rodents. Several other vertebrate species including man may become infected by ingestion of the third stage larvae produced by the intermediate hosts, usually slugs from the family Veronicellidae. There is a report of the diagnosis of abdominal angiostrongyliasis in Canis familiaris with lesions resembling those found in human disease. As a preliminar evaluation of the adequacy of a canine model for pathogenetic studies, a dog was inoculated with 75 L3 of A. costaricensis. Infection was established and fist stage larvae were found in feces up to 88 days post infection, sometimes in very large numbers (9.5 x 10(4) L1/g). No clinical manifestations or significant lesions were detected. These are indications that dog may play a role as a reservoir host for A. costaricensis.

Clinicopathologic findings and BRAF mutation in cutaneous melanoma in young adults.

Cutaneous melanoma in young patients is rare with increasing incidence. It is not clear whether the etiology and clinical outcome are similar to cutaneous melanoma in the elderly. Mutations in BRAF gene in patients with cutaneous melanoma, in general, range in frequency from 20% to 80%; however, the status and clinical significance of BRAF mutations in the young population have not been evaluated. We investigated 132 cases of primary cutaneous melanoma in patients aged between 18 and 30 years with emphasis on clinical characteristics, pathologic features, and molecular evaluation of mutation in the BRAF gene (BRAF(V600E)). It was predominantly seen in female individuals (61.4%), trunk was the most common site of involvement (40.4%), and superficially spreading melanoma was the predominant histologic type (79.5%). Mutation in BRAF(V600E) was analyzed successfully in 93 cases using an RT-PCR. The BRAF(V600E) mutation was identified in 38.7% (36/93) and was associated with vertical growth phase (P=0.01) and mild inflammatory infiltrate (P=0.02). No case of melanoma with regression phenomenon presented with BRAF(V600E) mutation (P<0.05). There was no significant association between BRAF(V600E) mutation and sex, histologic type, the Clark level, the Breslow index, solar elastosis, angiolymphatic and perineural invasion, satellitosis, and coexisting nevus. As in melanomas in older patients, these results probably indicate that BRAF mutation may not be the only key factor in melanoma tumorigenesis, and that there should be multiple alternative genetic pathways related to melanoma.

PCR for the diagnosis of abdominal angiostrongyliasis in formalin-fixed paraffin-embedded human tissue.

To date the diagnosis of abdominal angiostrongyliasis (AA) depends on the histological identification of Angiostrongylus costaricensis (AC) in surgical specimens. However, microscopic evaluation is time consuming and often fails in identifying the parasite. We tested whether PCR might help in the diagnosis of AA by identifying parasite DNA in formalin-fixed paraffin-embedded (FFPE) tissue. We used primers based on DNA from Angiostrongilus cantonensis. Four groups of FFPE intestinal tissue were tested: (1) confirmed cases (n = 20), in which AC structures were present in the target tissue; (2) presumptive cases (n = 20), containing changes secondary to AC infection in the absence of AC structures; (3) negative controls (n = 3), consisting of normal colonic tissue; and (4) tissue affected by other parasitoses (n = 7), including strongyloidiasis, ascaridiasis, schistosomiasis, and enterobiasis. Most lesions of confirmed cases were located in small and/or large bowel (90%), as compared with presumptive cases, in which 70% of lesions were in appendix (P = 0.0002). When confronted with cases of other parasitoses, PCR showed sensitivity of 55%, specificity of 100% and positive predictive value of 100%. In presumptive cases PCR was positive in 4 (20%). All specimens from negative controls and other parasitoses were negative. In conclusion, the PCR technique showed intermediate sensitivity and optimal specificity, being clinically relevant when positive for abdominal angiostrongyliasis. It allowed a 20% gain in diagnosis of presumptive cases. PCR might help in the diagnosis of abdominal angiostrongyliasis, particularly when the pathologists are not experienced with such disease.