Molecular and pathological insights into Chlamydia pecorum-associated sporadic bovine encephalomyelitis (SBE) in Western Australia

Background Despite its global recognition as a ruminant pathogen, cases of Chlamydia pecorum infection in Australian livestock are poorly documented. In this report, a C. pecorum specific Multi Locus Sequence Analysis scheme was used to characterise the C. pecorum strains implicated in two cases of sporadic bovine encephalomyelitis confirmed by necropsy, histopathology and immunohistochemistry. This report provides the first molecular evidence for the presence of mixed infections of C. pecorum strains in Australian cattle. Case presentation Affected animals were two markedly depressed, dehydrated and blind calves, 12 and 16 weeks old. The calves were euthanized and necropsied. In one calf, a severe fibrinous polyserositis was noted with excess joint fluid in all joints whereas in the other, no significant lesions were seen. No gross abnormalities were noted in the brain of either calf. Histopathological lesions seen in both calves included: multifocal, severe, subacute meningoencephalitis with vasculitis, fibrinocellular thrombosis and malacia; diffuse, mild, acute interstitial pneumonia; and diffuse, subacute epicarditis, severe in the calf with gross serositis. Immunohistochemical labelling of chlamydial antigen in brain, spleen and lung from the two affected calves and brain from two archived cases, localised the antigen to the cytoplasm of endothelium, mesothelium and macrophages. C. pecorum specific qPCR, showed dissemination of the pathogen to multiple organs. Phylogenetic comparisons with other C. pecorum bovine strains from Australia, Europe and the USA revealed the presence of two genetically distinct sequence types (ST). The predominant ST detected in the brain, heart, lung and liver of both calves was identical to the C. pecorum ST previously described in cases of SBE. A second ST detected in an ileal tissue sample from one of the calves, clustered with previously typed faecal bovine isolates. Conclusion This report provides the first data to suggest that identical C. pecorum STs may be associated with SBE in geographically separated countries and that these may be distinct from those found in the gastrointestinal tract. This report provides a platform for further investigations into SBE and for understanding the genetic relationships that exist between C. pecorum strains detected in association with other infectious diseases in livestock.

Determinants of vascular cognitive impairment : White matter lesions, brain atrophy and their neuropsychological correlates

The concept of vascular cognitive impairment (VCI) covers a wide spectrum of cognitive dysfunctions related to cerebrovascular disease. Among the pathophysiological determinants of VCI are cerebral stroke, white matter lesions and brain atrophy, which are known to be important risk factors for dementia. However, the specific mechanisms behind the brain abnormalities and cognitive decline are still poorly understood. The present study investigated the neuropsychological correlates of particular magnetic resonance imaging (MRI) findings, namely, medial temporal lobe atrophy (MTA), white matter hyperintensities (WMH), general cortical atrophy and corpus callosum (CC) atrophy in subjects with cerebrovascular disease. Furthermore, the cognitive profile of subcortical ischaemic vascular disease (SIVD) was examined. This study was conducted as part of two large multidisciplinary study projects, the Helsinki Stroke Aging Memory (SAM) Study and the multinational Leukoaraiosis and Disability (LADIS) Study. The SAM cohort consisted of 486 patients, between 55 and 85 years old, with ischaemic stroke from the Helsinki University Hospital, Helsinki, Finland. The LADIS Study included a mixed sample of subjects (n=639) with age-related WMH, between 65 and 84 years old, gathered from 11 centres around Europe. Both studies included comprehensive clinical and neuropsychological assessments and detailed brain MRI. The relationships between the MRI findings and the neuropsychological test performance were analysed by controlling for relevant confounding factors such as age, education and other coexisting brain lesions. The results revealed that in elderly patients with ischaemic stroke, moderate to severe MTA was specifically related to impairment of memory and visuospatial functions, but mild MTA had no clinical relevance. Instead, WMH were primarily associated with executive deficits and mental slowing. These deficits mediated the relationship between WMH and other, secondary cognitive deficits. Cognitive decline was best predicted by the overall degree of WMH, whereas the independent contribution of regional WMH measures was low. Executive deficits were the most prominent cognitive characteristic in SIVD. Compared to other stroke patients, the patients with SIVD also presented more severe memory deficits, which were related to MTA. The cognitive decline in SIVD occurred independently of depressive symptoms and, relative to healthy control subjects, it was substantial in severity. In stroke patients, general cortical atrophy also turned out to be a strong predictor of cognitive decline in a wide range of cognitive domains. Moreover, in elderly subjects with WMH, overall CC atrophy was related to reduction in mental speed, while anterior CC atrophy was independently associated with frontal lobe-mediated executive functions and attention. The present study provides cross-sectional evidence for the involvement of WMH, MTA, general cortical atrophy and CC atrophy in VCI. The results suggest that there are multifaceted pathophysiological mechanisms behind VCI in the elderly, including both vascular ischaemic lesions and neurodegenerative changes. The different pathological changes are highly interrelated processes and together they may produce cumulative effects on cognitive decline.

Arresting occlusal enamel caries lesions with pit and fissure sealants

The aim of this study was to evaluate the feasibility of pit and fissure sealants and the effectiveness of the two sealant methods applied in every-day practice in public dental health care in Finland. Two sealant methods were evaluated according to their effectiveness in preventing dentin caries and sealant retention. Application time with these sealant methods was compared. The survival rate of sealed first and second molars was followed for nine and 13 year periods, respectively. Caries risk evaluation and observed increased caries risk were the basis for considering sealant application. A questionnaire, sent to all public dental health centers in Finland, monitored the attitudes of the dental profession towards sealant application and explored the current policies used as well as changes noted in the sealant application protocol. DMFT (Decayed, Missing or Filled Teeth) index values collected from the health centers were evaluated. The difference in caries rate between two methods investigated was highly significant. When compared to the glass ionomer sealant method (GIC), the effectiveness of the resin-based method (RB) in preventing dentin caries was 74% and the rate difference 3%. The relative risk for RB-sealed surfaces vs. GIC-sealed surfaces of having detectable dentin caries was 0.3 (95% CI 0.12, 0.57). The retention rate of sealants was higher with RB than GIC (P<0.001). Application of RB sealant material was less time-consuming than application of GIC sealant. Occlusal dentin caries lesions were found in 4% and proximal caries in less than 2% of sealed teeth. The majority of respondents reported application of sealants on a systematic basis along with caries-risk evaluation. Those health centers sealing over suspected or detected enamel caries had lower average DMFT index values (1.0) when compared to DMFT values (1.2) of health centers applying sealants by alternative criteria. It is concluded that the RB sealant method is more effective than the GIC sealant method in preventing dentin caries. Sealant maintenance may increase the costs of a sealant program. Occlusal caries management may be improved if the applied sealant policies are changed towards an interceptive approach i.e. applying the sealants over detected or suspected enamel caries lesions instead of sealing sound teeth in a preventive manner.

Neuromagnetic studies on cortical somatosensory functions in infants and children : Normal development and effect of early brain lesions

Until recently, objective investigation of the functional development of the human brain in vivo was challenged by the lack of noninvasive research methods. Consequently, fairly little is known about cortical processing of sensory information even in healthy infants and children. Furthermore, mechanisms by which early brain insults affect brain development and function are poorly understood. In this thesis, we used magnetoencephalography (MEG) to investigate development of cortical somatosensory functions in healthy infants, very premature infants at risk for neurological disorders, and adolescents with hemiplegic cerebral palsy (CP). In newborns, stimulation of the hand activated both the contralateral primary (SIc) and secondary somatosensory cortices (SIIc). The activation patterns differed from those of adults, however. Some of the earliest SIc responses, constantly present in adults, were completely lacking in newborns and the effect of sleep stage on SIIc responses differed. These discrepancies between newborns and adults reflect the still developmental stage of the newborns’ somatosensory system. Its further maturation was demonstrated by a systematic transformation of the SIc response pattern with age. The main early adult­like components were present by age two. In very preterm infants, at term age, the SIc and SIIc were activated at similar latencies as in healthy fullterm newborns, but the SIc activity was weaker in the preterm group. The SIIc response was absent in four out of the six infants with brain lesions of the underlying hemisphere. Determining the prognostic value of this finding remains a subject for future studies, however. In the CP adolescents with pure subcortical lesions, contrasting their unilateral symptoms, the SIc responses of both hemispheres differed from those of controls: For example the distance between SIc representation areas for digits II and V was shorter bilaterally. In four of the five CP patients with cortico­subcortical brain lesions, no normal early SIc responses were evoked by stimulation of the palsied hand. The varying differences in neuronal functions, underlying the common clinical symptoms, call for investigation of more precisely designed rehabilitation strategies resting on knowledge about individual functional alterations in the sensorimotor networks.

Microhomology-mediated end joining is the principal mediator of double-strand break repair during mitochondrial DNA lesions

Mitochondrial DNA (mtDNA) deletions are associated with various mitochondrial disorders. The deletions identified in humans are flanked by short, directly repeated mitochondrial DNA sequences; however, the mechanism of such DNA rearrangements has yet to be elucidated. In contrast to nuclear DNA (nDNA), mtDNA is more exposed to oxidative damage, which may result in double-strand breaks (DSBs). Although DSB repair in nDNA is well studied, repair mechanisms in mitochondria are not characterized. In the present study, we investigate the mechanisms of DSB repair in mitochondria using in vitro and ex vivo assays. Whereas classical NHEJ (C-NHEJ) is undetectable, microhomology-mediated alternative NHEJ efficiently repairs DSBs in mitochondria. Of interest, robust microhomology-mediated end joining (MMEJ) was observed with DNA substrates bearing 5-, 8-, 10-, 13-, 16-, 19-, and 22-nt microhomology. Furthermore, MMEJ efficiency was enhanced with an increase in the length of homology. Western blotting, immunoprecipitation, and protein inhibition assays suggest the involvement of CtIP, FEN1, MRE11, and PARP1 in mitochondrial MMEJ. Knock-down studies, in conjunction with other experiments, demonstrated that DNA ligase III, but not ligase IV or ligase I, is primarily responsible for the final sealing of DSBs during mitochondrial MMEJ. These observations highlight the central role of MMEJ in maintenance of mammalian mitochondrial genome integrity and is likely relevant for deletions observed in many human mitochondrial disorders.

Report on Marine Mammal Unusual Mortality Event UMESE0501Sp: Multispecies Mass Stranding of Pilot Whales (Globicephala macrorhynchus), Minke Whale (Balaenoptera acutorostrata), and Dwarf Sperm Whales (Kogia sima) in North Carolina on 15-16 January 2005

On 15-16 January 2005, three offshore species of cetaceans (33 short-finned pilot whales, Globicephala macrorhynchus, one minke whale, Balaenoptera acutorostrata, and two dwarf sperm whales, Kogia sima) stranded alive on the beaches of North Carolina. The pilot whales stranded near Oregon Inlet, the minke whale in northern North Carolina, and the dwarf sperm whales near Cape Hatteras. Live strandings of three species in one weekend was unique in North Carolina and qualified as an Unusual Mortality Event. Gross necropsies were conducted on 16-17 January 2005 on 27 pilot whales, two dwarf sperm whales, and the minke whale. Samples were collected for clinical pathology, parasitology, gross pathology, histopathology, microbiology and serology. There was variation in the number of animals sampled for each collection type, however, due to carcasses washing off the beach or degradation in carcass condition during the course of the response. Comprehensive histologic examination was conducted on 16 pilot whales, both dwarf sperm whales, and the minke whale. Limited organ or only head tissue suites were obtained from nine pilot whales. Histologic examination of tissues began in February 2005 and concluded in December 2005 when final sampling was concluded. Neither the pilot whales nor dwarf sperm whales were emaciated although none had recently ingested prey in their stomachs. The minke whale was emaciated; it was likely a dependent calf that became separated from the female. Most serum biochemistry abnormalities appear to have resulted from the stranding and indicated deteriorating condition from being on land for an extended period. Three pilot whales had clinical evidence of pre-existing systemic inflammation, which was supported by histopathologic findings. Although gross and histologic lesions involving all organ systems were noted, consistent lesions were not observed across species. Verminous pterygoid sinusitis and healed fishery interactions were seen in pilot whales but neither of these changes were causes of debilitation or death. In three pilot whales and one dwarf sperm whale there was evidence of clinically significant disease in postcranial tissues which led to chronic debilitation. Cardiovascular disease was present in one pilot whale and one dwarf sperm whale; musculoskeletal disease and intra-abdominal granulomas were present in two pilot whales. These lesions were possible, but not definitive, causal factors in the stranding. Remaining lesions were incidental or post-stranding. The minke whale and three of five tested pilot whales had positive morbillivirus titers (≥1:8 with one at >1:256), but there was no histologic evidence of active viral infection. Parasites (nematodes, cestodes, and trematodes) were collected from 26 pilot whales and two dwarf sperm whales. Sites of collection included stomach, nasal/pterygoid, peribullar sinuses, blubber, and abdominal cavity. Parasite species, locations and loads were within normal limits for free-ranging cetaceans and were not considered causative for the stranding event. Gas emboli lesions which were considered consistent with or diagnostic of sonarassociated strandings of beaked whales or small cetaceans were not found in the whales stranded as part of UMESE0501Sp. Twenty-five heads were examined with nine specific anatomic locations of interest: extramandibular fat, intramandibular fat, auditory meatus, peribullar acoustic fat, peribullar soft tissue, peribullar sinus, pterygoid sinus, melon, and brain. The common finding in all examined heads was verminous pterygoid sinusitis. Intramandibular adipose tissue reddening, typically adjacent to the vascular plexus, was observed in some individuals and could represent localized hemorrhage resulting from vascular rete rupture, hypostatic congestion, or erythrocyte rupture during the freeze/thaw cycle. One cetacean had peracute to acute subdural hemorrhage that likely occurred from thrashing on the beach post-stranding, although its occurrence prior to stranding cannot be excluded. Information provided to NMFS by the U.S. Navy indicated routine tactical mid-frequency sonar operations from individual surface vessels over relatively short durations and small spatial scales within the area and time period investigated. No marine mammals were detected by marine mammal observers on operational vessels; standard operating procedure for surface naval vessels operating mid-frequency sonar is the use of trained visual lookouts using high-powered binoculars. Sound propagation modeling using information provided to NMFS indicated that acoustic conditions in the vicinity likely depended heavily on position of the receivers (e.g., range, bearing, depth) relative to that of the sources. Absent explicit information on the location of animals meant that it was not possible to estimate received acoustic exposures from active sonar transmissions. Nonetheless, the event was associated in time and space with naval activity using mid-frequency active sonar. It also had a number of features in common (e.g., the “atypical” distribution of strandings involving multiple offshore species, all stranding alive, and without evidence of common infectious or other disease process) with other sonar-related cetacean mass stranding events. Given that this event was the only stranding of offshore species to occur within a 2-3 day period in the region on record (i.e., a very rare event), and given the occurrence of the event simultaneously in time and space with a naval exercise using active sonar, the association between the naval sonar activity and the location and timing of the event could be a causal rather than a coincidental relationship. However, evidence supporting a definitive association is lacking, and, in particular, there are differences in operational/environmental characteristics between this event and previous events where sonar has apparently played a role in marine mammal strandings. This does not preclude behavorial avoidance of noise exposure. No harmful algal blooms were present along the Atlantic coast south of the Chesapeake Bay during the months prior to the event. Environmental conditions, including strong winds, changes in upwelling- to downwelling-favorable conditions, and gently sloping bathymetry, were consistent with conditions which have been correlated with other mass strandings. In summary, we did not find commonality in gross and histologic lesions that would indicate a single cause for this stranding event. Three pilot whales and one dwarf sperm whale had debilitating conditions identified that could have contributed to stranding, one pilot whale had a debilitating condition (subdural hemorrhage) that could have been present prior to or resulting from stranding. While the pilot and dwarf sperm whale strandings may have had a common cause, the minke whale stranding was probably just coincidental. On the basis of examination of physical evidence in the affected whales, however, we cannot definitively conclude that there was or was not a causal link between anthropogenic sonar activity or environmental conditions (or a combination of these factors) and the strandings. Overall, the cause of UMESE0501Sp in North Carolina is not and likely will not be definitively known. (PDF contains 240 pages)

A histopathologic evaluation of gross lesions excised from commercially important North Altantic marine fishes

Histopathologic studies of lesions found in commercially important North Atlantic marine fishes are uncommon. As part of a comprehensive Northeast Fisheries Center program ("Ocean Pulse") to evaluate environmental and resource health on the U.S. Continental Shelf from Cape Hatteras to Nova Scotia, grossly visible lesions of the gills, integument, muscle, and viscera of primarily bottom-dwelling fishes were excised and examined using light microscopy. Several gadid and pleuronectid fishes accounted for most of the lesions observed. Most pathological examinations were incidental to samples taken for age and growth determination and evaluation of predator/prey relationships. Several gadids, with either gill, heart, or spleen lesions, were sampled more intensively. Gill lesions principally affected gadids and were caused by either microsporidans or an unidentified oocyte-like cell. The majority of gastrointestinal lesions consisted of encapsulated or encysted larval worms or microsporidan-induced cysts. Few heart lesions were found. Integumental lesioos included ulcers, lymphocystis, and trematode metacercariae. Liver lesions almost always consisted of encapsulated or encysted larval helminths. Necrotic granulomata were seen in muscle and microsporidan-induced granulomata in spleen. Although not numerous, histologically interesting lesions were noted in integument, heart, liver, spleen, and muscle of several fish species. Histologic study of tissues excised from a variety of demersal and pelagic fishes from the eastern North Atlantic (France, Germany, Spain) revealed assorted integumental, renal, hepatic, and splenic lesions. Small sample size and non-random sampling precluded obtaining a meaningful quantitative estimate of the prevalence of the observed lesions in the population at risk; however, a useful census has been made of the types of lesions present in commercially important marine fishes. (PDF file contains 20 pages.)

Prevalência de doença celíaca em pacientes dispépticos sem diarreia atendidos na Disciplina de Gastroenterologia e Endoscopia Digestiva do Hospital Universitário Pedro Ernesto da UERJ

A Doença Celíaca (DC) é uma doença autoimune que afeta o intestino delgado de indivíduos geneticamente susceptíveis após contato com o glúten. Diversos estudos têm relatado aumento da prevalência ao longo dos anos. Objetivo: Determinar a prevalência de DC em pacientes adultos sem diarreia encaminhados à Disciplina de Gastroenterologia do HUPE da UERJ para serem submetidos a Endoscopia Digestiva Alta (EDA).Comparar os resultados do histopatológico das biópsias duodenais com os resultados sorológicos, utilizando o anticorpo antitransglutaminase tecidual IgA (ATGt IgA). Métodos: Pacientes que foram encaminhados ao nosso serviço para serem submetidos a EDA entre Julho de 2008 e Julho de 2010, com idade entre 18 e 85 anos foram aceitos no estudo. Critérios de exclusão foram cirrose, neoplasias do trato gastrointestinal, HIV, uso de imunossupressores e anticoagulantes, diarreia, hemorragia digestiva e DC. Coleta de sangue para pesquisa do anticorpo ATGt IgA (utilizando KIT ORGENTEC - Alemanha), avaliação endoscópica e exame histopatológico das biópsias de segunda porção duodenal foram feitos para cada paciente. Biópsias foram avaliadas de acordo com o critério de Marsh modificado. Resultados: Trezentos e noventa e nove pacientes consecutivos (112 homens, 287 mulheres), média de idade 49,616,4 anos, variando de 18-85 anos, sem diarreia, foram prospectivamente aceitos. Os sintomas clínicos mais prevalentes foram dor abdominal em 99,5%, pirose em 41,1%, plenitude pós prandial em 30,6%, náuseas e vômitos em 21,3%. Os achados endoscópicos foram: normais em 41,6%, lesões pépticas (esofagite, gastrite, duodenite e úlceras) em 41,6%, hérnia hiatal em 5,5%, pólipos gástricos em 3%, neoplasias em 1,3% e miscelânea em 7%. DC foi endoscopicamente diagnosticada em 13 pacientes (3,3%) com mucosa duodenal exibindo serrilhamento das pregas em 8 (2%), diminuição do pregueado em 2 (0,5%) e mucosa exibindo padrão nodular e mosaico em 3 (0,75%). Os achados histopatológicos de duodeno foram normais em 96,7%, duodenites inespecíficas em 2,7% e 3 pacientes (0,75%) confirmaram DC pelos critérios de Marsh modificado (IIIa, IIIb e IIIc). O anticorpo ATGt IgA foi positivo (>10 U/ml) em 1,3% (5/399). Conclusão: Este estudo mostrou que a prevalência de DC em pacientes dispépticos sem diarreia atendidos na Disciplina de Gastroenterologia e Endoscopia do HUPE/UERJ foi de 0,75% (1:133). A acurácia diagnóstica do anticorpo ATGt IgA é boa para pacientes com Marsh III e achados endoscópicos sugestivos. Nenhum dos pacientes tinha alterações Marsh I ou II. A EDA se mostrou um excelente método de triagem para definir os pacientes com graus mais acentuados de atrofia e que se beneficiariam de biópsia e sorologia para confirmação diagnóstica. Os resultados obtidos neste trabalho não justificam uma triagem rotineira de DC.

Hidatidose policística no norte do Brasil: abordagem parasitológica e histopatológica de pacas e casos humanos

A hidatidose policística é uma zoonose causada pelo cestóide Echinococcus vogeli, amplamente distribuído no norte do Brasil. Os hospedeiros definitivos são Speothos venaticus (cachorro-vinagre) e Canis familiaris (cães domésticos), enquanto Agouti paca (paca) é hospedeiro intermediário. Tanto as pacas quanto o homem (hospedeiro acidental) desenvolvem a forma larvar (metacestóide), principalmente na superfície e no interior do fígado. Esta tese tem como objetivo geral estudar as características parasitológicas e histopatológicas de metacestóides de E. vogeli, originários de pacas e humanos da região norte do Brasil, visto o conhecimento insuficiente ou mesmo o seu desconhecimento. Os fígados e mesentérios foram obtidos de oito pacientes com hidatidose policística durante ato cirúrgico na Fundação Hospital Estadual do Acre. Pacas foram capturadas no Município de Bujari, Floresta Estadual do Antimary, Acre. Durante a necropsia das pacas, foram observadas lesões macroscópicas (massas esbranquiçadas ou amareladas, semelhantes a bolhas na superfície dos fígados). Para a análise parasitológica foram aplicadas as microscopias de luz, contraste interferencial de Normaski (DIC) e varredura laser confocal. A análise morfométrica foi realizada com o auxílio do Programa Image Pro Plus Media Cybernetics. Os órgãos de pacas e humanos foram submetidos à análise histopatológica. Os pequenos e grandes ganchos rostelares apresentaram polimorfismo morfológico, enquanto a organização dos protoescólices acompanhou o padrão descrito para Echinococcus sp. Todas as pacas apresentavam cistos hepáticos, porém em apenas duas encontramos líquido hidático, comprovados pela presença dos ganchos e protoescólices. A análise histopatológica dos tecidos hepáticos das pacas confirmou a hidatidose policística e evidenciou, pela presença de agrupamentos de ovos, a coinfecção com Calodium hepaticum. As características morfológicas dos ganchos rostelares dos casos humanos não diferiram do descrito para as pacas, entretanto, os ganchos dos helmintos provenientes do fígado foram maiores que os mesentéricos. Já em relação aos protoescólices, os mesentéricos foram maiores do que os hepáticos. Cistos mesentéricos e hepáticos apresentaram protoescólices em diferentes estágios de desenvolvimento, com coroas de ganchos, formadas por grandes e pequenos ganchos, e dois pares de ventosas, além dos corpúsculos calcários. Os cistos hepáticos apresentaram as três membranas características (adventícia, anista e germinativa), enquanto os cistos mesentéricos não apresentaram a membrana adventícia, sendo a anista aquela que mais se destacou neste órgão. No mesentério, as células mononucleares foram os principais constituintes do infiltrado leucocitário, cuja intensidade foi relacionada à capacidade proliferativa da membrana germinativa. Além de cistos hepáticos típicos de fase crônica, dependendo da resposta inflamatória, foram observados cistos na fase aguda e sub-aguda. Foi encontrado um caso de coinfecção com vírus (HIV, HCB e HCV) e outro com envolvimento da vesícula biliar. Em suma, confirma-se hidatidose policística em pacas no Acre e são apresentados novos casos de infecção humana no Acre e Amazonas. Pela primeira vez, é demonstrado polimorfismo, padrão diferente de desenvolvimento dos cistos de acordo com o órgão, coinfecção e envolvimento da vesícula biliar

Hanseníase neural, aspectos diagnósticos da forma neural pura e mecanismos imunopatogênicos da lesão do nervo na doença. Participação de quimiocinas CCL2 e CXCL10 e metaloproteinases 2 e 9

O diagnóstico da hanseníase neural pura baseia-se em dados clínicos e laboratoriais do paciente, incluindo a histopatologia de espécimes de biópsia de nervo e detecção de DNA de Mycobacterium leprae (M. leprae) pelo PCR. Como o exame histopatológico e a técnica PCR podem não ser suficientes para confirmar o diagnóstico, a imunomarcação de lipoarabinomanana (LAM) e/ou Glicolipídio fenólico 1 (PGL1) - componentes de parede celular de M. leprae foi utilizada na primeira etapa deste estudo, na tentativa de detectar qualquer presença vestigial do M. leprae em amostras de nervo sem bacilos. Além disso, sabe-se que a lesão do nervo na hanseníase pode diretamente ser induzida pelo M. leprae nos estágios iniciais da infecção, no entanto, os mecanismos imunomediados adicionam severidade ao comprometimento da função neural em períodos sintomáticos da doença. Este estudo investigou também a expressão imuno-histoquímica de marcadores envolvidos nos mecanismos de patogenicidade do dano ao nervo na hanseníase. Os imunomarcadores selecionados foram: quimiocinas CXCL10, CCL2, CD3, CD4, CD8, CD45RA, CD45RO, CD68, HLA-DR, e metaloproteinases 2 e 9. O estudo foi desenvolvido em espécimes de biópsias congeladas de nervo coletados de pacientes com HNP (n=23 / 6 BAAR+ e 17 BAAR - PCR +) e pacientes diagnosticados com outras neuropatias (n=5) utilizados como controle. Todas as amostras foram criosseccionadas e submetidas à imunoperoxidase. Os resultados iniciais demonstraram que as 6 amostras de nervos BAAR+ são LAM+/PGL1+. Já entre as 17 amostras de nervos BAAR-, 8 são LAM+ e/ou PGL1+. Nas 17 amostras de nervos BAAR-PCR+, apenas 7 tiveram resultados LAM+ e/ou PGL1+. A detecção de imunorreatividade para LAM e PGL1 nas amostras de nervo do grupo HNP contribuiu para a maior eficiência diagnóstica na ausência recursos a diagnósticos moleculares. Os resultados da segunda parte deste estudo mostraram que foram encontradas imunoreatividade para CXCL10, CCL2, MMP2 e MMP9 nos nervos da hanseníase, mas não em amostras de nervos com outras neuropatias. Além disso, essa imunomarcação foi encontrada predominantemente em células de Schwann e em macrófagos da população celular inflamatória nos nervos HNP. Os outros marcadores de ativação imunológica foram encontrados em leucócitos (linfócitos T e macrófagos) do infiltrado inflamatório encontrados nos nervos. A expressão de todos os marcadores, exceto CXCL10, apresentou associação com a fibrose, no entanto, apenas a CCL2, independentemente dos outros imunomarcadores, estava associada a esse excessivo depósito de matriz extracelular. Nenhuma diferença na frequência da imunomarcação foi detectada entre os subgrupos BAAR+ e BAAR-, exceção feita apenas às células CD68+ e HLA-DR+, que apresentaram discreta diferença entre os grupos BAAR + e BAAR- com granuloma epitelioide. A expressão de MMP9 associada com fibrose é consistente com os resultados anteriores do grupo de pesquisa. Estes resultados indicam que as quimiocinas CCL2 e CXCL10 não são determinantes para o estabelecimento das lesões com ou sem bacilos nos em nervo em estágios avançados da doença, entretanto, a CCL2 está associada com o recrutamento de macrófagos e com o desenvolvimento da fibrose do nervo na lesão neural da hanseníase.

Assessment of contaminant body burdens and histopathology of fish and shellfish species frequently used for subsistence food by Alaskan Native communities

Subsistence food items can be a health concern in rural Alaska because community members often rely on fish and wildlife resources not routinely monitored for persistent bioaccumulative contaminants and pathogens. Subsistence activities are a large part of the traditional culture, as well as a means of providing protein in the diets for Tribal members. In response to the growing concerns among Native communities, contaminant body burden and histopathological condition of chum and sockeye salmon (Oncorhynchus keta and Oncorhynchus nerka) and the shellfish cockles and softshell clams (Clinocardium nuttallii and Mya arenaria) were assessed. In the Spring of 2010, the fish and shellfish were collected from traditional subsistence harvest areas in the vicinity of Nanwalek, Port Graham, and Seldovia, AK, and were analyzed for trace metals and residues of organic contaminants routinely monitored by the NOAA National Status & Trends Program (NS&T). Additionally, the fish and shellfish were histologically characterized for the presence, prevalence and severity of tissue pathology, disease, and parasite infection. The fish and shellfish sampled showed low tissue contamination, and pathologic effects of the parasites and diseases were absent or minimal. Taken together, the results showed that the fish and shellfish were healthy and pose no safety concern for consumption. This study provides reliable chemistry and histopathology information for local resource managers and Alaska Native people regarding subsistence fish and shellfish use and management needs.