Toxicity of three rare earth elements, and their combinations to algae, microcrustaceans, and fungi.

Rare earth elements (REEs) are naturally distributed in the environment, and are increasingly being used in agriculture and high technology materials worldwide, thereby increasing anthropogenic contamination and environmental risks. There exists scarce and contradictory toxicity information about REEs; hence, more studies are required, especially on their mixtures. Thus, this study aimed to assess the toxicities of La3+, Nd3+, Sm3+, and the combinations of these elements (binary 1:1 and ternary 1:1:1), to organisms from different trophic levels: producers (the microalgae Chlorella vulgaris and Raphidocelis subcapitata), primary consumers (the microcrustaceans Daphnia similis and Artemia salina), and decomposers (the fungi Penicillium simplicissimum and Aspergillus japonicus). Ecotoxicological bioassays were performed, and toxic concentrations were determined. Thereafter, toxicities of single and mixture REEs were classified as slightly to highly toxic according to their toxic units. Finally, a concentration addition (CA) model was used to estimate how REEs interact upon combining. Nd3+ was the most toxic element for all organisms, especially D. similis (48 h LC50 9.41 mg.L-1), and was therefore classified as highly toxic. Sm3+ promoted cell agglomeration in Chlorella vulgaris and was the most toxic of the tested elements for this organism (72 h IC50 25.78 mg.L-1). The CA model revealed synergistic responses for most of the combinations, principally Nd3+ + Sm3+, which was the most toxic combination for the tested organisms. Both fungi were the most resistant organisms, and A. japonicus produced exudate and sclerotia, which help in the detoxification of chemicals. Owing not only to the fact that fungi displayed a higher resistance to REEs, but also due to the absence of regulations for REEs released from the agricultural or industrial sector, and the lack of methods to treat effluents or to dispose of technological items containing REEs, these organisms should be considered as a model for the biosorption or bioremediation of REEs. Finally, the toxic effects of REEs, particularly Nd3+, on the biota and human health should be the focus of future studies due to their increased use in technology.

Biological stenciling of mineralization in the skeleton: Local enzymatic removal of inhibitors in the extracellular matrix.

Biomineralization is remarkably diverse and provides myriad functions across many organismal systems. Biomineralization processes typically produce hardened, hierarchically organized structures usually having nanostructured mineral assemblies that are formed through inorganic-organic (usually protein) interactions. Calcium­carbonate biomineral predominates in structures of small invertebrate organisms abundant in marine environments, particularly in shells (remarkably it is also found in the inner ear otoconia of vertebrates), whereas calcium-phosphate biomineral predominates in the skeletons and dentitions of both marine and terrestrial vertebrates, including humans. Reconciliation of the interplay between organic moieties and inorganic crystals in bones and teeth is a cornerstone of biomineralization research. Key molecular determinants of skeletal and dental mineralization have been identified in health and disease, and in pathologic ectopic calcification, ranging from small molecules such as pyrophosphate, to small membrane-bounded matrix vesicles shed from cells, and to noncollagenous extracellular matrix proteins such as osteopontin and their derived bioactive peptides. Beyond partly knowing the regulatory role of the direct actions of inhibitors on vertebrate mineralization, more recently the importance of their enzymatic removal from the extracellular matrix has become increasingly understood. Great progress has been made in deciphering the relationship between mineralization inhibitors and the enzymes that degrade them, and how adverse changes in this physiologic pathway (such as gene mutations causing disease) result in mineralization defects. Two examples of this are rare skeletal diseases having osteomalacia/odontomalacia (soft bones and teeth) - namely hypophosphatasia (HPP) and X-linked hypophosphatemia (XLH) - where inactivating mutations occur in the gene for the enzymes tissue-nonspecific alkaline phosphatase (TNAP, TNSALP, ALPL) and phosphate-regulating endopeptidase homolog X-linked (PHEX), respectively. Here, we review and provide a concept for how existing and new information now comes together to describe the dual nature of regulation of mineralization - through systemic mineral ion homeostasis involving circulating factors, coupled with molecular determinants operating at the local level in the extracellular matrix. For the local mineralization events in the extracellular matrix, we present a focused concept in skeletal mineralization biology called the Stenciling Principle - a principle (building upon seminal work by Neuman and Fleisch) describing how the action of enzymes to remove tissue-resident inhibitors defines with precision the location and progression of mineralization.

Do Riparian Buffers Protect Stream Invertebrate Communities in South American Atlantic Forest Agricultural Areas?

We investigated the influence and relative importance of insecticides and other agricultural stressors in determining variability in invertebrate communities in small streams in intensive soy-production regions of Brazil and Paraguay. In Paraguay we sampled 17 sites on tributaries of the Pirapó River in the state of Itapúa and in Brazil we sampled 18 sites on tributaries of the San Francisco River in the state of Paraná. The riparian buffer zones generally contained native Atlantic forest remnants and/or introduced tree species at various stages of growth. In Brazil the stream buffer width was negatively correlated with sediment insecticide concentrations and buffer width was found to have moderate importance in mitigating effects on some sensitive taxa such as mayflies. However, in both regions insecticides had low relative importance in explaining variability in invertebrate communities, while various habitat parameters were more important. In Brazil, the percent coverage of soft depositional sediment in streams was the most important agriculture-related explanatory variable, and the overall stream-habitat score was the most important variable in Paraguay streams. Paraguay and Brazil both have laws requiring forested riparian buffers. The ample forested riparian buffer zones typical of streams in these regions are likely to have mitigated the effects of pesticides on stream invertebrate communities. This study provides evidence that riparian buffer regulations in the Atlantic Forest region are protecting stream ecosystems from pesticides and other agricultural stressors. Further studies are needed to determine the minimum buffer widths necessary to achieve optimal protection.

Fermentation for Disinfesting Fruit Waste From Drosophila Species (Diptera: Drosophilidae).

Economic losses in a range of fruit crops due to the Drosophila suzukii (Matsumura) have become severe. Removal and treatment of fruit waste, which may harbor D. suzukii, is a key step in preventing reinfestation of fruit production. Natural fermentation for disinfesting fruit wastes from D. suzukii was examined at ambient air temperatures of 12-20 °C. Soft and stone fruit wastes infested with eggs, larvae, and pupae of Drosophila melanogaster (Meigen) or D. suzukii were placed in sealed vessels containing fruit wastes, and samples were retrieved at intervals and tested for the emergence of adults. Mean temperatures of the fruit waste in the sealed vessels during fermentation were 15-23 °C. Fermentation for 3 d was effective in disinfesting waste from different life stages of D. suzukii. Treatment for 4 d also ensured that the waste was free of viable life stages of D. melanogaster, which could be used as an indicator species for disinfestation of waste from D. suzukii owing to its greater tolerance of fermentation. The O2 concentration of the headspace air in the vessels became undetectable after 13-16 h, with a corresponding increase in CO2 concentration, which exceeded 80% vol/vol. The resulting hypoxia and hypercapnia may explain the efficacy of the fermentation treatment in disinfesting the waste. Fermented fruit remained attractive to D. suzukii and retained its capacity to rear a life cycle. Covering or mixing fermented fruit with a sufficient depth (0.1 m) or volume (×9) of soil or coir prevented the reinfestation of treated waste.

Development of a benthic multimetric index for the Serra da Bocaina bioregion in Southeast Brazil.

Brazil faces a challenge to develop biomonitoring tools to be used in water quality assessment programs, but few multimetric indices were developed so far. This study is part of an effort to test and implement programs using benthic macroinvertebrates as bioindicators in Rio de Janeiro State. Our aim was first to test the Multimetric Index for Serra dos Órgãos (SOMI) for a different area--Serra da Bocaina (SB)--in the same ecoregion. We sampled 27 streams of different sizes and altitudes in the SB region. Despite the environmental similarities, results indicated biological differences between reference sites of the two regions. Considering these differences, we decided to develop an index specific for the SB region, the Serra da Bocaina Multimetric Index (MISB). We tested twenty-two metrics for sensitivity to impairment and redundancy, and six metrics were considered valid to integrate the MISB: Family Richness, Trichoptera Richness, % Coleoptera, % Diptera, IBE-IOC index, EPT / Chironomidae ratio. A test of the MISB in eleven sites indicated it was more related to land-use and water physico-chemical parameters than with altitude or stream width, being a useful tool for the monitoring and assessment of streams in the bioregion.

A three-year follow-up on the efficacy of psychosocial interventions for patients with mild dementia and their caregivers: the multicentre, rater-blinded, randomised Danish Alzheimer Intervention Study (DAISY).

OBJECTIVES: To examine the long-term efficacy at the 36-month follow-up of an early psychosocial counselling and support programme lasting 8-12 months for community-dwelling patients with mild Alzheimer's disease and their caregivers. DESIGN: Multicentre, randomised, controlled, rater-blinded trial. SETTING: Primary care and memory clinics in five Danish districts. PARTICIPANTS: 330 home-dwelling patients with mild Alzheimer's disease and their primary caregivers (dyads). INTERVENTIONS: Dyads were randomised to receive intervention during the first year after diagnosis. Both intervention and control groups had follow-up visits at 3, 6, 12 and 36 months. MAIN OUTCOME MEASURES: Primary outcomes for the patients assessed at 36-month follow-up were changes from baseline in global cognitive function (Mini-Mental State Examination), depressive symptoms (Cornell Depression Scale) and proxy-rated EuroQoL quality of life on visual analogue scale. The primary outcomes for the caregivers were changes from baseline in depressive symptoms (Geriatric Depression Scale) and self-rated EuroQoL quality of life on a visual analogue scale. The secondary outcome measures for the patient were proxy-rated Quality of Life Scale for Alzheimer's disease (QoL-AD), Neuropsychiatric Inventory-Questionnaire, Alzheimer's disease Cooperative Study Activities of Daily Living Scale, all-cause mortality and nursing home placement. RESULTS: At a 36-month follow-up, 2 years after the completion of the Danish Alzheimer Intervention Study (DAISY), the unadjusted positive effects previously detected at the 12-month follow-up in one patient primary outcome (Cornell depression score) and one patient secondary outcome (proxy-rated QoL-AD) disappeared (Cornell depression score, p=0.93; proxy-rated QoL-AD, p=0.81). No long-term effect of DAISY intervention on any other primary and secondary outcomes was found at the 36-month follow-up. CONCLUSIONS: For patients with very mild Alzheimer's disease and their caregivers, an intensive, multi-component, semitailored psychosocial intervention programme with counselling, education and support during the first year after diagnosis did not show any positive long-term effect on primary and secondary outcomes. TRIAL REGISTRATION: The study was registered in the Clinical Trial Database (http://www.controlled-trials.com/ISRCTN74848736).

The influence of connectivity in forest patches, and riparian vegetation width on stream macroinvertebrate fauna.

We assessed two dimensions of stream connectivity: longitudinal (between forest patches along the stream) and lateral (riparian vegetation), using macroinvertebrate assemblages as bioindicators. Sites representing different land-uses were sampled in a lowland basin that holds a mosaic of protected areas. Land-use analysis, forest successional stages and riparian zone widths were calculated by the GIS analysis. Macroinvertebrate fauna was strongly affected by land-use. We observed a continuous decrease in the number of sensitive species, %Shredders and IBE-IOC biotic index from the upstream protected area to highly deforested sites, increasing again where the stream crosses a Biological Reserve. When analysing buffer strips, we found aquatic fauna responding to land-use alterations beyond the 30 m riparian corridor (60 m and 100 m wide). We discussed the longitudinal connectivity between forest patches and the riparian vegetation buffer strips necessary to hold high macroinvertebrate diversity. We recommend actions for the increase/maintenance of biodiversity in this and other lowland basins.

Influence of agricultural land-use and pesticides on benthic macroinvertebrate assemblages in an agricultural river basin in southeast Brazil.

Land-use alterations and pesticide run-offs are among the main causes for impairment in agricultural areas. We evaluated the influence of different land-uses (forest, pasture and intensive agriculture) on the water quality and on benthic macroinvertebrate assemblages on three occasions: in the dry season, wet season and at the end of the wet season. Macroinvertebrates responded to this gradient of impairment: agricultural sites had significantly lower richness numbers than forested and pasture sites, and all major invertebrate groups were significantly affected. Most taxa found in forested sites were found in pasture sites, but often with lower densities. In this case, the loss of habitats due to sedimentation and the lower complexity of substrates seem to be the disruptive force for the macroinvertebrate fauna.

Efficacy of psychosocial intervention in patients with mild Alzheimer's disease: the multicentre, rater blinded, randomised Danish Alzheimer Intervention Study (DAISY).

OBJECTIVE: To assess the efficacy at 12 months of an early psychosocial counselling and support programme for outpatients with mild Alzheimer's disease and their primary care givers. DESIGN: Multicentre, randomised, controlled, rater blinded trial. SETTING: Primary care and memory clinics in five Danish districts. PARTICIPANTS: 330 outpatients with mild Alzheimer's disease and their 330 primary care givers. INTERVENTIONS: Participating dyads (patient and primary care giver) were randomised to control support during follow-up or to control support plus DAISY intervention (multifaceted and semi-tailored counselling, education, and support). MAIN OUTCOME MEASURES: Primary outcomes at 12 months for patients were change from baseline in mini mental state examination (MMSE) score, Cornell depression scale score, and proxy rated European quality of life visual analogue scale (EQ-VAS) score. For care givers, outcomes were change from baseline in geriatric depression scale (GDS 30 items) score and EQ-VAS score. RESULTS: Because of multiple testing, statistical significance was set at an adjusted P limit of <0.0005. At 12 months there were no significant differences between the two allocation groups in changes from baseline in the primary and secondary outcomes. However, although non-significant with the adjusted P limit, a small difference was observed for one of the primary patient outcomes (Cornell depression scale score) in patients in favour of the DAISY intervention group before and after adjusting for attrition (P = 0.0146 and P = 0.0103 respectively). CONCLUSIONS: The multifaceted, semi-tailored intervention with counselling, education, and support for patients with mild Alzheimer's disease and their care givers did not have any significant effect beyond that with well structured follow-up support at 12 months after adjustment for multiple comparisons. The small positive effect found in the unadjusted primary outcome addressing depressive symptoms in patients may call for further research focusing on patients with Alzheimer's disease and comorbid depression. TRIAL REGISTRATION: ISRCTN74848736.

The Danish Alzheimer intervention study: rationale, study design and baseline characteristics of the cohort.

BACKGROUND: There is a lack of appropriately designed trials investigating the efficacy of psychosocial interventions for patients with mild dementia and their family caregivers. This paper reports the rationale and design of the Danish Alzheimer Disease Intervention Study and baseline characteristics of the cohort. METHODS: The study was a 1-year multicentre randomized controlled rater-blinded trial with randomization to follow-up and a multifaceted semitailored intervention programme or to follow-up only (with extension of follow-up to 3 years). The intervention included a counselling programme, teaching courses, written information and logbooks. The outcomes included clinical efficacy parameters, patient satisfaction and health economic consequences. RESULTS: A total of 330 patients and their 330 caregivers were included during a period of 18 months. The majority (65.2 %) of the caregivers were spouses. At inclusion the mean age of the patients and caregivers was 76.2 and 66.0 years, respectively. CONCLUSION: The study will explore the added value of a multifaceted intervention programme and contribute to the design of future interventions for patients with mild dementia and their caregivers.

Impact of anti-cancer drugs and other determinants on serum protein binding of morphine 6-glucuronide.

BACKGROUND AND THE PURPOSE OF THE STUDY: The aim of the present study was to examine factors that may influence the protein binding of morphine 6-glucuronide (M6G), the most active metabolite of morphine. METHODS: An enzyme-linked immunoabsorbent assay technique was used to measure the M6G concentration in serum of 18 healthy adults, 18 neonatal and 7 children with cancer. Total and free M6G concentrations were measured following equilibrium dialysis for 3 hrs and at physiological pH at 37°C. The influence of vincristine, methotrexate, 6-mercaptopurine, morphine, human albumin, alpha-1-acid glycoprotein, palmitic acid, oleic acid and pH on M6G protein binding was examined. RESULTS: M6G was 66.87±0.73 percent free in human serum at physiological pH and temperature. The percentage free (unbound) was increased significantly by vincristine (4.33%) and methotrexate (9.68%), but 6- mercaptopurine and morphine had no significant effect on it. Free percentages of M6G was reduced by decreasing serum albumin concentration but was unaffected by the presence of alpa-1-acid glycoprotein (AAG) or changes in serum pH. Similar results were obtained in human serum albumin (HAS) solutions. Addition of palmitic acid and oleic acid reduced protein binding significantly by 6.3% and 7.4%, respectively. MAJOR CONCLUSION: Although M6G in this study was not highly bounded, but because of its high analgesic potency, any change in its free concentration due to concurrent medication or disease caused significant changes in its effects. This dearth of evidence has been implicated in the reluctance of professionals to be cautious in prescribing them to children, particularly in the neonatal period.

[Development of a glaucoma microstent with drainage into the suprachoroidal space: fluid mechanical model approach]. / Entwicklung eines Glaukommikrostents mit Drainage in den suprachoroidalen Raum: Fluidmechanische Modellbetrachtungen.

BACKGROUND: The purpose of this study was to develop a microstent with valve function, which normalizes the intraocular pressure (IOP) and drains into the suprachoroidal space. In comparison to the subconjunctival space the suprachoroidal space is attributed with less fibroblast colonization and activity. METHODS: Different glaucoma drainage devices were idealized as tubes and the flow rates were calculated according to Hagen-Poiseuille. The dimensions of the ideal glaucoma implant were modified with respect to an aqueous humor production of 2 microl/min and the different outflow pathways. Specific components of glaucoma drainage devices at the inlet and outlet were not included. RESULTS: The volume flow calculation of the tested glaucoma implants showed that the dimensions of all lumina were too large to prevent postoperative hypotension. A maximum inner tube diameter of 53 microm was calculated for drainage into the suprachoroidal space based on an intra-ocular pressure (IOP) of 20 mmHg. CONCLUSION: The glaucoma microstent has to guarantee an aqueous humor flow for physiological IOP. An increase of IOP has to be regulated to physiological pressure conditions by the microvalve.

Spatial and temporal distribution of benthic macroinvertebrates in a Southeastern Brazilian river.

Benthic macroinvertebrate assemblages are structured according to physical and chemical parameters that define microhabitats, including food supply, shelter to escape predators, and other biological parameters that influence reproductive success. The aim of this study is to investigate spatial and temporal distribution of macroinvertebrate assemblages at the Macaé river basin, in Rio de Janeiro state, Southeastern Brazil. According to the "Habitat Assessment Field Data Sheet--High Gradient Streams" (Barbour et al., 1999), the five sampling sites are considered as a reference condition. Despite the differences in hydrological parameters (mean width, depth and discharge) among sites, the physicochemical parameters and functional feeding groups' general structure were similar, except for the less impacted area, which showed more shredders. According to the Detrended Correspondence Analysis based on substrates, there is a clear distinction between pool and riffle assemblages. In fact, the riffle litter substrate had higher taxa in terms of richness and abundance, but the pool litter substrate had the greatest number of exclusive taxa. A Cluster Analysis based on sampling sites data showed that temporal variation was the main factor in structuring macroinvertebrate assemblages in the studied habitats.

Localization of methyl benzoate synthesis and emission in Stephanotis floribunda and Nicotiana suaveolens flowers.

The emission of fragrances can qualitatively and quantitatively differ in different parts of flowers. A detailed analysis was initiated to localize the floral tissues and cells which contribute to scent synthesis in STEPHANOTIS FLORIBUNDA (Asclepiadaceae) and NICOTIANA SUAVEOLENS (Solanaceae). The emission of scent compounds in these species is primarily found in the lobes of the corollas and little/no emission can be attributed to other floral organs or tissues. The rim and centre of the petal lobes of S. FLORIBUNDA contribute equally to scent production since the amount of SAMT (salicylic acid carboxyl methyltransferase) and specific SAMT activity compensate each other in the rim region and centre region. IN SITU immunolocalizations with antibodies against the methyl benzoate and methyl salicylate-synthesizing enzyme indicate that the adaxial epidermis with few subepidermal cell layers of S. FLORIBUNDA is the site of SAMT accumulation. In N. SUAVEOLENS flowers, the petal rim emits twice as much methyl benzoate due to higher total protein concentrations in the rim versus the petal centre; and, both the adaxial and abaxial epidermis house the BSMT (salicylic acid/benzoic acid carboxyl methyltransferase).

Spatial and temporal distribution of benthic macroinvertebrates in a Southeastern Brazilian river

A fauna de macroinvertebrados bentônicos é estruturada por fatores físicos e químicos que determinam os microhábitats, incluindo a disponibilidade de alimento, a existência de refúgios contra predadores e tempestades, o sucesso reprodutivo e outros parâmetros biológicos. O objetivo deste estudo foi investigar a distribuição espaço-temporal da comunidade de macroinvertebrados bentônicos na bacia do rio Macaé, Estado do Rio de Janeiro, Brasil. De acordo com o Habitat Assessment Field Data Sheet – High Gradient Streams (Barbour et al., 1999), os cinco locais coletados são considerados como referência para propósito de biomonitoramento. Apesar das diferenças em parâmetros hidrográficos (largura, profundidade e vazão médias), os parâmetros físico-químicos e a composição dos grupos de alimentação funcional foram similares entre os locais, com exceção da área mais preservada, onde a porcentagem de organismos cortadores foi maior. De acordo com a Análise de Correspondência, há uma clara distinção entre áreas de remanso e de correnteza. De fato, o substrato folhiço de correnteza apresentou a fauna mais rica e abundante, enquanto o substrato folhiço de fundo apresentou o maior número de táxons exclusivos. A Análise de Agrupamento considerando os dados dos locais de coleta indicou que a variação temporal foi o fator mais importante na estruturação das comunidades nos hábitats estudados.

Spatial and temporal distribution of benthic macroinvertebrates in a Southeastern Brazilian river

Benthic macroinvertebrate assemblages are structured according to physical and chemical parameters that define microhabitats, including food supply, shelter to escape predators, and other biological parameters that influence reproductive success. The aim of this study is to investigate spatial and temporal distribution of macroinvertebrate assemblages at the Macaé river basin, in Rio de Janeiro state, Southeastern Brazil. According to the "Habitat Assessment Field Data Sheet - High Gradient Streams" (Barbour et al., 1999), the five sampling sites are considered as a reference condition. Despite the differences in hydrological parameters (mean width, depth and discharge) among sites, the physicochemical parameters and functional feeding groups' general structure were similar, except for the less impacted area, which showed more shredders. According to the Detrended Correspondence Analysis based on substrates, there is a clear distinction between pool and riffle assemblages. In fact, the riffle litter substrate had higher taxa in terms of richness and abundance, but the pool litter substrate had the greatest number of exclusive taxa. A Cluster Analysis based on sampling sites data showed that temporal variation was the main factor in structuring macroinvertebrate assemblages in the studied habitats.

A fauna de macroinvertebrados bentônicos é estruturada por fatores físicos e químicos que determinam os microhábitats, incluindo a disponibilidade de alimento, a existência de refúgios contra predadores e tempestades, o sucesso reprodutivo e outros parâmetros biológicos. O objetivo deste estudo foi investigar a distribuição espaço-temporal da comunidade de macroinvertebrados bentônicos na bacia do rio Macaé, Estado do Rio de Janeiro, Brasil. De acordo com o Habitat Assessment Field Data Sheet - High Gradient Streams (Barbour et al., 1999), os cinco locais coletados são considerados como referência para propósito de biomonitoramento. Apesar das diferenças em parâmetros hidrográficos (largura, profundidade e vazão médias), os parâmetros físico-químicos e a composição dos grupos de alimentação funcional foram similares entre os locais, com exceção da área mais preservada, onde a porcentagem de organismos cortadores foi maior. De acordo com a Análise de Correspondência, há uma clara distinção entre áreas de remanso e de correnteza. De fato, o substrato folhiço de correnteza apresentou a fauna mais rica e abundante, enquanto o substrato folhiço de fundo apresentou o maior número de táxons exclusivos. A Análise de Agrupamento considerando os dados dos locais de coleta indicou que a variação temporal foi o fator mais importante na estruturação das comunidades nos hábitats estudados.

Application of biological measures for stream integrity assessment in south-east Brazil.

The sensitivity of eleven metrics using macroinvertebrate assemblages were evaluated in an environmental gradient in a tropical river in south-east Brazil. Eight sites were sampled in an altitudinal range of 160-650 m.a.s.l. during 1999 (April and August) and 2000 (February). Four substrates were sampled at each site: riffle litter, pool litter, stony substrates and sediment. Correspondence Analysis indicated that assemblages were primarily more influenced by physical changes (like deforestation and erosion processes) than the water chemistry. The sensitivity of each metric was evaluated through the application of box-and-whisker plot method by its power to assess impairment (metrics should be able to discriminate reference sites from impaired sites) and natural variability (metrics should not discriminate two reference sites). Metrics that failed in at least one of the above premises were not considered as sensitive. In this study, the most sensitive metrics were Shannon index, BMWP-ASPT, %_EPT, and relative abundance of EPT to Chironomidae.

Nicotine enemas for treatment of ulcerative colitis: a study of the pharmacokinetics and adverse events associated with three doses of nicotine.

BACKGROUND: Transdermal nicotine is of value in active ulcerative colitis but causes adverse events because of systemic absorption. Nicotine enemas may give rise to fewer adverse events. AIM: To assess the pharmacokinetics of nicotine enemas in three doses. METHODS: Thirteen volunteers, all non-smokers but three ex-smokers, were given enemas on separate occasions containing 3, 6 and 9 mg of nicotine, in ascending dose order. Adverse events were recorded and blood samples taken over 8 h for measurement of serum nicotine and cotinine. RESULTS: Enemas were retained by most subjects. Eleven of 14 adverse events were 'early'--30-105 min after the enema, corresponding to maximum plasma nicotine concentrations; three events were later, 4-8 h after the enema and unrelated to the tmax. 'Early' adverse events occurred in eight subjects--six with 9 mg. The three highest plasma nicotine concentrations were with 9 mg and associated with headache, nausea and sweating. Only one had adverse events with 3 mg and withdrew from the study. Nicotine Cmax with 6 and 9 mg doses were respectively two and three times the value with 3 mg. Peak nicotine concentrations occurred 44-50 min after the enema. CONCLUSION: The 6 mg dose of nicotine probably represents the dose to use in clinical practice - for the highest therapeutic dose with a low risk of adverse events.

Molecular comparisons of the Culex pipiens (L.) complex esterase gene amplicons.

The amplification of carboxylesterase genes is a mechanism of organophosphate resistance in Culex mosquitoes. Amplified carboxylesterase genes from an insecticide resistant Culex pipiens strain collected in Cyprus were analysed and compared to other Culex amplified carboxylesterase alleles. A 12 kb section of genomic DNA containing two gene loci coding for carboxylesterase alleles A5 and B5 was cloned and sequenced. A comparison between this amplicon and one from a strain with co-amplified carboxylesterase alleles A2 and B2 revealed a number of differences. The intergenic spacer was 3.7 kb in length in the A5-B5 amplicon (2.7 kb in A2-B2) and contained putative Juan and transposable elements upstream of B5. A fragment of a gene with high homology to aldehyde oxidase was also present immediately downstream of A5. The comparison revealed no differences that would explain the successful spread of the A2-B2 amplicon worldwide whilst the A5-B5 amplicon is restricted to the Mediterranean.

Fatal EBV-related post-transplant lymphoproliferative disorder (LPD) after matched related donor nonmyeloablative peripheral blood progenitor cell transplant.

A 39-year-old male underwent a nonmyeloablative stem cell transplant (NMAPBPCT) from his HLA-matched sister for recurrent anaplastic large cell lymphoma in CR-2, receiving fludarabine, cyclophosphamide, and rabbit antithymocyte globulin for the preparative therapy. The patient was readmitted on day+33 for persistent culture-negative fevers. He rapidly developed marked elevations of alkaline phosphatase and bilirubin. Liver biopsy showed a periportal infiltrate of large immunoblastic appearing cells. The tumor cells did not stain for CD3/CD20/CD30 and alk protein, but did stain for CD79a/LCA and CD43. In situ hybridization for Epstein-Barr virus (EBV) RNA (EBER 1) was strongly positive in the periportal infiltrating lymphocytes. Fluorescence in situ hybridization (FISH) studies revealed female (XX) cells in the tumor cells and male (XY) in the surrounding hepatic parenchymal cells. The patient developed severe lactic acidosis, oliguric renal failure and expired on day+44. Both donor and patient had positive IgG serologies for EBV VCA and EBNA pretransplant. The donor also had a positive IgM titer for EBV VCA in the pretransplant specimen. The LPD may have been related to the intense immunosuppression of the preparative therapy and the presence of recent EBV infection in the donor.