Aerobic digestibility of waste aerobic granular sludge (AGS) assessed by respirometry, physical-chemical analyses, modeling and 16S rRNA gene sequencing.

Research has evolved on aerobic granular sludge (AGS) process, but still there are very few studies on the treatment of excess AGS sludge, with almost none considering its aerobic digestion. Here therefore, the aerobic digestibility of typical AGS sludge was assessed. Granules were produced from acetate-based synthetic wastewater (WW) and were subjected to aerobic digestion for 64 d. The stabilization process was monitored over time through physical-chemical parameters, oxygen uptake rates (OUR) and 16S rRNA gene sequencing. The microbial analyses revealed that the cultivated granules were dominated by slow-growing bacteria, mainly ordinary heterotrophic organisms with potential for polyhydroxyalkanoates (PHA) aerobic storage (PHA-OHOs), polyphosphate and glycogen accumulating organisms (PAOs and GAOs), fermentative anaerobes and nitrifiers (AOB and NOB). Differential abundance analysis of the bacterial data (before versus after digestion) discriminated between the most vulnerable microbiome genera and those most resistant to aerobic digestion. Furthermore, modeling of the stabilization process determined that the endogenous decay rate constant (bH) for the heterotrophs present in the granules was notably low; bH = 0.05 d-1 (average), four times less than for common activated sludge (AS), which is rated at 0.2 d-1. For first time, the research reveals another important feature of AGS sludge, i.e. the slow-decaying character of its bacteria (along with their known slow-growing character). This results in slower stabilization, need of bigger digesters and reconsideration of the specific OUR limits in biosolids regulations (SOUR limit of 1.5 mg/gTSS.h), for waste AGS compared to conventional waste AS. The study suggests that aerobic digestion of waste AGS (fully-granulated) could differ from that of conventional AS. Future work is needed on aerobic digestibility of real AGS sludges from municipal and industrial WWs, compared to synthetic WWs.

Ordinary heterotrophic organisms with aerobic storage capacity provide stable aerobic granular sludge for C and N removal.

The study investigated the mechanisms and microbial communities underlying the long-term stability and removal performances shown by aerobic granular sludge (AGS) reactor involving polyhydroxyalkanoates (PHA) aerobic-storing bacteria. The characteristics of the sludge, removal performances and bacterial community structure were determined. The prevailing metabolic phenotype was similar in the parent conventional activated sludge (CAS) reactor and its upgraded AGS version, showing high COD and NH4 uptake, versus low P and N reduction. Polyphosphate and glycogen accumulating organisms, PAO and GAO, were not enriched in the reactors despite initial targeting of anaerobic-aerobic cycle. Instead, PHA-aerobic storing bacteria (Thauera and Paracoccus) were dominant, but revealing a stable AGS system for BOD and N removal. The PAO/GAO failed selection and Thauera overgrowth were analyzed for beneficial use in developing alternative AGS technology for BOD and N removal applications.

Primary Hyperparathyroidism and Pancreatitis: A Rare Association with Multiple Facets.

Primary hyperparathyroidism (PHPT) is rarely associated with the occurrence of acute or chronic pancreatitis. Hypercalcemia plays a major role in the pathogenesis. We report five cases of pancreatitis revealing PHPT. Patients and Methods. This is a retrospective study of 4 years, including all patients admitted to intensive care unit or gastroenterology department, for an acute or chronic pancreatitis revealing primary hyperparathyroidism. Results. We included 5 patients, all female, with mean age 54 years [40-76 years]. The PHPT was in all cases revealed by acute pancreatitis (AP). This one was oedematous in four cases and severe in one case. It occurred twice in calcified chronic pancreatitis (CCP). There was hypercalcemia in all cases. The PHPT was associated with a high rate of parathyroid hormone in 4 cases. The secreting lesion was an adenoma in 5 cases. Two patients had in addition bilateral renal calcifications. The outcome was favorable in 4 patients among whom 3 have had parathyroid surgery. A death was noted by superinfection of necrosis in the case of severe AP. Conclusion. The occurrence of pancreatitis during hyperparathyroidism is rare. Normal or elevated calcemia during acute or chronic pancreatitis should always get attention.

An in silico model to demonstrate the effects of Maspin on cancer cell dynamics.

Most cancer treatments efficacy depends on tumor metastasis suppression, where tumor suppressor genes play an important role. Maspin (Mammary Serine Protease Inhibitor), an non-inhibitory serpin has been reported as a potential tumor suppressor to influence cell migration, adhesion, proliferation and apoptosis in in vitro and in vivo experiments in last two decades. Lack of computational investigations hinders its ability to go through clinical trials. Previously, we reported first computational model for maspin effects on tumor growth using artificial neural network and cellular automata paradigm with in vitro data support. This paper extends the previous in silico model by encompassing how maspin influences cell migration and the cell-extracellular matrix interaction in subcellular level. A feedforward neural network was used to define each cell behavior (proliferation, quiescence, apoptosis) which followed a cell-cycle algorithm to show the microenvironment impacts over tumor growth. Furthermore, the model concentrates how the in silico experiments results can further confirm the fact that maspin reduces cell migration using specific in vitro data verification method. The data collected from in vitro and in silico experiments formulates an unsupervised learning problem which can be solved by using different clustering algorithms. A density based clustering technique was developed to measure the similarity between two datasets based on the number of links between instances. Our proposed clustering algorithm first finds the nearest neighbors of each instance, and then redefines the similarity between pairs of instances in terms of how many nearest neighbors share the two instances. The number of links between two instances is defined as the number of common neighbors they have. The results showed significant resemblances with in vitro experimental data. The results also offer a new insight into the dynamics of maspin and establish as a metastasis suppressor gene for further molecular research.

Intuitive wireless control of a robotic arm for people living with an upper body disability.

Assistive Technologies (ATs) also called extrinsic enablers are useful tools for people living with various disabilities. The key points when designing such useful devices not only concern their intended goal, but also the most suitable human-machine interface (HMI) that should be provided to users. This paper describes the design of a highly intuitive wireless controller for people living with upper body disabilities with a residual or complete control of their neck and their shoulders. Tested with JACO, a six-degree-of-freedom (6-DOF) assistive robotic arm with 3 flexible fingers on its end-effector, the system described in this article is made of low-cost commercial off-the-shelf components and allows a full emulation of JACO's standard controller, a 3 axis joystick with 7 user buttons. To do so, three nine-degree-of-freedom (9-DOF) inertial measurement units (IMUs) are connected to a microcontroller and help measuring the user's head and shoulders position, using a complementary filter approach. The results are then transmitted to a base-station via a 2.4-GHz low-power wireless transceiver and interpreted by the control algorithm running on a PC host. A dedicated software interface allows the user to quickly calibrate the controller, and translates the information into suitable commands for JACO. The proposed controller is thoroughly described, from the electronic design to implemented algorithms and user interfaces. Its performance and future improvements are discussed as well.

A hybrid computational model for the effects of maspin on cancer cell dynamics.

Cancer metastasis is a complex multistep process which allows cancer cells to establish new tumours in distant organs. The process of metastasis involves cell migration and invasion; it is what makes cancer a fatal disease. The efficiency of most cancer treatments depends on metastasis suppression. Maspin is a type II tumour metastasis suppressor which has multiple cellular effects. It has been described as a key regulatory protein in both the intracellular and extracellular environments. Maspin has been shown to reduce cell migration, invasion, proliferation and angiogenesis, and increase apoptosis and cell-cell adhesion in in vitro and in vivo experiments. The clinical data regarding the predictive effects of maspin expression are variable. To date, the whole cellular mechanisms that maspin uses to influence tumour cell behaviours have not been clearly defined. The diversity of the effects of maspin motivated us to develop an intelligent model to investigate its effects on cellular proliferation and migration. This paper reports a hybrid model of solid tumour growth in order to investigate the impact of maspin on the growth and evolutionary dynamics of the cancer cell. A feed-forward neural network was used to model the behaviours (proliferation, quiescence, apoptosis and/or movement) of each cell, which has been suggested as a suitable model of cell signalling pathways. Results show that maspin reduces migration by 10-40%, confirmed by published in vitro data. The model also shows a reduction in cell proliferation by 20-30% in the presence of maspin. So far, this is the first attempt to model the effect of maspin in a computational model to verify in vitro data. This will provide new insights into the tumour suppressive properties of maspin and inform the development of novel cancer therapies.

As(III) and As(V) sorption on iron-modified non-pyrolyzed and pyrolyzed biomass from Petroselinum crispum (parsley).

The sorption of As(III) and As(V) from aqueous solutions onto iron-modified Petroselinum crispum (PCFe) and iron-modified carbonaceous material from the pyrolysis of P. crispum (PCTTFe) was investigated. The modified sorbents were characterized with scanning electron microscopy. The sorbent elemental composition was determined with energy-dispersive X-ray spectroscopy (EDS). The principal functional groups from the sorbents were determined with FT-IR. The specific surfaces and points of zero charge (pzc) of the materials were also determined. As(III) and As(V) sorption onto the modified sorbents were performed in a batch system. After the sorption process, the As content in the liquid and solid phases was determined with atomic absorption and neutron activation analyses, respectively. After the arsenic sorption processes, the desorption of Fe from PCFe and PCTTFe was verified with atomic absorption spectrometry. The morphology of PC changed after iron modification. The specific area and pzc differed significantly between the iron-modified non-pyrolyzed and pyrolyzed P. crispum. The kinetics of the arsenite and arsenate sorption processes were described with a pseudo-second-order model. The Langmuir-Freundlich model provided the isotherms with the best fit. Less than 0.02% of the Fe was desorbed from the PCFe and PCTTFe after the As(III) and As(V) sorption processes.

Adsorption of cadmium by Na and Fe modified zeolitic tuffs and carbonaceous material from pyrolyzed sewage sludge.

In the present study the process of adsorption of cadmium from aqueous solutions using Na-zeolitic tuff, Fe-zeolitic tuff and carbonaceous material from pyrolyzed sewage sludge treated with HCl was investigated. The cadmium removal efficiency was studied as a function of contact time, adsorbate concentration, pH and adsorbent dose. The results showed that removal of cadmium was best described by the Langmuir-Freundlich isotherm. The kinetic experimental results were best described by the pseudo-first order model. The results indicate that the adsorption mechanism is physical and chemical sorption on heterogeneous materials. The maximum retention of cadmium was at pH around 6.0 for the materials. In kinetic studies, the Na and Fe modified zeolitic tuffs showed similar sorption capacities for cadmium and they were higher than the capacity found for carbonaceous material.

Clinical relevance of cagA and vacA gene polymorphisms in Helicobacter pylori isolates from Senegalese patients.

The molecular epidemiology of Helicobacter pylori in Africa is poorly documented. From January 2007 to December 2008, we investigated 187 patients with gastric symptoms in one of the main tertiary hospitals in Dakar, Senegal. One hundred and seventeen patients were culture-positive for H. pylori. Polymorphisms in vacA and cagA status were investigated by PCR; the 3'-region of cagA was sequenced, and EPIYA motifs were identified. Bacterial heterogeneity within individuals was extensively assessed by using an approach based on vacA and cagA heterogeneity. Fourteen per cent of H. pylori-positive patients displayed evidence of mixed infection, which may affect disease outcome. Patients with multiple vacA alleles were excluded from subsequent analyses. Among the final study population of 105 patients, 29 had gastritis only, 61 had ulcerated lesions, and 15 had suspicion of neoplasia based on endoscopic findings. All cases of suspected neoplasia were histologically confirmed as gastric cancer (GC). The cagA gene was present in 73.3% of isolates. CagA proteins contained zero (3.7%), one (93.9%) or two (2.4%) EPIYA-C segments, and all were western CagA. Most of the isolates possessed presumed high-vacuolization isotypes (s1i1m1 (57.1%) or s1i1m2 (21.9%)). Despite the small number of cases, GC was associated with cagA (p 0.03), two EPIYA-C segments in the C-terminal region of CagA (p 0.03), and the s1 vacA allele (p 0.002). Multiple EPIYA-C segments were less frequent than reported in other countries, possibly contributing to the low incidence of GC in Senegal.

[Helicobacter pylori and gastroduodenal lesions in Dakar, Senegal]. / Helicobacter pylori et pathologie gastroduodenale a Dakar (Sénégal).

INTRODUCTION: The bacteriological characteristics of Helicobacter pylori (HP) vary in function of time and place. The aim of this study was to update histological and bacteriological feature of HP infection in patients presenting gastroduodenal lesions in Dakar, Senegal. PATIENTS AND METHOD: This prospective study included patients with gastroduodenal lesions managed over a 6-month period in a digestive endoscopy center in Dakar. In all cases gastric biopsy was performed to obtain specimens for histological diagnosis according Sydney modified classification and HP culture with antibiogram. RESULTS: A total of 158 patients were included. Mean patient age was 48.7 years and the sex-ratio was 1.2. Endoscopic lesions were ulcer in 88 cases, gastritis in 54 cases and tumors in 16. Histological examination demonstrated chronic gastritis in 100% of cases, inflammatory activity in 79.1%, metaplasia in 78.5%, gastric atrophy in 41.1%, adenocarcinoma in 7.6%, dysplasia in 5.7%, and MALT lymphoma in 2.5% with presence of HP in 72.8% of cases. Cultures were positive for HP in 65.8% of cases. Antibiograms indicated that HP was sensitive to amoxicilline in 100% of cases, clarithromycine in 96.6%, ciprofloxacine in 84.1%, and métronidazole in 29.5%. CONCLUSION: Chronic gastritis is a constant feature of gastroduodenal lesions in Dakar. Histology combined with culture showed HP infection in 78.5% of cases. The antibiotic sensitivity of HP in Dakar has changed over the past decade.

Relationship between physical activity measured using accelerometers and energy expenditure measured using doubly labelled water in Indian children.

BACKGROUND/OBJECTIVES: To test the association between physical activity measured using accelerometer counts (Actigraph) and energy expenditure (EE) measured using the doubly labelled water (DLW) method in free-living children in India. The aim of this study was to explore the usefulness of Actigraphs in estimating EE. SUBJECTS/METHODS: Total EE (TEE) was measured in 58 children aged 8-9 years over a period of 2 weeks using the DLW technique. Physical activity level (PAL) was estimated from TEE, and the basal metabolic rate was predicted from weight. Physical activity was measured simultaneously using the Actigraph accelerometers (MTI AM7164 and GT1M). TEE was also calculated from the Actigraph counts using a published equation. RESULTS: TEE (mean: 6.6 vs 5.7 MJ, P=0.04) and Actigraph counts (counts/minute: 557 vs 465, P=0.02; total counts: 445 534 vs 354 748, P=0.004) were higher in boys than in girls. There were no significant correlations between either total Actigraph counts (r=0.15, P=0.3) or counts/minute (r=0.18, P=0.2), and TEE estimated using DLW. Similarly, there were no significant correlations between Actigraph counts and PAL (r=0.10, P=0.5; r=0.17, P=0.2, respectively). The Bland-Altman analysis showed poor agreement between TEE estimated using the DLW method and TEE derived from the Actigraph equation. CONCLUSIONS: Activity measured using Actigraph accelerometers was not related to TEE and PAL derived using the DLW technique in children in Mysore. Actigraphs may not be useful in predicting EE in this setting, but may be better used for judging activity patterns.

Rapid assessment of maternal activity among rural Indian mothers (Pune Maternal Nutrition Study).

OBJECTIVE: To develop an activity questionnaire for objective and rapid assessment of maternal habitual physical activity with the aim of describing its relationship with birth size. DESIGN: Prospective observational study. SETTING: Six villages near Pune, Maharashtra, India. SUBJECTS: Seven hundred and ninety-seven rural Indian mothers were studied after excluding abortions and termination of pregnancies, foetal anomalies, etc. METHOD: Principal components analysis (PCA) was used to identify a few leading questions from an elaborate questionnaire involving 36 questions related to 14 typical maternal activities. RESULTS: On the basis of high loading (>0.8), PCA identified three activities, i.e. farming, fetching water and washing clothes. Questions and sub-questions related to these activities only were considered for defining an activity score for rapid assessment. Validation of this new activity score, with the score based on the elaborate questionnaire, showed 70% sensitivity as well as specificity. New activity scores showed strong inverse relationships similar to those observed using the elaborate questionnaire, at early as well as late gestation, with neonatal head circumference (P=0.001 and 0.055) and mid-arm circumference (P=0.02 and 0.03). CONCLUSIONS: Simple questionnaires, based on leading activities identified by PCA, can be as informative as longer and detailed questionnaires. This method has potential for adaptation, especially in rural communities in developing countries.

Plasma leptin concentration and change in bone density among elderly men and women: the Hertfordshire Cohort Study.

Several studies have shown an association between circulating leptin concentration and bone mineral density. but most studies are cross-sectional in design and report findings in women only. We per-formed a population-based longitudinal study relating baseline plasma leptin concentration to bone mass at the lumbar spine and femoral neck and to change in bone density at these sites over four years in a cohort of 302 men and women aged 60 75 years born and still resident in Hertfordshire, UK. Baseline plasma leptin concentration was strongly positively correlated with body mass index (men: r = 0.71, P 0.0001; women: r = 0.79, P < 0.0001) and with bone mineral content,bone mineral density, and volumetric bone mineral density at all sites (r = 0.24-0.36, P < 0.001) in both sexes: associations with change in bone density were markedly weaker and inconsistent. Adjustment for adult lifestyle determinants of osteoporosis made little difference to our results, but the associations of leptin with bone mass were no longer significant after adjustment for body mass index. These results suggest that the relationship between plasma leptin and bone mass is similar in men and women and that it is mediated through the strong association of both variables with adiposity, rather than through a direct association of leptin on bone cell function.

Growth hormone predicts bone density in elderly women.

Evidence is accumulating that the risk of osteoporosis may be influenced by environmental factors during intrauterine and early postnatal life; such programming might be mediated through modification of the GH/IGF-1 axis during critical periods in its development. To address this issue, we explored the relationships among birth weight, circulating GH profile, bone density, and bone loss rate in a group of British women. The study population consisted of 38 women 60-75 years old resident in Hertfordshire for whom detailed birth records were available. Twenty-four-hour circulating GH profiles were obtained during an inpatient stay on a metabolic ward, after an overnight rest. The circulating profile of GH was characterised by estimating the peak, median, trough, and total concentrations from 72 samples measured sequentially over 24 h in each subject. Bone mineral density was assessed at the lumbar spine and femoral neck at baseline and at follow-up 4 years later. Lumbar spine bone mineral content (BMC) and density (BMD) were positively associated with all measures of GH concentration, although relationships were strongest for BMC with trough GH (r = 0.47, P < 0.01). Associations persisted after adjustment for age, body mass index, smoking, alcohol consumption, physical activity, and osteoarthritis score in multiple regression models. However, associations of GH concentration with femoral neck BMC were weak, and there was no association between any measure of GH concentration and bone loss at either site. Total (integrated) daily GH concentration tended to increase (P = 0.08) with rising birth weight, while IGF-1 concentration fell (P = 0.05) with rising birth weight, suggesting a role for the GH/IGF-1 axis in the programming of adult bone mass among women.

Disrupted mitochondrial electron transport function increases expression of anti-apoptotic bcl-2 and bcl-X(L) proteins in SH-SY5Y neuroblastoma and in Parkinson disease cybrid cells through oxidative stress.

Death of dopamine neurons in Parkinson disease (PD) may arise from consequences of the complex I (C-I) defect in the mitochondrial electron transport chain (ETC). Whether cells activate programmed death (apoptosis) pathways derives, in part, from relative activities of proteins such as bcl-2 and bcl-X(L), that have anti-apoptotic actions. We studied the responses of bcl-2 and bcl-X(L) genes in pharmacologic (acute incubation with methylpyridinium (MPP+)) and mitochondrial transgenic ("cybrid") models of Parkinson disease C-I defects. MPP+ incubation increased levels of bcl-2 and bcl-X(L) proteins in native SH-SY5Y cells but not in rho(0) cells devoid of ETC activity. MPP+ increased bcl-2 mRNA levels by 40% at 8 hr. Confocal microscopic imaging showed that the intracellular distribution of immunoreactive bcl-2 was not significantly associated with mitochondrial membranes at baseline but was associated with mitochondria after 12 hr of MPP+. Immunoreactive bcl-X(L) protein was significantly and equally associated with mitochondrial membranes both at baseline and after MPP+. PD cybrids showed increased basal levels of bcl-2 and bcl-X(L) proteins, similar to the maximum levels found after MPP+ treatment of control SY5Y cells. After MPP+ exposure, bcl-2 protein levels increased in control cybrids but did not increase further in PD cybrids. Both pharmacologically generated and transgenically induced C-I inhibition increases levels of anti-apoptotic bcl proteins, possibly from increased gene transcription. Augmentation of bcl-2 and bcl-X(L) expression may delay neurodegeneration in PD.

Determinants of bone loss in elderly men and women: a prospective population-based study.

While several studies have described the rate and pattern of involutional bone loss in women, far less information is available for men. Furthermore, the roles of lifestyle and body build in determining bone loss rate in both sexes have been largely extrapolated from cross-sectional studies. We addressed this issue in a population-based longitudinal study which sought to ascertain rates of bone loss at the femoral neck and lumbar spine in a cohort of men and women aged 60-75 years at baseline, and to relate this loss to anthropometric and lifestyle variables. We additionally investigated the capacity of biochemical markers of bone turnover to predict bone loss rates in these subjects. Women lost bone at all sites; this ranged from 0.20%/year at the lumbar spine to 1.43%/year at the femoral trochanteric region. By contrast, men lost only 0. 20%/year at the trochanteric region, and gained at the lumbar spine (0.33%/year) and at Ward's triangle (0.27%/year) over the 4-year period. Anthropometric measurements were associated with bone loss in both sexes; lower baseline body mass index (BMI) and a greater rate of loss of adiposity over the follow-up period were both associated with greater bone loss at all proximal femoral sites. These attained statistical significance after Bonferroni correction at the total proximal femur among both men (r = 0.29), p<0.01) and women (r = 0.31, p<0.05). Lifestyle factors associated with lower rates of bone loss (after adjustment for BMI) included alcohol consumption at the femoral neck among women (p = 0.007) and physical activity at the lumbar spine among men (p = 0.05). Serum parathyroid hormone, 25-hydroxyvitamin D and biochemical markers of bone turnover did not predict bone loss after adjustment for adiposity.

Profiles of endogenous circulating cortisol and bone mineral density in healthy elderly men.

Exogenous glucocorticoids are known to increase the risk of osteoporosis. However, the contribution made by endogenous circulating cortisol concentrations to adult skeletal status remains unknown. We examined this issue in a sample of 34 healthy men, aged 61-72 yr. Venous blood samples were obtained under standard conditions every 20 min over a 24-h period. Measurements were made of serum cortisol and cortisol-binding globulin. Bone mineral density was measured at the lumbar spine and proximal femur using dual energy x-ray absorptiometry. Measurements were made at baseline and 4 yr later. There was a weak negative association between integrated cortisol concentration and lumbar spine bone density (r = -0.37; P < 0.05); similar relationships (P < 0.05) existed at three of five proximal femoral sites. There were also statistically significant positive associations between the trough cortisol concentration and bone loss rate at the lumbar spine (r = 0.38; P < 0.05), femoral neck (r = 0.47; P < 0.001), and the trochanteric region (r = 0.41; P = 0.02) over the 4-yr follow-up period. The cross-sectional relationships between cortisol concentration and bone density were removed by adjustment for body mass index, but the influence on bone loss rate remained significant after adjusting for adiposity, cigarette smoking, alcohol consumption, dietary calcium intake, physical activity, and serum testosterone and estradiol levels. These observations suggest that the endogenous cortisol profile of healthy elderly men is a determinant of their bone mineral density and their rate of involutional bone loss.

Visualization of cyclosporin A and Ca2+-sensitive cyclical mitochondrial depolarizations in cell culture.

Mitochondria not only facilitate chemiosmotic energy transduction, but also are excitable organelles that are important participants in intracellular Ca2+ signaling and are obligate participants in the active cell death cascade known as apoptosis. Underlying these functions is the cyclosporin A (CSA)-sensitive mitochondrial permeability transition pore (MTP), which can open transiently in a low conductance mode (MTPL) to relieve excess Ca2+, and irreversibly during the initiation of apoptosis. Here we image for the first time CSA- and Ca2+-sensitive cyclical mitochondrial depolarizations in cultures of the SH-SY5Y human neuroblastoma cell. In addition, we show that mitochondrial transmembrane potential (DeltaPsi) increases in response to CSA, indicating a baseline channel activity. Moreover, networks of mitochondria are shown to behave as an excitable system that may use Ca2+ as a diffusible messenger to recruit neighboring mitochondria to depolarize. We propose that these depolarizations represent MTPL activity. Our data further reinforce the notion that mitochondria are excitable organelles and suggest coordinated activation of MTPL.

Characterization and time course of MPP+ -induced apoptosis in human SH-SY5Y neuroblastoma cells.

A genetic defect in complex I of the mitochondrial electron transport chain (ETC) is implicated in the etiology of Parkinson's disease (PD), and has been studied in hybrid mitochondrial transgene cells based on the SH-SY5Y neuroblastoma. We sought to characterize further the mechanisms and time course of cell death in cultures of human SH-SY5Y neuroblastoma cells exposed to the ETC complex I inhibitor methylpyridinium ion (MPP+). We verify previous reports that apoptosis occurs after MPP+ exposure in SH-SY5Y cells. Nuclear pyknosis, the end stage of apoptosis, is evident after 18-hr exposure to 5 mM MPP+ and reversible until 10 hr, providing a temporal window within which to look for molecular and physiological correlates of MPP+-induced apoptosis. We then looked for mitochondrial correlates of MPP+ induced apoptosis in SH-SY5Y cells. Using flow cytometry, we found that MPP+ -induced increased reactive oxygen species (ROS) and lactate production consistent with inhibition of the ETC. Rho(o) cells, lacking a functional ETC, showed no ROS production, compensatory lactate production or apoptosis after exposure to MPP+. Finally, we show a collapse in ROS production and mitochondrial potential that is temporally correlated with irreversibility of MPP+ -induced apoptosis.