Microanatomical Changes in the Leaves of Arundo donax (L.) Caused by Potentially Toxic Elements from Municipal Sewage Sediment.

An open-field 3-year-long microplot experiment was set up with three micropropagated lines (SC Blossom, BFT Indiana, and STM Hajdúsági) of giant reed (Arundo donax L.). Plants were grown on a soil cover of a former sewage settling pond located in Debrecen Lovász-Zug, Hungary. Soil cover of the sewage sediment was moderately contaminated with various toxic elements (As, Ba, Cd, Cr, Cu, Mn, Ni, Pb, and Zn). The highest total concentration of examined toxic elements in leaves was found in the BFT Indiana line (∑326 mg/kg), while in the SC Blossom and STM Hajdúsági lines, ∑210 mg/kg and ∑182 mg/kg were measured, respectively. The highest Zn concentration (117 mg/kg) was found in the leaves of in BFT Indiana line and was 67% higher than that in SC Blossom and 95% more than in the STM Hajdúsági line. The BFT Indiana leaves showed typical signs of adaptation to heavy metal stress in the case of numerous micromorphometric characteristics. The extent of leaf mesophylls decreased, and the number of bulliform cells and phytoliths, as well as the sclerenchymatous stock, increased. The size of the vascular bundles was reduced. The size of the stomata decreased while the stomatal density increased. It can be concluded that the BFT Indiana line had the best adaptational response to heavy metal stress.

Herbivorous Juvenile Grass Carp (Ctenopharyngodon idella) Fed with Genetically Modified MON 810 and DAS-59122 Maize Varieties Containing Cry Toxins: Intestinal Histological, Developmental, and Immunological Investigations.

Feeding experiments with juvenile grass carp (Ctenopharyngodon idella) fed with genetically modified maize MON 810 or DAS-59122 dried leaf biomass were carried out with 1-, 3- and 6-month exposures. Dosages of 3-7 µg/fish/day Cry1Ab or 18-55 µg/fish/day Cry34Ab1 toxin did not cause mortality. No difference occurred in body or abdominal sac weights. No differences appeared in levels of inorganic phosphate, calcium, fructosamine, bile acids, triglycerides, cholesterol, and alanine and aspartame aminotransferases. DAS-59122 did not alter blood parameters tested after 3 months of feeding. MON 810 slightly decreased serum albumin levels compared to the control, only in one group. Tapeworm (Bothriocephalus acheilognathi) infection changed the levels of inorganic phosphate and calcium. Cry34Ab1 toxin appeared in blood (12.6 ± 1.9 ng/mL), but not in the muscle. It was detected in B. acheilognathi. Cry1Ab was hardly detectable in certain samples near the limit of detection. Degradation of Cry toxins was extremely quick in the fish gastrointestinal tract. After 6 months of feeding, only mild indications in certain serum parameters were observed: MON 810 slightly increased the level of apoptotic cells in the blood and reduced the number of thrombocytes in one group; DAS-59122 mildly increased the number of granulocytes compared to the near-isogenic line.

Spiegelmer-Based Sandwich Assay for Cardiac Troponin I Detection.

Two subunits of the ternary troponin complex, I and C, have cardiac muscle specific isoforms, and hence could be applied as highly-selective markers of acute coronary syndrome. We aimed at paving the way for the development of a robust cardiac troponin I-detecting sandwich assay by replacing antibodies with nuclease resistant aptamer analogues, so-called spiegelmers. To complement the previously generated spiegelmers that were specific for the N-terminus of cTnI, spiegelmers were selected for an amino acid stretch in the proximity of the C-terminal part of the protein by using a D-amino acid composed peptide. Following the selection, the oligonucleotides were screened by filter binding assay, and surface plasmon resonance analysis of the most auspicious candidates demonstrated that this approach could provide spiegelmers with subnanomolar dissociation constant. To demonstrate if the selected spiegelmers are functional and suitable for cTnI detection in a sandwich type arrangement, AlphaLisa technology was leveraged and the obtained results demonstrated that spiegelmers with different epitope selectivity are suitable for specific detection of cTnI protein even in human plasma containing samples. These results suggest that spiegelmers could be considered in the development of the next generation cTnI monitoring assays.

Synthesis of TiO2/WO3 Composite Nanofibers by a Water-Based Electrospinning Process and Their Application in Photocatalysis.

TiO2/WO3 nanofibers were prepared in a one-step process by electrospinning. Titanium(IV) bis(ammonium lactato)dihydroxide (TiBALDH) and ammonium metatungstate (AMT) were used as water-soluble Ti and W precursors, respectively. Polyvinylpyrrolidone (PVP) and varying ratios of TiBALDH and AMT were dissolved in a mixture of H2O, EtOH and CH3COOH. The as-spun fibers were then heated in air at 1 °C min-1 until 600 °C to form TiO2/WO3 composite nanofibers. Fiber characterization was done using TG/DTA, SEM-EDX, FTIR, XRD, and Raman. The annealed composite nanofibers had a diameter range of 130-1940 nm, and the results showed a growth in the fiber diameter with an increasing amount of WO3. The photocatalytic property of the fibers was also checked for methyl orange bleaching in visible and UV light. In visible light, the photocatalytic activity increased with an increase in the ratio of AMT, while 50% TiBALDH composite fibers showed the highest activity among the as-prepared fibers in UV light.

Multiplexed assessment of the surface density of DNA probes on DNA microarrays by surface plasmon resonance imaging.

In terms of hybridization assays surface plasmon resonance imaging (SPRi) offers high throughput, label-free and real-time monitoring of the binding kinetics. This requires DNA microarrays on bare or modified gold SPRi chips, which are generally premade by an off-line microspotting procedure. Therefore, the surface density of the immobilized probes is not known although it is an essential quality control parameter, especially, when it can vary in a broad range as in case of self-assembled thiol-labeled DNAs on gold surface. Here we show that the small molecular weight ruthenium(III) hexamine complex (RuHex) introduced earlier for electrochemical quantitation of DNA coverage on gold electrodes can be used also in SPRi to assess the surface density of DNA probes in DNA microarrays. A single injection of RuHex solution allows the simultaneous visualization and quantification of the surface density of DNA probes (ranging in this study from 4 × 1011 to 1.7 × 1013 molecules cm-2) on all spots of a microarray made by microspotting thiol labeled short DNA probes both in prehybridized and single-stranded form on a gold SPRi chip. The methodology was applied to determine the effect of the surface density of DNA probes on the hybridization efficiency and kinetics of complementary microRNAs, using hsa-miR-208a-3p as model. Single mismatch duplexes were found to be more effectively destabilized than fully complementary duplexes by steric hindrance at large surface densities of the DNA probes, which offers an effective mean to increase single mismatch selectivity.

Gestational Exposure to the Synthetic Cathinone Methylenedioxypyrovalerone Results in Reduced Maternal Care and Behavioral Alterations in Mouse Pups.

The member of synthetic cathinone family, methylenedioxypyrovalerone (MDPV), is a frequently used psychoactive drug of abuse. The objective of our study was to determine the effect of MDPV (administered from the 8th to the 14th day of gestation) on the behavior of neonatal and adolescent mice, as well as its effect on maternal care. We measured maternal care (pup retrieval test, nest building), locomotor activity (open field test), and motor coordination (grip strength test) of dams, whereas on pups we examined locomotor activity at postnatal day 7 and day 21 (open field test) and motor coordination on day 21 (grip strength test). On fresh-frozen brain samples of the dams we examined the expression of two important peptides implicated in the regulation of maternal behavior and lactation: tuberoinfundibular peptide 39 (TIP39) mRNA in the thalamic posterior intralaminar complex, and amylin mRNA in the medial preoptic nucleus. We detected decreased birth rate and survival of offspring, and reduced maternal care in the drug-treated animals, whereas there was no difference between the motility of treated and control mothers. Locomotor activity of the pups was increased in the MDPV treated group both at 7 and 21 days of age, while motor coordination was unaffected by MDPV treatment. TIP39 and amylin were detected in their typical location but failed to show a significant difference of expression between the drug-treated and control groups. The results suggest that chronic systemic administration of the cathinone agent MDPV to pregnant mice can reduce birth rate and maternal care, and it also enhances motility (without impairment of motor coordination) of the offspring.

[Comparative evaluation of vertigo in patients after stapedotomy and stapedectomy]. / Posztoperatív vertigo vizsgálata stapedotomián és stapedectomián átesett betegeknél.

INTRODUCTION AND AIM: The reason of gradually developing conductive hearing loss in otosclerotic patients is the ossification of the stapes footplate to the surrounding bony structures and the therapy of stapes fixation is mainly surgical. In stapedotomy the footplate of the stapes is fenestrated with laser and microdrill in a diameter of 0.8 mm, whereas in stapedectomy there is complete removal of the footplate followed by the reconstruction of the ossicular chain. In the early postoperative period, temporary vertigo is frequently recorded which significantly influences the recovery. METHOD: In the Department of Otorhinolaryngology, University of Pécs both stapedectomy and stapedotomy were performed on a daily basis between 01.02.2010 and 15.03.2012. Our study focused on comparing the degree of postoperative vertigo after the two types of surgery. We hypothesized that the smaller fenestration of the stapes footplate during stapedotomy limits exposure to the inner ear reducing the severity of dizziness. Vertigo was evaluated subjectively with a retrospective questionnaire and objectively with static posturography. RESULTS: On the 1st postoperative day, significantly fewer patients reported vertigo in the stapedotomy group and with significantly lower intensity. Results of the questionnaire regarding the later postoperative period showed no significant differences between the groups. Based on the analysis of the posturography test results, no significant difference was detected between the postoperative stability of the two groups. Results of the questionnaire and the posturography showed no correlation. Posturography test results did not confirm the presence of subjective vertigo. CONCLUSION: Many factors may play a role in the development of vertigo after stapes surgery, but the type of intervention does not influence it. Orv Hetil. 2017; 158(38): 1503-1511.

The combination of trastuzumab and pertuzumab administered at approved doses may delay development of trastuzumab resistance by additively enhancing antibody-dependent cell-mediated cytotoxicity.

Although the recently concluded CLEOPATRA trial showed clinical benefits of combining trastuzumab and pertuzumab for treating HER2-positive metastatic breast cancer, trastuzumab monotherapy is still the mainstay in adjuvant settings. Since trastuzumab resistance occurs in over half of these cancers, we examined the mechanisms by which treatment of intrinsically trastuzumab-resistant and -sensitive tumors can benefit from the combination of these antibodies. F(ab')2 of both trastuzumab and pertuzumab were generated and validated in order to separately analyze antibody-dependent cell-mediated cytotoxicity (ADCC)-based and direct biological effects of the antibodies. Compared to monotherapy, combination of the two antibodies at clinically permitted doses enhanced the recruitment of natural killer cells responsible for ADCC, and significantly delayed the outgrowth of xenografts from intrinsically trastuzumab-resistant JIMT-1 cells. Antibody dose-response curves of in vitro ADCC showed that antibody-mediated killing can be saturated, and the two antibodies exert an additive effect at sub-saturation doses. Thus, the additive effect in vivo indicates that therapeutic tissue levels likely do not saturate ADCC. Additionally, isobole studies with the in vitro trastuzumab-sensitive BT-474 cells showed that the direct biological effect of combined treatment is additive, and surpasses the maximum effect of either monotherapy. Our results suggest the combined therapy is expected to give results that are superior to monotherapy, whatever the type of HER2-positive tumor may be. The combination of both antibodies at maximum clinically approved doses should thus be administered to patients to recruit maximum ADCC and cause maximum direct biological growth inhibition.

V-ATPase inhibition overcomes trastuzumab resistance in breast cancer.

The HER2 oncogene targeting drug trastuzumab shows remarkable efficacy in patients overexpressing HER2. However acquired or primary resistance develops in most of the treated patients why alternative treatment strategies are strongly needed. As endosomal sorting and recycling are crucial steps for HER2 activity and the vacuolar H⁺-ATPase (V-ATPase) is an important regulator of endocytotic trafficking, we proposed that targeting V-ATPase opens a new therapeutic strategy against trastuzumab-resistant tumor cells in vitro and in vivo. V-ATPase inhibition with archazolid, a novel inhibitor of myxobacterial origin, results in growth inhibition, apoptosis and impaired HER2 pro-survival signaling of the trastuzumab-resistant cell line JIMT-1. This is accompanied by a decreased expression on the plasma membrane and accumulation of HER2 in the cytosol, where it colocalizes with endosomes, lysosomes and autophagosomes. Importantly, microscopic analysis of JIMT-1 xenograft tumor tissue of archazolid treated mice confirms the defect in HER2-recycling which leads to reduced tumor growth. These results suggest that V-ATPase inhibition by archazolid induces apoptosis and inhibits growth of trastuzumab-resistant tumor cells by retaining HER2 in dysfunctional vesicles of the recycling pathway and consequently abrogates HER2-signaling in vitro as well as in vivo. V-ATPase inhibition is thus suggested as a promising strategy for treatment of trastuzumab-resistant tumors.

The real face of juvenile polyposis syndrome.

Colorectal cancers are mostly sporadic; some cases of familial clustering and autosomal dominant conditions are also known to occur. Juvenile polyposis syndrome (JPS) is an autosomal dominant condition caused by the mutation of the SMAD4 or the BMPR1A genes. JPS is characterized by hamartomatous polyps developing in the upper and lower intestine. Contradicting previous studies, many of these polyps can go through malignant transformation.This paper reports the case of a male patient who was continuously treated for juvenile polyposis. During the eighteen years of treatment, more than hundred polyps were endoscopically removed from his gastrointestinal tract. The patient's care was interrupted for eight years due to insufficient compliance. He was subsequently referred to our Department of Gastroenterology in severe clinical condition caused by metastatic colorectal cancer. He died after a short palliative therapy at the age of 31. His first-degree accessible relatives were further examined for juvenile polyposis syndrome. Several gastrointestinal polyps of different histological origin were observed in the deceased patient's brother, who subsequently had to undergo a left lateral hemicolectomy. Genetic analyses revealed mutations of the BMPR1A gene in the clinically affected brother, the brother's daughter, and in the deceased proband's daughter.Indebt genetic analyses helped customize and deliver care to a very specific group of individuals. We were able to identify potential family members on whom preventive care and treatment could be focused and simultaneously prevented unnecessary clinical and invasive procedures on those who were healthy. Furthermore, these analyses helped prevent future unnecessary trauma or distress on the analyzed family.

MRI analysis of mGluR5 and mGluR1 antagonists, MTEP and R214127 in the cerebral forebrain of awake, conscious rats.

Metabotropic glutamate receptors mGluR5 and mGluR1 mediate key neuropsychiatric functions in health and disease and their antagonists hold promise to treat anxiety, depression, inflammation, and neuropathic pain. Pharmacological magnetic resonance imaging (phMRI) using a functional MRI approach in awake, conscious rodents can determine the activities of receptor ligands without the potential interference of anesthetics and independent of the specific biochemical mechanism of action of the candidate molecule. In this study we determined the neuronal activation patterns of 3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine (MTEP) and 1-(3,4-dihydro-2H-pyrano[2,3-b]quinolin-7-yl0-2phenyl-1-ethanone (R214127), antagonists of mGluR5 and mGluR1 receptors by phMRI. We found that MTEP and R214127 activated specific primary somatosensory, piriform, entorhinal and motor cortices and the caudateputamen each to a different extent and in partly overlapping manners. Additional analysis of the activation data indicated that these brain regions and their connections are involved in mediating neuropathic pain and also, reward and olfaction. Using awake, conscious animals in phMRI can be a useful approach in characterizing candidate mGluR5 and mGlR1 antagonists also allowing a more direct comparison of animal and human phMRI studies.

Sitting-up vertigo and trunk retropulsion in patients with benign positional vertigo but without positional nystagmus.

BACKGROUND: Presently, the unambiguous diagnosis of benign paroxysmal positioning vertigo (BPPV) requires the detection of positioning or positional nystagmus provoked by Dix-Hallpike (for vertical semicircular canals) or supine roll (for horizontal semicircular canals) manoeuvres, which indicates canalo- or cupolithiasis of affected semicircular canals. There are patients, however, in whom--despite typical complaints of BPPV--no positional nystagmus can be documented; this is called 'subjective BPPV' (sBPPV). These patients usually complain of short vertigo spells during and after sitting up, sometimes with abnormal retropulsion of the trunk. AIM: In this study, the authors aimed to ascertain whether these patients in fact demonstrate abnormal sitting-up trunk oscillations when measured by posturography. Of 200 unselected patients with vertigo or dizziness, 43% had sBPPV with vertigo spells while sitting up, and 20% classical BPPV. METHODS: Posturographic recordings were performed in 20 patients with sBPPV and sitting-up vertigo. RESULTS AND DISCUSSION: Seven of the 20 patients had trunk oscillations during the act of sitting up and for a short time immediately afterwards. Based on their findings, the authors propose a new type of BPPV, the so-called Type 2 BPPV (typical complaints of BPPV, no nystagmus in Dix-Hallpike positions but short vertigo spell while sitting up), which may be the result of chronic canalolithiasis within the short arm of a posterior canal. Furthermore, the authors suggest that Type 2 BPPV, which could be identical to sBPPV or constitute a major subgroup of it, occurs frequently among patients with vertigo. For therapy, the authors recommend repetitive sit-ups from the Dix-Hallpike positions to liberate the short arm of the posterior canal from canaloliths.

Fontolizumab in moderate to severe Crohn's disease: a phase 2, randomized, double-blind, placebo-controlled, multiple-dose study.

BACKGROUND: The safety and efficacy of fontolizumab, a humanized anti-interferon gamma antibody, was investigated in patients with Crohn's disease (CD). Elevated gut mucosal levels of interferon gamma, a key cytokine involved in the inflammatory process of CD, are associated with disease symptoms. METHODS: A total of 201 patients with Crohn's Disease Activity Index (CDAI) scores between 250 and 450 were randomized to receive an initial intravenous dose of 1.0 or 4.0 mg/kg fontolizumab or placebo, followed by up to 3 subcutaneous doses of 0.1 or 1.0 mg/kg fontolizumab or placebo every 4 weeks. Clinical response at day 29, the primary efficacy endpoint, was defined as a decrease in the CDAI of at least 100 points from baseline levels. RESULTS: Of 201 patients, 135 (67%) completed the study. Day 29 response rates were similar in all treatment groups (31%-38%). At subsequent timepoints a significantly greater proportion of patients in the 1.0 mg/kg intravenous / 1.0 mg/kg subcutaneous fontolizumab group had clinical response and significantly greater improvement in the CDAI score compared with patients who received placebo. All fontolizumab groups had significant improvement in C-reactive protein levels. The overall frequency of adverse events was similar in all groups (58%-75%); most events were related to exacerbation of CD. There was a low frequency (5.2%) of neutralizing antibodies to fontolizumab. CONCLUSIONS: Although a strong clinical response to fontolizumab was not observed, significant decreases in C-reactive protein levels suggest a biological effect. Fontolizumab was well tolerated, and further studies to assess its efficacy are warranted.

Efficacy and safety of infliximab induction therapy in Crohn's Disease in Central Europe--a Hungarian nationwide observational study.

BACKGROUND: Infliximab (IFX) has proven to be an effective addition to the therapeutic arsenal for refractory, fistulizing, and steroid dependent Crohn's disease (CD), with efficacy in the induction and maintenance of clinical remission of CD. Our objective in this study is to report the nationwide, multicenter experience with IFX induction therapy for CD in Hungary. METHODS: During a 6-year-period, beginning in 2000, a total of 363 CD patients were treated with IFX as induction therapy (5 mg/kg IFX infusions given at week 0, 2 and 6) at eleven centers in Hungary in this observational study. Data analysis included patient demographics, important disease parameters and the outcome of IFX induction therapy. RESULTS: Three hundred and sixty three patients (183 women and 180 men) were treated with IFX since 2000. Mean age was 33.5 +/- 11.2 years and the mean duration of disease was 6.7 +/- 6.1 years. The population included 114 patients (31.4%) with therapy-refractory CD, 195 patients (53.7%) with fistulas, 16 patients (4.4%) with both therapy-refractory CD and fistulas, and 26 patients (7.2%) with steroid dependent CD. Overall response rate was 86.2% (313/363). A higher response rate was observed in patients with shorter disease duration (p = 0.05, OR:0.54, 95%CI:0.29-0.99) and concomitant immunosuppressant therapy (p = 0.05, OR: 2.03, 95%CI:0.165-0.596). Concomitant steroid treatment did not enhance the efficacy of IFX induction therapy. Adverse events included 34 allergic reactions (9.4%), 17 delayed type hypersensitivity (4.7%), 16 infections (4.4%), and 3 malignancies (0.8%). CONCLUSION: IFX was safe and effective treatment in this cohort of Hungarian CD patients. Based on our experience co-administration of immunosuppressant therapy is suggested in patients receiving IFX induction therapy. However, concomitant steroid treatment did not enhanced the efficacy of IFX induction therapy.

(-)Deprenyl-N-oxide, a (-)deprenyl metabolite, is cytoprotective after hypoxic injury in PC12 cells, or after transient brain ischemia in gerbils.

BACKGROUND AND AIMS: (-)-Deprenyl (selegiline) possesses cyto-protective effect in a much lower concentration, than it is needed to inhibit MAO-B activity. In permanent MCA occlusion stroke model in rats, the infarct volume and the number of apoptotic neurons in the penumbra region were decreased by low concentration (-)deprenyl treatment. Augmented Bcl-2 protein expression was documented as the responsible factor of this effect. The stabilization of mitochondrial membrane and diminished ROS production are the further possible consequences of (-)deprenyl treatment. It is not clear however that (-)deprenyl, or its metabolites are the acting neuroprotective molecules in the hypoxic/ischemic conditions. We report here the possible cyto-protective effect of deprenyl-N-oxide (DNO), a recently synthesized (-)deprenyl metabolite. METHODS: DNO in a very low dose (10(-5,-8,-12) M) was tested in PC12 cell culture after hypoxia and in gerbils after transient occlusion of bilateral common carotid artery. In PC12 culture the cell death was visualized by PI staining. The level of reactive oxygen species was measured by the Cerium method, and the mitochondrial membrane integrity was labeled by JC1 staining. Apoptotic neurons were counted on formaldehyde fixed gerbil brain slices after TUNEL and caspase-3 immune-staining - NIKON/BIORAD confocal microscopy was used for the quantitative analysis. RESULTS: DNO treatment significantly decreased the frequency of cell death in PC12 cultures after hypoxia, increased the mitochondrial transmembrane potential (DeltaY(m)) and decreased the ROS production. In the CA2 regions of gerbil hippocampus, we found significantly less apoptotic neurons than in the untreated controls. CONCLUSION: Transient hypoxia or ischemia induced cell damage could be diminished by DNO. This (-)deprenyl metabolite is an active cell protective molecule.

Ultrastructural localization of calcyon in the primate cortico-basal ganglia-thalamocortical loop.

Recent observations suggest that calcyon, a novel single transmembrane protein implicated in schizophrenia and attention-deficit/hyperactivity disorder, regulates clathrin-mediated endocytosis in brain. To explore the role of calcyon in neurotransmission, we investigated its distribution in the neuropil of the primate prefrontal cortex (PFC), striatum (STR) and mediodorsal thalamic nucleus (MD), three brain regions implicated in these neuropsychiatric disorders. Calcyonimmunoreactivity revealed by immunoperoxidase technique, was localized in both pre- and postsynaptic structures including axons, spines and dendrites, as well as myelinated fibers and astroglial processes in all the three brain regions. The morphological diversity of immunopositive boutons suggest that in addition to glutamatergic, calcyon could regulate GABAergic as well as monoaminergic neurotransmission. Consistent with the role of calcyon in endocytosis, calcyon-immunoreactivity was rarely found at the synaptic membrane specializations proper, although it was present in distal compartments of neuronal processes establishing synapses. Given the widespread upregulation of calcyon in schizophrenic brain, these findings underscore a potential association with deficits in a range of neurotransmitter systems in the cortico-basal ganglia-thalamic loop.

A family with two consecutive nonsense mutations in BMPR1A causing juvenile polyposis.

We describe a novel germline mutation of BMPR1A in a family with juvenile polyposis and colon cancer. This mutation consists of two consecutive substitutions (735-6 TG>AT) that cause two nonsense mutations (Y245X, G246X), inherited in an autosomal dominant fashion, on one parental chromosome. This mutation caused protein truncation, and represents a novel case of consecutive nonsense mutations in human disease.

Optimal Alignment novel software procedure for 3D reconstruction of electronmicroscopic serial sections.

3D reconstruction from electronmicroscopic (EM) serial sections substantially differs from modeling body parts by linking convoluted planes delivered by CT and NMR. Namely, variations both in relative X-Y position and rotation of the target elements between the adjacent images and also additional problems caused by deformed, deteriorated or missing sections can only be overruled by an aligning paradigm, which exploits all the pixel-level information, and results in an optimal fitting with selected precision. This paper presents a complex computer program called Optimal Alignment, which performs the precise elaboration of X-Y shift and relative rotation of two consecutive images. The required searching process will be customized by setting four independent parameters which relate the span and density of the pixel-scanning basic process. Optimalization of fitting accuracy versus running time can be achieved by a rather short training period. The potential precision of Optimal Alignment based on complex algorithms is far superior to manual aligning of EM photographs with the eye-wrist-mouse facility. The resulted database of alignment orientation parameters can serve as an advanced source for the 3D reconstructing programs. Optimal Alignment software tool (supported by Hungarian Space Office grant TP 138) will be demonstrated on a basal forebrain NPY+ axonal reconstruction, performed in L. Záborszky's laboratory (supported by NIH grant NSO23945).

Prevention of NSAID-associated gastrointestinal lesions: a comparison study pantoprazole versus omeprazole.

AIM: To investigate tolerability and efficacy of pantoprazole 20 mg, once daily (o.d.), pantoprazole 40 mg o.d., and omeprazole 20 mg o.d., in patients taking nonsteroidal anti-inflammatory drug(s) (NSAIDs). METHODS: Included in this randomized, double-blind, multicenter, parallel-group study were rheumatic patients (>55 yr) on continual NSAIDs and with at least one more recognized risk factor that contributes to the development of gastrointestinal (GI) injury. Study duration was 6 months, and the treatment consisted of pantoprazole 20 mg o.d. (N = 196), pantoprazole 40 mg o.d. (N = 199), or omeprazole 20 mg o.d. (N = 200). Patients took NSAID(s) (except COX-2 inhibitors), had no more than five erosions/petechiae in the upper GI tract, no current peptic ulcers or reflux esophagitis, and had at most moderate intensity GI symptoms. Endoscopy was performed at baseline, 3, and 6 months. The primary end points were lack of "therapeutic failure" and lack of "endoscopic failure" at 6 months. RESULTS: After 6 months, the probabilities to remain in remission were 90% pantoprazole 20 mg o.d., 93% pantoprazole 40 mg o.d., and 89% omeprazole 20 mg o.d. for lack of "therapeutic failure;" 91% pantoprazole 20 mg o.d., 95% pantoprazole 40 mg o.d., and 93% omeprazole 20 mg o.d. for lack of "endoscopic failure." CONCLUSIONS: For patients taking NSAIDs continually, pantoprazole 20 mg o.d., pantoprazole 40 mg o.d., or omeprazole 20 mg o.d. provide equivalent, effective, and well-tolerated prophylaxis against GI lesions, including peptic ulcers.

Visualization of mitochondrial membrane potential and reactive oxygen species via double staining.

Quantitative and qualitative analysis of both generated reactive oxygen species (ROS) and mitochondrial membrane potential cannot be detected simultaneously. We here introduce a simple, new double staining method. We have successfully used this for several years utilizing cerium for ROS detection and JC-1 staining to assess the mitochondrial membrane potential. The resultant signals on laser confocal images can be localized in the same cells and can easily quantify them. We used a confocal microscope along with our new, combined staining method to both visualize mitochondrial membrane potential (DeltaPsim) and imaged ROS. These were quantified by JC-1 staining and by cerium ions with reflectance in a method modified in our laboratory. To test this double labeling technique we used PC 12 cells subjected to 1 h hypoxia and 24h re-oxygenization. We are able to produce a quantitative analysis of red/green signals of JC-1 that reflected the energy state of the cells. Cerium reflectance correlates with the amount of ROS release in the same cells. Significant differences have been calculated after hypoxia and re-oxygenation in both modality of the cell staining. The red/green ratio was 18.2+/-9.3 (n=30) in normoxic cells versus 1.65+/-0.9 (n=30) in the hypoxia/re-oxygenation group (p<0.05). In the same randomly selected cells the average cerium reflectance signal intensity was 2.5+/-1.2 (n=30) in the control group while 5.8+/-3.1 (n=30) in the hypoxia/re-oxygenation group (p<0.05). This assay, by characterizing hypoxic injury and re-oxygenization induced ROS production, offers a qualitative and quantitative method to detect the consequences of oxidative stress in experimental conditions and to detect different cell protective strategies.