INCT-MM_Activity report2009
Transcrição
INCT-MM_Activity report2009
National Institute of Science and Technology of Molecular Medicine Instituto Nacional de Ciência e Tecnologia de INCT-MM Medicina Molecular 2009 Activity Report : INCT Medicina Molecular 2009 Activity Report Marco Aurélio Romano-Silva Director #$$%$ /%"##% "$', ""#$$%$# #+# #&&$$ 0 !.1% "#!%##$#"#01- $ & '$%.#:<8;9;2766=37 33666<:36> INCT-MM Marcus Vinícius Gomez Vice-director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rof. Marco A. Romano-Silva, Director, INCT de Medicina Molecular $&!$/% &$!'&! equipment, such as the PET/CT Discovery D690 from GE Healthcare. At the time we were writing this report, civil construction work was under way. The INCT-MM will be able to provide its users with first-class tools in response to our researchers’ needs. Meanwhile, the research groups have been developing their specific protocols, pre-clinical and clinical, to be ready when the PET/CT systems become operational. Educational activities are also being developed, with several training courses and scientific activities to enlight the public, specialized and lay, about our Institute. INCT-MM This is the first Annual Activity Report of the Instituto Nacional de Ciência e Tecnologia – Medicina Molecular (INCTMM). The feeling at this moment is still of the excitement, having our research program being recognized as of such excellence that was selected to be one of the National Institutes by a high level international committee. In 2009, our main task was to select the equipments to setup our main facility, the Centro de Excelência em Imagem Molecular (Center for Excellence in Molecular Imaging). Although we had a budget constraint, intense negotiations allowed us to purchase state-of-the-art > %&$&(!&& INCT-MM Prof. Marco Aurélio Romano-Silva President PI Administrative Committee Prof. Marcus Vinícius Gomez Preclinical Prof. Luiz A. De Marco Molecular Genetics and Cancer Pro. Carlos Jorge Simal Nuclear Medicine - Define infrastructure policy of use; -Expenditure policy; -Protocols approval; - Autorship and co-autorship policy - Extramural interface: private and public sectors; - Ethics; - Attraction and integration of new groups; - Reports; Dr. Carlos Malamut Cyclotron Meetings December 9th 2008 February 6th 2009 May 5th 2009 May 12th 2009 August 17th 2009 INCT-MM September 17th 2009 ? INCT-MM $%&$'&'$ @ !,&! Funded by a joint agreement between agencies from the Brazilian Federal and State of Minas Gerais Governments, the INCT-MM will operate multiuser facilities for research in Molecular Medicine. The headquarter of the INCT-MM is located at the School of Medicine (Faculdade de Medicina) of Universidade Federal de Minas Gerais, in Belo Horizonte, State of Minas Gerais, Brazil. INCT-MM A &&3!3&3$&& !!+ INCT-MM 98 +!&$! The PETtrace tracer production system is a compact, automated cyclotron designed for the fast, easy and efficient production of PET radiotracers. This is a fixed energy isochronous accelerator, manufactured by General Electric (GE PETtrace8). The cyclotron is installed in a bunker with walls and ceiling of 1.90 meters of concrete. The PETtrace is an integrated system of production of PET tracers that the target. This facilitates the expansion of target systems and processing for the production of oxygen-15, carbon-11, nitrogen-13 and fluorine-18. The CDTN cyclotron has an external beam placed in separate concrete bunker. The Cyclotron Sector includes, besides the two concrete bunkers, a Control Room, Technical/Electric and Target Preparation Areas. The Laboratory for the production of the radiopharmaceuticals has three hot cells with walls of 75 mm of lead, and with installed equipment for synthesis, splitting, filling and closing of the vials. PETtrace. The system can be configured with different target systems and processing for the production of PET radioisotopes common. The basic configuration provides six positions of The equipment used for FDG synthesis is a TRACERLAB MX GE HEALTHCARE. The Lab Production has two synthesizers TRACERLAB installed in a Hot Cell Dual COMECER, BBS-2 model. The laboratory has another dual hot cell model BBS-2 for future INCT-MM comprise a negative ion cyclotron control system with a workstation operator, target systems and synthesis systems. It operates with the efficiency required to match clinical schedules, the flexibility needed for research protocols and the performance necessary to meet regional distribution demands. 99 use in the radiopharmaceuticals. synthesis of The splitting, filling and closing of the vials are performed by automated system, called THEODORICO, made also by COMECER. THEODORICO is mounted inside a hot cell wall. INCT-MM 9: 2 PET/CT, even in areas subject to motion. Advanced processing techniques allow to reach full clinical potential with the ability to reconstruct 3D imaging and gated studies at incredible speeds. 4x faster VUE Point HD and motion reconstruction times VUE Point HD ensures intelligent reconstruction technology, a GE exclusive, providing outstanding image quality for most patient types. By combining the LightSpeed VCT with the breakthrough motion management capabilities of the Discovery PET system, the new Discovery PET/CT platform provides the next evolution PET/CT system, bringing clinicallyrelevant innovations designed to open doors to new and advanced procedure possibilities in non-invasive diagnostic imaging. INCT-MM The INCT-MM has acquired a state-ofthe-art PET/CT system from GE Healthcare, at a cost of approximately USD2 million. The D690 with time-offlight technology delivers truly powerful options for rserach. From the highest sensitivity in the industry, the next generation of MotionFree technology, advanced information processing and faster reconstruction with the IBM BladeCenter™ this system delivers. As a key component of the image chain, the Discovery PET/CT 690 includes the scintillator with a proven track record and the highest sensitivity in the industry improving lesion detectably, potentially reducing the dose requirement and allowing for faster scans and increased throughput. GE Healthcare has developed a way to capture detail and precision with 9; INCT-MM Layout of the Center of Excellence in Molecular Imaging of the INCT-MM (Centro de Excelência em Imagem Molecular) 9< $3 %+%& !'$ &% LabPET 4 PET scanner. The preclinical PET system will be used in development of new tracers, and for that will be installed in a facility adjacent to the cyclotron, located at the main Campus of UFMG. The Institute has a full molecular biology infrastructure including DNA sequencer, thermocyclers, two real time PCR systems. LabPET™: APD based PET Sub-System INCT-MM LYSO and LGSO scintillators individually coupled to Avalanche Photo Diode (APD) detectors Advanced detector design and parallel signal processing digital electronics to achieve high count rate performance. Digital detector technology delivers high spatial resolution. 9= $!%!"+ Leica Confocal Microscopy System Our Leica TCS SP5 Confocal fully covers a broad range of requirements in confoca imaging with excellent overall performance. The system provides the full range of scan speeds at the highest resolution. With its highefficiency SP detection (five channels simultaneously) and AOBS (Acousto Optical Bream Splitter), the Leica TCS SP delivers bright, noise-free images with minimal photo damage at high speed. Carl Zeiss ApoTome system In traditional microscopy, ApoTome allows the very fast production of extremely high-quality optical sections through fluorescence-labeled biological specimens – e.g. for the threedimensional visualization of nerve cells. The technique provides microscope images with a level of quality rivaling that of existing methods, but with lower costs and less equipment. Therefore, it offers a large number of users in biomedical research new possibilities for more exact and more reliable results that have been available exclusively to special research institutes and large imaging centers until now. Nancy Scardua Binda, PhD student INCT-MM 9> !)+&!&$+ Guava Cytometer System Karen Torres, post-doctoral fellow &% The new Animal Experiment Core has just been finished, and will be a facility with capacity for maintenance rooms experimentation rooms, including primary cell culturing and a Fluorescence In vivo imaging system for small animals. In addition to mice, rats and rabbits, the core has a zebrafish (Danio rerio) facility, which is part of the strategy of the INCTMM for developing animal models of complex diseases, in addition to our existing C. elegans culture facility at the Neuroscience lab. Our Guava PCA-96 System provides automated, higher throughput screening of 96-well microplates or a tray of microcentrifuge tubes. This flexible sample format makes the Guava PCA-96 System ideal for screening in basic research, target validation, assay development, and clinical trials. Like its smaller cousin, the Guava PCA System uses a green laser for excitation, two fluorescence detectors, and a forward scatter detector for assessing relative size. The walkaway automated sampling of 96-well plates makes it easy to screen large numbers of samples and plates in one day. INCT-MM Daniela Valadão, post-doc. Zebrafish aquaria. Zebrafish (Danio rerio) 9? Hospital das Clínicas-UFMG The HC/UFMG hospital complex is a large university hospital, consisting of a central hospital and several annexes. Hospitalization: offers capacity of 505 beds, providing care to almost all medical specialties and subspecialties. The hospital performs around 2100 surgeries monthly and serves 40 250 patients between emergency consultations, hospitalization s and outpatient visits. The buildings were built over the years as the needs of the time, demand and adequacy (the first block of the hospital dates from 1950). INCT-MM 9@ &% INCT-MM 9A '$!&!* %%$"'& &%- accomplish these objectives we have done: (1) in vitro experiments with isolated organs, tissue slices, purified nervous terminations (sinaptosomes) and cell culture, and (2) in vivo tests with laboratory animals. International contributions as existing with the Dr. Michael John Brammer, Director of the Brain Image Analysis Unit, Institute Psychiatry, University of London, were important in the field of confocal microscopy and image analysis. This contribution is being extended to the functional imaging analysis in animals. The project was extremely successful, with several international publications, two registered patents and one in progress, as well as several PhD thesis. Prof. Marcus V. Gomez, Vice-Director, INCT de Medicina Molecular Phα1β toxin prevents peripheral and spinal capsaicin-induced nociception in rats: role of direct blocking of TRPV1 receptor Previously, we have shown that the peptide toxin Phα1β blocked high-threshold voltagesensitive calcium channels (VSCCs), especially from the N-type, and induced antinociceptive effect by controlling nociceptive processes in the spinal cord. In the present study, we demonstrated that the previous treatment with Phα1β effectively reduced spontaneous nociception and mechanical allodynia induced by peripheral or spinal capsaicin injection in rats. On the other hand, the selective N-type calcium channel blocker ω-conotoxin MVIIA failed to reduce the nociception induced by capsaicin. Similar to Phα1β, the selective antagonist of the TRPV1 receptor SB366791 was also able to prevent capsaicin-trigged spontaneous nociception and mechanical allodynia. Capsaicin-induced calcium influx in rat DRG neurons or in HEK cells expressing TRPV1 was also inhibited by both Phα1β and SB366791. Moreover, there was no addictive effect on the inhibitory action of Phα1β and INCT-MM Prof. Gomez and his group have been working with peptide neurotoxins from scorpions and spiders for more than 40 years. The initial studies started with the scorpion neurotoxins (Gomez & Diniz, 1966; Gomez et al., 1973, 1975). During last the fifteen years, we have studied and characterized, in different types of ionic channels, the pharmacological actions of the neurotoxins Tx3-3 spider, Tx3-4 and Tx3-6 as well as their effect in the release of neurotransmitters (Romano-Silva et al. 1993, 1994) and acetylcholine (Moura et al., 1998). The group constituted, in 2006, the Institute of Millennium, aiming to integrate highly competent researchers to identify new toxins, to produce functional toxins for pharmacological study and, mainly, to carry out studies to investigate the potential therapeutical use of toxins to treat cerebral ischemia, pain and cardiac arrhythmias. Thus, a group with great experience in molecular and cellular studies together with several other researchers with experience in functional assays and in vivo studies analyzed the therapeutical toxin action of the spider Phoneutria nigriventer. The next step is to obtain data that prove the effectiveness of these toxins, and to show that the use of this new therapeutical armory will not produce adverse effects in the patients. To :8 SB366791 on capsaicin-stimulated calcium transients in DRG neurons, suggesting an overlap of action. Neither Phα1β nor SB 366791 altered the specific binding of [3H]resiniferatoxin in rat DRG membranes, indicating that either substances did not bind at the vanilloid site in TRPV1. Taken together, our results demonstrated that the peptide Phα1β is capable of directly blocking the TRPV1 receptor, an effect that may be critical for its antinociceptive action. INCT-MM Calcium increase in HEK293VR1 cells is sensitive to Phα1β and SB366791. Transmitted light merged with fluorescent images (488 nm excitation, 530–570 emission) of VR1 transfected cells staining with fluo-4. (A) before and (B) after 0.1 nM capsaicin addition. Scale bar 100 μm. VR1 cells were stimulated twice with 0.1 nM capsaicin with a 12-min interval between stimuli, (C) Control, (D) 10 μM SB366791, and (E) 100nM Phα1β were added at the interval between stimuli. (F) Amplitude ratio of second and first stimulus *p <0.05 compared to control statistically different . :9 Phoneutria spider toxins block ischemia-induced glutamate release and neuronal death of cells layers of the retina. In this study was investigated the effect of calcium channel blockers spider toxins on cell viability and the glutamate content of ischemic retinal slices. Rat retinal slices were subjected to ischemia via exposure to oxygen-deprived low glucose (ODLG) medium for 45 min. Slices were either treated or not treated with the toxins PhTx3, Tx3-3 and Tx3-4. After ODLG insult, glutamate content and cell viability were assessed in the slices by confocal and optic microscopy. In the retinal ischemic slices that were treated with PhTx3, Tx3–3 and Tx3–4, confocal imaging showed a decrease in cell death of 79.5% ± 3.1, 75.5% ± 5.8 and 61% ± 3.8, respectively. Neuroprotective effects were also observed 15, 30, 60 and 90 min after the onset of the retinal ischemia injury. As a result of the ischemia, glutamate increased from 6.2 ± 1.0 to 13.2 ± 1.0 nMol/mg protein and was inhibited by PhTx3, Tx3–3 and Tx3–4 to 8.6 ± 0.7, 8.8 ± 0.9 and 7.4 ± 0.8 nMol/mg protein, respectively. Histological analysis of the live cells in the outer, inner and ganglion cell layers of the ischemic slices showed a considerable reduction in cell death by the toxins treatment. Spider toxins reduced glutamate content and cell death of retinal ischemic slices. INCT-MM Confocal images (10x magnification) of representative retinal slices showing a control retinal slice submitted to ischemia (A); a retinal slice that was neither submitted to ischemia nor treated with spider toxins (B); a retinal slice submitted to ischemia in the presence of 1.0 mg/mL PhTx3 (C); a retinal slice submitted to ischemia in the presence of 8.0 nM Tx3-3 (D); and a retinal slice submitted to ischemia in the presence of 8.0 nM Tx3-4 (E). Dead cells appear red, and live cells appear green. The graph (F) shows the percentage of dead cells in retinal slices that were submitted to ischemia insult. One group was not treated with spider toxins (black bar), and the others (open bars) were treated with 1.0 mg/mL PhTx3, 8.0 nM Tx3-3 or 8.0 nM Tx3-4. Dead cells in the treated slices are given as a percentage of their respective ischemic untreated slices (A). The results are expressed as the mean ± standard error of mean (SEM) of dead cells in ten fields of at least three different experiments. For other details, see the Materials and Methods section. *There was a significant difference compared ischemic slices not treated with toxins treated, p < 0.05. **There was difference compared ischemic slices treated by toxins PhTx3 and Tx3-4, p < 0.05. :: !!+!$ 1. Production and characterization of radiopharmaceuticals useful for research and diagnosis using PET. First, we developed a target port (FIG. 1) for solid materials, to engage outside line of the cyclotron, for research. In 2009 a project was developed to evaluate the stability of FDG and the use of an autoclave in the production process of FDG. The work was published in the Annals of the International Nuclear Atlantic Conference (INAC), held in Rio de Janeiro in 2009. Was obtained and characterized, also, the sodium fluoride (Na18F), a radioisotope used for studies of tumors and nononcological diseases of the bone. Interest in Na18F has been resumed, mainly due to the formation of high-resolution images. The group UPPR / CDTN produced Na18F injectable solution with guaranteed quality from the points of microbiological, physicochemical and biological. Showed a biodistribution profile in mice similar to that available in literature, with clearance equal to 0.29 mL min -1, volume of distribution of 14.0 7g.mL-1 and primarily renal elimination. The maximum concentration of 5.0 ± 0.5% DI.g-1 was observed in the bone 30 minutes after injection. The results strongly indicate that the Na18F produced in the UPPR CDTN can be used safely for clinical use in bone scintigraphy. This paper has been accepted for publication in the journal "Brazilian Journal of Pharmaceutical Sciences." Figure 1 - Port-targets for irradiation of solid targets Production of 64 coper The process for obtaining 64Cu through the nickel-enriched, was developed with electrodeposition 64Ni (FIG. 2) on a tablet of silver. Then insert the electrodeposited material is placed inside the holder and irradiated targets under specific conditions. After the irradiated material is dissolved in acid and strong 64Cobre is separated using ion exchange resins. INCT-MM For 2010 the goal is the extraction and characterization of 18F-choline in addition to implementation of the synthesis module for radiopharmaceuticals labeled with 11C. 2. Study for the deployment of new targets This project's main objective is to obtain radionuclides using solid targets, needed for the development of new PET radiopharmaceuticals that do not involve 18F. Figure 2 - System electrodeposition :; Production of nanoparticles. 67Gálio from ZnO Gallium 67 can be obtained from the Znenriched and it is necessary to obtain pellets of zinc oxide with porosity, density, mechanical strength and morphology specific to be irradiated. This work is part of PhD project in collaboration with the UFMG. Some work was described in the article published in "Brazilian Journal of Physics." And published in the Annals of the International Conference on Advanced Materials, 2009, RJ and International Conference on Nanostructured Materials, 2008, RJ. Figure 2 - electron microscopy image of ZnO pellets with satisfactory characteristics can be used as target 8 Production of Radiopharmaceuticals for the production of fluorine-18. On this subject was done a preliminary study, published in the Annals of the International Nuclear Atlantic Conference (INAC), held in Rio de Janeiro in 2009. The study of the field of neutron and gamma radiation around the cyclotron will provide important data to: (A) optimization of radiological protection of the individual worker and the public; (B) optimization / improvement of the shield so as to increase the life of the bunker (eg placement of borated polyethylene plates to reduce the activation of concrete); (C) use of the radiation field for various applications: materials science, dosimetry, etc.. 3. Neutron dosimetry in UPPR / CDTN In the year 2009 were measured and studies to evaluate the radiation field around the cyclotron PETtrace GE Section- INCT-MM :< !'$ !!"*%%% analysis of the NFATc1 protein showed a predominant nuclear staining in the multinucleated giant cells. CONCLUSION: The development of giant cells lesions of the jaws and cherubism are possibly mediated by overexpression of NFAT in the nucleus of the multinucleated cells. Amaral FR, Brito JA, Perdigão PF, Carvalho V, Souza PE, Gomez MV, De Marco L, Gomez RS. Journal of Oral Pathology and Medicine 39: 269-274, 2010. Prof. Luiz A. De Marco, Laboratory of Molecular Genetics, INCT-MM Novel mutations of the BSCL2 and AGPAT2 genes in 10 families with Berardinelli Seip congenital generalized lipodystrophy syndrome. CONTEXT: Congenital generalized lipodystrophy, or Berardinelli-Seip syndrome, is a rare INCT-MM NFATc1 and TNFalpha expression in giant cell lesions of the jaws. BACKGROUND: Activation mutations of SH3BP2 gene have been demonstrated in cherubism and central giant cell lesion (CGCL). In the present study we first attempted to investigate the SH3BP2 gene in peripheral giant cell lesion (PGCL). The effect of SH3BP2 gene mutations on the transcription of the downstream genes nuclear factor of activated T cells (NFATc1) and the cytokine tumor necrosis factoralpha (TNF-alpha) was also investigated together with the immunolocalization of NFATc1 protein in a set of cases of PGCL, CGCL and cherubism with and without SH3BP2 mutation. METHOD: Fresh samples of five PGCL, five CGCL and one cherubism cases were included in this study. One of the samples of CGCL presented a somatic heterozygous mutation c.1442A>T in exon 11. The cherubism case showed a heterozygotic substitution c.320C>T in both blood and lesion. These mutations were previously published. All coding and flanking regions of the SH3BP2 gene were sequenced in the cases of PGCL. The realtime polymerase chain reaction (RT-PCR) was performed to analyze the transcription of NFATc1 and TNF-alpha genes. The immunohistochemical analysis of the NFATc1 protein was also performed. RESULTS: No SH3BP2 gene mutation was found in PGCL. The RT-PCR showed increased expression of NFATc1 and decreased transcription of TNF-alpha in all the samples. The immunohistochemical Novel mutations in the gene associated with sporadic central giant cell lesions and cherubism. Central giant cell lesion (CGCL) is a reactive bone lesion that occurs mainly in the mandible, characterized by the multinucleated osteoclast-like giant cells in a background of oval to spindle-shaped mononuclear cells. The etiology is unknown and occurs more commonly in young adults. Cherubism, a rare disease found predominantly in females has histologic characteristics indistinguishable from those of CGCL and is caused by mutations mostly present in exon 9 of the SH3BP2 gene. In this study, we investigated four cases of CGCL and one case of cherubism. DNA was extracted from peripheral blood and tumor tissue and all coding and flanking regions of the SH3BP2 amplified by PCR and directly sequenced to identify underlying mutations. Two novel mutations were found; a heterozygous missense mutation c.1442A>T (Q481L) in exon 11 in one sporadic case of CGCL and a heterozygous germline and tumor tissue missense mutation c.320C>T (T107M) in exon 4 in one patient with cherubism. These findings open a new window to investigate the possible relationship between the pathogenesis of the cherubism and CGCL. Carvalho VM, Perdigão PF, Amaral FR, Souza PEA, De Marco L, Gomez RS. Oral Diseases 15: 106-110, 2009 := autosomal recessive disease caused by mutations in either the BSCL2 or AGPAT2 genes. This syndrome is characterized by an almost complete loss of adipose tissue usually diagnosed at birth or early infancy resulting in apparent muscle hypertrophy. Common clinical features are acanthosis nigricans, hepatomegaly with or without splenomegaly and high stature. Acromegaloid features, cardiomyopathy and mental retardation can also be present. DESIGN: We investigated 11 kindreds from different geographical areas of Brazil (northeast and southeast). All coding regions as well as flanking intronic regions of both genes were examined. Polymerase chain reaction (PCR) amplifications were performed using primers described previously and PCR products were sequenced directly. RESULTS: Four AGPAT2 and two BSCL2 families harboured the same set of mutations. BSCL2 gene mutations were found in the homozygous form in four kindreds (c.412C>T c.464T>C, c.518-519insA, IVS5-2A>G), and in two kindreds compound mutations were found (c.1363C>T, c.424A>G). In the other four families, one mutation of the AGPAT2 gene was found (IVS3-1G>C and c.299G>A). CONCLUSIONS: We have demonstrated four novel mutations of the BSCL2 and AGPAT2 genes responsible for Berardinelli-Seip syndrome and Brunzell syndrome (AGPAT2related syndrome). Miranda D, Wajchenberg BL, Calsolari MR, Aguiar MJ, Silva JMCL, Ribeiro MG, Fonseca C, Amaral D, Boson W, Resende BA, De Marco L. Clinical Endocrinology 71: 512-517, 2009. INCT-MM Familial hyperparathyroidism: Surgical outcome after 30 years of follow-up in 3 families with germline HRPT2 mutations. It is still debatable which is the best management to familial forms of hyperparathyroidism. Conservative, minimally invasive or aggressive surgical approaches have been proposed from different groups around the world. Our objective was to study the gene mutation, expression of HRPT2 and the clinical outcome after 32 years of follow-up in one Brazilian kindred with familial isolated hyperparathyroidism (FIHP). Clinical and biochemical data, direct sequencing of the HRPT2 gene, analysis of parafibromin expression using RT-PCR, and immunohistochemistry were done. A nonsense mutation was found in exon 1 (c.96G>A)(p.Trp32X) in all affected members studied. Using RT-PCR, mRNA transcription was altered with complete absence of both transcripts in tumor tissue. Immunohistochemical analysis of tumors showed loss of parafibromin immunoreactivity. In this kindred there was a high prevalence of recurrence (75%), or persistence after less than subtotal parathyroidectomy that led us to consider a more aggressive surgical approach should be discussed among the affected family members, once surgical criteria was met. We concluded that it is necessary to individualize the surgical approach for HRPT2-related hyperparathyroidism until we can gather a better phenotype-genotype correlation in larger series, to best define their treatment. Sarquis M, Dias EP, Eng C, De Marco L. Surgery 145: 249-250, 2009. :> Silva JM, Ribeiro MG, Fonseca C, Amaral D, Boson WL, Resende BA, De Marco L. Clin Endocrinol (Oxf). 2009 Oct;71(4):512-7. Prof. Débora M. Miranda, Laboratory of Molecular Genetics INCT-MM Novel mutations of the BSCL2 and AGPAT2 genes in 10 families with Berardinelli-Seip congenital generalized lipodystrophy syndrome. Congenital generalized lipodystrophy, or BerardinelliSeip syndrome, is a rare autosomal recessive disease caused by mutations in either the BSCL2 or AGPAT2 genes. This syndrome is characterized by an almost complete loss of adipose tissue usually diagnosed at birth or early infancy resulting in apparent muscle hypertrophy. Common clinical features are acanthosis nigricans, hepatomegaly with or without splenomegaly and high stature. Acromegaloid features, cardiomyopathy and mental retardation can also be present. DESIGN: We investigated 11 kindreds from different geographical areas of Brazil (northeast and southeast). All coding regions as well as flanking intronic regions of both genes were examined. Polymerase chain reaction (PCR) amplifications were performed using primers described previously and PCR products were sequenced directly. RESULTS: Four AGPAT2 and two BSCL2 families harboured the same set of mutations. BSCL2 gene mutations were found in the homozygous form in four kindreds (c.412C>T c.464T>C, c.518-519insA, IVS5-2A>G), and in two kindreds compound mutations were found (c.1363C>T, c.424A>G). In the other four families, one mutation of the AGPAT2 gene was found (IVS3-1G>C and c.299G>A). CONCLUSIONS: We have demonstrated four novel mutations of the BSCL2 and AGPAT2 genes responsible for Berardinelli-Seip syndrome and Brunzell syndrome (AGPAT2related syndrome). Miranda DM, Wajchenberg BL, Calsolari MR, Aguiar MJ, Association of SLITRK1 to Gilles de la Tourette Syndrome. Previously the Slit and Trk-like family member 1 (SLITRK1) gene was identified as a candidate gene for Gilles de la Tourette Syndrome (GTS) based on a patient that carried a chromosomal inversion on 13q, as well as the identification of two rare DNA variants in the SLITRK1 gene. Since that report, studies have tested for the two rare variants in GTS and either did not find them, or when found, they did not segregate with the disorder in families, casting doubt on the relationship of this gene to GTS. We tested for these two rare variants and genotyped three polymorphisms that tag the currently identified major haplotypes of this gene in a sample of 154 nuclear families with GTS. In addition, the entire coding region was screened for novel DNA variants. We did not find the two reported rare variants in any of the probands or siblings in these families. We did however find significant evidence for association of a single polymorphism and of haplotypes of the three tagging polymorphisms. These findings provide the first support for the original finding indicating SLITRK1 as a susceptibility gene for GTS and indicate that further study of this gene in GTS is warranted. Miranda DM, Wigg K, Kabia EM, Feng Y, Sandor P, Barr CL. Am J Med Genet B Neuropsychiatr Genet. 2009 Jun 5;150B(4):483-6. :? Prof. Ana Cristina Simões e Silva, Pediatric Nephrology Genetic deletion of the angiotensin-(1-7) receptor Mas leads to glomerular hyperfiltration and microalbuminuria.. Angiotensin-(1–7), an active fragment of both angiotensins I and II, generally opposes the vascular and proliferative actions of angiotensin II. Here we evaluated effects of the angiotensin-(1–7) Molecular Pathophysiology of Renal Tubular Acidosis. Renal tubular acidosis (RTA) is characterized by metabolic acidosis due to renal impaired acid excretion. Hyperchloremic acidosis with normal anion gap and normal or minimally affected glomerular filtration rate defines this disorder. RTA can also present with hypokalemia, medullary nephrocalcinosis and nephrolitiasis, as well as growth retardation and rickets in children, or short stature and osteomalacia in adults. In the past decade, remarkable progress has been made in our understanding of the molecular pathogenesis of RTA and the fundamental molecular physiology of renal tubular transport processes. This review summarizes hereditary diseases caused by mutations in genes encoding transporter or channel proteins operating along the renal tubule. Review of the molecular basis of hereditary tubulopathies reveals various loss-of-function or gain-of-function mutations in genes encoding cotransporter, exchanger, or channel proteins, which are located in the luminal, basolateral, or endosomal membranes of the tubular cell or in paracellular tight junctions. These gene mutations result in a variety of functional defects in transporter/channel INCT-MM Hemostatic changes in patients with end stage renal disease undergoing hemodialysis.. Patients undergoing hemodialysis may show both thrombotic complications and bleeding abnormalities. Hemostatic changes in patients on hemodialysis may result from alterations in vessel wall integrity and platelet function, and reduced blood flow in the native arteriovenous fistula. Vascular complications represent 20–25% of all hospitalizations of patients on hemodialysis. Literature survey revealed that changes in the hemostatic system may play a major role in vascular complications observed in these patients. Thus, it is essential to investigate hemostatic alterations in patients on hemodialysis so that adequate regimes for anticoagulant therapy could be implemented. In this review we discuss hemostatic abnormalities in end stage renal disease patients undergoing hemodialysis. Clinica Chimica Acta 2010; 411: 135 - 139 receptor Mas on renal physiology and morphology using Mas-knockout mice. Compared to the wild-type animals, Mas knockout mice had significant reductions in urine volume and fractional sodium excretion without any significant change in free-water clearance. A significantly higher inulin clearance and microalbuminuria concomitant with a reduced renal blood flow suggest that glomerular hyperfiltration occurs in the knockout mice. Histological analysis found reduced glomerular tuft diameter and increased expression of collagen IV and fibronectin in the both the mesangium and interstitium, along with increased collagen III in the interstitium. These fibrogenic changes and the renal dysfunction of the knockout mice were associated with an upregulation of angiotensin II AT1 receptor and transforming growth factor-b mRNA. Our study suggests that Mas acts as a critical regulator of renal fibrogenesis by controlling effects transduced through angiotensin II AT1 receptors in the kidney. Kidney International 2009;75:1184 - 1193 :@ proteins, including decreased activity, impaired gating, defective trafficking, impaired endocytosis and degradation, or defective assembly of channel subunits. Further molecular studies of inherited tubular transport disorders may shed more light on the molecular pathophysiology of these diseases and may significantly improve our understanding of the mechanisms underlying renal salt homeostasis, urinary mineral excretion, and blood pressure regulation in health and disease. The identification of the molecular defects in inherited tubulopathies may provide a basis for future design of targeted therapeutic interventions and, possibly, strategies for gene therapy of these complex disorders. Current Genomics 2009; 10: 51 - 59. The Vasoactive Peptide Angiotensin-(1 7), Its Receptor Mas and the Angiotensin-converting Enzyme Type 2 are Expressed in the Human Endometrium. Angiotensin (Ang)-(1-7) is INCT-MM Citocinas e quimiocinas no transplante renal.. Renal transplantation is currently the best modality of renal substitutive therapy. Unfortunately graft survival is interrupted by episodes of acute rejection or even interstitial fibrosis/tubular atrophy. The measurement of urinary chemokines and cytokines as an alternative tool to diagnose these complications has been reported in past years. Those substances are clearly related to the immunoinflammatory mechanisms of renal transplantation, and can be detected in renal tissue, plasma, and urine samples of transplant recipients. Anti-inflammatory drugs, renin-angiotensin system inhibitors, and some antagonists of cytokines’ receptors, although used only experimentally, can interfere with the expression of these immune system mediators and consequently alter renal transplantation outcome. This article reviewed studies on the measurement of cytokines/ chemokines and their receptors in urine, plasma, and renal tissue of transplant recipients, aiming at evaluating a possible association of the levels of those mediators with renal transplantation complications and allograft survival. Jornal Brasileiro de Nefrologia 2009; 31: 286 – 296. one of the major active components of the renin-angiotensin system, produced from cleavage of Ang II by angiotensinconverting-enzyme type 2 (ACE2), which acts through a specific G protein-coupled receptor, Mas. We have investigated whether the human endometrium expresses these components during menstrual cycle. By radioimmunoassay, Ang-(1-7) was detected in endometrial wash fluid at picomolar concentrations. Using immunofluorescence, both the peptide and its receptor were identified in cultured endometrial epithelial and stromal cells. By immunohistochemistry, Ang(1-7) was localized in the endometrium throughout menstrual cycle, being more concentrated in the glandular epithelium of mid- and late secretory phase. This pattern corresponded to the ACE2 mRNA, which was more abundant in epithelial cells than in stromal cells (2-fold increase, p < 0.05) and in the secretory vs. proliferative phase (6.6-fold increase, p < 0.01). The receptor Mas was equally distributed between epithelial and stromal cells and did not change during menstrual cycle. The physiological role of this peptide system in normal and pathological endometrium warrants further investigation. Reproductive Sciences 2009; 16: 247-256. :A personality disorder, panic disorder and alcoholism). The frequency of S allele carriers was higher only in those patients who had made a violent suicide attempt in their lifetime (x(2)=16.969; p=0.0001). In a logistic regression model including these factors, S allele carrier (5-HTTLPR) was the only factor associated with violent suicide attempt. Sample size and retrospective assessment of suicide behavior history are the limitations of this study. Our study showed that serotonin polymorphism (5HTTLPR) is strongly associated with violent suicidal behavior in BD patients. If confirmed, our results could be an important step to create a genetic tool for long-term suicide prediction. Neves FS, Malloy-Diniz LF, Romano-Silva MA, Aguiar GC, de Matos LO, Correa H. J Affect Disord. 2010 Jan 20. Prof. Humberto Correa, Laboratory of Neuroscience, Clinical Psychiatry Expression of neuronal calcium sensor-1 (NCS-1) is decreased in leukocytes of schizophrenia and bipolar disorder patients. Schizophrenia (SCZ) and bipolar disorder (BPD) are severe illnesses representing an enormous social, familiar and individual burden that affect 1% of the population world-wide. Several evidences indicate abnormalities of the dopamine system in both SCZ and BPD. Neuronal calcium sensor-1 (NCS-1) is a protein that has many functions in neurotransmission such as inhibition of dopamine D(2) receptor desensitization, regulation of ionic channels and INCT-MM Is the serotonin transporter polymorphism (5-HTTLPR) a potential marker for suicidal behavior in bipolar disorder patients? Suicide prediction is a huge challenge for mental health workers. Structured interviews based on epidemiological and clinical factors don't show effectiveness for suicide prevention. Biological markers, such as 5-HTTLPR, could help for identification of potential suicide attempters. METHODS: We evaluated 198 bipolar patients and 103 health controls, using a structured interview according to DSM-IV criteria. Genotyping, blind of clinical assessment for identification of S carriers and structured interviews were performed in order to describe clinical and epidemiological factors which could be associated with suicide behavior. Statistical analyses were calculated by the x(2) test and logistic regression model. We found that 26.77% and 16.67% had a lifetime history of non violent suicide attempt and violent suicide attempt, respectively. The clinical factors associated with violent and non violent suicide attempt had several differences. Violent suicide attempters had an earlier illness onset and had a higher number of psychiatric comorbidities (borderline ;8 enhancement of exocytosis of neurotransmitters. In addition, NCS-1 protein expression and mRNA levels were found increased in pre-frontal cortex (PFC) of SCZ and BPD patients. NCS-1 expression in neural and neuroendocrine cells is well documented and, recently, it was shown that NCS-1 is also expressed in mast cells and neutrophils. NCS-1 has important functions in mast cells since it stimulates Fc epsilon RI-triggered exocytosis and the release of arachidonic acid metabolites. Then, due to the known close relation between the nervous and immune systems, we sought to investigate the NCS-1 expression in lymphocytes and monocytes (CD4+ T lymphocytes, CD56+ NK cells, CD19+ B lymphocytes and CD14+ monocytes) of SCZ and BPD patients. Using flow cytometry, our results have shown that NCS-1 expression was diminished in CD4+T lymphocytes, CD19+ B lymphocytes and CD14+ monocytes of BPD patients and also decreased in CD4+ T lymphocytes and CD56+ NK cells of SCZ patients. Results suggest that immune cells might be a cellular model for studies with SCZ and BPD patients considering NCS-1 functions. Efforts need to be done to investigate the motive of the decreased percentage of immune cells expressing NCS-1 in patients with SCZ and BPD. Torres KC, Souza BR, Miranda DM, Sampaio AM, Nicolato R, Neves FS, Barros AG, Dutra WO, Gollob KJ, Correa H, Romano-Silva MA. Prog Neuropsychopharmacol Biol Psychiatry. 2009 Mar 17;33(2):229-34 The leukocytes expressing DARPP-32 are reduced in patients with schizophrenia and bipolar disorder. Bipolar disorder (BPD) and schizophrenia (SCZ) are severe disorders representing an enormous social, familiar and individual burden, being SCZ the most disabling psychiatric disorder characterized by psychosis and cognitive impairment. It is well known that SCZ and BPD are associated with abnormalities in dopamine signaling pathway. Recent data in the literature have demonstrated altered expression levels of some proteins involved in the modulation of this pathway in both brain and peripheral tissues. It was shown that protein and mRNA levels of dopamine and cAMP regulated phosphoprotein (DARPP-32) were downregulated in dorsolateral prefrontal cortex (DLPFC) of patients with SCZ or BPD when compared to controls. Due to the difficulty to access brain tissue and the absence of objective laboratory tests for bio-markers, we measured DARPP-32 expression in blood INCT-MM Representative dot-plots from PBMC. The NCS-1 expression by lymphocytes and monocyteswas verified by flowcytometry assays. PBMC from control individuals or SCZ or BPD patients were stained to surface subpopulation markers and intracellular NCS-1 granularity versus size from PBMC showing the region 1 (R1) for lymphocyte gate and region 2 (R2) for monocyte gate (A). Isotype control of Abs utilized to extracellular or intracellular stain cells (B). Percentage of CD4+ T cells expressing NCS-1 from control individual (C), CD19+ B cells expressing NCS-1 (D), CD14+ monocytes expressing NCS-1 (E) and CD56+ NK cells expressing NCS-1 (F). The dot-plots (B–F) demonstrate the frequencies of cells expressing the indicated molecules as detected using antibodies conjugated with PE or ALEXA 488 as described in Material and methods. ;9 cell sub-populations (CD4+ T lymphocytes, CD56+ NK cells, CD19+ B lymphocytes and CD14+ monocytes) taking advantage of the close relation of nervous and immune systems. Using flow cytometry as the analytical method, our results have shown that the DARPP-32 expression was diminished in CD4+ T lymphocytes, CD19+ B lymphocytes and CD14+ monocytes of BPD patients and was also decreased in CD4+ T lymphocytes and CD56+ NK cells of SCZ patients. These results showed that DARPP-32 expression in immune cells agrees with reports of reduced DARPP-32 protein in the DLPFC of BPD or SCZ patients. Our data suggest that DARPP-32 expression in PBMC could be used as a source of bio-markers to help in the treatment response of neuropsychiatry disorders as a window to the changes in the brain of those patients. Torres KC, Souza BR, Miranda DM, Nicolato R, Neves FS, Barros AG, Dutra WO, Gollob KJ, Correa H, Romano-Silva MA. Prog Neuropsychopharmacol Biol Psychiatry. 2009 Mar 17;33(2):214-9. '$ cells, the rest of the regional lymph nodes may not need to be removed. Because fewer lymph nodes are removed, there may be fewer side effects. When multiple regional lymph nodes are removed, the patient may experience side effects such as lymphedema (swelling caused by excess fluid build-up), numbness, a persistent burning sensation, infection, and difficulty moving the affected body area. INCT-MM Standard lymph node removal involves surgery to remove most of the lymph nodes in the area of the tumor (regional lymph nodes). For example, breast cancer surgery may include removing most of the axillary lymph nodes, the group of lymph nodes under the arm. This is called axillary lymph node dissection (ALND). If Sentinel Lymph Node biopsy is done and the sentinel node does not contain cancer A sentinel lymph node biopsy is done in two stages. In the first part of the procedure, which takes one to two hours, the patient goes to the nuclear medicine department of the hospital for an injection of a radioactive tracer known as technetium 99. A doctor who specializes in nuclear medicine first numbs the area around the tumor with a local anesthetic and then injects the radioactive technetium. He or she usually injects a blue dye as well. The doctor will then use a gamma camera to take pictures of the lymph nodes before surgery. This type of imaging study is ;: called lymphoscintigraphy. After the lymphoscintigraphy, the patient must wait several hours for the dye and the radioactive material to travel from the tissues around the tumor to the sentinel lymph node. He or she is then taken to the operating room and put under general anesthesia. Next, the surgeon injects more blue dye into the area around the tumor. The surgeon then uses a hand-held probe connected to a gamma ray counter to scan the area for the radioactive technetium. The sentinel lymph node can be pinpointed by the sound made by the gamma ray counter. The surgeon makes an incision about 0.5 in long t o remove the sentinel node. The blue dye that has been injected helps to verify that the surgeon is removing the right node. The incision is then closed and the tissue is sent to the hospital laboratory for examination. INCT-MM ;; '&! $ INCT-MM ;< $$'&!'$% & !!+ Through the course of Technology in Radiology and Diagnostic Imaging, UFMG inaugurates its performance in the business of technology degrees. Such courses also form high level professionals, who can even academic career, studying Masters or Doctorate. However, this type of degree is more focused to meet specific demands from the market. The Technology course in Radiology and Diagnostic Imaging will be offered at night, with admission of 80 students per year, 40 per semester. The student must deal with sophisticated equipment in the field of diagnostic imaging, widely used in medicine. The career demands affinities with the areas of Health Informatics and The course includes, besides technical knowledge determined to humanistic education. Students will have the infrastructure of Medical College and Hospital. The technologist in Radiology and Diagnostic Imaging will be able to work with equipment in the areas of radiology, computed tomography, magnetic resonance imaging, radiotherapy and nuclear medicine. The spread of courses offered by UFMG is to form professionals with more extensive than that offered by other courses in the area.'ll Be able to work with the most sophisticated imaging equipment, and also develop an academic career. INCT-MM ;= $'&!'$%% % + Education and Training Training CAMH Canadá Training Center IOP Reino Unido Child Health Women's Health CDTN Foreign collaborators Medical Residencies Internal Medicin Graduate Programs PUC-MG Faculdade de Medicina da UFMG Surgery INCT-MM Molecular Medicine Neuroscience Graduate Programs ICBUFMG Instituto Milênio de Neurotoxinas SCM-MG UFSM-RS Graduate Course Graduate course The INCT-MM has a net of participating graduate programs integrated with medical residencies in the various participating institutions. See above. INCT-MM ;> 0 !&)1 The website “Image of the Week” was created to complement the teaching of imaging disciplines. It follows the principles of “Image Challenge” of The New England Journal of Medicine, available at imagechallenge.nejm.org. In addition, the site will have links and presentations regarding principles and use of imaging technology in molecular medicine. It is coordinated by Dr. Viviane Parisotto Marino, from the Nuclear Medicine Service of Hospital das Clínicas –UFMG, and one of the associated laboratories of the INCT-MM. INCT-MM ;? '&! % INCT-MM ;@ '&! %766> 1. 13. de Sá MS, Costa JF, Krettli AU, Zalis MG, Maia GL, Sette IM, Câmara Cde A, Filho JM, GiuliettiHarley AM, Ribeiro Dos Santos R, Soares MB. Antimalarial activity of betulinic acid and derivatives in vitro against Plasmodium falciparum and in vivo in P. berghei-infected mice. Parasitol Res. 2009 Jul;105(1):275-9 1.818 14. DIAS, C. S. ; SILVA, José Maria Penido ; DINIZ, José Silvério Santos ; LIMA, Eleonora Moreira ; MARCIANO, R C ; LANA, L. G. ; TRIVELATO, Ana Luiza L. ; LIMA, M. S. ; Simoes e Silva, Ana Cristina ; OLIVEIRA, Eduardo Araújo . Risk factors for recurrent urinary tract infections in a cohort of patients with primary vesicoureteral reflux. The Pediatric Infectious Disease Journal, 2009. 15. Dias-Lopes C.; G. Guimarães ; L. Felicori ; KALAPOTHAKIS, E. ; Paula Fernandes ; NGUYEN, C. ; MOLINA, Franck ; GRANIER, C. ; Chávez-Olórtegui, Carlos . A protective immune response against lethal, dermonecrotic and hemorrhagic effects of Loxosceles intermedia venom elicited by a 27residue peptide. Toxicon (Oxford), 2009. INCT-MM Almeida, L ; CAROLINO, Rachel Melilo ; Sperandio, D ; Nehemy, M.B. ; DE MARCO, L . A importância dos fatores genéticos na degeneração macular relacionada à idade (DMRI). Arquivos Brasileiros de O ftalmologia, 2009. 2. Amaral E, Leite LF, Gomez MV, Prado MA, Guatimosim C. Ouabain evokes exocytosis dependent on ryanodine and mitochondrial calcium stores that is not followed by compensatory endocytosis at the neuromuscular junction. Neurochem Int. 2009 55(6):406-13 3. Campos SB, Miranda DM, Souza BR, Pereira PA, Neves FS, Bicalho MA, Melillo PH, Tramontina J, Kapczinski F, Romano-Silva MA, Correa H. Association of polymorphisms of the tryptophan hydroxylase 2 gene with risk for bipolar disorder or suicidal behavior. J Psychiatr Res. 2009 Sep 30. 4. Carvalho, VM ; Perdigão, PF ; Amaral, FR ; de Souza, PEA ; DE MARCO, L ; Gomez, RS . Novel mutations in the gene associated with sporadic central giant cell lesions and cherubism. Oral Diseases, v. 15, p. 106-110, 2009 5. COSTA, José Eustáquio da ; GOMES, Carolina Cavaliéri ; COTA, L. O. ; PATARO, A. L. ; SILVA, Jeane de Fátima Correia ; GOMEZ, R. S. ; COSTA, Fernando de Oliveira . Polymorphisms in the promoter region of the gene for 5-HTT in individuals with aggresive periodontitis. Journal of Oral Science, v. 50, p. 193-198, 2008. 6. Cunico W, Gomes CR, Facchinetti V, Moreth M, Penido C, Henriques MG, Varotti FP, Krettli LG, Krettli AU, da Silva FS, Caffarena ER, de Magalhães CS. Synthesis, antimalarial evaluation and molecular modeling studies of hydroxyethylpiperazines, potential aspartyl protease inhibitors, Part 2. Eur J Med Chem. 2009 v. 44, p. 1363-1368 Apr 8. 7. Cunico, Wilson ; Gomes, Claudia R.B. ; Facchinetti, Victor ; Moreth, Marcele ; Penido, Carmen ; Henriques, Maria G.M.O. ; Varotti, Fernando P. ; Krettli, Luisa G. ; Krettli, Antoniana U. ; da Silva, Franklin S. . Synthesis, antimalarial evaluation and molecular modeling studies of hydroxyethylpiperazines, potential aspartyl protease inhibitors, Part 2. European Journal of Medicinal Chemistry, v. 44, p. 3816-3820, 2009. . 2.882 8. DAROCHA, F ; CORREA, H ; TEIXEIRA, A . Obsessive compulsive disorder and immunology: A review. Progress in Neuro-Psychopharmacology & Biological Psychiatry, v. 32, p. 1139-1146, 2008. 2.638 9. de Castro BM, De Jaeger X, Martins-Silva C, Lima RD, Amaral E, Menezes C, Lima P, Neves CM, Pires RG, Gould TW, Welch I, Kushmerick C, Guatimosim C, Izquierdo I, Cammarota M, Rylett RJ, Gomez MV, Caron MG, Oppenheim RW, Prado MA, Prado VF. The vesicular acetylcholine transporter is required for neuromuscular development and function. Mol Cell Biol. 2009 Oct;29(19):5238-50 10. de Castro, Bráulio M. ; Pereira, Grace S. ; Magalhães, Vinicius ; Rossato, Janine I. ; De Jaeger, Xavier ; Martins-Silva, Cristina ; Leles, Bruno ; Lima, Patricia ; Gomez, Marcus V. ; Gainetdinov, Raul R. ; Caron, Marc G. ; IZQUIERDO, Ivan ; Cammarota, Martin ; Prado, Vania F. ; Prado, Marco A.M. . Reduced expression of the vesicular acetylcholine transporter causes learning deficits in mice. Genes, Brain and Behavior, 2009 Feb;8(1):23-35 11. de Oliveira MV, de Carvalho Fraga CA, Gomez RS, Paula AM. Immunohistochemical expression of interleukin-4, -6, -8, and -12 in inflammatory cells in surrounding invasive front of oral squamous cell carcinoma Head Neck. 2009 May 7 12. De Paula AM, Souza LR, Farias LC, Corrêa GT, Fraga CA, Eleutério NB, Silveira AC, Santos FB, Haikal DS, Guimarães AL, Gomez RS. Analysis of 724 cases of primary head and neck squamous cell carcinoma (HNSCC) with a focus on young patients and p53 immunolocalization Oral Oncol. 2009 Apr 7 ;A 16. DINIZ, Marina Gonçalves ; Borges, Érica Rievrs ; GUIMARÃES, André Luiz Sena ; MOREIRA, Paula Rocha ; Brito, João Artur Ricieri ; GOMEZ, Marcus Vinícius ; MARCO, Luiz de ; Gomez, Ricardo Santiago . PTCH1 isoforms in odontogenic keratocysts. Oral Oncology, v. 45, p. 291-295, 2009. 17. Diniz, MG ; Borges ER ; Guimaraes, A.L.S. ; Moreira, PR ; Brito, JA ; GOMEZ, Marcus Vinicius ; DE MARCO, L ; GOMEZ, Ricardo S . PTCH 1 isoforms in odontogenic keratocysts. Oral Oncology, 45:291-295, 2009 18. 19. Dutra WO, Moreira PR, Souza PE, Gollob KJ, Gomez RS. Implications of cytokine gene polymorphisms on the orchestration of the immune response: lessons learned from oral diseases. Cytokine Growth Factor Rev. 2009 Jun;20(3):223-32. 20. Felicori L, Fernandes PB, Giusta MS, Duarte CG, Kalapothakis E, Nguyen C, Molina F, Granier C, Chávez-Olórtegui C. An in vivo protective response against toxic effects of the dermonecrotic protein from Loxoscelesintermedia spider venom elicited by synthetic epitopes. Vaccine 2009 Jun 24;27(31):4201-8 21. Ferreira AD ; NEVES FS ; ROCHA FF ; MARCO, L. ; MIRANDA, D. M. ; ROMANO-SILVA, M A ; CORREA, H . The role of 5-HTTLPR polymorphism in antidepressant-associated mania in bipolar disorder. Journal of Affective Disorders, v. 112, p. 267-272, 2009 . 22. FILARDI, F. ; SOARES, F. M. ; ROCHA, R. F. ; CORREA, H, Humberto Correa ; Teixeira JR, AL . Increased surgical morbidity of psychiatric patients submitted to appendectomy. Revista Brasileira de Psiquiatria (São Paulo), v. 29, p. 192-193, 2007. 1.318 23. FILARDI, F. ; SOUZA, K. C. A. ; PAULINO, N. J. ; CASTRO, J. O. ; CORREA, H, Humberto Correa . Suicide in belo Horizonte between 2004 and 2006. Revista Brasileira de Psiquiatria (São Paulo), v. 29, p. 190-191, 2007. 1.318 24. Fortes-Dias, Consuelo Latorre ; Santos, Roberta Márcia Marques dos ; Magro, Angelo José ; Fontes, Marcos Roberto de Mattos ; Chávez-Olórtegui, Carlos ; GRANIER, Claude . Identification of continuous interaction sites in PLA2-based protein complexes by peptide arrays. Biochimie (Paris. Print), p. 1-11, 2009. 3.071 25. Gomes CC ; Oliveira, CS ; Pimenta, L.G. ; DE MARCO, L ; GOMEZ, Ricardo S . Immunolocalization of DNMT1 and DNMT3a in salivary gland neoplasms. Pathobiology (Basel), 2009 26. Gomes CC, Oliveira Cda S, Castro WH, de Lacerda JC, Gomez RS. Clonal nature of odontogenic tumours. J Oral Pathol Med. 2009 Apr;38(4):397-400. 27. Gomez RS, Dutra WO, Moreira PR. Epigenetics and periodontal disease: future perspectives. Inflamm Res. 2009 May 8 28. GUIMARÃES, André Luiz Sena ; GOMES, Carolina Cavaliéri ; SILVA, Luciano Marques da ; SILVA, Jeane de Fátima Correia ; VICTÓRIA, Junia Maria Netto ; GOMEZ, R. S. ; BITTENCOURT, Henrique . Association between oral HSV-1 and survival in allogeneic hematopoietic stem cell transplanted patients. Medicina Oral, Patología Oral y Cirugía Bucal, v. 14, p. E62-E68, 2009 29. 30. Hell RC, Amim P, de Andrade HM, de Avila RA, Felicori L, Oliveira AG, Oliveira CA, Nascimento E, Tavares CA, Granier C, Chávez-Olórtegui C. Immunodiagnosis of human neurocysticercosis using a synthetic peptide selected by phage-display. Clin Immunol. 2009 Apr;131(1):129-38 INCT-MM 31. Krettli AU, Adebayo JO, Krettli LG. Testing of natural products and synthetic molecules aiming at new antimalarials. Curr Drug Targets. 2009 Mar;10(3):261-70 32. Krettli AU. The utility of anti-trypomastigote lytic antibodies for determining cure of Trypanosoma cruzi infections in treated patients: an overview and perspectives. Mem Inst Oswaldo Cruz. 2009 Jul;104 Suppl 1:142-51 33. KRETTLI, A. U. . Antimalarial drug discovery: screening of Brazilian medicinal plants and purifi ed compounds. Expert Opinion on Drug Discovery, 2009. v. 4, p. 95-108 34. Miranda DM, Wajchenberg BL, Calsolari MR, Aguiar MJ, Silva JM, Ribeiro MG, Fonseca C, Amaral D, Boson WL, Resende BA, De Marco L. . Novel mutations of the BSCL2 and AGPAT2 genes in 10 families with Berardinelli-Seip congenital generalized lipodystrophy syndrome. Clinical Endocrinology 71(4):512-7 35. Miranda DM, Wigg K, Kabia EM, Feng Y, Sandor P, Barr CL. Association of SLITRK1 to Gilles de la Tourette Syndrome. Am J Med Genet B Neuropsychiatr Genet. 2009 Jun 5;150B(4):483-6. 36. Miranda, G.V. ; MIRANDA, D. ; Costa, E. ; CORREA, Humberto ; BOSON, Wolfanga ; DE MARCO, L ; SILVA, Marco Aurelio Romano . Estudo sobre o Transtorno Disfórico Pré-menstrual em uma população de mulheres em Belo Horizonte. Revista Médica de Minas Gerais, 2009. 37. MOREIRA, Paula Rocha ; GUIMARAES, M. M. ; GUIMARÃES, André Luiz Sena ; DINIZ, Marina Gonçalves ; GOMES, Carolina Cavaliéri ; BRITO, J. A. R. ; GOMEZ, R. S. . Methylation of P16, P21, P27, RB1 and P53 genes in odontogenic keratocysts. Journal of Oral Pathology and Medicine, v. 38, p. 99-103, 2009. <8 INCT-MM 38. MOURA, Mariela Dutra Gontijo ; NOVAES JR, João Batista ; GOMEZ, R. S. ; SOUTO, G. R. ; MESQUITA, Ricardo Alves de . PERIPHERAL DENTINOGENIC GHOST CELL TUMOUR IN CHILD: A CASE REPORT. International Journal of Pediatric Otorhinolaryngology Extra, v. 2, p. 250-253, 2007 1.118 39. Navarro KCL ; MIRANDA, D. M. ; Resende B ; ROMANO-SILVA, M A . The leukocytes expressing DARPP-32 is reduced in patients with schizophrenia and bipolar disorder. Progress in NeuroPsychopharmacology & Biological Psychiatry, 2009. 33(2):214-9. 40. Navarro KCL ; Souza BR ; MIRANDA, D. M. ; ROMANO-SILVA, M A . Expression of Neuronal Calcium Sensor-1 (NCS-1) is decreased in leukocytes of schizophrenia and bipolar disorder patients. Progress in Neuro-Psychopharmacology & Biological Psychiatry, 2009. 17;33(2):22941. PERDIGÃO, Paolla Freitas ; Carvalho, VM ; DE MARCO, L ; GOMEZ, Ricardo S . Mutation of ameloblastin gene in calcifying epithelial odontogenic tumor. Anticancer Research, 2009. 42. Pereira PCB; MIRANDA, DM; Oliveira EA; Simões e Silva AC . Molecular Pathophysiology of Renal Tubular Acidosis. Current Genomics, 2009. 10:51-9. 43. Pereira RM, dos Santos RA, da Costa Dias FL, Teixeira MM, Simões e Silva AC. Renin-angiotensin system in the pathogenesis of liver fibrosis. . World J Gastroenterol. 2009 Jun 7;15(21):2579-86. 44. PEREIRA, André Barreto ; REZENDE, Nilton Alver ; TEIXEIRA, ANTÔNIO L. ; TEIXEIRA, Mauro Martins ; Simoes e Silva, Ana Cristina . Citocinas e quimiocinas no transplante renal. Jornal Brasileiro de Nefrologia, 2009 45. PICARRO, C. ; TATSUO, E. S. ; AMARAL, V. F. ; GOMEZ, R. S. ; LANNA, J. . Morphological comparison of processus vaginalis from boys with undescended testis and hernia sacs from boys with inguinal hernia. European Journal of Pediatric Surgery, 2009. 19(3):145-7 46. Pinheiro AC, da Silva AJ, Prado MA, Cordeiro MD, Richardson M, Batista MC, de Castro Junior CJ, Massensini AR, Guatimosim C, Romano-Silva MA, Kushmerick C, Gomez MV. Phoneutria spider toxins block ischemia-induced glutamate release, neuronal death, and loss of neurotransmission in hippocampus Hippocampus. 2009 47. PINHEIRO, Sérgio Veloso Brant ; FERREIRA, Anderson José ; KITTEN, Gregory ; SILVEIRA, Katia Daniela da ; SILVA, Deivid Augusto da ; SANTOS, Sergio H S ; GAVA, Elizandra ; CASTRO, Carlos Henrique ; MAGALHÃES, Junio Alves ; MOTA, Renata Koza da ; BOTELHO-SANTOS, Giancarla A ; BADER, Michael ; ALENINA, Natalia ; SANTOS, Robson Augusto Souza dos ; SIMÕES E. SILVA, ANA C. . Genetic deletion of the angiotensin-(1-7) receptor Mas leads to glomerular hyperfiltration and microalbuminuria. Kidney International, 2009; v. 75, p. 1184-1193. 48. REIS, Helton Jose ; GUATIMOSIM, C ; Paquet M ; Magad Santos ; Fabiola Ribeiro ; Artur Kummer ; PEREIRA, Grace S ; SALGADO, J. V. ; Antonio Teixeira ; Andras Palotas . Neurotransmitters in the Central Nervous System.. Current Medicinal Chemistry (Hilversum), 2009. 16(7):796-840 49. Sarquis MMS, Dias EP, Eng C, De Marco LA (2009) Familial hyperparathyroidism: Surgical outcome after 30 years of follow-up in 3 families with germline HRPT2 mutations. Surgery 145: 249-250 (doi:10.1016/j.surg.2008.09.002 50. SILVA, José Maria Penido ; DINIZ, José Silvério Santos ; LIMA, Eleonora Moreira ; PINHEIRO, Sérgio Veloso Brant ; PARIZZOTO, V. S. ; CARDOSO, L. S. B. ; COLOSIMO, Enrico A ; SIMÕES e SILVA AC ; OLIVEIRA, Eduardo Araújo . Independent risk factors for renal damage in a series of primary vesicoureteral reflux: a multivariate analysis. Nephrology (Carlton), 2009; 14, p. 198-204. 51. SOARES, C. M. B. M. ; DINIZ, José Silvério Santos ; LIMA, Eleonora Moreira ; OLIVEIRA, Gilce R ; CANHESTRO, Mônica R ; COLOSIMO, Enrico A ; SIMÕES e SILVA AC ; OLIVEIRA, Eduardo Araújo . Predictive factors of progression to chronic kidney disease stage 5 in a predialysis interdisciplinary programme. Nephrology Dialysis Transplantation,. 2009 Mar;24(3):848-55. 52. SOUZA, A ; FERREIRA, J ; CORDEIRO, M ; VIEIRA, L ; DECASTRO, C ; TREVISAN, G ; REIS, H ; SOUZA, I ; RICHARDSON, M ; PRADO, M ; PRADO, V F ; GOMEZ, M. V. . Analgesic effect in rodents of native and recombinant Ph?1? toxin, a high-voltage-activated calcium channel blocker isolated from armed spider venom. Pain (Amsterdam), Nov 15;140(1):115-26 53. Souza RP, Tampakeras M, Basile V, Shinkai T, Rosa DV, Potkin S, Meltzer HY, Lieberman JA, Romano-Silva MA, Kennedy JL. Hum Psychopharmacol. 2009 Dec;24(8):676-9. 54. SOUZA, R P ; Soares EC ; ROSA, D V ; SOUZA, B R ; GOMES KM ; VALVASSORI, S. S. ; REUS GZ ; INACIO CG ; MARTINS MR ; GOMEZ, M V ; QUEVEDO, J. ; ROMANO-SILVA, M.A. . Cerebral DARPP-32 expression after methylphenidate administration in young and adult rats. International Journal of Developmental Neuroscience, Int J Dev Neurosci. 2009 Feb;27(1):1-7. . 55. TORRES, K. C. ; SOUZA, B R ; MIRANDA, D M ; NICOLATO, R ; NEVES, F S ; GUIMARÃES, Alexandre Barros ; DUTRA, W O ; GOLLOB, K J ; ROMANO-SILVA, M.A. . The leukocytes expressing DARPP-32 is reduced in patients with schizophrenia and bipolar disorder. Progress in Neuro-Psychopharmacology & Biological Psychiatry, 2009 Mar 17;33(2):214-9 56. TORRES, K. C. ; SOUZA, B R ; MIRANDA, D M ; SAMPAIO, A.M. ; NICOLATO, R ; NEVES, F S ; BARROS, A. G. ; DUTRA, W O ; GOLLOB, K J ; CORREA, H ; ROMANO-SILVA, M.A. . Expression of Neuronal Calcium Sensor-1 (NCS-1) is decreased in leukocytes of schizophrenia and bipolar <9 disorder patients. Progress in Neuro-Psychopharmacology & Biological Psychiatry, 2009 Mar 17;33(2):229-34.. 57. Vaisbich. M.H. ; CARNEIRO, Juliana Garcia ; BOSON, Wolfanga ; RESENDE, Bruna Araujo ; DE MARCO, L ; Honjo, R.S. ; Kim, C.A. ; KOCH, V. H. . Nephrogenic Diabetes Insipidus (NDI): clinical, laboratorial and genetic characterization of five Brazilian patients. Clinics (São Paulo), 2009. 58. Vilas-Boas WW, Ribeiro-Oliveira A Jr, Pereira RM, Ribeiro R da C, Almeida J, Nadu AP, Simões e Silva AC, dos Santos RA. Relationship between angiotensin-(1-7) and angiotensin II correlates with hemodynamic changes in human liver cirrhosis. . World J Gastroenterol. 2009 May 28;15(20):2512-9. 59. Westphalen RI, Gomez RS, Hemmings HC Jr. Nicotinic receptor-evoked hippocampal norepinephrine release is highly sensitive to inhibition by isoflurane. Br J Anaesth. 2009 Mar;102(3):355-60. INCT-MM