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Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique

Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第2期10卷 112-119页

作者： Manuel TORALIBAEZ Margarita CARU Miguel A. HERRERA Luis GONZALEZ Luis M. MARTIN jorge miranda Rafael M. NAVARRO-CERRILLO Department of Forest Management Faculty of ForestryUniversity of Chile Santa Rosa 11315La PintanaSantiagoChile Department of Ecological Sciences Faculty of SciencesUniversity of ChileSanta Rosa 11315La PintanaSantiagoChile Department of Forestry School of Agriculture and Forestry University of Córdoba14080 CórdobaSpain Department of Genetic School of Agriculture and ForestryUniversity of Córdoba14080 CórdobaSpain Department of Forest Management Faculty of ForestryUniversity of ChileSanta Rosa 11315La PintanaSantiagoChile

A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D. Don) Endl. in Chile. Ten (out of 34) clones from introduction trial located in Voipir-Villarrica, Chile, were studied. The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction. The PCR tests were carried out employing 10-mer random primers. The amplification products were detected by electrophoresis in agarose gels. Forty nine polymorphic bands were obtained with the selected primers (BG04, BF07, BF12, BF13, and BF14) and were ordered according to their molecular size. The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was con- structed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA). Of the primers tested, 5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism. A total of 49 polymorphic RAPD bands were detected out of 252 bands. The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones, suggesting a geographic relationship. The results indicate that the RAPD markers permitted the identification of the assayed clones, although they are derived from the same geo- graphic origin.

关键词： RAPD标记技术 PCR检测基因型鉴定北美红杉无性系随机扩增多态性DNA 智利基因组DNA提取

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First Detection of Human Polyomaviruses in HIV Patients with Suspected Neurological Disease in Montería, Colombia

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Modern Research in Inflammation 2015年第3期4卷 25-31页

作者： Vaneza Tique Samia Barrera Salim Mattar jorge miranda Francisco Camargo University of Cordoba Instituto de Investigaciones Biologicas del Tropico Córdoba Colombia Clinica Salud Social Sincelejo Sucre Clinica Zayma Intensive Care Unit Monteria Cordoba

Objective: The objective is to establish the presence of JC and/or BK polyomavirus in HIV patients with symptoms of encephalitis and/or meningitis. Methodology: From September 2009 to December 2011, a prospective study was conducted. 34 HIV patients with symptoms consistent with encephalitis and/or meningitis were included. The work was conducted in 3 hospitals in the city of Monteria. Viral DNA extraction was performed on samples of cerebrospinal fluid (CSF) using a commercial kit (Quiagen, USA). The detection of BKV and JCV was performed by multiplex real-time PCR (LightMix?, Roche Diagnostics, Germany) with primers specific for the short t antigen gene fragment, labeled probes and one internal control. Results: In 9 (26%) of 34 patients included in the study, JCV virus was detected;only 1 (3%) patient had coinfection with JCV/BKV. The mortality rate was 3%. The cytochemical examination of CSF in positive patients presented average values: 40.7 mg/dL glucose, 171.66 mg/dL protein, 19.8 mm3 leukocytes, and 109.8 mm3 erythrocytes. Conclusion: The findings demonstrate that JCV and BKV have a significant occurrence in HIV patients with CSF infections in Monteria.

关键词： Progressive Multifocal Leukoencephalopathy Meningitis Encephalopathy AIDS

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Tamoxifen:an FDA approved drug with neuroprotective effects for spinal cord injury recovery

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Neural Regeneration Research 2016年第8期11卷 1208-1211页

作者： Jennifer M.Colón jorge D.miranda Department of Physiology School of MedicineUniversity of Puerto Rico Medical Sciences Campus

Spinal cord injury（SCI） is a condition without a cure,affecting sensory and/or motor functions.The physical trauma to the spinal cord initiates a cascade of molecular and cellular events that generates a non-permissive environment for cell survival and axonal regeneration.Among these complex set of events are damage of the blood-brain barrier,edema formation,inflammation,oxidative stress,demyelination,reactive gliosis and apoptosis.The multiple events activated after SCI require a multi-active drug that could target most of these events and produce a permissive environment for cell survival,regeneration,vascular reorganization and synaptic formation.Tamoxifen,a selective estrogen receptor modulator,is an FDA approved drug with several neuroprotective properties that should be considered for the treatment of this devastating condition.Various investigators using different animal models and injury parameters have demonstrated the beneficial effects of this drug to improve functional locomotor recovery after SCI.Results suggest that the mechanism of action of Tamoxifen administration is to modulate anti-oxidant,anti-inflammatory and anti-gliotic responses.A gap of knowledge exists regarding the sex differences in response to Tamoxifen and the therapeutic window available to administer this treatment.In addition,the effects of Tamoxifen in axonal outgrowth or synapse formation needs to be investigated.This review will address some of the mechanisms activated by Tamoxifen after SCI and the results recently published by investigators in the field.

关键词： selective estrogen receptor modulator trauma antioxidant anti-inflammatory regeneration reactive gliosis demyelination, estradiol

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Tamoxifen and Src kinase inhibitors as neuroprotective/neuroregenerative drugs after spinal cord injury

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Neural Regeneration Research 2015年第3期10卷 385-390页

作者： Iris K.Salgado Aranza I.Torrado Jose M.Santiago jorge D.miranda Department of Physiology School of MedicineUniversity of Puerto Rico Medical Sciences Campus University of Puerto Rico Carolina Campus Department of Natural Sciences

Spinal cord injury （SCI） is a devastating condition that produces significant changes in the life- style of patients. Many molecular and cellular events are triggered after the initial physical impact to the cord. Two major phases have been described in the field of SCI： an acute phase and late phase. Most of the therapeutic strategies are focused on the late phase because this provides an opportunity to target cellular events like apoptosis, demyelination, scar formation and axonal outgrowth. In this mini-review, we will focus on two agents （tamoxifen and a Src kinase family inhibitor known as PP2） that have been shown in our laboratory to produce neuroprotective （increase cell survival） and/or regenerative （axonal outgrowth） actions. The animal model used in our laboratory is adult female rat （N250 g） with a moderate contusion （12.5 mm） to the spinal cord at the T10 level, using the MASCIS impactor device. Tamoxifen or PP2 was administered by implantation of a 15 mg pellet （Innovative Research of America, Sarasota, FL, USA） or by intraperitoneal injections （1.5 mg/kg, every 3 days）, respectively, to produce a long-term effect （28 days）. Tamoxifen and the Src kinase inhibitor, PP2, are drugs that in rats with a moderate spinal cord injury promote functional locomotor recovery, increase spared white matter tissue, and stimulate axonal outgrowth. Moreover, tamoxifen reduces the formation of reactive oxygen species. Therefore, these drugs are possible therapeutic agents that have a neuroprotective/regen- erative activity in vertebrates with SCI.

关键词： tamoxifen Src kinase PP2 trauma regeneration neuroprotection

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Long-term treatment with PP2 after spinal cord injury resulted in functional locomotor recovery and increased spared tissue

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Neural Regeneration Research 2014年第24期9卷 2164-2173页

作者： Odrick R.Rosas Aranza I.Torrado Jose M.Santiago Ana E.Rodriguez Iris K.Salgado jorge D.miranda Department of Physiology School of Medicine University of Puerto Rico Medical Sciences Campus Department of Natural Sciences University of Puerto Rico Carolina Campus

The spinal cord has the ability to regenerate but the microenvironment generated after trauma reduces that capacity. An increase in Src family kinase （SFK） activity has been implicated in neuropathological conditions associated with central nervous system trauma. Therefore, we hypothesized that a decrease in SFK activation by a long-term treatment with 4-amino-5-（4- chlorophenyl）-7-（t-butyl）pyrazolo[3,4-d]pyramidine （PP2）, a selective SFK inhibitor, after spinal cord contusion with the New York University （NYU） impactor device would generate a permissive environment that improves axonal sprouting and/or behavioral activity. Results demonstrated that long-term blockade of SFK activation with PP2 increases locomotor activity at 7, 14, 21 and 28 days post-iniury in the Basso, Beattie, and Bresnahan open field test, round and square beam crossing tests. In addition, an increase in white matter spared tissue and serotonin fiber density was observed in animals treated with PP2. However, blockade of SFK activity did not change the astrocytic response or infiltration of cells from the immune system at 28 days post-injury. Moreover, a reduced SFK activity with PP2 diminished Ephexin （a guanine nudeotide exchange factor） phosphorylation in the acute phase （4 days post-injury） after trauma. Together, these findings suggest a potential role of SFK in the regulation of spared tissue and/or axonal outgrowth that may result in functional locomotor recovery during the pathophysiology generated after spinal cord injury. Our study also points out that ephexinl phosphorylation （activation） by SFK action may be involved in the repulsive microenvironment generated after spinal cord injury.

关键词： nerve regeneration trauma regeneration Src family kinase Eph receptors ephexin spared tissue locomotor recovery GFAP ED1 serotonin fibers neural regeneration

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Application of a Pulsed Electric Field to Modify the Free Energy of the Active Site of an Azoreductase

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Journal of Applied Mathematics and Physics 2024年第1期12卷 249-255页

作者： Jesus Magyber de Jesus miranda Pedro Guillermo Reyes-Romero jorge Mulia-Rodríguez Daniel Osorio-González Molecular Biophysics Laboratory Faculty of Sciences Autonomous University of State of Mexico Toluca Mexico Advanced Physics Laboratory Faculty of Sciences Autonomous University of State of Mexico Toluca Mexico

The Gibbs free energy is strongly related to the stability and catalytic function of an enzyme through the energetic changes that occur in the chemical reactions the enzyme catalyzes. In this in silico study, a pulsed electric field was applied to an azoreductase, and its effect on the Gibbs free energy of molecular docking with two dyes was measured. We propose that certain stimuli from a pulsed electric field favor the structural stability of the enzyme by promoting an arrangement in the active site, potentially leading to an enhancement of enzymatic activity overall.

关键词： Pulsed Electric Field Molecular Dynamics Docking Azoreductase

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Development of a Ready-to-Use PSMA-11 Kit Formulation and Biological Evaluation of Binding Affinity

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Advances in Molecular Imaging 2024年第2期13卷 7-22页

作者： Maria Renata Valente Brandão Freire Natália Marchesan Bexiga Marco Antonio Stephano João Ezequiel Oliveira Carlos Roberto jorge Soares Ana Cláudia Camargo miranda Anna Carolina de Alcântara Barbosa Marycel Figols de Barboza Laura Fernanda Garcia Luiz Felipe S. Teixeira Maria Helena Marumo Bellini Natanael Gomes da Silva Luiza Mascarenhas Balieiro Joel Mendes Elaine Bortoleti de Araújo Centro de Radiofarmcia Instituto de Pesquisas Energticas e Nucleares (IPEN) Cidade Universitria So Paulo Brasil Departamento de Tecnologia Farmacutica Faculdade de Cincias Farmacuticas (USP) Cidade Universitria So Paulo Brasil Centro de Biotecnologia Instituto de Pesquisas Energticas e Nucleares (IPEN) Cidade Universitria So Paulo Brasil Departamento de Medicina Nuclear do Hospital Israelita Albert Einstein So Paulo/SP Brasil

Introduction: 68Ga-PSMA-11 is considered the gold standard in detection of micro and oligometastases in advanced prostate cancer, being used for therapeutic planning, as well as, potentially, for evaluating response to treatment. The development of ready-to-use lyophilized kit of PSMA-11 adds quality and safety to the routine use of this radiopharmaceutical and represents a pharmacotechnical challenge as it must preserve the integrity and specificity of the ligand. Methods: PSMA-11 kit formulation was proposed, considering radiolabeling parameters and the preservation of the peptide during the lyophilization process, using mannitol as an excipient. Critical temperature characterization studies were carried out using DSC equipment and the freeze-drying process was developed. The direct radiolabeling conditions were evaluated and standardized using 68Ge/68Ga generator eluate from two different manufacturers (ITG and Eckert & Ziegler). The radiochemical purity was evaluated by TLC and HPLC. Biological evaluation was carried out with lyophilized PSMA-11 to demonstrate the integrity of the peptide and preservation of biological activity after the lyophilization process. Results: Based on critical temperature characterization studies, the freeze-drying cycle was designed to reach a freezing temperature of around −40˚C and primary drying at 2˚C. Using 20 mg of mannitol, an intact and elegant lyophilized cake was obtained. PSMA-11 lyophilized kit was directly labeled with 68Ga eluate from 68Ge/68Ga GMP generators (ITG and Eckert & Ziegler) resulting in % RP > 95% at pH 4.0 to 4.5. The results obtained from in vitro and in vivo biological competition studies confirmed the preservation of PSMA-11 affinity for the receptor after lyophilization. Conclusion: A lyophilized formulation (Kit) of PSMA-11 was successfully obtained, which preserved the integrity and biological activity of the peptide and guaranteed radiolabeling efficiency.

关键词： 68Ga-PSMA-11 Prostate-Specific Membrane Antigen (PSMA) Ready-to-Use Kit GMP-Grade Generators Metastatic Prostate Cancer PET Imaging

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Agronomic evaluation of eight 41 B×110 richter grapevine genotypes as rootstock candidates for mediterranean viticulture

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Horticultural Plant Journal 2023年第4期9卷 720-728页

作者： Diana Marín Carlos miranda Francisco Javier Abad jorge Urrestarazu Blanca Mayor Ana Villa-Llop Luis Gonzaga Santesteban Department of Agronomy Biotechnology and Food SciencePublic University of NavarreCampus ArrosadiaPamplona 31006NavarraSpain Institute for Multidisciplinary Research in Applied Biology(IMAB-UPNA) Public University of NavarreCampus ArrosadiaPamplona 31006NavarraSpain Institute for Agri-food Technology and Infrastructure of Navarre(INTIA) Edificio de PeritosAvda.Serapio Huici 22Villava 31610NavarraSpain

Choosing the most appropriate rootstock(s)is a key decision for the profitability of vineyards;therefore,there must be a sufficient range of rootstocks in the market adapted to different environmental conditions and production objectives.However,rootstock-breeding programs have been scarce in recent decades,and most of the rootstocks used today were bred a century ago,when the needs of the sector were very different from today.In this work,we aimed to evaluate new rootstock candidates before their introduction in the market.An agronomic evaluation was conducted on eight novel rootstock genotypes obtained from the first generation of the cross-pollination of 41 B Millardet et de Grasset(41 B)and 110 Richter(110 R)grafted with‘Syrah’and‘Tempranillo’and planted in a typical vineyard of the Ebro Valley in Spain.During the four consecutive growing seasons(2016–2019),growth,yield and berry composition parameters at harvest were collected.A linear mixed-effects model was constructed,considering year and block as random effects.Multiple factor analysis and hierarchical clustering on principal components were performed to establish clusters of genotypes with similar behaviour.The rootstock candidates showed a very wide performance range compared to their parents.The trial allowed us to identify two very promising candidates(RG8 and RG10),whose registration as commercial rootstocks is already in progress.

关键词： Vitis vinifera Graft Phenolic quality Vigour Yield

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Ventral phalloplasty

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Asian Journal of Andrology 2008年第1期10卷 155-157页

作者： jorge Caso Michael Keating Alejandro miranda-Sousa Rafael Carrion Division of Urology Department of Interdisciplinary Oncology University of South Florida Tampa FL 33612 USA

Aim： To present a simple technique during penile prosthesis implantation that promotes the perception of increased phallic length. Methods： The penoscrotal web is defined. A ＂check mark＂ incision is made with excision of scrotal tissue. Excellent exposure is provided for implantation of the cylinders, pump and reservoir. Wound closure is performed longitudinally. Results： This technique is a modified extension of surgeries described in the pediatric literature for webbed penis. Loss of penile length following penile implantation surgery is worrisome for patients suffering from erectile dysfunction （ED）. This technique helps with patient satisfaction, cosmetic results, and improves perception of penile length. Conclusion： Ventral phalloplasty is a safe, technically simple procedure that may be performed in concert with penile prosthesis implantation or as a stand alone procedure under＇certain circumstances.

关键词： penile implantation impotence phalloplasty genital lengthening

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Prevalence of type 2 diabetes mellitus in the pediatric population of a third-level care hospital in Mexico City in 2013 and 2018

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World Journal of Diabetes 2023年第3期14卷 290-298页

作者： jorge Mario Molina-Díaz Blanca Estela Vargas-Terrez Patricia Guadalupe Medina-Bravo Antonia Martínez-Ambrosio América Liliana miranda-Lora Miguel Klünder-Klünder Department of Child Endocrinology Children’s Hospital Federico GomezAutonomous University of MexicoMexico City 06250Mexico Faculty of Business La Salle UniversityMexico City 06250Mexico Department of Endocrinology Children’s Hospital Federico GomezMexico City 06720Mexico Department of Nursing and Endocrinology Children’s Hospital Federico GómezMexico City 06720Mexico Epidemiological Research Unit in Endocrinology and Nutrition Children’s Hospital Federico GomezMexico City 06720Mexico Division of Research Children’s Hospital Federico GomezMexico City 06720Mexico

BACKGROUND Diabetes mellitus type 2(T2DM)is a state of hyperglycemia due to a defect in the secretion and/or action of insulin,and it represents the most common form of diabetes worldwide.In Mexico,10.3%of the adult population have been diagnosed with T2DM and it is expanding to the pediatric population.AIM To evaluate and compare the prevalence of T2DM in the pediatric population at the Children’s Hospital,“Hospital Infantil de México Federico Gómez(HIMFG)”,at two time periods:2013 vs 2018.METHODS A comparison of two cross-sectional studies was done(2013 and 2018).The study population was comprised of children and adolescents 8-17 years old,from the Diabetes Clinic at the aforementioned institution.A comprehensive interrogation regarding family history and perinatal antecedents was performed.Complete blood work after 12 h of fasting was obtained to determine serum levels of glucose,glycated hemoglobin,lipid profile,C-peptide,and insulin.The data were analysed using the statistical software package SPSS v.23.0.A P value of<0.05 was considered statistically significant.RESULTS A total of 151 patients were included:47 from 2013,and 104 from 2018.There were age differences noted between the two periods with younger patients presenting T2DM in 2013.Also,T2DM predominated in the male sex in 55.36%in 2013 vs 32.7%in 2018.An increased prevalence of T2DM was noted from 2013 to 2018(20.2%vs 33.0%,respectively),which was a statistically significant 12.8%increase(P<0.0001).The illness phenotype was more aggressive in the 2018 group with the presence of a higher proportion of obesity,insulin resistance,and adverse lipid profiles.CONCLUSION The prevalence of T2DM at the HIMFG institution from 2013 to 2018 increased by 12.8%(20.2%vs 33.0%,respectively).The study results demonstrate the need for vigilance in T2DM trends,and to strengthen programs of healthy nutrition and physical activity as well as early detection and risk factors for obesity,data on insulin resistance,and metabolic syndrome,with the aim of preventing the deve

关键词： Diabetes Children adolescents Epidemiology Etiology Risk factors and complications

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