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1.
Niger. j. med. (Online) ; 30(4): 356-361, 2021.
Article in English | AIM, AIM | ID: biblio-1290631

ABSTRACT

Coronavirus disease 2019 (COVID­19) accounts for over 92 million confirmed cases worldwide, with over 2.06 million deaths. In the past year, more than 290 candidate vaccines have been tested; COVID-19 vaccine development was sped up, with shortened timeline, due to the urgent global need in the face of the pandemic. In addition, people with the highest risk of contracting the disease, such as health workers with a high risk of exposure, elderlies, and people with underlying comorbidities, were prioritized with vaccination rollout. The article narratively reviewed original and review articles available on PubMed and Google Scholar related to the theme to provide up­to­date information. The different templates developed and studied for COVID-19 vaccines include the whole-virus vaccine, viral vector vaccine, nucleic acid (deoxyribonucleic acid and ribonucleic acid), and protein subunit vaccine. Myths impede vaccine uptake in this part of the globe. Adopting these myths leads to sharing and spreading, which negatively impacts the prevention of COVID-19 and vaccine uptake. Adverse event following immunizations (AEFIs) is classified based on severity, from minor to severe. The minor ones are common events that pose no potential health risks to the receiver of the vaccine. The type determines the safety profile, severity, and frequency of AEFIs observed with the vaccine administration. Overall, this pandemic has heightened the global level of threat awareness; it has also provided motivation to prepare for future pandemics by developing new vaccines.


Subject(s)
Humans , COVID-19 Vaccines , COVID-19 , Vaccines, DNA , Protein Subunits , Nigeria
2.
Article in English | WPRIM | ID: wpr-922255

ABSTRACT

The N-methyl-D-aspartate receptor (NMDAR) in central nerve system is mostly composed of GluN1 and GluN2 subunits. The classical NMDAR has been intensively studied. However, GluN3‑containing NMDAR is much less expressed and have atypical channel properties. Recently, accumulating evidences have revealed two types of GluN3‑containing NMDAR: glutamate-gated GluN1/GluN2/GluN3 NMDAR and glycine-gated GluN1/GluN3 NMDAR. The former may play important roles in regulating synapse maturation and pruning non-used synapses, and its elevated expression at the adult stage may alter synaptic reorganization in some neuropsychiatric disorders. The latter is expressed in the medial habenula and involves in control of aversion. This article reviews the recent progresses on the expression, functional properties of GluN3‑containing atypical NMDARs and the physiological and pathological relevance.


Subject(s)
Central Nervous System/metabolism , Protein Subunits/metabolism , Receptors, N-Methyl-D-Aspartate , Synapses
3.
Neuroscience Bulletin ; (6): 507-518, 2019.
Article in English | WPRIM | ID: wpr-775416

ABSTRACT

Myasthenia gravis (MG) is a prototypical antibody-mediated neurological autoimmune disease with the involvement of humoral immune responses in its pathogenesis. T follicular helper (Tfh) cells have been implicated in many autoimmune diseases. However, whether and how Tfh cells are involved in MG remain unclear. Here, we established and studied a widely-used and approved animal model of human MG, the rat model with acetylcholine receptor alpha (AChRα) subunit (R-AChR)-induced experimental autoimmune myasthenia gravis (EAMG). This model presented mild body-weight loss 10 days after the first immunization (representing the early stage of disease) and more obvious clinical manifestations and body-weight loss 7 days after the second immunization (representing the late stage of disease). AChR-specific pre-Tfh cells and mature Tfh cells were detected in these two stages, respectively. In co-cultures of Tfh cells and B cells, the number of IgG2b-secreting B cells and the level of anti-AChR antibodies in the supernatant were higher in the cultures containing EAMG-derived Tfh cells. In immunohistochemistry and immunofluorescence assays, a substantial number of CD4/Bcl-6 T cells and a greater number of larger germinal centers were observed in lymph node tissues resected from EAMG rats. Based on these results, we hypothesize that an AChR-specific Tfh cell-mediated humoral immune response contributes to the development of EAMG.


Subject(s)
Animals , B-Lymphocytes , Allergy and Immunology , Disease Models, Animal , Female , Immunity, Humoral , Lymph Nodes , Allergy and Immunology , Myasthenia Gravis, Autoimmune, Experimental , Allergy and Immunology , Protein Subunits , Allergy and Immunology , Proto-Oncogene Proteins c-bcl-6 , Allergy and Immunology , Rats, Inbred Lew , Receptor Cross-Talk , Receptors, Cholinergic , Allergy and Immunology , T-Lymphocytes, Helper-Inducer , Allergy and Immunology
4.
Protein & Cell ; (12): 553-567, 2018.
Article in English | WPRIM | ID: wpr-757973

ABSTRACT

ATP-sensitive potassium channels (K) are energy sensors on the plasma membrane. By sensing the intracellular ADP/ATP ratio of β-cells, pancreatic K channels control insulin release and regulate metabolism at the whole body level. They are implicated in many metabolic disorders and diseases and are therefore important drug targets. Here, we present three structures of pancreatic K channels solved by cryo-electron microscopy (cryo-EM), at resolutions ranging from 4.1 to 4.5 Å. These structures depict the binding site of the antidiabetic drug glibenclamide, indicate how Kir6.2 (inward-rectifying potassium channel 6.2) N-terminus participates in the coupling between the peripheral SUR1 (sulfonylurea receptor 1) subunit and the central Kir6.2 channel, reveal the binding mode of activating nucleotides, and suggest the mechanism of how Mg-ADP binding on nucleotide binding domains (NBDs) drives a conformational change of the SUR1 subunit.


Subject(s)
Adenosine Triphosphate , Metabolism , Amino Acid Sequence , Animals , Binding Sites , Cryoelectron Microscopy , Ligands , Mesocricetus , Mice , Models, Molecular , Nucleotides , Metabolism , Pancreas , Metabolism , Potassium Channels, Inwardly Rectifying , Chemistry , Metabolism , Protein Binding , Protein Multimerization , Protein Structure, Quaternary , Protein Subunits , Chemistry , Metabolism , Sf9 Cells , Spodoptera , Sulfonylurea Receptors , Chemistry , Metabolism
5.
Ann. hepatol ; 16(2): 297-303, Mar.-Apr. 2017. tab, graf
Article in English | LILACS | ID: biblio-887236

ABSTRACT

ABSTRACT Introduction and aim. The inability to distinguish cancer (CSCs) from normal stem cells (NSCs) has hindered attempts to identify safer, more effective therapies for hepatocellular carcinoma (HCC). The aim of this study was to document and compare cell membrane potential differences (PDs) of CSCs and NSCs derived from human HCC and healthy livers respectively and determine whether altered GABAergic innervation could explain the differences. Material and methods. Epithelial cell adhesion molecule (EpCAM) positive stem cells were isolated from human liver tissues by magnetic bead separations. Cellular PDs were recorded by microelectrode impalement of freshly isolated cells. GABAA receptor subunit expression was documented by reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence. Results. CSCs were significantly depolarized (-7.0 ± 1.3 mV) relative to NSCs (-23.0 ± 1.4 mV, p < 0.01). The depolarized state was associated with different GABAA receptor subunit expression profiles wherein phasic transmission, represented by GAGAA α3 subunit expression, was prevalent in CSCs while tonic transmission, represented by GABAA α6 subunit expression, prevailed in NSCs. In addition, GABAA subunits α3, β3, γ3 and δ were strongly expressed in CSCs while GABAA π expression was dominant in NSCs. CSCs and NSCs responded similarly to GABAA receptor agonists (ΔPD: 12.5 ± 1.2 mV and 11.0 ± 3.5 mV respectively). Conclusion. The results of this study indicate that CSCs are significantly depolarized relative to NSCs and these differences are associated with differences in GABAA receptor subunit expression. Together they provide new insights into the pathogenesis and possible treatment of human HCC.


Subject(s)
Humans , Neoplastic Stem Cells/metabolism , Receptors, GABA-A/metabolism , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/metabolism , GABA-A Receptor Agonists/pharmacology , Epithelial Cell Adhesion Molecule/metabolism , Liver/cytology , Liver Neoplasms/metabolism , Phenotype , Stem Cells/drug effects , Neoplastic Stem Cells/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Biomarkers/metabolism , Fluorescent Antibody Technique , Immunomagnetic Separation , Receptors, GABA-A/drug effects , Receptors, GABA-A/genetics , Reverse Transcriptase Polymerase Chain Reaction , Protein Subunits , Liver Neoplasms/genetics , Membrane Potentials/drug effects
6.
Rev. Soc. Bras. Med. Trop ; 50(1): 99-103, Jan.-Feb. 2017. graf
Article in English | LILACS | ID: biblio-1041391

ABSTRACT

Abstract: INTRODUCTION: Infection with all serotypes of dengue virus (DV) results in augmented antigen presentation by MHC class I molecules. However, the upregulation of immunoproteasome subunits only results from infection with two serotypes. This study aims to elucidate changes in the expression of immunoproteasome subunits resulting from infection with DV, particularly DV serotype 2 (DV2). METHODS: HepG2 cells were grown in various culture milieu. Total cellular RNA and proteins were extracted and quantified. RESULTS: Results demonstrated sequestration of immunoproteasome subunits LMP2 and LMP7 in DV2-infected cells. CONCLUSIONS: This study provides insights into the mechanisms underlying immune evasion by DV.


Subject(s)
Humans , Dengue Virus/metabolism , Proteasome Endopeptidase Complex/metabolism , Gene Expression Regulation , Protein Subunits , Dengue Virus/classification , Hep G2 Cells , Serogroup
7.
Pakistan Journal of Pharmaceutical Sciences. 2017; 30 (3 [Special]): 1063-1067
in English | IMEMR | ID: emr-189311

ABSTRACT

The aim of this study was to observe the effect of Zhuang medicine medicated thread moxibustion on protomics in serum of postherpetic neuralgia patient with herpes zoster and discuss the action mechanism of Zhuang medicine medicated thread moxibustion in treating postherpetic neuralgia [PHN]


Methods: patients were divided into three groups in clinic, namely the pre-treatment group [n=20], post-treatment group [n=20] and healthy group [n=20], patients meeting corresponding conditions were recruited in the voluntary principle to accept PHN pain evaluation and protomics tests respectively


Results: compared with the pre-treatment group, visual analogue scale [VAS] in posttreatment group obviously reduced, protomics indexes like MFG-ES, Lymphotoxin beta/TNFSF3, IL-19, Neuritin, NCAM-1/CD56 and PECAM-1/CD31 obviously changed


Conclusions: the Zhuang medicine medicated thread moxibustion can treat the postherpetic neuralgia patient with herpes zoster, its internal mechanism is to possibly change the protomics indexes like MFG-ES, Lymphotoxin beta/TNFSF3, IL-19, Neuritin, NCAM-1/CD56 and PECAM-1/CD31


Subject(s)
Humans , Moxibustion , Neuralgia, Postherpetic , Herpes Zoster , Protein Subunits
8.
Electron. j. biotechnol ; 19(4): 44-51, July 2016. ilus
Article in English | LILACS | ID: lil-793952

ABSTRACT

Background: The acidic subunit of amarantin (AAC)-the predominant amaranth seed storage protein-has functional potential and its third variable region (VR) has been modified with antihypertensive peptides to improve this potential. Here, we modified the C-terminal in the fourth VR of AAC by inserting four VY antihypertensive peptides. This modified protein (AACM.4) was expressed in Escherichia coli. In addition, we also recombinantly expressed other derivatives of the amarantin protein. These include: unmodified amarantin acidic subunit (AAC); amarantin acidic subunit modified at the third VR with four VY peptides (AACM.3); and amarantin acidic subunit doubly modified, in the third VR with four VY peptides and in the fourth VR with the RIPP peptide (AACM.3.4). Results: E. coli BL21-CodonPlus (DE3)-RIL was the most favorable strain for the expression of proteins. After 6 h of induction, it showed the best recombinant protein titer. The AAC and AACM.4 were obtained at higher titers (0.56 g/L) while proteins modified in the third VR showed lower titers: 0.44 g/L and 0.33 g/L for AACM.3 and AACM.3.4, respectively. As these AAC variants were mostly expressed in an insoluble form, we applied a refolding protocol. This made it possible to obtain all proteins in soluble form. Modification of the VR 4 improves the thermal stability of amarantin acidic subunit; AAC manifested melting temperature (Tm) at 34°C and AACM.4 at 37.2°C. The AACM.3 and AACM.3.4 did not show transition curves. Conclusions: Modifications to the third VR affect the thermal stability of amarantin acidic subunit.


Subject(s)
Plant Proteins/metabolism , Amaranthus , Plant Proteins/isolation & purification , Plant Proteins/chemistry , Temperature , Protein Engineering , Blotting, Western , Bioreactors , Protein Subunits , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Protein Stability , Fermentation , Globulins
9.
Article in English | WPRIM | ID: wpr-285228

ABSTRACT

The ubiquitin-proteasome system plays a pivotal role in breast tumorigenesis by controlling transcription factors, thus promoting cell cycle growth, and degradation of tumor suppressor proteins. However, breast cancer patients have failed to benefit from proteasome inhibitor treatment partially due to proteasome heterogeneity, which is poorly understood in malignant breast neoplasm. Chemical crosslinking is an increasingly important tool for mapping protein three-dimensional structures and proteinprotein interactions. In the present study, two cross-linkers, bis (sulfosuccinimidyl) suberate (BS(3)) and its water-insoluble analog disuccinimidyl suberate (DSS), were used to map the subunit-subunit interactions in 20S proteasome core particle (CP) from MDA-MB-231 cells. Different types of gel electrophoresis technologies were used. In combination with chemical cross-linking and mass spectrometry, we applied these gel electrophoresis technologies to the study of the noncovalent interactions among 20S proteasome subunits. Firstly, the CP subunit isoforms were profiled. Subsequently, using native/SDSPAGE, it was observed that 0.5 mmol/L BS(3) was a relatively optimal cross-linking concentration for CP subunit-subunit interaction study. 2-DE analysis of the cross-linked CP revealed that α1 might preinteract with α2, and α3 might pre-interact with α4. Moreover, there were different subtypes of α1α2 and α3α4 due to proteasome heterogeneity. There was no significant difference in cross-linking pattern for CP subunits between BS(3) and DSS. Taken together, the gel-based characterization in combination with chemical cross-linking could serve as a tool for the study of subunit interactions within a multi-subunit protein complex. The heterogeneity of 20S proteasome subunit observed in breast cancer cells may provide some key information for proteasome inhibition strategy.


Subject(s)
Amino Acid Sequence , Breast Neoplasms , Drug Therapy , Genetics , Pathology , Cell Line, Tumor , Cross-Linking Reagents , Electrophoresis, Gel, Two-Dimensional , Female , Humans , Mass Spectrometry , Proteasome Endopeptidase Complex , Protein Binding , Protein Isoforms , Genetics , Protein Subunits , Genetics , Proteomics , Succinimides
10.
Article in English | WPRIM | ID: wpr-8374

ABSTRACT

PURPOSE: Dengue virus infection is now a global problem. Currently, there is no licensed vaccine or proven antiviral treatment against this virus. All four serotypes (1-4) of dengue virus can infect human. An effective dengue vaccine should be tetravalent to induce protective immune responses against all four serotypes. Most of dengue vaccine candidates are monovalent, or in the form of physically mixed multivalent formulations. Recently envelope protein domain III of virus is considered as a vaccine candidate, which plays critical roles in the most important viral activities. Development of a tetravalent protein subunit vaccine is very important for equal induction of immune system and prevention of unbalanced immunity. Here, we have presented and used a rational approach to design a tetravalent dengue vaccine candidate. MATERIALS AND METHODS: We designed a multi domain antigen by fusing four consensus domain III sequences together with appropriate hydrophobic linkers and used several types of bioinformatics software and neural networks to predict structural and immunological properties of the designed tetravalent antigen. RESULTS: We designed a tetravalent protein (EDIIIF) based on domain III of dengue virus envelope protein. According to the results of the bioinformatics analysis, the constructed models for EDIIIF protein were structurally stable and potentially immunogenic. CONCLUSION: The designed tetravalent protein can be considered as a potential dengue vaccine candidate. The presented approach can be used for rational design and in silico evaluation of chimeric dengue vaccine candidates.


Subject(s)
Computational Biology , Computer Simulation , Consensus , Dengue Virus , Dengue , Humans , Immune System , Protein Structure, Tertiary , Protein Subunits , Staphylococcal Protein A
11.
Rev. bras. anestesiol ; 65(3): 207-212, May-Jun/2015. tab, graf
Article in English | LILACS | ID: lil-748922

ABSTRACT

BACKGROUND AND OBJECTIVE: Postoperative pain treatment in mastectomy remains a major challenge despite the multimodal approach. The aim of this study was to investigate the analgesic effect of intravenous lidocaine in patients undergoing mastectomy, as well as the postoperative consumption of opioids. METHODS: After approval by the Human Research Ethics Committee of the Instituto de Medicina Integral Prof. Fernando Figueira in Recife, Pernambuco, a randomized, blind, controlled trial was conducted with intravenous lidocaine at a dose of 3 mg/kg infused over 1 h in 45 women undergoing mastectomy under general anesthesia. One patient from placebo group was. RESULTS: Groups were similar in age, body mass index, type of surgery, and postoperative need for opioids. Two of 22 patients in lidocaine group and three of 22 patients in placebo group requested opioid (p = 0.50). Pain on awakening was identified in 4/22 of lidocaine group and 5/22 of placebo group (p = 0.50); in the post-anesthetic recovery room in 14/22 and 12/22 (p = 0.37) of lidocaine and placebo groups, respectively. Pain evaluation 24 h after surgery showed that 2/22 and 3/22 patients (p = 0.50) of lidocaine and placebo groups, respectively, complained of pain. CONCLUSION: Intravenous lidocaine at a dose of 3 mg/kg administered over a period of an hour during mastectomy did not promote additional analgesia compared to placebo in the first 24 h, and has not decreased opioid consumption. However, a beneficial effect of intravenous lidocaine in selected and/or other therapeutic regimens patients cannot be ruled out. .


JUSTIFICATIVA E OBJETIVO: O tratamento da dor pós-operatória em mastectomia continua sendo um grande desafio apesar da abordagem multimodal. O objetivo deste estudo foi investigar o efeito analgésico da lidocaína intravenosa em pacientes submetidas a mastectomia, como também, o consumo de opioide pós-operatório. MÉTODOS: Após aprovação pelo comitê de ética e pesquisa em seres humanos do Instituto de Medicina Integral Prof. Fernando Figueira em Recife - Pernambuco foi realizado ensaio clínico aleatório encoberto placebo controlado com lidocaína intravenosa na dose de 3 mg/kg infundida em uma hora, em 45 mulheres submetidas a mastectomia sob anestesia geral. Excluída uma paciente do grupo placebo. RESULTADOS: Os grupos foram semelhantes quanto à idade, índice de massa corpórea, tipo de intervenção cirúrgica e necessidade de opioide no pós-operatório. Solicitaram opioide 2/22 pacientes nos grupos da lidocaína e 3/22 placebo (p = 0,50). Identificada a dor ao despertar em 4/22 no grupo lidocaína e 5/22 (p = 0,50) no grupo placebo; na sala de recuperação pós-anestésica em 14/22 e 12/22 (p = 0,37) nos grupos lidocaína e placebo respectivamente. Ao avaliar a dor 24 horas após o procedimento cirúrgico 3/22 e 2/22 (p = 0,50) das pacientes relataram dor em ambos os grupos respectivamente. CONCLUSÃO: A lidocaína intravenosa na dose de 3mg/kg administrada em um período de uma hora no transoperatório de mastectomia não promoveu analgesia adicional em relação ao grupo placebo nas primeiras 24 horas e não diminuiu o consumo de opioide. Contudo, um efeito benéfico da lidocaína intravenosa em pacientes selecionadas e/ou em outros regimes terapêuticos não pode ser descartado. .


JUSTIFICACIÓN Y OBJETIVO: El tratamiento del dolor postoperatorio en la mastectomía continúa siendo un gran reto a pesar del abordaje multimodal. El objetivo de este estudio fue investigar el efecto analgésico de la lidocaína intravenosa en pacientes sometidas a mastectomía, así como el consumo postoperatorio de opiáceos. MÉTODOS: Después de la aprobación por el Comité de Ética e Investigación en seres humanos del Instituto de Medicina Integral Prof. Fernando Figueira, en Recife, Pernambuco, se realizó un ensayo clínico aleatorizado, encubierto, placebo controlado con lidocaína intravenosa en una dosis de 3 mg/kg infundida en una hora, en 45 mujeres sometidas a mastectomía bajo anestesia general. Una paciente del grupo placebo fue excluida. RESULTADOS: Los grupos fueron similares en cuanto a la edad, índice de masa corporal, tipo de intervención quirúrgica y necesidad de opiáceos en el postoperatorio. Solicitaron opiáceos 2/22 pacientes en los grupos de la lidocaína y 3/22 placebo (p = 0,50). Fue identificado el dolor al despertar en 4/22 en el grupo lidocaína y 5/22 (p = 0,50) en el grupo placebo; en la sala de recuperación postanestésica en 14/22 y 12/22 (p = 0,37) en los grupos lidocaína y placebo, respectivamente. Al calcular el dolor 24 h después del procedimiento quirúrgico 3/22 y 2/22 (p = 0,50) de las pacientes relataron dolor en ambos grupos respectivamente. CONCLUSIÓN: La lidocaína intravenosa en una dosis de 3 mg/kg administrada en un período de una hora en el transoperatorio de mastectomía no generó analgesia adicional con relación al grupo placebo en las primeras 24 h y no disminuyó el consumo de opiáceos. Sin embargo, no puede ser descartado un efecto beneficioso de la lidocaína intravenosa en pacientes seleccionadas y/o en otros regímenes terapéuticos. .


Subject(s)
Humans , Metapneumovirus/genetics , Transcription, Genetic , Viral Proteins/chemistry , Amino Acid Sequence , Adenosine Monophosphate/metabolism , Crystallography, X-Ray , DNA , Edetic Acid/pharmacology , Molecular Dynamics Simulation , Molecular Sequence Data , Protein Binding , Protein Conformation , Protein Multimerization , Protein Stability , Protein Subunits/chemistry , RNA, Viral/metabolism , RNA, Viral/ultrastructure , Scattering, Small Angle , Solutions , Solvents , Viral Proteins/metabolism , Viral Proteins/ultrastructure , Zinc Fingers
12.
Braz. dent. j ; 26(2): 105-109, Mar-Apr/2015. tab, graf
Article in English | LILACS | ID: lil-741217

ABSTRACT

Dried, fresh and glycolic extracts of Zingiber officinale were obtained to evaluate the action against G. mellonella survival assay against Enterococcus faecalis infection. Eighty larvae were divided into: 1) E. faecalis suspension (control); 2) E. faecalis + fresh extract of Z. officinale (FEO); 3) E. faecalis + dried extract of Z. officinale (DEO); 4) E. faecalis + glycolic extract of Z. officinale (GEO); 5) Phosphate buffered saline (PBS). For control group, a 5 μL inoculum of standardized suspension (107 cells/mL) of E. faecalis (ATCC 29212) was injected into the last left proleg of each larva. For the treatment groups, after E. faecalis inoculation, the extracts were also injected, but into the last right proleg. The larvae were stored at 37 °C and the number of dead larvae was recorded daily for 168 h (7 days) to analyze the survival curve. The larvae were considered dead when they did not show any movement after touching. E. faecalis infection led to the death of 85% of the larvae after 168 h. Notwithstanding, in treatment groups with association of extracts, there was an increase in the survival rates of 50% (GEO), 61% (FEO) and 66% (DEO) of the larvae. In all treatment groups, the larvae exhibited a survival increase with statistically significant difference in relation to control group (p=0.0029). There were no statistically significant differences among treatment groups with different extracts (p=0.3859). It may be concluded that the tested extracts showed antimicrobial activity against E. faecalis infection by increasing the survival of Galleria mellonella larvae.


Extratos seco, fresco e glicólico de Zingiber officinale foram obtidos para avaliar suas ações por meio de ensaio de sobrevivência em G. mellonella contra infecção por Enterococcus faecalis. Oitenta larvas foram divididas em: 1) Suspensão de E. faecalis (controle); 2) E. faecalis + extrato fresco de Z. officinale (FEO); 3) E. faecalis + extrato seco de Z. officinale (DEO); 4) E. faecalis + extrato glicólico de Z. officinale (GEO); 5) Solução tampão fosfato salina (PBS). Para o grupo de controle, 5 µL de inóculo de suspensão padronizada (107 células/mL) de E. faecalis (ATCC 29212) foi injetado na última proleg esquerda de cada lagarta. Para os grupos com tratamento, após a injeção de E. faecalis, os extratos foram injetados na última proleg direita. Após as injeções, as lagartas foram armazenadas a 37 °C e o número de animais mortos foi registrado diariamente em 168 h (7 dias) para analisar a curva de sobrevivência. As lagartas foram consideradas mortas quando elas não mostraram qualquer movimento após o toque. A infecção por E. faecalis levou à morte de 85% das lagartas após 168 h. Não obstante, nos grupos de tratamento com associação dos extratos, houve um aumento nas taxas de sobrevivência de 50% (GEO), 61% (FEO) e 66% (DEO) das lagartas. Em todos os grupos com tratamento, as lagartas apresentaram um aumento na sobrevivência, com diferença estatisticamente significativa em relação ao grupo controle (p=0,0029). Não houve diferença estatisticamente significativa entre os tratamentos com os diferentes extratos (p=0,3859). Pode concluir-se que os extratos testados mostraram atividade antimicrobiana contra a infecção por E. faecalis, aumentando a sobrevivência das lagartas de G. mellonella.


Subject(s)
Humans , Receptors, GABA-A/chemistry , Binding Sites , Benzamidines/chemistry , Benzamidines/metabolism , Benzamidines/pharmacology , Conserved Sequence , Crystallography, X-Ray , Cell Membrane/chemistry , Cell Membrane/metabolism , Drug Design , GABA-A Receptor Agonists/chemistry , GABA-A Receptor Agonists/metabolism , GABA-A Receptor Agonists/pharmacology , Genetic Predisposition to Disease , Glycosylation , Models, Molecular , Mutation/genetics , Protein Structure, Quaternary , Protein Structure, Tertiary , Protein Subunits , Polysaccharides/chemistry , Polysaccharides/metabolism , Receptors, GABA-A/genetics , Synaptic Transmission
13.
Protein & Cell ; (12): 275-287, 2015.
Article in English | WPRIM | ID: wpr-757603

ABSTRACT

Pyruvate kinase isoform M2 (PKM2) converts phosphoenolpyruvate (PEP) to pyruvate and plays an important role in cancer metabolism. Here, we show that post-translational modifications and a patient-derived mutation regulate pyruvate kinase activity of PKM2 through modulating the conformation of the PKM2 tetramer. We determined crystal structures of human PKM2 mutants and proposed a "seesaw" model to illustrate conformational changes between an inactive T-state and an active R-state tetramers of PKM2. Biochemical and structural analyses demonstrate that PKM2(Y105E) (phosphorylation mimic of Y105) decreases pyruvate kinase activity by inhibiting FBP (fructose 1,6-bisphosphate)-induced R-state formation, and PKM2(K305Q) (acetylation mimic of K305) abolishes the activity by hindering tetramer formation. K422R, a patient-derived mutation of PKM2, favors a stable, inactive T-state tetramer because of strong intermolecular interactions. Our study reveals the mechanism for dynamic regulation of PKM2 by post-translational modifications and a patient-derived mutation and provides a structural basis for further investigation of other modifications and mutations of PKM2 yet to be discovered.


Subject(s)
Acetylation , Allosteric Regulation , Carrier Proteins , Chemistry , Genetics , Metabolism , Crystallography, X-Ray , Fructosediphosphates , Chemistry , Metabolism , Gene Expression , Humans , Kinetics , Membrane Proteins , Chemistry , Genetics , Metabolism , Models, Molecular , Mutation , Neoplasms , Genetics , Pathology , Phosphorylation , Protein Conformation , Protein Multimerization , Protein Processing, Post-Translational , Protein Subunits , Chemistry , Genetics , Metabolism , Thyroid Hormones , Chemistry , Genetics , Metabolism , Tumor Cells, Cultured
14.
Article in English | WPRIM | ID: wpr-264616

ABSTRACT

By now, the digestive stability experiments provided by most authoritative organizations are in vitro tests. Evaluating the protein digestive stability with in vivo models should be more objective. The present study aimed to verify the in vivo digestibility of soybean β-conglycinin β-subunit in Wuzhishan (WZS) minipigs. Three minipigs were surgically fitted with O-stomach and T-ileum cannulae and fed with soybean meals. According to SDS-PAGE, the 50 kD fraction of soybean β-conglycinin β-subunit persisted in the gastric fluid until 6 h after feeding, which was detected at 3 h and clearly visible at 4-6 h in the intestinal fluid. Western blot with anti-β-conglycinin β-subunit McAb confirmed it.


Subject(s)
Animals , Antigens, Plant , Chemistry , Metabolism , Digestion , Physiology , Globulins , Chemistry , Metabolism , Male , Protein Subunits , Chemistry , Metabolism , Seed Storage Proteins , Chemistry , Metabolism , Soybean Proteins , Chemistry , Metabolism , Swine , Swine, Miniature , Physiology
15.
Biomédica (Bogotá) ; 34(4): 598-604, oct.-dic. 2014. graf, tab
Article in Spanish | LILACS | ID: lil-730944

ABSTRACT

Institución donde se ejecutó el trabajo: Programa de Estudio y Control de Enfermedades Tropicales, PECET, Unidad de Malacología Médica y Trematodos (UMMT), Sede de Investigación Universitaria, Universidad de Antioquia, Medellín, Colombia Introducción. La fasciolosis es la enfermedad transmitida por vectores con mayor distribución latitudinal, longitudinal y altitudinal, debido a la capacidad colonizadora del parásito Fasciola hepatica y de sus huéspedes intermediarios, los moluscos limneidos. Estos caracoles se investigan por su importancia epidemiológica, pero su identificación taxonómica es difícil por la similitud fenotípica entre especies. En este sentido, con respecto a Lymnaea cousini , un huésped de F. hepatica en Colombia, existe incertidumbre en razón de su similitud morfológica con L. meridensis , descrita recientemente en Venezuela. Objetivo. Confirmar con el marcador del gen de la citocromo oxidasa I en el ADN mitocondrial COI (ADNmt), el estatus taxonómico de ejemplares morfológicamente caracterizados como L. cousini provenientes de Nariño, Norte de Santander y Santander (Colombia), depositados en la Colección de Moluscos Vectores de la Universidad de Antioquia, VHET N° 37. Materiales y métodos. Para la amplificación del COI mitocondrial, se extrajo ADN total del pie de cada ejemplar con el estuche DNeasy Blood and Tissue (Qiagen ® ). Los productos amplificados se enviaron a secuenciar a Macrogen Inc., Corea. Las 27 secuencias generadas en esta investigación se compararon con secuencias publicadas en el GenBank, incluidas las secuencias de la localidad tipo de L. cousini. Resultados. Se encontraron dos nuevos haplotipos de L. cousini para Colombia. Los especímenes de Nariño correspondían al haplotipo A, referenciado en Ecuador, y los especímenes de Santander y Norte de Santander, a un nuevo haplotipo al que se denominó D. Conclusión. Mediante el marcador mitocondrial del COI , se confirmó que los especímenes pertenecían a la especie L. cousini . Con el hallazgo se duplicó el número de haplotipos conocidos de la especie en Colombia y se amplió su distribución geográfica al suroeste y nordeste de la región altoandina colombiana.


Introduction: Fasciolosis is the disease transmitted by vectors with the highest latitudinal, longitudinal, and altitudinal distribution due to the colonizing capacity of the parasite Fasciola hepatica and its intermediate hosts, Lymnaeidae mollusks. These snails are under research due to their epidemiological importance, but their taxonomic identification is difficult given their interspecific phenotypical similarity. For this reason, there is uncertainty regarding Lymnaea cousini -a host of F. hepatica in Colombia- due to the morphological similarity it has with Lymnaea meridensis , recently described for Venezuela. Objective: To confirm with the COI marker (ADNmt) the taxonomic status of individuals morphologically identified as L. cousini from Nariño, Norte de Santander, and Santander (Colombia), deposited in the Vector Mollusks Collection VHET No. 37 of Universidad de Antioquia. Materials and methods: The amplification of the mitochondrial COI required total DNA extraction of each individual´s foot using the DNeasy Blood and Tissue Kit (Qiagen®). Products amplified were sent for sequencing to Macrogen Inc., Korea. Twenty seven sequences generated in this research were compared to sequences published in the GenBank, including sequences of the type locality of L. cousini . Results: Two new haplotypes of L. cousini were obtained for Colombia. Specimens from Nariño correspond to haplotype A, referenced for Ecuador, and specimens from Santander and Norte de Santander belong to a new haplotype we called haplotype D. Conclusion : By using the mitochondrial COI marker, we confirmed that the species under study did correspond to L. cousini . The number of known haplotypes of the species for Colombia has been duplicated and its geographical distribution has been extended to the southwest and northeast of the Colombian high Andean region.


Subject(s)
Animals , Disease Vectors/classification , Electron Transport Complex IV/analysis , Fasciola hepatica , Lymnaea/classification , Base Sequence , Biomarkers , Colombia , DNA , DNA, Mitochondrial/genetics , Electron Transport Complex IV/genetics , Haplotypes/genetics , Lymnaea/enzymology , Lymnaea/genetics , Molecular Sequence Data , Phylogeny , Protein Subunits , Sequence Alignment , Sequence Homology, Nucleic Acid
16.
J. bras. nefrol ; 36(4): 512-518, Oct-Dec/2014. tab
Article in English | LILACS | ID: lil-731151

ABSTRACT

Introduction: Tuberculosis is a common opportunistic infection in renal transplant patients. Objective: To obtain a clinical and laboratory description of transplant patients diagnosed with tuberculosis and their response to treatment during a period ranging from 2005 to 2013 at the Pablo Tobón Uribe Hospital. Methods: Retrospective and descriptive study. Results: In 641 renal transplants, tuberculosis was confirmed in 12 cases. Of these, 25% had a history of acute rejection, and 50% had creatinine levels greater than 1.5 mg/dl prior to infection. The disease typically presented as pulmonary (50%) and disseminated (33.3%). The first phase of treatment consisted of 3 months of HZRE (isoniazid, pyrazinamide, rifampicin and ethambutol) in 75% of the cases and HZME (isoniazid, pyrazinamide, moxifloxacin and ethambutol) in 25% of the cases. During the second phase of the treatment, 75% of the cases received isoniazid and rifampicin, and 25% of the cases received isoniazid and ethambutol. The length of treatment varied between 6 and 18 months. In 41.7% of patients, hepatotoxicity was associated with the beginning of anti-tuberculosis therapy. During a year-long follow-up, renal function remained stable, and the mortality rate was 16.7%. Conclusion: Tuberculosis in the renal transplant population studied caused diverse nonspecific symptoms. Pulmonary and disseminated tuberculosis were the most frequent forms and required prolonged treatment. Antituberculosis medications had a high toxicity and mortality. This infection must be considered when patients present with a febrile syndrome of unknown origin, especially during the first year after renal transplant. .


Introdução: A tuberculose é uma infecção oportunista comum em pacientes transplantados renais. Objetivo: Oferecer uma descrição clínica e laboratorial de pacientes transplantados com diagnóstico de tuberculose e sua resposta ao tratamento durante o período entre 2005 e 2013 no Hospital Pablo Tobón Uribe. Métodos: Estudo retrospectivo descritivo. Resultados: Em 641 transplantes renais, a tuberculose foi confirmada em 12 pacientes. Destes, 25% tinham histórico de rejeição aguda e 50% apresentaram níveis de creatinina superiores a 1,5 mg/dl antes da infecção. A patologia geralmente se apresentava como pulmonar (50%) e disseminada (33,3%). A primeira fase do tratamento consistiu de três meses de HZRE (isoniazida, pirazinamida, rifampicina e etambutol) em 75% dos casos e HZME (isoniazida, pirazinamida, moxifloxacina e etambutol) em 25% dos pacientes. Durante a segunda fase do tratamento, 75% dos pacientes receberam isoniazida e rifampicina e 25% isoniazida e etambutol. A duração do tratamento variou entre seis e 18 meses. Em 41,7% dos pacientes, hepatotoxicidade foi associada ao início do tratamento da tuberculose. Durante o seguimento de um ano a função renal manteve-se estável e a taxa de mortalidade foi de 16,7%. Conclusão: A tuberculose foi responsável por diversos sintomas inespecíficos na população de transplantados renais estudada. Tuberculose pulmonar e disseminada foram as formas mais frequentes de acometimento e necessitaram de tratamento prolongado. Medicamentos contra a tuberculose apresentaram alta toxicidade e mortalidade. Esta infecção deve ser considerada quando o paciente apresenta síndrome febril de origem desconhecida, especialmente durante o primeiro ano após o transplante renal. .


Subject(s)
Animals , Female , Male , Mice , Locus Coeruleus/drug effects , Narcotics/pharmacology , Neural Inhibition/drug effects , Neurons/drug effects , Potassium Channels/metabolism , Barium/pharmacology , Calcium/metabolism , Enkephalin, Methionine/pharmacology , G Protein-Coupled Inwardly-Rectifying Potassium Channels , GTP-Binding Proteins/metabolism , Heterozygote , Homozygote , Ion Channel Gating/drug effects , Ion Channel Gating/physiology , Locus Coeruleus/cytology , Locus Coeruleus/physiology , Mice, Knockout , Membrane Potentials/drug effects , Membrane Potentials/physiology , Neural Inhibition/physiology , Neurons/physiology , Patch-Clamp Techniques , Protein Subunits , Potassium Channel Blockers/pharmacology , Potassium Channels, Inwardly Rectifying/antagonists & inhibitors , Potassium Channels, Inwardly Rectifying/deficiency , Potassium Channels, Inwardly Rectifying/genetics , Potassium Channels, Inwardly Rectifying/metabolism , Potassium Channels/deficiency , Potassium Channels/genetics
17.
Article in English | WPRIM | ID: wpr-14498

ABSTRACT

Several studies have reported that the citrus red mites Panonychus citri were an important allergen of citrus-cultivating farmers in Jeju Island. The aim of the present study was to purify and assess properties of a cysteine protease from the mites acting as a potentially pathogenic factor to citrus-cultivating farmers. A cysteine protease was purified using column chromatography of Mono Q anion exchanger and Superdex 200 HR gel filtration. It was estimated to be 46 kDa by gel filtration column chromatography and consisted of 2 polypeptides, at least. Cysteine protease inhibitors, such as trans poxy-succinyl-L-leucyl-amido (4-guanidino) butane (E-64) and iodoacetic acid (IAA) totally inhibited the enzyme activities, whereas serine or metalloprotease inhibitors did not affect the activities. In addition, the purified enzyme degraded human IgG, collagen, and fibronectin, but not egg albumin. From these results, the cysteine protease of the mites might be involved in the pathogenesis such as tissue destruction and penetration instead of nutrient digestion.


Subject(s)
Animals , Chromatography, Gel , Chromatography, Ion Exchange , Collagen/metabolism , Cysteine Proteases/chemistry , Cysteine Proteinase Inhibitors/metabolism , Fibronectins/metabolism , Humans , Immunoglobulin G/metabolism , Molecular Weight , Protein Subunits/chemistry , Proteolysis , Substrate Specificity , Tetranychidae/enzymology
18.
Chinese Journal of Virology ; (6): 554-560, 2014.
Article in Chinese | WPRIM | ID: wpr-280328

ABSTRACT

To compare different adjuvant formulation and explore the impact of Calcineurin B subunit(CnB) as adjuvant with a novel HBV protein particle (HBSS1) vaccine in mice, female C57BL/6 mice were immunized HBSS1 with Al(OH)3 only, or a normal dose (5 μg) CnB only, or (CnB+ Al(OH)3) mixture as the adjuvant. All immunized groups were primed twice at 4-week intervals; followed by boosting with recombinant adenoviral based HBV vaccine(rAdSS1) at 10-week intervals. We detected the antigen specific humoral response in mice, including total IgG antibody and IgG subtyping. Then, we characterized the specific cell-mediated immune (CMI) response by detection of γ-interferon secreting splenocytes after stimulaton with S or PreS1 peptide pools. No enhancement of immunity was found among the mice with 5 μg of CnB alone or combined with Al(OH), adjuvanted vaccine,which could not induce higher level of anti-PreS1 and anti-S antibodies and CMI than that of HBSS1 alone or Al(OH)3 adjuvanted vaccines. We concluded that CnB is not an effective adjuvant for a novel HBV subunit vaccine.


Subject(s)
Adjuvants, Immunologic , Pharmacology , Animals , Calcineurin , Pharmacology , Female , Hepatitis B Vaccines , Allergy and Immunology , Mice , Mice, Inbred C57BL , Protein Subunits
19.
Article in English | WPRIM | ID: wpr-103954

ABSTRACT

A total of 16 Taenia multiceps isolates collected from naturally infected sheep or goats in Gansu Province, China were characterized by sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The complete cox1 gene was amplified for individual T. multiceps isolates by PCR, ligated to pMD18T vector, and sequenced. Sequence analysis indicated that out of 16 T. multiceps isolates 10 unique cox1 gene sequences of 1,623 bp were obtained with sequence variation of 0.12-0.68%. The results showed that the cox1 gene sequences were highly conserved among the examined T. multiceps isolates. However, they were quite different from those of the other Taenia species. Phylogenetic analysis based on complete cox1 gene sequences revealed that T. multiceps isolates were composed of 3 genotypes and distinguished from the other Taenia species.


Subject(s)
Animals , China , Cluster Analysis , Cysticercosis/parasitology , DNA, Helminth/chemistry , DNA, Mitochondrial/chemistry , Electron Transport Complex IV/genetics , Genetic Variation , Goat Diseases/parasitology , Goats , Phylogeny , Polymerase Chain Reaction , Protein Subunits/genetics , Sequence Analysis, DNA , Sheep , Sheep Diseases/parasitology , Taenia/classification
20.
Protein & Cell ; (12): 911-920, 2013.
Article in English | WPRIM | ID: wpr-757559

ABSTRACT

Mediator is a highly conserved large protein complex (25 proteins, >1000 kDa) and preeminently responsible for eukaryotic transcription, which contains a dissociable 'Cdk8 module'. Although increasing evidence demonstrates that Cdk8 module plays both positive and negative roles in transcription regulation, the detailed structure, and subunit organization, molecular mechanism how it regulates transcription remain elusive. Here we used single-particle electron microscopy to characterize the structure and subunit organization of the Cdk8 module and illuminated the substantial mobility of the Med13 subunit results in the structural flexibility. The Cdk8 module interaction with core Mediator is concurrent with active transcription in vivo. An interaction with the Cdk8 module induces core Mediator into very extended conformation in vitro, which is presumed to be an active functional state of Mediator. Taken together, our results illuminated the detailed architecture of Cdk8 module, and suggested the Cdk8 module could positively regulate transcription by modulating Mediator conformation.


Subject(s)
Cyclin-Dependent Kinase 8 , Chemistry , Humans , Mediator Complex , Chemistry , Models, Molecular , Protein Conformation , Protein Subunits , Chemistry , Saccharomyces cerevisiae Proteins , Chemistry
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