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1.
Biochem Mol Biol Educ ; 47(3): 249-256, 2019 05.
Article in English | MEDLINE | ID: mdl-30779452

ABSTRACT

The bibliometric analysis of the Brazilian periodical Journal of Biochemistry Education (JBE) covered the 117 articles published in 15 volumes in the period 2001-2017. Our results showed a positive trend in JBE publications with a significant increase in the number of articles since 2014, which can be related to the increase in research groups working in this area. The Southeast region of Brazil was the most productive one mainly due to the contribution of papers from institutions located in the State of São Paulo. Only four articles aimed the undergraduate courses (87.0%) showing methodological approaches to teach biochemistry (51.3%) and laboratory exercises (18.8%) among others. Most of the 332 authors contributed to a single article (87.7%) and just 3% of them published more than twice in JBE. The majority of the JBE articles had at least one citation in Google Scholar. There is also a great variety in the references used by the authors. Our analysis showed that JBE is an important peer reviewed publication aimed to improve teaching and learning of Biochemistry in Brazil. © 2019 International Union of Biochemistry and Molecular Biology, 47(3):249-256, 2019.


Subject(s)
Bibliometrics , Biomedical Research/education , Learning , Publications , Teaching/education , Brazil , Laboratories
2.
PeerJ ; 4: e2670, 2016.
Article in English | MEDLINE | ID: mdl-27867765

ABSTRACT

BACKGROUND: Hepatitis C virus (HCV) core protein, in addition to its structural role to form the nucleocapsid assembly, plays a critical role in HCV pathogenesis by interfering in several cellular processes, including microRNA and mRNA homeostasis. The C-terminal truncated HCV core protein (C124) is intrinsically unstructured in solution and is able to interact with unspecific nucleic acids, in the micromolar range, and to assemble into nucleocapsid-like particles (NLPs) in vitro. The specificity and propensity of C124 to the assembly and its implications on HCV pathogenesis are not well understood. METHODS: Spectroscopic techniques, transmission electron microscopy and calorimetry were used to better understand the propensity of C124 to fold or to multimerize into NLPs when subjected to different conditions or in the presence of unspecific nucleic acids of equivalent size to cellular microRNAs. RESULTS: The structural analysis indicated that C124 has low propensity to self-folding. On the other hand, for the first time, we show that C124, in the absence of nucleic acids, multimerizes into empty NLPs when subjected to a pH close to its isoelectric point (pH ≈ 12), indicating that assembly is mainly driven by charge neutralization. Isothermal calorimetry data showed that the assembly of NLPs promoted by nucleic acids is enthalpy driven. Additionally, data obtained from fluorescence correlation spectroscopy show that C124, in nanomolar range, was able to interact and to sequester a large number of short unspecific nucleic acids into NLPs. DISCUSSION: Together, our data showed that the charge neutralization is the major factor for the nucleocapsid-like particles assembly from C-terminal truncated HCV core protein. This finding suggests that HCV core protein may physically interact with unspecific cellular polyanions, which may correspond to microRNAs and mRNAs in a host cell infected by HCV, triggering their confinement into infectious particles.

3.
Methods Enzymol ; 567: 237-56, 2016.
Article in English | MEDLINE | ID: mdl-26794357

ABSTRACT

Isothermal titration calorimetry (ITC) is a label-free technique that allows the direct determination of the heat absorbed or released in a reaction. Frequently used to determining binding parameters in biomolecular interactions, it is very useful to address enzyme-catalyzed reactions as both kinetic and thermodynamic parameters can be obtained. Since calorimetry measures the total heat effects of a reaction, it is important to consider the contribution of the heat of protonation/deprotonation that is possibly taking place. Here, we show a case study of the reaction catalyzed by the glucose-6-phosphate dehydrogenase (G6PD) from Leuconostoc mesenteroides. This enzyme is able to use either NAD(+) or NADP(+) as a cofactor. The reactions were done in five buffers of different enthalpy of protonation. Depending on the buffer used, the observed calorimetric enthalpy (ΔH(cal)) of the reaction varied from -22.93 kJ/mol (Tris) to 19.37 kJ/mol (phosphate) for the NADP(+)-linked reaction, and -11.67 kJ/mol (Tris) to 7.32 kcal/mol or 30.63 kJ/mol (phosphate) for the NAD(+) reaction. We will use this system as an example of how to extract proton-independent reaction enthalpies from kinetic data to ensure that the reported accurately represent the intrinsic heat of reaction.


Subject(s)
Calorimetry/methods , Glucosephosphate Dehydrogenase/metabolism , Entropy , Leuconostoc/enzymology
4.
Biochim Biophys Acta ; 1864(4): 359-71, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26773352

ABSTRACT

Membrane fusion is a highly regulated process that allows enveloped viruses to enter cells and replicate. Viral glycoproteins trigger membrane fusion by means of internal sequences known as fusion peptides. The hepatitis C virus (HCV) genome encodes the envelope glycoproteins E1 and E2, but their specific roles in the fusion step and the localization of the fusion peptide remain uncharacterized. Here, we studied the biophysics of the interactions between the glycoprotein E2 peptide HCV421-445 and four different micellar systems providing ionic, non-ionic and zwitterionic surfaces to investigate the importance of electrostatic interactions for peptide-membrane binding. Circular dichroism, fluorescence spectroscopy and calorimetry were used to characterize peptide-micelle interactions and structural changes. Fluorescence quenching showed that HCV421-445 interacts with SDS or CTAB ionic, n-OGP non-ionic and DPC zwitterionic micelles. The indole ring of Trp seems to anchor the peptide in micelles. Trp residues seem to be more deeply inserted in ionic and non-ionic micelles where peptide interactions are more stable than with DPC zwitterionic micelles. The interaction with zwitterionic micelles appears to occur at the surface. Both interaction types are exothermic because of peptide-micelle interactions and a gain of secondary structure in the helical conformation. HCV421-445 interacts with detergent monomers and micelles. Peptide-micelle interaction is pH-independent. HCV421-445 interacts with membranes, promoting aggregation and coalescence of vesicles with content leakage, suggesting that HCV421-445 may participate in membrane fusion. This structural characterization contributes to our understanding of the molecular process that promotes fusion, which is important in the further development of new antiviral therapies.


Subject(s)
Hepacivirus/chemistry , Viral Envelope Proteins/chemistry , Amino Acid Sequence , Lipid Bilayers/chemistry , Micelles , Molecular Sequence Data , Protein Structure, Secondary , Spectrometry, Fluorescence , Thermodynamics
5.
Immunotherapy ; 7(7): 721-38, 2015.
Article in English | MEDLINE | ID: mdl-26250414

ABSTRACT

In recent years, the treatment of metastatic colorectal cancer (mCRC) has evolved significantly with the increase of new therapeutic options, leading to an improved median survival for these patients. In particular, the identification of molecular targets in tumor cells has led to the introduction of biological drugs for the treatment of mCRC. Panitumumab is a fully human monoclonal antibody that binds the EGF receptor of tumor cells and inhibits downstream cell signaling with antitumor effect on inhibition of tumor growth. Its use has been approved by randomized clinical trials as monotherapy in chemorefractory patients or combined with chemotherapy in the treatment of RAS wild-type mCRC, where it demonstrated a significant improvement in survival and response rate. The purpose of this review is to analyze the use and efficacy profile of panitumumab, particularly focusing on recently reported data on its use, and future perspectives in patients with mCRC.


Subject(s)
Antibodies, Monoclonal/therapeutic use , Colorectal Neoplasms/drug therapy , ErbB Receptors/antagonists & inhibitors , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/pathology , ErbB Receptors/metabolism , Female , Humans , Male , Neoplasm Metastasis , Panitumumab , Randomized Controlled Trials as Topic
6.
Arch Insect Biochem Physiol ; 90(2): 104-15, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26111116

ABSTRACT

Lipid peroxidation is promoted by the quasi-lipoxygenase (QL) activity of heme proteins and enhanced by the presence of free calcium. Unlike mammalian plasma, the hemolymph of Rhodnius prolixus, a vector of Chagas disease, contains both a free heme-binding protein (RHBP) and circulating lipoproteins. RHBP binds and prevents the heme groups of the proteins from participating in lipid peroxidation reactions. Herein, we show that despite being bound to RHBP, heme groups promote lipid peroxidation through a calcium-dependent QL reaction. This reaction is readily inhibited by the presence of ethylene glycol tetraacetic acid (EGTA), the antioxidant butylated hydroxytoluene or micromolar levels of the main yolk phosphoprotein vitellin (Vt). The inhibition of lipid peroxidation is eliminated by the in vitro dephosphorylation of Vt, indicating that this reaction depends on the interaction of free calcium ions with negatively charged phosphoamino acids. Our results demonstrate that calcium chelation mediated by phosphoproteins occurs via an antioxidant mechanism that protects living organisms from lipid peroxidation.


Subject(s)
Calcium/metabolism , Carrier Proteins/metabolism , Hemeproteins/metabolism , Lipid Peroxidation , Rhodnius/metabolism , Vitellins/metabolism , Animals , Female , Heme-Binding Proteins , Hemolymph/metabolism , Insect Proteins/metabolism , Rabbits
7.
Springerplus ; 4: 255, 2015.
Article in English | MEDLINE | ID: mdl-26085975

ABSTRACT

PURPOSE: Evidences have shown that neutrophil-to-lymphocyte ratio (NLR) has a prognostic value in patients with cancer. We wanted to test the prognostic significance of NLR in prostatic cancer of patients who are candidate to radical prostatectomy. METHODS: We have considered 731 patients. Complete demographic data including age, tumor stage, Gleason score, complete blood count and serum biochemical profile were collected. Pre-treatment percentage of neutrophils and NLR were considered, and correlated with patients data and recurrence free survival. RESULTS: 389 patients were evaluated, mean age 65 years, mean follow-up 51.5 months, mean recurrence free survival 51.3 months. Total neutrophil count does not correlate with biochemical recurrence and disease free survival. Patients with a value higher of 60% of neutrophils are more likely to have a recurrence. Patients with a total lymphocyte count <1,500 have a higher rate of relapse. NLR was not correlated with baseline total PSA, with Gleason score and with pathological stage; patients with a NLR >3 has a higher incidence of recurrence. In multivariate analysis including age, total PSA and NLR, NLR is the most important factor able to predict recurrence. There are some limitations to this study; first, this is a retrospective study, and the total number of patients analyzed is relatively small. CONCLUSIONS: Our study suggests that pre-treatment NLR may be associated with disease free survival in patients with prostate cancer, and could be introduced in clinical practice. NLR has the advantage of low economic cost and wide availability.

8.
Med. intensiva ; 32(4): [1-7], 20150000. tab, fig
Article in Spanish | LILACS | ID: biblio-884187

ABSTRACT

La parálisis periódica tirotóxica hipocalémica es una patología de muy baja frecuencia en la Argentina; no obstante, es muy importante tenerla en cuenta como urgencia, ya que puede ser fatal si el diagnóstico y el tratamiento correctos no son oportunos. El objetivo de esta investigación bibliográfica es destacar el aumento de la incidencia de una enfermedad potencialmente fatal y, a la vez, fácilmente reversible con el tratamiento adecuado y oportuno, en las Salas de Guardia de nuestro país, a fin de que se la tenga en cuenta en las sospechas diagnósticas de emergencia y el aporte que puede hacer el bioquímico desde un Servicio de Urgencia, para ayudar al diagnóstico en forma rápida y certera.(AU)


Hypokalemic thyrotoxic periodic paralysis is a disease of very low frequency in Argentina; however, it is very important to take it into account as an emergency, because it can be fatal without a timely diagnosis and treatment. The objective of this research is to highlight its increased incidence in the emergency rooms of our country (it is a potentially fatal disease, but at the same time it is easily reversible with a proper and timely treatment), in order to include it in the emergency diagnostic suspicion and the biochemist contribution in an emergency service to reach a quick and accurate diagnosis.(AU)


Subject(s)
Humans , Thyroiditis , Hypokalemic Periodic Paralysis , Hyperthyroidism , Hypokalemia
9.
Biochim Biophys Acta ; 1828(11): 2419-27, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23792068

ABSTRACT

rBPI21 belongs to the antimicrobial peptide and protein (AMP) family. It has high affinity for lipopolysaccharide (LPS), acting mainly against Gram-negative bacteria. This work intends to elucidate the mechanism of action of rBPI21 at the membrane level. Using isothermal titration calorimetry, we observed that rBPI21 interaction occurs only with negatively charged membranes (mimicking bacterial membranes) and is entropically driven. Differential scanning calorimetry shows that membrane interaction with rBPI21 is followed by an increase of rigidity on negatively charged membrane, which is corroborated by small angle X-ray scattering (SAXS). Additionally, SAXS data reveal that rBPI21 promotes the multilamellarization of negatively charged membranes. The results support the proposed model for rBPI21 action: first it may interact with LPS at the bacterial surface. This entropic interaction could cause the release of ions that maintain the packed structure of LPS, ensuring peptide penetration. Then, rBPI21 may interact with the negatively charged leaflets of the outer and inner membranes, promoting the interaction between the two bacterial membranes, ultimately leading to cell death.


Subject(s)
Antimicrobial Cationic Peptides/chemistry , Cell Membrane/drug effects , Recombinant Proteins/chemistry , Antimicrobial Cationic Peptides/pharmacology , Calorimetry , Gram-Negative Bacteria/drug effects , Lipopolysaccharides/pharmacology , Recombinant Proteins/pharmacology , Scattering, Small Angle , X-Ray Diffraction
10.
Br J Cancer ; 108(5): 1126-32, 2013 Mar 19.
Article in English | MEDLINE | ID: mdl-23511629

ABSTRACT

BACKGROUND: Currently, sunitinib represents one of the therapeutic strongholds for renal cell carcinoma, but the criteria for treatment selection are lacking. We assessed the role of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) polymorphisms in the prediction of the clinical outcome in metastatic renal cell carcinoma (mRCC) patients. METHODS: A total of 84 tumour samples from mRCC patients receiving first-line sunitinib were tested for VEGF and VEGFR single-nucleotide polymorphisms (SNPs). The SNP results were correlated with progression-free survival (PFS) and overall survival (OS). RESULTS: Median PFS was 8.22 months, although whereas median OS was 32.13 months. The VEGF A rs833061 resulted significant in PFS (17 vs 4 months; P<0.0001) and OS (38 vs 10 months; P<0.0001). The VEGF A rs699947 was significant for PFS (18 vs 4 months; P=0.0001) and OS (37 vs 16 months; P<0.0001). The VEGF A rs2010963 was significant in PFS (18 vs 8 vs 2 months; P=0.0001) and OS (31 vs 36 vs 9 months; P=0.0045). The VEGR3 rs6877011 was significant in PFS (12 vs 4 months; P=0.0075) and OS (36 vs 17 months; P=0.0001). At multivariate analysis, rs833061, rs2010963 and rs68877011 were significant in PFS, and rs833061 and rs68877011 were independent factors in OS. CONCLUSIONS: In our analysis, patients with TT polymorphism of rs833061, CC polymorphism of rs699947, CC polymorphism of rs2010963 and CG polymorphism of rs6877011 seem to have a worse PFS and OS when receiving first-line sunitinib.


Subject(s)
Antineoplastic Agents/therapeutic use , Carcinoma, Renal Cell/genetics , Indoles/therapeutic use , Kidney Neoplasms/genetics , Polymorphism, Single Nucleotide , Pyrroles/therapeutic use , Receptors, Vascular Endothelial Growth Factor/genetics , Vascular Endothelial Growth Factor A/genetics , Aged , Aged, 80 and over , Carcinoma, Renal Cell/drug therapy , Carcinoma, Renal Cell/mortality , Carcinoma, Renal Cell/pathology , Disease-Free Survival , Female , Humans , Kidney Neoplasms/drug therapy , Kidney Neoplasms/mortality , Kidney Neoplasms/pathology , Male , Middle Aged , Neoplasm Metastasis , Sunitinib , Treatment Outcome
11.
PLoS One ; 7(10): e47596, 2012.
Article in English | MEDLINE | ID: mdl-23094066

ABSTRACT

Membrane fusion is a crucial step in flavivirus infections and a potential target for antiviral strategies. Lipids and proteins play cooperative roles in the fusion process, which is triggered by the acidic pH inside the endosome. This acidic environment induces many changes in glycoprotein conformation and allows the action of a highly conserved hydrophobic sequence, the fusion peptide (FP). Despite the large volume of information available on the virus-triggered fusion process, little is known regarding the mechanisms behind flavivirus-cell membrane fusion. Here, we evaluated the contribution of a natural single amino acid difference on two flavivirus FPs, FLA(G) ((98)DRGWGNGCGLFGK(110)) and FLA(H) ((98)DRGWGNHCGLFGK(110)), and investigated the role of the charge of the target membrane on the fusion process. We used an in silico approach to simulate the interaction of the FPs with a lipid bilayer in a complementary way and used spectroscopic approaches to collect conformation information. We found that both peptides interact with neutral and anionic micelles, and molecular dynamics (MD) simulations showed the interaction of the FPs with the lipid bilayer. The participation of the indole ring of Trp appeared to be important for the anchoring of both peptides in the membrane model, as indicated by MD simulations and spectroscopic analyses. Mild differences between FLA(G) and FLA(H) were observed according to the pH and the charge of the target membrane model. The MD simulations of the membrane showed that both peptides adopted a bend structure, and an interaction between the aromatic residues was strongly suggested, which was also observed by circular dichroism in the presence of micelles. As the FPs of viral fusion proteins play a key role in the mechanism of viral fusion, understanding the interactions between peptides and membranes is crucial for medical science and biology and may contribute to the design of new antiviral drugs.


Subject(s)
Flavivirus/chemistry , Lipid Bilayers/chemistry , Peptides/chemical synthesis , Tryptophan/chemistry , Viral Fusion Proteins/chemistry , Amino Acid Sequence , Circular Dichroism , Hydrogen-Ion Concentration , Hydrophobic and Hydrophilic Interactions , Membrane Fusion , Micelles , Molecular Dynamics Simulation , Molecular Sequence Data , Protein Conformation , Spectrometry, Fluorescence , Static Electricity
12.
Opt Express ; 20(4): 4444-53, 2012 Feb 13.
Article in English | MEDLINE | ID: mdl-22418204

ABSTRACT

The refractive indexes, material attenuation and damage fractions of a multi-step ion implanted Lithium Niobate (LiNbO3) waveguide were analyzed as functions of the annealing temperatures. An almost flat damage depth profile was designed to reduce the uncertainties related to the indexes profile shape, thus providing a better test-case for the characterizations. The measurements performed on the fabricated optical waveguides confirmed the predicted step-index profiles showing that the light is confined inside the damaged layer. The low measured attenuation (less than 0.8 dB/cm @ 632.8 nm) makes the obtained waveguide attractive for device fabrication.

13.
Br J Cancer ; 106(5): 799-804, 2012 Feb 28.
Article in English | MEDLINE | ID: mdl-22315053

ABSTRACT

BACKGROUND: Lactate dehydrogenase (LDH) represents a predictive factor in colorectal cancer patients treated with the angiogenesis inhibitor PTK/ZK. We explored the role of pre-treatment LDH serum levels in colorectal cancer patients receiving first-line bevacizumab. METHODS: Metastatic colorectal cancer treated with first-line bevacizumab was eligible. A control group including all consecutive patients treated with chemotherapy alone was also considered. Pre-treatment LDH serum levels were collected for all cases. RESULTS: Median progression-free survival (PFS) in the control group for patients with high and low LDH levels was 4.2 and 8 months, respectively (P=0.0003). Median overall survival (OS) was 19.6 and 34.9 months for patients with high and low LDH levels, respectively (P=0.0014). In the bevacizumab group, partial responses were seen in 14 (58%) high-LDH and 8 (14%) low-LDH patients (P=0.0243), respectively, median PFS was 7.3 and 8.5 months, respectively (P=0.2), and median OS was 22 and 26.6 months, respectively (P=0.7). CONCLUSION: High LDH levels correlated with worse prognosis. Bevacizumab seemed capable of improving clinical outcome in this specific group of patients who usually present with an adverse natural history. The improved response rate also suggests a role for LDH as a predictive marker.


Subject(s)
Angiogenesis Inhibitors/therapeutic use , Antibodies, Monoclonal, Humanized/therapeutic use , Colorectal Neoplasms/drug therapy , L-Lactate Dehydrogenase/blood , Adult , Aged , Angiogenesis Inhibitors/adverse effects , Antibodies, Monoclonal, Humanized/adverse effects , Bevacizumab , Biomarkers, Tumor/blood , Colorectal Neoplasms/enzymology , Colorectal Neoplasms/mortality , Colorectal Neoplasms/pathology , Disease-Free Survival , Female , Humans , Male , Middle Aged , Prognosis , Treatment Outcome , Vascular Endothelial Growth Factor A/antagonists & inhibitors , Vascular Endothelial Growth Factor A/immunology
14.
Langmuir ; 27(13): 8248-56, 2011 Jul 05.
Article in English | MEDLINE | ID: mdl-21657261

ABSTRACT

Surface tension and isothermal titration calorimetry (ITC) were used to determine the critical micelle concentration (cmc) of the zwitterionic amidosulfobetaine surfactants ASB-14 and ASB-16 (linear-alkylamidopropyldimethylammoniopropanosulfonates) at 25 °C. The cmc and the heat of micellization were determined from 15 to 75 °C by ITC for both surfactants. The increase in temperature caused significant changes in the enthalpy and in the entropy of micellization, with small changes in the standard Gibbs energy (ΔG(mic)), which is consistent to an enthalpy−entropy compensation with a compensatory temperature of 311 K (ASB-14) and 314 K (ASB-16). In the studied temperature range, the heat capacity of micellization (ΔC(p)(mic)) was essentially constant. The experimental ΔC(p)(mic) was lower than that expected if only hydrophobic interactions were considered, suggesting that polar interactions at the head groups are of significant importance in the thermodynamics of micelle formation by these surfactants. Indeed, a NMR NOESY spectrum showed NOEs that are improbable to occur within the same monomer, resulting from interactions at the polar head groups involving more than one monomer. The ITC and NMR results indicate a tilt in the polar headgroup favoring the polar interactions. We have also observed COSY correlations typical of dipolar interactions that could be recovered with the partial alignment of the molecule in solution, which results in an anisotropic tumbling. The anisotropy suggested an ellipsoidal shape of the micelles, which results in a positive magnetic susceptibility, and ultimately in orientation induced by the magnetic field. Such an ellipsoidal shape was confirmed from results obtained by SAXS experiments that revealed aggregation numbers of 108 and 168 for ASB-14 and ASB-16 micelles, respectively. This study characterizes an interesting micelle system that can be used in the study of membrane proteins by solution NMR spectroscopy.


Subject(s)
Betaine/analogs & derivatives , Membrane Proteins/chemistry , Surface-Active Agents/chemistry , Thermodynamics , Betaine/chemistry , Calorimetry , Magnetic Resonance Spectroscopy , Micelles , Models, Molecular , Molecular Structure , Solubility , Surface Tension
15.
PLoS One ; 6(1): e15756, 2011 Jan 13.
Article in English | MEDLINE | ID: mdl-21249196

ABSTRACT

The Ebola fusion peptide (EBO16) is a hydrophobic domain that belongs to the GP2 membrane fusion protein of the Ebola virus. It adopts a helical structure in the presence of mimetic membranes that is stabilized by the presence of an aromatic-aromatic interaction established by Trp8 and Phe12. In spite of its infectious cycle becoming better understood recently, several steps still remain unclear, a lacuna that makes it difficult to develop strategies to block infection. In order to gain insight into the mechanism of membrane fusion, we probed the structure, function and energetics of EBO16 and its mutant W8A, in the absence or presence of different lipid membranes, including isolated domain-resistant membranes (DRM), a good experimental model for lipid rafts. The depletion of cholesterol from living mammalian cells reduced the ability of EBO16 to induce lipid mixing. On the other hand, EBO16 was structurally sensitive to interaction with lipid rafts (DRMs), but the same was not observed for W8A mutant. In agreement with these data, W8A showed a poor ability to promote membrane aggregation in comparison to EBO16. Single molecule AFM experiments showed a high affinity force pattern for the interaction of EBO16 and DRM, which seems to be a complex energetic event as observed by the calorimetric profile. Our study is the first to show a strong correlation between the initial step of Ebola virus infection and cholesterol, thus providing a rationale for Ebola virus proteins being co-localized with lipid-raft domains. In all, the results show how small fusion peptide sequences have evolved to adopt highly specific and strong interactions with membrane domains. Such features suggest these processes are excellent targets for therapeutic and vaccine approaches to viral diseases.


Subject(s)
Ebolavirus/pathogenicity , Hemorrhagic Fever, Ebola/etiology , Membrane Fusion , Membrane Microdomains/metabolism , Viral Fusion Proteins/metabolism , Cholesterol/metabolism , Protein Binding , Viral Envelope Proteins , Virus Diseases
16.
Biophys Chem ; 152(1-3): 99-108, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20843595

ABSTRACT

The XIAP-BIR3 domain blocks a substantial portion of the apoptosis pathway and is an attractive target for novel anticancer agents. The tetrapeptide AVPI, from the protein Smac/DIABLO, binds to the XIAP-BIR3 domain, allowing the cancer cells to die. Here we characterize the binding parameters of AVPI to XIAP-BIR3 and analyze its effects on the thermodynamic stability of this domain. XIAP-BIR3 was exceptionally stable against physical and chemical treatments and became even more stable by interaction with AVPI. Nuclear magnetic resonance experiments demonstrated that conformational selection is taking place upon AVPI interaction with XIAP-BIR3. Molecular dynamics simulations corroborate that the flexibility of XIAP-BIR3 is significantly reduced. The positive binding entropy associated with a loss of conformational entropy involved in the binding indicates that hydrophobic interactions play an important role in the interaction and domain stabilization. The mechanism of XIAP-BIR3 stabilization and its implications for drug affinity optimization are discussed.


Subject(s)
Oligopeptides/chemistry , X-Linked Inhibitor of Apoptosis Protein/chemistry , Amino Acid Sequence , Apoptosis , Calorimetry, Differential Scanning , Circular Dichroism , Humans , Hydrophobic and Hydrophilic Interactions , Molecular Dynamics Simulation , Protein Binding , Protein Stability , Protein Structure, Tertiary , Spectrometry, Fluorescence , Thermodynamics , X-Linked Inhibitor of Apoptosis Protein/metabolism
17.
Gynecol Oncol ; 106(1): 268-71, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17493670

ABSTRACT

BACKGROUND: Although most patients at high risk of Gestational Trophoblastic Neoplasia (GTN) respond to standard treatments, there is a group of patients that will die because of it. The use of new single or combination drugs in this population has become a priority. CASE REPORT: We present the case of a relapsed high risk choriocarcinoma patient who did not respond to several chemotherapy treatments nor to PET guided salvage surgery. Because of treatment toxicity, the patient was started on Capecitabine, with which she achieved total remission, still present after 15 months of starting treatment. CONCLUSIONS: The use of Capecitabine and the multidisciplinary management of this population should be taken into account for patients at high risk of relapsing to EP/EMA because of its efficacy and little toxicity.


Subject(s)
Antimetabolites, Antineoplastic/therapeutic use , Choriocarcinoma/drug therapy , Deoxycytidine/analogs & derivatives , Fluorouracil/analogs & derivatives , Salvage Therapy , Trophoblastic Neoplasms/drug therapy , Adult , Capecitabine , Deoxycytidine/therapeutic use , Female , Fluorouracil/therapeutic use , Humans , Pregnancy , Risk Factors
18.
Biophys Chem ; 126(1-3): 59-64, 2007 Mar.
Article in English | MEDLINE | ID: mdl-16824668

ABSTRACT

This mini-review shows the valuable contributions of Professor Julian Sturtevant to the current applications of calorimetry to the study of enzyme-catalyzed reactions. The more recent applications of calorimetric techniques such as isothermal titration calorimetry and flow calorimetry to the study of enzyme kinetics, as well as the advantages on using calorimetric techniques in the determination of kinetic parameters of enzymes, is also discussed here.


Subject(s)
Calorimetry/methods , Enzymes/chemistry , Thermodynamics , Catalysis , Kinetics
19.
Orig Life Evol Biosph ; 36(5-6): 597-603, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17136430

ABSTRACT

Absorption or emission spectroscopy is a powerful tool for detecting chemical compounds, diluted in fluid media: the sensitivity of this technique depends on the optical path of the source radiation, on the spectral window used for analysis and on the spectrometer performances. In this view, we designed and produced the first prototypes of an integrated scanning Fourier Transform Microinterferometer with Mach-Zehnder geometry, by using MEOS (Micro Electro Optical Systems) technologies. The microdevice, obtained by fabricating integrated optical waveguides on LiNbO(3) (LN) crystals, is electrically driven, without moving parts, by exploiting the electrooptical properties of the material. The microdevice operates the Fourier Transform of the input radiation spectral distribution, which can be reconstructed starting from the output signal by means of Fast Fourier Transform (FFT) techniques. The microinterferometer weights few grams, the power consumption is of a few mW and, in principle, can operate in the LN transmittance range (0.36-4.5 microm). The microinterferometer performances were preliminary tested in the (0.4-1.7 microm) spectral window. In the Visible region (0.4-0.7 mum) this microsystem demonstrated a spectral resolution suitable for detecting the characteristic lines of the solar spectrum together with the absorption bands of common gases present in Earth's atmosphere. In a further experiment we tested its performances for gas trace detection by using a calibrated NO(2) optical gas cell, showing the possibility to reveal up to 10 ppb, when suitable optical paths are used. Finally, colorimetry tests for the titration of an organic dye (E131) in alcohol solution are presented.


Subject(s)
Interferometry/instrumentation , Niobium/chemistry , Oxides/chemistry , Spectroscopy, Fourier Transform Infrared/instrumentation , Crystallization , Equipment Design , Gases/analysis , Gases/chemistry , Interferometry/methods , Miniaturization , Nitrogen Dioxide/chemistry , Reproducibility of Results , Spectroscopy, Fourier Transform Infrared/methods
20.
Biochemistry ; 45(31): 9540-9, 2006 Aug 08.
Article in English | MEDLINE | ID: mdl-16878988

ABSTRACT

The gene Aedes aegypti intestinal mucin 1 (AeIMUC1) encodes a putative peritrophic matrix (PM) protein that is expressed in the midgut of mosquito larvae and adults and is upregulated in response to exposure to heavy metals. The AeIMUC1 protein has a predicted secretory signal peptide and three putative chitin-binding domains (CBDs) with an intervening mucin-like domain. Immunofluorescence and immunoelectron microscopy experiments established that AeIMUC1 is a bona fide PM protein, and binding of the recombinant protein to chitin was demonstrated in vitro. Previous experiments suggested that the Ae. aegypti PM can bind toxic heme molecules generated during blood digestion. However, the identity of the binding molecule(s) was unknown. Using of heme-agarose beads and spectrophotometric and microcalorimetric titrations, we show that recombinant AeIMUC1 can bind large amounts of heme in vitro, suggesting for the first time a role for a PM protein in heme detoxification during blood digestion. Binding of heme to AeIMUC1 was accompanied by an altered circular dichroism spectrum indicating a change in protein conformation, consistent with an increase in secondary structure. Heme-binding activity was mapped to the AeIMUC1 CBDs, suggesting that these domains possess dual chitin- and heme-binding activity.


Subject(s)
Aedes/metabolism , Chitin/metabolism , Heme/metabolism , Insect Proteins/metabolism , Mucins/metabolism , Amino Acid Sequence , Animals , Chitin/chemistry , Circular Dichroism , Female , Heme/chemistry , Insect Proteins/analysis , Insect Proteins/chemistry , Intestinal Mucosa/metabolism , Intestines/chemistry , Intestines/ultrastructure , Microscopy, Immunoelectron , Molecular Sequence Data , Mucins/analysis , Mucins/chemistry , Protein Sorting Signals , Protein Structure, Tertiary , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism
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