Identification of human LDHC4 as a potential target for anticancer drug discovery

One of the distinct hallmarks of cancer cells is aerobic glycolysis (Warburg effect). Lactate dehydrogenase A (LDHA) is thought to play a key role in aerobic glycolysis and has been extensively studied, while lactate dehydrogenase C (LDHC), an isoform of LDHA, has received much less attention. Here we showed that human LDHC was significantly expressed in lung cancer tissues, overexpression of Ldhc in mice could promote tumor growth, and knock-down of LDHC could inhibit the proliferation of lung cancer A549 cells. We solved the first crystal structure of human LDHC4 and found that the active-site loop of LDHC4 adopted a distinct conformation compared to LDHA4 and lactate dehydrogenase B4 (LDHB4). Moreover, we found that (ethylamino) (oxo)acetic acid shows about 10 times selective inhibition against LDHC4 over LDHA4 and LDHB4. Our studies suggest that LDHC4 is a potential target for anticancer drug discovery and (ethylamino) (oxo)acetic acid provides a good start to develop lead compounds for selective drugs targeting LDHC4.

Growth hormone receptor gene polymorphism. Spontaneous catch up growth in small for gestational age patients / Polimorfismo del gen del receptor de la hormona de crecimiento. Crecimiento postnatal espontáneo en niños pequeños para la edad gestacional

Abstract The growth hormone receptor (GHR) mediates the effect of growth hormone (GH) on linear growth and metabolism. In humans, it exists as two isoforms differing by the retention or exclusion of exon 3; a full-length GHR isoform (GHRfl) and the exon 3-deleted isoform (GHRd3). The genotypic frequency of this polymorphism was analyzed in several studies and in different human populations. However scarce information in Argentinean population is available. Associations between GHRd3 and growth have been reported previously. Some studies have shown that the presence of GHRd3 polymorphism might be a potential variant that improves growth response to recombinant human GH (rhGH) therapy in patients born small for gestational age (SGA), among others. However, over the years the results have been controversial and inconclusive. Based on this, it would be proposed that variants at the genomic level are not completely reflected at the mRNA level. Our aim was to evaluate the genotypic frequencies (%) of the GHR gene polymorphism (GHRfl/GHRfl; GHRfl/GHRd3; GHRd3/GHRd3) in normal Argentinean population (n = 94) and SGA patients (n = 65), and the expression of these polymorphisms at mRNA level in the fetal side of placenta tissues was analyzed. In addition, their asso ciation with spontaneous postnatal catch-up growth in SGA patients was also evaluated. In this study, we show a significant increment of compensatory growth in small for gestational age children (SGA) associated to the presence of the GHRd3 allele polymorphism. In addition, the expression of GHR in healthy placentas revealed that no alternative splicing mechanism occurs.

Resumen El receptor de la hormona de creci miento (GHR) media la acción de la hormona de crecimiento (GH) en el crecimiento lineal y el metabolismo. En los seres humanos, existen dos isoformas que difieren en la retención (GHRfl) o exclusión del exón 3 (GHRd3). La frecuencia genotípica de este polimorfismo fue analizada en varios estudios y en diferentes poblaciones. Sin embargo, la información disponible en la población argentina es escasa. Se ha reportado anteriormente asociación entre el polimorfismo GHRd3 y el crecimiento. Varios estudios ha n demostrado que la presencia del polimorfismo GHRd3 podría mejorar, en pacientes nacidos pequeños para la edad gestacional, entre otros, la respuesta a la terapia con GH humana recombinante (rhGH). Sin embargo, a lo largo de los años los resultados han sido con trovertidos y no concluyentes. En base a esto, se propondría que las variantes a nivel genómico no se reflejan completamente a nivel del ARNm. Nuestro objetivo fue evaluar la frecuencia genotípica de los polimorfismos del gen del GHR (GHRfl/GHRfl; GHRfl/GHRd3; GHRd3/GHRd3) en la población argentina normal (n = 94) y en niños pequeños para la edad gestacional (n = 65), y se analizó la expresión de estos polimorfismos a nivel de ARNm en la porción fetal de placentas sanas. Además, se evaluó la asociación de este polimorfismo con el cre cimiento postnatal espontáneo en pacientes pequeños para la edad gestacional. En este estudio, mostramos un incremento significativo del crecimiento compensatorio en niños pequeños para la edad gestacional asociado a la presencia del polimorfismo del alelo GHRd3. Además, los ensayos de expresión de GHR en placentas sanas revelaron que no se produciría ningún mecanismo de splicing alternativo.

Value of subtypes of antimitochondrial antibodies in primary biliary cholangitis / 中华风湿病学杂志

Objective:To investigate the distribution and clinical significance of subtypes of antimitochondrial antibodies (AMA)-M2, M4, M9 in primary biliary cholangitis (PBC).Methods:A total of 1 367 patients were detected with AMA-M2, M4, M9 in Peking Union Medical College Hospital (PUMCH) from Jan 2014 to Dec 2019 and the clinical parameters were collected. The distribution patterns of AMA subtypes in different groups were analyzed and the diagnostic sensitivity and specificity of AMA subtypes in PBC were calculated. Chi-square test was used for statistical analysis.Results:In 1 367 patients, 236 of whom were positive for AMA subtypes. The positivity of AMA subtypes in female was significantly higher than in male (20.34% vs 9.41%, χ2=23.792, P<0.01). In addition, the positivity of AMA subtypes was significantly higher in 30-65 years old patients than in patients younger than 30 years old or older than 65 years old [(20.00%(193/965) vs 10.97%(17/155) vs 10.53%(26/247), χ2=17.209, P<0.01]. 110 patients with positive AMA subtypes were diagnosed with PBC. The diagnostic sensitivity and specificity of AMA-M2 were both desirable [94.64%(106/112) and 92.35%(1 159/1 255)]. Although the specificity of AMA-M4 was as high as 99.12%(1 244/1 255), its sensitivity was very low [15.18%(17/122)]. Combined detection of different AMA subtypes could not improve the diagnostic sensitivity and specificity significantly. Diseases other than PBC can be positive for AMA subtypes, predominantly for AMA-M2. Conclusion:Female and 30-65 years old patients were more frequently positive for AMA subtypes. AMA-M2 was the most valuable AMA subtype for diagnosing PBC.

Abnormal thymic stromal lymphopoietin expression in the gastrointestinal mucosa of patients with eosinophilic gastroenteritis / Expressão anormal da linfopoietina estromal tímica na mucosa gastrointestinal depacientes com gastroenterite e osinofílica

Abstract Objective: To investigate the differential expression of the thymic stromal lymphopoietin isoforms, short and long, and discern their biological implications under eosinophilic gastroenteritis. Methods: The expression of thymic stromal lymphopoietin and its two isoforms in tissues was assessed by quantitative RT-PCR in healthy controls (n = 24) and patients with eosinophilic gastroenteritis (n = 17). Results: Thymic stromal lymphopoietin mRNA was significantly reduced in eosinophilic gastroenteritis when compared with healthy controls (p < 0.0001). A significantly lower amount of short thymic stromal lymphopoietin mRNA was observed in eosinophilic gastroenteritis when compared with controls (p < 0.05), while a significantly higher amount of long thymic stromal lymphopoietin mRNA was observed in eosinophilic gastroenteritis when compared with controls (p < 0.05). Peak eosinophilic count is significantly positively correlated with the expression of long thymic stromal lymphopoietin mRNA in the gastrointestinal mucosal of patients with eosinophilic gastroenteritis (r s = 0.623, p < 0.005), while peak eosinophilic count is significantly negatively correlated with the expression of short thymic stromal lymphopoietin mRNA in the gastrointestinal mucosal of patients with eosinophilic gastroenteritis (r s = −0.4474, p < 0.05). Conclusions: Abnormal mucosal thymic stromal lymphopoietin expression may contribute to gastrointestinal mucosa damage in eosinophilic gastroenteritis.

Resumo Objetivo: Investigar a expressão diferencial das isoformas da linfopoietina estromal tímica, curta e longa, e discernir suas implicações biológicas na gastroenterite eosinofílica. Métodos: Avaliamos a expressão das isoformas da linfopoietina estromal tímica e suas duas isoformas através da técnica RT-PCR quantitativa em tecidos de controles saudáveis (n = 24) e pacientes com gastroenterite eosinofílica (n = 17). Resultados: Demonstramos que o RNAm das isoformas da linfopoietina estromal tímica estava significativamente reduzido na gastroenterite eosinofílica em comparação com os controles saudáveis (p < 0,0001). Também descobrimos uma quantidade significativamente menor de RNAm das isoformas da linfopoietina estromal tímica curta na gastroenterite eosinofílica em comparação com os controles (p < 0,05) e uma quantidade significativamente maior de RNAm das isoformas da linfopoietina estromal tímica longa na gastroenterite eosinofílica em comparação com os controles (p < 0,05). O pico da contagem eosinofílica está correlacionado positiva e significativamente com a expressão do RNAm das isoformas da linfopoietina estromal tímica longa na mucosa gastrointestinal de pacientes com gastroenterite eosinofílica (rs = 0,623, p < 0,005), enquanto o pico de contagem eosinofílica está negativa e significativamente correlacionado com a expressão do RNAm das isoformas da linfopoietina estromal tímica curta na mucosa gastrointestinal de pacientes com gastroenterite eosinofílica (rs = -0,4474, p < 0,05). Conclusões: A expressão anormal das isoformas da linfopoietina estromal tímica na mucosa pode contribuir para o dano da mucosa gastrointestinal na gastroenterite eosinofílica.

Histone H2A isoforms: Potential implications in epigenome plasticity and diseases in eukaryotes

Epigenetic mechanisms including the post-translational modifications of histones, incorporation of histonevariants and DNA methylation have been suggested to play an important role in genome plasticity by allowingthe cellular environment to define gene expression and the phenotype of an organism. Studies over the pastdecade have elucidated how these epigenetic mechanisms are significant in orchestrating various biologicalprocesses and contribute to different pathophysiological states. However, the role of histone isoforms and theirimpact on different phenotypes and physiological processes associated with diseases are not fully clear. Thisreview is focussed on the recent advances in our understanding of the complexity of eukaryotic H2A isoformsand their roles in defining nucleosome organization. We elaborate on their potential roles in genomic complexity and regulation of gene expression, and thereby on their overall contribution towards cellular phenotypeand development of diseases

Progress of effects of 14-3-3 protein on gastric cancer and its mechanism / 解放军医学杂志

14-3-3 proteins are a highly conserved family of cellular proteins, and widely expressed in all eukaryotes. Because they can bind to a variety of functional signal proteins, so be regarded as "bridge proteins" in protein interaction. Studies have shown that the expression level of 14-3-3 protein family in malignant tumors is closely related to tumor size, depth of invasion and metastasis. Some other studies have shown that 14-3-3 protein plays a pivotal role in cell signal transduction, control of cell cycle process and regulation of apoptosis, and plays an important role in the development of gastric cancer, proliferation, invasion, prognosis and drug resistance, so is expected to become a novel therapeutic target for gastric cancer. The present paper aims to expound the role and relevant mechanisms of 14-3-3 protein in gastric cancer.

Lipoprotein(a): An independent, genetic, and causal factor for cardiovascular disease and acute myocardial infarction

Lipoprotein(a) [Lp(a)] is a circulating lipoprotein, and its level is largely determined by variation in the Lp(a) gene (LPA) locus encoding apo(a). Genetic variation in the LPA gene that increases Lp(a) level also increases coronary artery disease (CAD) risk, suggesting that Lp(a) is a causal factor for CAD risk. Lp(a) is the preferential lipoprotein carrier for oxidized phospholipids (OxPL), a proatherogenic and proinflammatory biomarker. Lp(a) adversely affects endothelial function, inflammation, oxidative stress, fibrinolysis, and plaque stability, leading to accelerated atherothrombosis and premature CAD. The INTER-HEART Study has established the usefulness of Lp(a) in assessing the risk of acute myocardial infarction in ethnically diverse populations with South Asians having the highest risk and population attributable risk. The 2018 Cholesterol Clinical Practice Guideline have recognized elevated Lp(a) as an atherosclerotic cardiovascular disease risk enhancer for initiating or intensifying statin therapy.

Diagnostic Significance of p38 Isoforms (p38α, p38β, p38γ, p38δ) in Head and Neck Squamous Cell Carcinoma: Comparative Serum Level Evaluation and Design of Novel Peptide Inhibitor Targeting the Same / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The p38 mitogen-activated protein kinase (MAPKs) play a crucial role in the production of pro-inflammatory cytokines and over-expression of it increase cytokines which promote cancer. Among four isoforms, p38α has been well studied in head and neck squamous cell carcinoma (HNSCC) and other cancers as a therapeutic target. p38δ has recently emerged as a potential disease-specific drug target. Elevated serum p38α level in HNSCC was reported earlier from our lab. This study aims to estimate the levels of p38 MAPK-isoforms in the serum of HNSCC and design peptide inhibitor targeting the same. MATERIALS AND METHODS: Levels of p38 MAPK isoforms in the serum of HNSCC and healthy controls were quantified by surface plasmon resonance technology. The peptide inhibitor for p38 MAPK was designed by molecular modeling using Grid-based Ligand Docking with Energetics tools and compared with known specific inhibitors. RESULTS: We have observed highly elevated levels of all four isoforms of p38 MAPK in serum of HNSCC patients compared to the control group. Further, serum p38α, p38β, and p38δ levels were down regulated after therapy in follow-up patients, while p38γ showed no response to the therapy. Present study screened designed peptide WFYH as a specific inhibitor against p38δ. The specific inhibitor of p38δ was found to have no effect on p38α due to great structural difference at ATP binding pocket. CONCLUSION: In this study, first time estimated the levels of p38 MAPK isoforms in the serum of HNSCC. It can be concluded that p38 MAPK isoforms can be a diagnostic and prognostic marker for HNSCC and p38δ as a therapeutic target.

A research for a newly discovered TMPRSS4-isoforms and its biological charcteristics in transfected colon cancer cells / 安徽医科大学学报

Objective To explore a newly discovered transmembrane protease serine 4 (TMPRSS4) isoforms and its molecular charcteristics in transfected colon cancer cells. Methods The named T4-1A and T4-1B of the TMPRSS4-isoforms were authenticated by the RT-PCR and Western blot,and then transfected to human colon cancer cells (DLD-1). Those stable transfected cells of migration and invasion were illustrated using wound healing assays and matrigel invasion assays. Results Successfully constructed T4-1A and T4-1B recombinant vectors,and obtained T4-1A and T4-1B transfected cell lines,and their T4-1A and T4-1B were highly expressed in stable DLD-1. Compared to cells transfected with empty vector of pcDNA6,the transfected DLD-1 with T4-1A enhanced migration and invasion were statistical significance (P < 0. 05). However,compared to pcDNA6 no significant1 difference was found for T4-1B. Moreover,the biological characteristics of T4-1A and TMPRSS4 were very similar. Conclusion The T4-1A and T4-1B is newly TMPRSS4-isoforms,and protease domain included to a T4-1A has further facilitated the migration and invasion of colon cancer cells,and further studies provide a theoretic base in the molecular biomedical characteristics of TMPRSS4.

Advances in NMDA receptors-related antidepressants / 中国药理学通报

Recent studies have demonstrated that ketamine , an N-methyl-D-aspartate ( NMDA ) receptor antagonist , can pro-duce rapid and effective antidepressant actions , especially for patients with treatment-resistant depression , bipolar disorder and suicide idea.However, the clinical application of ketamine is limited by its psychotomimetic adverse reactions and potential a-buse.Therefore, development of novel ketamine-like antidepres-sant agents with fewer side effects is actively investigated currently.The latest researches indicate that the metabolites of ket-amine, the S-ketamine/R-ketamine and the modulators of NM-DA receptor subunits can exert safe and effective antidepressant actions.In this article, the research advance in NMDA recep-tors-related antidepressants mentioned above will be reviewed .

Expression of RUNX1 isoformsand its clinical significance in acute leukemia / 中国肿瘤生物治疗杂志

@# Objective: : To investigate the relationship between the expression of RUNX1 isoforms and the clinical curative effect and the prognosis of acute leukemia (AL), in order to provide valuable experimental data for the individualized treatment, MRD (minimal residual disease) monitoring and prognosis prediction of AL. Methods: AL patients with primary treatment (PT, n=88) and recrudescence (RC, n=10) that treated at the Department of Hematology of Affiliated Hospital of Guangdong Medical University from April, 2012 to April, 2013 were included in this study. Real-time PCR was used to examine the mRNA expression of RUNX1 isoforms (RUNX1a and RUNX1b/c) in PT patients, RC patients and controls (non-malignant hematological disease patients).The changes in mRNA expression of RUNX1a and RUNX1b/c in patients before and after the chemotherapy were also observed. Results： ： (1)The expression levels of RUNX1a mRNAinAML andALL PT group andAML RC group was significantly higher than those of control group (P<0.05); The expression level of RUNX1a mRNAinAMLPT group was increased compared withALLPT group (P<0.05). (2) The expression levels of RUNX1a and RUNX1b/c mRNAinAML andALL patients at initial treatment were significantly higher than those after complete remission (CR) (P<0.05). (3) By comparing the expression levels of RUNX1a and RUNX1b/c mRNA at initial diagnosis, there was no significant difference between 6-month death group and survival group, CR group and NCR (non-complete remission) group after first cycle of chemotherapy, or the high leukocyte group and non-high leukocyte group (all P>0.05).The expression level of RUNX1a mRNA in AML-ETO positive group was higher than that of negative group (P<0.05). (4) The expression levels of RUNX1a and RUNX1b/c mRNA in patients with acute leukemia decreased with the increasing chemotherapy cycle, and significantly increased when had a relapse, which may even succeed the initial level.Conclusion: RUNX1a isoforms participate in the pathogenesis of acute leukemia, and isrelated to the relapse ofAML. The expression levels of RUNX1a and RUNX1b/c mRNAare related to the clinical efficacy that can be used as an indicator of curative effect, but have no significant correlation with the prognosis of the disease.Dynamic monitor of theexpression levels of RUNX1a and RUNX1b/c isomers can be used as an effective indicator of MRD monitoring after chemotherapy, which can be used to evaluate the efficacy and identify the risk of recurrence at early stage.

Polimorfismos del receptor D2 / D2 receptor polymorphisms

Los prolactinomas son tumores bien diferenciados que se originan en las células lactotropas pituitarias, una línea celular que secreta fisiológicamente prolactina (PRL). A nivel hipofisario, la dopamina está implicada en la regulación de la secreción de PRL por las células lactotropas y este efecto inhibitorio está mediado por la activación del receptor de prolactina tipo 2 (DRD2). Hay varios polimorfismos del DRD2, el primero y más estudiado es TaqI A1; está demostrado que este alelo se encuentra asociado a una reducción de la actividad cerebral dopaminérgica, además de observarse una reducción en su capacidad de unión de aproximadamente un 30%. Este alelo se ha vinculado con una menor densidad de DRD2 en el cuerpo estriado, especialmente en el putamen y caudado ventral, y la cantidad de DRD2 en portadores del alelo A1 fue un 30-40% más bajo que en los no portadores (es decir, TaqI A2 homocigotos). En la literatura, hay evidencia que apoya la posible participación de los polimorfismos DRD2 en la regulación de la secreción hormonal.

Prolactinomas are well differentiated tumours that originate in the pituitary lactotrope cells, a cell line that physiologically secretes prolactin (PRL). At pituitary level, dopamine is involved in the regulation of PRL secretion by lactotropes, and this inhibitory effect is mediated by activation of prolactin type 2 receptor (DRD2). Of the several DRD2 polymorphisms, the first and most studied is TaqI A1. It has been demonstrated that this allele is associated with a reduced dopaminergic brain activity, and a reduction in its binding capacity of approximately 30% also being observed. This allele was associated with a lower density of DRD2 in the striatum, especially in the putamen and ventral caudate. The amount of DRD2 in A1 allele carriers was 30 - 40% lower than in non-carriers (this is, TaqI A2 Homozygotes). There is evidence in the literature, that supports the possible involvement of DRD2 polymorphisms in the regulation of hormonal secretion.

Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions

Abstract Background Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results The RP-HPLC profile of the isolated crotapotin chains already indicated that the chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the chain, at positions 31 and 40. Moreover, substitutions could also be observed in and chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.

Protocolos de estimulación ovárica en inseminación artificial según el patrón utilizado de glicosilación de la hormona folículo estimulante / The impact of different ovarian stimulating protocols in artificial insemination based on the glycosylation pattern of the follicle stimulating hormone

Resumen ANTECEDENTES: los estudios clínicos que comparan diferentes protocolos de estimulación ovárica no son concluyentes. Nuestro estudio compara tres protocolos según el patrón de glicosilación de la hormona folículo estimulante (FHS) que imita el ciclo natural para inseminación artificial. OBJETIVO: proponer un protocolo de estimulación ovárica secuencial con FSH urinaria (isoforma ácida) seguido de FSH recombinante (isoforma menos ácida) y evaluar su efectividad respecto de los esquemas tradicionales de estimulación con FSH recombinante (FSHr) y FSH urinaria (FSHu). MATERIALES Y MÉTODOS: estudio de cohortes, retrospectivo, efectuado entre mayo de 2012 y mayo 2015 en parejas infértiles en protocolo de inseminación artificial. El grupo de estudio se dividió en tres: 1) FSHr, 2) FSHhp y 3) estimulación sequencial: FSHhp + FSHr. Para el análisis estadístico se utilizaron χ2, ANOVA o test Mann-Whitney. Los resultados se reportan con límite de significación de p < 0.05. RESULTADOS: se estudiaron 178 parejas infértiles en protocolo de inseminación artificial y se efectuaron 299 ciclos de estimulación ovárica con inseminación artificial; grupo 1) 99 con FSHr, 2)100 con FSHhp y 3) 100 con estimulación sequencial: FSHhp + FSHr. Los resultados gestacionales fueron mayores en el grupo FSHu y secuencial. En el análisis costo-efectividad se observó menor tasa con el esquema secuencial en comparación con los otros dos. Al comparar los tres protocolos según los intentos previos de inseminación, en la serie con intentos previos, se encontraron diferencias estadísticamente significativas en βHCG y gestación al comparar FSHr versus secuencial. El protocolo FSHr fue menos efectivo que los otros dos tratamientos. CONCLUSIONES: se planteó la estimulación ovárica controlada con protocolo secuencial en pacientes que recibirían inseminación artificial y, aunque no se encontraron diferencias significativas, sí se observó una tendencia en la que tanto la dosis como los días de estimulación requerida son menores con este protocolo que con los clásicos con FSHr o FSHu.

Abstract BACKGROUND: Earlier trials comparing different ovarian stimulating protocols in assisted reproduction found no significant differences in the results. OBJECTIVE: To evaluate the effect of three different stimulation protocols based on the glycosylation pattern of the follicle stimulating hormone (FSH) in an intrauterine insemination (IUI) program. The stimulating protocols are: human FSH (acidic isoform), recombinant FSH (less acidic isoform) and combined urinary and recombinant FSH. MATERIALS AND METHODS: Cohort study, retrospective, conducted between May 2012 and May 2015 in infertile couples in protocol of artificial insemination. The study group was divided into three: 1) FSHr, 2) FSHhp and 3) sequential pacing: FSHhp + FSHr. For statistical analysis, χ2, ANOVA or Mann-Whitney test were used. The results are reported with a significance limit of p < 0.05. RESULTS: 178 infertile couples were studied in an artificial insemination protocol and 299 cycles of ovarian stimulation were performed with artificial insemination; Group 1) 99 with FSHr, 2) 100 with FSHhp and 3) 100 with sequential stimulation: FSHhp + FSHr. CONCLUSION: In this study, controlled ovarian stimulation with sequential protocol was considered in patients who would receive artificial insemination and, although no significant differences were found, a trend was observed in which both the dose and the days of stimulation required are lower with this protocol than with the classics with FSHr or FSHu.

Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions

Background: Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods: The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results: The RP-HPLC profile of the isolated crotapotin chains already indicated that the α chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion: It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the α chain, at positions 31 and 40. Moreover, substitutions could also be observed in ß and γ chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.(AU)

Real Time Cell Analysis of Model Target Cell Lines Exposed to Purified Lipoprotein (a).

Lipoprotein (a) [Lp(a)] is a novel independent cardiovascular risk factor and it includes, beyond apoB100, apolipoprotein (a), whose molecular weight is dependent on the number of genetically encoded kringle IV type 2 repeats and inversely related with Lp(a) plasma concentration. Risk thresholds for molecular weights have been proposed, but there is not a full consensus and the role of the different isoforms in pathogenesis has not yet been clarified. The aim of the present work is to explore the biological effect of low and high molecular weight Lp(a) isoforms on cultured cells. Real-time impedance analysis has been performed on model cell lines of atherogenesis and Lp(a) metabolism (THP-1, HUVEC, HASMC and HepG2) using affinity purified Lp(a) with 22 (low number) and 31 (high number) kringle IV type 2 repeats, respectively. Normalized Cell Index data show that all the cell lines tested are modified by Lp(a), though with a variable intensity. Low and high molecular weight Lp(a) isoforms at similar concentrations can exert opposite modifications on the impedance kinetics of different cell lines. These data suggest that purified Lp(a) can modify the behaviour of adherent cell lines, an effect which can be detected as impedance variation and which is influenced by its specific isoform.

Recent advances in breast cancer stem cell phenotypes and clinical application / 中国癌症杂志

The cancer stem cell theory suggests that cancer develops from a subset of tumor cells that possess characteristics of stem cells. Breast cancer stem cells comprise a sub-population, which possesses the capacity of self-renewal and the potential for differentiation and high tumorigenicity. Evidence from bothin vitro andin vivo studies demonstrates breast cancer stem cells are responsible for tumor relapse, invasion and metastasis, chemo- and radio-resistance and epithelial-mesenchymal transition (EMT). Herein, this review highlighted the recent advances in breast cancer stem cells.

The Expression of Plasma Membrane Ca2 + -ATPase Isoforms 1~4 and the Splice Variants at Sites A and C in the Neonatal Rat Vestibular Organ / 听力学及言语疾病杂志

Objective To study the expression of plasma membrane Ca2 + -ATPase isoforms 1 -4 and the splice variants at sites A and C in the neonatal rat vestibular organ.Methods Ten rats at postnatal 2 days (P2 ) were decapitated and their vestibular organs (macula utriculi and macula sacculi)were isolated.The total proteins of the vestibular organs were extracted.The expression of PMCA1-4 splice variants at sites A and C was detected by RT-PCR.Results The splice variants of PMCA1-4 at sites A and C in macula utriculi and macula sacculi of neo-natal rat vestibular organs were PMCA1x/b,PMCA2w/(a,b),PMCA3z/(a,b,c)and PMCA4 (x,z)/b.Conclusion The splice variants at sites A and C among PMCA1,PMCA2,PMCA3 and PMCA4 were different in the vestibu-lar organs of neonatal rats,which could be explained that macula utriculi and macula sacculi had different require-ments of Ca2 + turning for these PMCA isoforms.

The Expression of Plasma Membrane Ca2 + -ATPase Isoforms 1 ~3in the Basi lar Membrane of Neonatal Rat Cochlea / 听力学及言语疾病杂志

Objective To study the expression of plasma membrane Ca2+-ATPase isoforms 1~3 (PMCA 1~3 )in the basilar membrane (BM)of the neonatal rat cochlea by Western blot.The PMCA2 content in single BM of the neonatal rat was also examined.Methods Four rats at postnatal 2 days (P2)and 8 days (P8)were respective-ly decapitated and their BMs were isolated.The total proteins of BMs were extracted.The 20μg total proteins were respectively loaded to the gel.The expression of PMCA1-3 was detected by Western blot.Likewise,3μg total proteins from P2 and P8 rat BM were loaded.The expression of PMCA2 was detected by Western blot.Four rats at P8 were decapitated and their BM was isolated.The 5μg,10μg and 20μg total proteins of P8 rat BM were added to the gel and 100 ng,400 ng and 800 ng bovine serum albumin (BSA)were also loaded as reference.After electro-phoresis,the gel was separated into two parts.One part was used for SYPRO staining and the other part was used for PMCA2 detection by Western blot.Results In the 20μg BM total proteins of P2 and P8 rats,the expression of PMCA1 was weak (0.126±0.024,0.131±0.012,respectively),PMCA2 was strong (4.16±0.528,4.25±0.319, respectively),and PMCA3 was barely expressed (0 ).There was a statistical difference among PMCA1 ,PMCA2 and PMCA3(P<0.05).In the 3μg BM total proteins of P2 and P8 rats,the expression of PMCA2 in P8 (4.571± 0.336)was higher than P2 (3.622±0.285).There was a statistical difference(P<0.05).The PMCA2 content in the BM of a P8 rat was about 2 .5 ng.Conclusion There was a different-level expression of PMCA1~3 in the neonatal rat BM with highest expression of PMCA2 ,which could be explained that cochlear hair cells had different requirements of Ca2+ turning for these PMCA isoforms.

Polimorfismos del receptor de GH en pacientes acromegálicos / GH Receptor Polymorphisms in Acromegalic Patients

El receptor de la hormona de crecimiento (GHR) (hormona de crecimiento. media los efectos de GH. Se identificaron varias isoformas de dicho receptor, producto de polimorfismos en las secuencias codificantes del gen del GHR. La isoforma (d3-GHR) carece de una secuencia de 22 aminoácidos codificada por el exón 3, y si bien su capacidad de unión al ligando se mantiene intacta, afectaría las propiedades funcionales del GHR, y podría facilitar la activación del GHR. Se observó que el 50 % de los pacientes acromegálicos tienen al menos un alelo d3. Se reportó que el alelo d3 contribuyó como predictor de respuesta posquirúrgica con niveles significativamente menores de hormona de crecimiento (GH) a diferencia de los fl-GHR (forma completa), con similares niveles de IGF-1 (insulin-likegrowth factor 1). La ausencia del exón 3 se asoció además con mayor probabilidad de no alcanzar IGF-I normal después del tratamiento tanto quirúrgico como médico. Los portadores d3-GHR se asocian con menor dosis requerida de PEG (Pegvisomant), siendo el mecanismo de esta mayor sensibilidad desconocido. Estos pacientes también podrían tener mayor prevalencia de comorbilidades acromegálicas. Estos estudios demuestran la importancia del genotipo del GHR, no solo como variable predictora de evolución clínica, sino que su evaluación podría ser útil para individualizar la terapia óptima en cada paciente.

The growth hormone receptor mediates the effects of GH. Several isoforms of this receptor, as a product of polymorphisms in the coding sequences of GHR gene, have been identified. The d3-GHR isoform lacks a 22-amino-acid sequence encoded by exon 3, and although its ligand binding capacity remains intact, its deletion would affect the functional properties of GHR, and could facilitate the activation of the GHR. It has been reported that 50 % of acromegalic patients have at least one d3 allele. The d3 allele has been shown to contribute to predict postoperative response presenting significantly lower levels of GH in contrast to fl-GHR (full-length), with similar levels of IGF-1. The absence of exon 3 was also associated with abgreater likelihood of failing to achieve normal IGF-I after both surgical and medical treatment. In carriers of d3-GHR, lower doses of PEG (Pegvisomant) were required; the mechanism of this increased sensitivity is unknown.These patients may also have a higher prevalence of acromegalic comorbidities. These studies demonstrate the importance of the GHR genotype, which may not only contribute to predict clinical outcome, but also help to identify the optimal therapy for each patient.