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Genome size (GS) or DNA nuclear content is considered a useful index for making inferences about evolutionary models and life history in animals, including taxonomic, biogeographical, and ecological scenarios. However, patterns of GS variation and their causes in crustaceans are still poorly understood. This study aimed to describe the GS of five Neotropical Synalpheus non-gambarelloides shrimps (S. apioceros, S. minus, S. brevicarpus, S. fritzmueller, and S. scaphoceris) and compare the C-values of all Caridea infraorder in terms of geography and phylogenetics. All animals were sampled in the coast of Sao Paulo State, Brazil, and GS was assessed by flow cytometry analysis (FCA). The C-values ranged from 7.89 pg in S. apioceros to 12.24 pg in S. scaphoceris. Caridean shrimps had higher GS than other Decapoda crustaceans. The results reveal a tendency of obtaining larger genomes in species with direct development in Synalpheus shrimps. In addition, a tendency of positive biogeographical (latitudinal) correlation with Caridea infraorder was also observed. This study provides remarkable and new protocol for FCA (using gating strategy for the analysis), which led to the discovery of new information regarding GS of caridean shrimps, especially for Neotropical Synalpheus, which represents the second-largest group in the Caridea infraorder. Key words: C-value, snapping shrimp, Decapoda, DNA content La taille du genome ou le contenu en ADN nucleaire est considere comme un indice utile pour faire des inferences au sujet des modeles evolutifs et de l'histoire de vie chez les animaux, incluant des scenarios taxonomiques, biogeographiques et ecologiques. Cependant, les types de variation quant a la taille des genomes ainsi que leurs causes sont encore mal compris chez les crustacees. Cette etude visait a decrire la variation pour la taille du genome chez cinq especes de crevettes neotropicales Synalpheus nongambarelloides (S. apioceros, S. minus, S. brevicarpus, S. fritzmueller et S. scaphoceris) et de comparer les valeurs C au sein du sous-ordre Caridea en termes de geographie et de phyllogenetique. Tous les specimens ont ete captures sur la cote de l'etat de Sao Paulo au Bresil et la taille du genome a ete mesuree par cytometrie en flux (FCA). Les valeurs C variaient entre 7,89 pg chez le S. apioceros et 12,24 pg chez le S. scaphoceris. Les crevettes carideennes presentaient des genomes de plus grande taille que les autres crustacees de l'ordre des Decapoda. Les resultats ont revele une tendance a l'accroissement de la taille des genomes chez les crevettes du genre Synalpheus presentant un developpement direct. De plus, les auteurs ont egalement note une correlation biogeographique positive (latitudinale) au sein du sous-ordre Caridea. Cette etude rapporte un nouveau protocole remarquable pour l'analyse FCA (faisant appel a une strategie d'analyse hierarchique), lequel a permis de faire des observations inedites au sujet de la taille du genome chez les crevettes carideennes, specifiquement chez des especes neotropicales du genre Synalpheus, lequel represente le second plus grand groupe au sein du sous-ordre Caridea. [Traduit par la Redaction] Mots-cles : valeur C, crevettes pistolet, Decapoda, contenu en ADN, 1. Introduction The order Decapoda constitutes a large representative group in the extensive and specious subphylum Crustacea. Chace (1951), Martin and Davis (2001), De Grave et al. (2009), and De [...]

The A-kinase anchoring protein 5 (AKAP5) has a variety of biological activities. This study explored whether AKAP5 was involved in cardiomyocyte apoptosis induced by hypoxia and reoxygenation (H/R) and its possible mechanism. H9C2 cells were used to construct an H/R model in vitro, followed by AKAP5 overexpression. Flow cytometry was performed to determine the rate of cardiomyocyte apoptosis. Phosphorylation of phospholamban (PLN), sarcoplasmic/endoplasmic reticulum calcium ATPase 2a (SERCA2a), and apoptosis-related proteins was determined by western blotting. Immunofluorescence staining and immunoprecipitation were performed to detect the distribution and interaction between AKAP5, protein kinase A (PKA), and PLN. After H/R induction, H9C2 cells exhibited significantly reduced AKAP5 protein expression. Upregulation of AKAP5 promotes cell survival and significantly reduces lactate dehydrogenase (LDH) levels and apoptosis rates in H9C2 cells. In addition, the overexpression of AKAP5 was accompanied by the activation of the PLN/SERCA2a signaling pathway and a reduction in apoptosis. Immunofluorescence staining and immunoprecipitation revealed that AKAP5 colocalized and interacted with PLN and PKA. Interestingly, St-Ht31, an inhibitory peptide that disrupts AKAP interactions with regulatory subunits, inhibits the effect of AKAP5 overexpression on H/R-induced apoptosis in H9C2 cardiomyocytes. AKAP5 overexpression alleviated H/R-induced cardiomyocyte apoptosis possibly by anchoring PKA to mediate the PLN/SERCA pathway, suggesting that AKAP5 is a potential therapeutic target for the prevention and treatment of ischemiareperfusion injury. Key words: A-kinase anchoring protein 5 (AKAP5), protein kinase A (PKA), sarcoendoplasmic reticulum calcium transport ATPase (SERCA), hypoxia and reoxygenation (H/R), apoptosis. La proteine AKAP5 (A-kinase anchoring protein 5) possede des activites biologiques variees. Cette etude a examine si AKAP5 est impliquee dans l'apoptose des cardiomyocytes induite par l'hypoxie/reoxygenation (H/R) ainsi que le mecanisme d'action sous-jacent. Des cellules H9C2 ont ete utilisees pour etablir un modele d'H/R in vitro, suivi par la surexpression d'AKAP5 dans les cellules. La cytometrie en flux a ete utilisee pour detecter le taux d'apoptose des cardiomyocytes. L'expression du phospholambane (PLN) phosphoryle, de SERCA2a et de proteines liees a l'apoptose a ete determinee par transfert Western. La coloration par immunofluorescence et l'immunoprecipitation ont ete utilisees pour detecter la distribution et l'interaction entre AKAP5, PKA et PLN. Apres l'induction de l'H/R, les cellules H9C2 montraient une expression significativement reduite de la proteine AKAP5. La regulation a la hausse d'AKAP5 favorisait la survie cellulaire et reduisait de maniere significative le niveau de LDH et le taux d'apoptose des cellules H9C2. En outre, la surexpression d'AKAP5 s'accompagnait de l'activation de la voie de signalisation PLN/SERCA2a et d'une reduction de l'apoptose. La coloration par immunofluorescence et l'immunoprecipitation ont revele que AKAP5 etait colocalisee et interagissait avec PLN et PKA. Il est interessant de noter que St-Ht31, un peptide inhibiteur qui perturbe les interactions entre AKAP et les sous-unites regulatrices, inhibait l'effet de la surexpression d'AKAP5 sur l'apoptose induite par l'H/R dans les cardiomyocytes H9C2. La surexpression d'AKAP5 attenuait l'apoptose des cardiomyocytes induite par l'H/R, possiblement par l'intermediaire de l'ancrage a la PKA pour medier la voie PLN/SERCA, ce qui suggere que AKAP5 est une cible therapeutique potentielle pour la prevention et le traitement des lesions d'ischemie-reperfusion. [Traduit par la Redaction] Mots-cles : AKAP5, proteine kinase A (PKA), ATPase de transport du calcium du SERCA (sarco/endoplasmic reticulum calcium ATPase), hypoxie et reoxygenation (H/R), apoptose., Introduction Cardiovascular diseases (CVDs) account for a large proportion of all human diseases. The incidence and mortality of acute myocardial infarction (AMI) remains high (Boateng and Sanborn 2013; Virani et [...]

Nisin, an antimicrobial peptide produced by Lactococcus lactis, is widely used as a safe food preservative and has recently attracted the attention of researchers as a potential anticancer agent. The cytotoxicity of nisin against human cervical cancer cell lines (HeLa), human ovarian carcinoma cell lines (OVCAR-3 and SK-OV-3), and human umbilical vein endothelial cells (HUVECs) was evaluated using an MTT assay. The apoptotic effect of nisin was identified by Annexin-V/propidium iodide assay, which was further confirmed by western blotting analysis, mitochondrial membrane potential ([DELTA][psi]Wm) analysis, and reactive oxygen species (ROS) assay. The MTT assay showed concentration-dependent cytotoxicity of nisin towards cancer cell lines, with [IC.sub.50] values of 11.5-23 [micro]M, but less toxicity against normal endothelial cells. Furthermore, the treatment of cervical cancer cells with 12 [micro]M nisin significantly (P < 0.05) increased the Bax/Bcl-2 ratio (4.9 fold), reduced [DELTA][psi]Wm (70%), and elevated ROS levels (1.7 fold). These findings indicate that nisin may have anticancer and apoptogenic activities through mitochondrial dysfunction and oxidative stress damage in cervical cancer cells. Key words: nisin, apoptosis, mitochondrial membrane potential, reactive oxygen species, uterine cervical neoplasms, ovarian neoplasms. La nisine, un peptide antimicrobien produit par Lactococcus lactis, est largement utilisee comme agent de conservation alimentaire securitaire et elle a recemment attire l'attention de nombreux chercheurs comme agent anticancereux potentiel. La cytotoxicite de la nisine envers les cellules HeLa, OVCAR-3, SK-OV-3 et HUVEC a ete evaluee a l'aide du test MTT. L'effet apoptotique de la nisine a ete identifie par le dosage de l'Annexin-V/iodure de propidium, puis il a ete confirme par transfert de type Western, l'analyse du potentiel de la membrane mitochondriale (DW m) et le dosage de derives reactifs de l'oxygene (DRO). Le test MTT a revele la cytotoxicite de la nisine envers les lignees cellulaires cancereuses de maniere dependante de la concentration, avec des valeurs CI50 de 11,5-23 [micro]M, mais une toxicite moindre envers les cellules endotheliales normales. En outre, le traitement des cellules cancereuses du col de l'uterus avec 12 lM de nisine augmentait de maniere significative (P < 0,05) le rapport Bax/Bcl-2 (4,9 fois), reduisait le DW m (70 %) et augmentait les niveaux de DRO (1,7 fois). Ces resultats indiquent que la nisine pourrait avoir des activites anticancereuses et apoptogenes par le biais d'un dysfonctionnement mitochondrial et de dommages lies au stress oxydatif dans les cellules cancereuses du col de l'uterus. [Traduit par la Redaction] Mots-cles : nisine, apoptose, potentiel de la membrane mitochondriale, derives reactifs de l'oxygene, neoplasmes du col de l'uterus, neoplasmes ovariens., Introduction Gynecologic cancers (e.g., cervical and ovarian cancers) are common cancers with high mortality rates among women worldwide. Cervical cancer is the fourth most common cancer among women and the [...]

Nasopharyngeal carcinoma (NPC) is a common tumor in the head and neck and is prevalent in China, especially in the southern regions. Molecular mechanisms have attracted much attention in NPC research. FOXD1 has been reported to be a tumor promoter in various cancers. The present study was designed to explore the function of FOXD1 in NPC cells. Functional analyses, including the trypan blue staining assay, EdU and JC-1 assay, and flow cytometry analysis, revealed that FOXD1 facilitated NPC cell proliferation and inhibited NPC cell apoptosis. Next, by means of 'starBase' database and mechanism analyses, such as RIP assay, RNA pull-down assay and luciferase reporter assay, miR-378a-3p was found to target FOXD1 and negatively regulate FOXD1 expression in NPC cells. Moreover, miR-378a-3p plays a suppressive role in NPC cells. LINC00641 was identified as a sponge of miR-378a-3p and positively modulated FOXD1 expression in NPC cells. Finally, a series of rescue assays indicated that LINC00641 accelerated NPC cell proliferation and hindered NPC cell apoptosis through FOXD1 upregulation. In conclusion, the present study demonstrated an innovative ceRNA mechanism of LINC00641/miR-378a-3p/FOXD1 in NPC cells, which might provide new insights into NPC treatment. Key words: FOXD1, LINC00641, nasopharyngeal carcinoma. Le carcinome du nasopharynx (CNP) est une tumeur fréquente de la tête et du cou et il est très répandu en Chine, notamment dans les régions du sud. Les mécanismes moléculaires attirent beaucoup d'attention dans la recherche sur le CNP. FOXD1 a été signalé comme un promoteur de tumeur dans divers cancers. La présente étude a été conçue pour explorer la fonction de FOXD1 dans les cellules du CNP. Des analyses fonctionnelles, dont le test de coloration au bleu de trypan, EdU et JC-1 ainsi que l'analyse par cytométrie en flux, ont révélé que FOXD1 facilite la prolifération des cellules de CNP et inhibe leur apoptose. Par la suite, à l'aide de la base de données << starBase >> et d'analyses des mécanismes comprenant l'immunoprécipitation d'ARN, la précipitation d'ARN et le dosage de l'activité d'un rapporteur luciférase, les auteurs ont découvert que le miR-378a-3p cible FOXD1 et régule négativement l'expression de FOXD1 dans les cellules de CNP. En outre, le miR-378a-3p jouait un rôle suppresseur dans les cellules de CNP. Le LINC00641 a été identifié comme << éponge >> du miR-378a-3p et pourrait moduler positivement l'expression de FOXD1 dans les cellules de CNP. Enfin, une série de tests de sauvetage a indiqué que le LINC00641 accélérait la prolifération des cellules de CNP et entravait l'apoptose des cellules de CNP par le biais de la régulation positive de FOXD1. En conclusion, la présente étude a démontré un mécanisme d'ARNce innovant de LINC00641/miR-378a-3p/FOXD1 dans les cellules de CNP, ce qui pourrait fournir un nouvel aperçu du traitement des NPC. [Traduit par la Rédaction] Mots-clés : FOXD1, LINC00641, carcinome du nasopharynx., Introduction Nasopharyngeal carcinoma (NPC) is the most frequent malignant head and neck epithelial tumor in southern China and Southeast Asia (Liao et al. 2018). Various factors including hereditary factor, environmental [...]

In our study, we explored the effect of astragaloside IV (AgIV) on carboplatin chemotherapy in prostate cancer cell lines in vitro and in vivo. Cell viability assay, colony formation assay, flow cytometry, Western blot, immunohistochemistry, immunofluorescence, and tumor xenograft growth assay were conducted. We found that AgIV significantly decreased the half-maximal inhibitory concentration of carboplatin in prostate cancer cell lines LNCap and PC-3. Moreover, AgIV enhanced the effect of carboplatin in suppressing colony formation and inducing cell apoptosis. A low-dose carboplatin treatment upregulated N-cadherin and Vimentin expression and downregulated E-cadherin expression, but this effect was abolished by combining with AgIV. Carboplatin treatment increased the levels of p-AKT and p-p65 and decreased p-I[kappa]B[alpha], but AgIV treatment suppressed this. In addition, AgIV synergized with carboplatin to suppress tumor xenograft growth of PC-3 cells, and decreased pAKT and p-p65 levels in vivo. Our results suggested that AgIV enhanced carboplatin sensitivity in prostate cancer cell lines by suppressing AKT/NF-[KAPPA]B signaling, thus suppressed epithelial-mesenchymal transition induced by carboplatin. Our findings provided a new mechanism for AgIV in overcoming drug resistance of platinum-based chemotherapy and suggested a potential combination therapy of AgIV and carboplatin in prostate cancer. Key words: carboplatin, astragaloside IV, AKT/NF-[KAPPA]B, prostate cancer. Dans cette etude, les auteurs ont explore l'effet de l'astragaloside IV (AgIV) sur la chimiotherapie au carboplatine dans des lignees cellulaires de cancer de la prostate in vitro et in vivo. Des mesures de viabilite cellulaire et de formation de colonies, des analyses en cytometrie en flux, par buvardage Western, en immunohistochimie et en immunofluorescence ainsi que des tests de croissance de xenogreffes tumorales ont ete realises. Ils ont constate que l'AgIV reduisait de maniere significative la concentration inhibitrice 50 % du carboplatine dans les lignees cellulaires de cancer de la prostate LNCap et PC-3. De plus, lAgIV renforcait l'effet du carboplatine en reprimant la formation de colonies et en induisant l'apoptose. Un traitement a faible dose de carboplatine regulait positivement l'expression de la N-cadherine et de la vimentine et regulait a la baisse l'expression de la E-cadherine, mais cet effet etait supprime par sa combinaison avec lAgIV Le traitement au carboplatine augmentait les niveaux de p-AKT et de p-p65 et diminuait le p-I[kappa]B[alpha], mais le traitement a l'AgIV supprimait cet effet. De plus, l'AgIV agissait en synergie avec le carboplatine pour supprimer la croissance des xenogreffes tumorales de cellules PC-3, diminuait les niveaux de p-AKT et de p-p65 in vivo. En resume, ces resultats suggerent que l'AgIV augmente la sensibilite au carboplatine des lignees cellulaires de cancer de la prostate en reprimant la signalisation par AKT/NF-[KAPPA]B, supprimant ainsi la transition epithelio-mesenchymateuse induite par le carboplatine. Ces resultats ont revele un nouveau mecanisme selon lequel l'AgIV permettrait de surmonter la resistance a la chimiotherapie a base de platine, et ont suggere l'application d'une therapie potentielle combinant lAgIV et le carboplatin dans le cancer de la prostate. [Traduit par la Redaction] Mots-cles: carboplatine, astragaloside IV, AKT/NF-[KAPPA]B, cancer de la prostate., Introduction Prostate cancer is the most commonly diagnosed malignancy in males of old age, with a peak incidence at ~70 years. Androgen receptor had played an important role in the [...]

We describe an automated multidimensional approach for the analysis of flow cytometry data based on pattern classification. Flow cytometry is a widely used technique both for research and clinical purposes where it has become essential for the diagnosis and follow up of a wide spectrum of diseases, such as HIV-infection and neoplastic disorders. Flow cytometry data sets are composed of quite a large number of observations that can be viewed as elements of a n-dimensional space. The aim of the analysis of such data files is typically to classify groups of cellular events as specific populations with biological meaning. Despite significant improvements in data acquisition capabilities of flow cytometers, data analysis is still based on bi-dimensional strategies which were defined a long time ago. These are strongly dependent on the expertise of an expert operator, this approach being relatively subjective and potentially leading to unreliable results. Automated analysis of flow cytometry data is an essential step to improve reproducibility of the results. The proposed automated analysis was implemented on peripherial blood lymphocyte subsets from 307 samples stained and prepared in an identical way and it was capable of identifying all cell subsets present in each sample studied that could also be detected in the same data files by an expert operator. A highly significant correlation was found between the results obtained by an expert operator using a conventional manual method of analysis and those obtained using the implemented automated approach. Index Terms--Automation, B-cell chronic lymphoproliferative disorders, cancer, flow cytometry, leukemia, lymphocytosis, pattern classification, vector quantization.

Flow cytometry with antibodies to bone-specific proteins was used to identify circulating osteoblast-lineage cells in 11 adolescent males and 11 adult males. Findings reveal that osteoblast-lineage cells circulate in physiologically significant numbers, correlate with markers of bone formation, and are markedly higher during pubertal growth.

Abstract Objective: To analyze the pattern of cell-surface expression of the triggering receptor expressed on myeloid cells (TREM) 1 during septic shock. Design and setting: Prospective clinical study in an [...]

A culture-independent approach is used to identify bacteria involved in dimethylsulfoniopropionate (DMSP) degradation at the species level. The analysis indicates that diverse bacterioplankton participate in the metabolism of DMSP in coastal marine systems and their relative importance varies temporally.

The quantitation of fluorescence radiance may at first suggest the need to obtain the number of fluorophore that are responsible for the measured fluorescence radiance. This goal is beset by […]

Research has been conducted on the rhodamine 123 used for the mitochondrial respiratory function evaluation. The optimization and validation of the low-concentration rhodamine 123 straining protocol for the flow-cytometric assessment of mitochondrial membrane potential change in whole yeast cells have been carried out and are reported.

The rate of detection and sizing of individual fluorescently labeled DNA fragments in conventional single-molecule flow cytometry (SMFC) is limited by optical saturation, photon-counting statistics, and fragment overlap to approximately 100 fragments/s. We have increased the detection rate for DNA fragment sizing in SMFC to approximately 2000 fragments/s by parallel imaging of the fluorescence from individual DNA molecules, stained with a fluorescent intercalating dye, as they passed through a planar sheet of excitation laser light, resulting in order of magnitude improvements in the measurement speed and the sample throughput compared to conventional SMFC. Fluorescence bursts were measured from a fM solution of DNA fragments ranging in size from 7 to 154 kilobase pairs. A data acquisition time of only a few seconds was sufficient to determine the DNA fragment size distribution. A linear relationship between the number of detected photons per burst and the DNA fragment size was confirmed. Application of this parallel fluorescence imaging method will lead to improvements in the speed, throughput, and sensitivity of other types of flow-based analyses involving the study of single molecules, chromosomes, cells, etc.

Information for the study of evolution is provided by variations in quantitative morphological traits of various species. Flow cytometry tests on hepatocytes reveal the presence of cell allomery. Large morphological traits have a low coefficient of variation.

Saccharomyces cerevisiae extends carboxyfluorescein (CF) in an energy dependent manner through a secondary transport system. Flow cytometry is used for rapid cell-by-cell analysis of efflux of CF from the cells, and the efflux is saturable and obeys Michaelis-Menten kinetics. Proton motive force is the driving force in the transport system of anionic compounds in S. cerevisiae.

Clathrin-coated vesicles (CCV) present in the renal cortex are abundant in apical proteins as confirmed by flow cytometry analysis. The apical enzymes are found to flow into endosomal elements of renal proximal tubular cells.

Fixed cells with different nucleic acid contents and scatter properties are sorted by flow cytometry (FCM). For each sort, 10,000 cells are captured on poly-L-lysine-coated microplates, which have resulted in efficient and reproducible PCR amplification.

Flow cytometry, fluorescence microscopy, and biochemical analysis were employed to study the use of green fluorescent protein (GFP) as a reporter for the magnetosomal localization and expression of fusion proteins in the microaerophilic Magnetospirillum gryphiswaldense. The data confirm the application of GFP as a reporter for protein localization under magnetite-forming conditions and the utility of MamC as an anchor for magnetosome-specific display of heterologous gene fusions.