Mechanisms for phase separation between TDP-43 and ubiquitin in vitro / 军事医学

Objective To explore the characteristics and mechanism of phase separation between TAR DNA binding protein-43(TDP-43)and ubiquitin.Methods The TARDBP gene and its truncated genes were inserted into vectors to construct recombinant plasmids for expression and protein purification.The phase separation system of ubiquitin and TDP-43 was constructed in vitro.The characteristics of the droplets formed via liquid-liquid phase separation were observed by fluorescence microscopy.The plasmids of ubiquitin and TDP-43 were co-transfected into HEK293T cells to observe aggregates containing TDP-43 and ubiquitin and find out whether TDP-43 could be ubiquitinated.Results The GFP-8Ub,TDP-43 full-length(FL)and truncated proteins were purified.TDP-43 FL and C-terminal domain(CTD)proteins were able to form droplets via phase separation with ubiquitin.The droplets changed into solid-like aggregates after prolonged incubation.Insolvable aggregates containing TDP-43 and ubiquitin were formed.TDP-43 was ubiquitinated under stress conditions in HEK293T cells after being co-transfected with ubiquitin and TDP-43 recombinant plasmids.Conclusion TDP-43 undergoes co-phase separation with ubiquitin,mainly driven by the multivalent interaction between TDP-43′s CTD structural domain and ubiquitin.The droplets finally form aggregates with solid-like properties.Under stress conditions,especially when the protein homeostasis is disrupted,TDP-43 and ubiquitin form aggregates while TDP-43 is ubiquitinated.This study reveals the basic mechanism of TDP-43 co-phase separation with ubiquitin and liquid-solid transformation.

Impact of Different Land Use Practices on Size of Soil Aggregates and Its Mean Weight Diameter under Vertisols of Central India

The present study was carried out at JNKVV, Jabalpur (23010’ N latitude, 79057’ E longitudes and at elevation 393.0 meters above mean sea level). This study was carried out in 2019 that laid out in split plot design with nine main treatments of land use practices (forest land, perennial forage land, uncultivated land, aonla orchard, rice-wheat system, soybean-wheat system, guava orchard, mango orchard and citrus orchard) and three sub-plot treatments of soil depths (0-20, 20-40 and 40-60 cm) which were replicated three times. A total of 81 soil samples were subjected to determination of different size (> 5.0, 2.0-5.0, 1.0-2.0, 0.50-1.0, 0.25-0.50, 0.10-0.25 and <0.10 mm) water stable soil aggregates and mean weight diameter of soil aggregates. Results revealed that land use practices and soil depths significantly affect the mean weight diameter of soil aggregates, distribution of different size soil aggregates. It was noted that irrespective of soil depths, proportions of macro-aggregates (>0.50 mm size) and mean weight diameter were highest under forest land and lowest in soybean-wheat system. However, number of micro-aggregates (< 0.50 mm) increased with soil depths and macro-aggregates and mean weight diameter of soil aggregates were highest at 0-20 cm depth. It can be concluded that extent of soil disturbance significantly alters the proportion of macro-aggregates (>0.50 mm size) and mean weight diameter of water stable soil aggregates with higher in undisturbed (forest, uncultivated and perennial forage) land uses and lower in crop lands which decreased with increase in soil depths.

Optimization and application of caprylic acid precipitation in the purification of monoclonal antibody / 生物工程学报

In response to the market demand for therapeutic antibodies, the upstream cell culture scale and expression titer of antibodies have been significantly improved, while the production efficiency of downstream purification process is relatively fall behind, and the downstream processing capacity has become a bottleneck limiting antibody production throughput. Using monoclonal antibody mab-X as experimental material, we optimized the caprylic acid (CA) precipitation process conditions of cell culture fluid and low pH virus inactivation pool, and studied two applications of using CA treatment to remove aggregates and to inactivate virus. Based on the lab scale study, we carried out a 500 L scale-up study, where CA was added to the low pH virus inactivation pool for precipitation, and the product quality and yield before and after precipitation were detected and compared. We found that CA precipitation significantly reduced HCP residuals and aggregates both before and after protein A affinity chromatography. In the aggregate spike study, CA precipitation removed about 15% of the aggregates. A virus reduction study showed complete clearance of a model retrovirus during CA precipitation of protein A purified antibody. In the scale-up study, the depth filtration harvesting, affinity chromatography, low pH virus inactivation, CA precipitation and depth filtration, and cation exchange chromatography successively carried out. The mixing time and stirring speed in the CA precipitation process significantly affected the CA precipitation effect. After CA precipitation, the HCP residue in the low pH virus inactivation solution decreased 895 times. After precipitation, the product purity and HCP residual meet the quality criteria of monoclonal antibodies. CA precipitation can reduce the chromatography step in the conventional purification process. In conclusion, CA precipitation in the downstream process can simplify the conventional purification process, fully meet the purification quality criterion of mab-X, and improve production efficiency and reduce production costs. The results of this study may promote the application of CA precipitation in the purification of monoclonal antibodies, and provide a reference for solving the bottleneck of the current purification process.

Biochemical and microscopy evidence on adverse effects of nitroxidized human serum albumin

Earlier researches have pointed about the accumulation of peroxynitrite modified proteins and their aggregates in the etiopathogenesis of many age-related neurodegenerative and several autoimmune diseases. Human serum albumin (HSA) is present in abundance in plasma and is susceptible to modification by peroxynitrite. In this study, HSA modified with peroxynitrite (nitroxidized-HSA) formed aggregate besides other gross structural changes. Aggregation or assembly of aberrant proteins is responsible for increase in production of reactive species and is often correlated with toxicity in neurodegenerative diseases. However, lack of literature on the cytotoxicity of aggregated nitroxidized-HSA led us to explore its toxicity using human peripheral blood lymphocytes. Elevated protein carbonyl coupled with decreased protein thiol, and release of antioxidant enzymes and lactate dehydrogenase (LDH) were observed upon incubation of lymphocytes with nitroxidized-HSA. Trypan blue exclusion and MTT assays indicated nitroxidized-HSA induced injury/death of lymphocytes. This may be attributed to the observed reactive oxygen species generation during the interaction of nitroxidized-HSA with lymphocytes. Moreover, the analysis of the cellular morphology by dual staining, fluorescence and confocal microscopy further confirms the cytotoxicity of nitroxidized-HSA. Since various age-related degenerative diseases are characterized by deposition of protein aggregates, the demonstrated toxicity of nitroxidized-HSA may be an important driver in the pathophysiology of neurodegenerative diseases.

In vivo self-aggregation and efficient preparation of recombinant lichenase based on ferritin / 生物工程学报

Enzyme separation, purification, immobilization, and catalytic performance improvement have been the research hotspots and frontiers as well as the challenges in the field of biocatalysis. Thus, the development of novel methods for enzyme purification, immobilization, and improvement of their catalytic performance and storage are of great significance. Herein, ferritin was fused with the lichenase gene to achieve the purpose. The results showed that the fused gene was highly expressed in the cells of host strains, and that the resulted fusion proteins could self-aggregate into carrier-free active immobilized enzymes in vivo. Through low-speed centrifugation, the purity of the enzymes was up to > 90%, and the activity recovery was 61.1%. The activity of the enzymes after storage for 608 h was higher than the initial activity. After being used for 10 cycles, it still maintained 50.0% of the original activity. The insoluble active lichenase aggregates could spontaneously dissolve back into the buffer and formed the soluble polymeric lichenases with the diameter of about 12 nm. The specific activity of them was 12.09 times that of the free lichenase, while the catalytic efficiency was 7.11 times and the half-life at 50 ℃ was improved 11.09 folds. The results prove that the ferritin can be a versatile tag to trigger target enzyme self-aggregation and oligomerization in vivo, which can simplify the preparation of the target enzymes, improve their catalysis performance, and facilitate their storage.

Avaliação da reatividade ao ataque químico de resíduo oriundo da mineração da scheelita utilizado como agregado em argamassas cimentícias / Evaluation of reactivity to the chemical attack of waste from the mining of scheelite used as aggregate in cementitious mortars

RESUMO A mineração de scheelita, localizada em Currais Novos, Rio Grande do Norte, tem gerado grandes volumes de resíduos sólidos que são destinados inadequadamente aos sistemas ambientais, causando impactos ambientais negativos ao município. Uma das alternativas encontradas para destinar adequadamente esses resíduos é o uso em argamassas de matriz cimentícia. No entanto, não há estudos consistentes sobre a viabilidade técnica quanto aos ataques químicos e à influência na sua durabilidade. Nesse contexto, este trabalho teve como objetivo verificar se as argamassas confeccionadas com resíduos da mineração de scheelita em substituição total ao agregado convencional são suscetíveis ao ataque por sulfato de sódio. A caracterização física, química e mineralógica do resíduo foi realizada fazendo uso das seguintes técnicas: análise granulométrica por peneiramento; determinação da massa unitária; obtenção do teor de material pulverulento; espectrometria de fluorescência de raios X, e difração de raios X. A investigação da ocorrência do ataque por sulfato de sódio foi avaliada pela variação dimensional dos corpos de prova quando imersos em solução de sulfato de sódio, de acordo com a metodologia da Associação Brasileira de Normas Técnicas NBR 13583. Os resultados do traço 1:3 indicaram que os agregados oriundos dos resíduos da mineração de scheelita apresentaram comportamento reativo ao ataque por sulfato de sódio (expansão maior que 0,06%), bem como houve aumento da resistência à compressão simples em 4,74%. Portanto, embora tenha sido constatado que os corpos de prova incorporados com resíduos de scheelita tenham sido reativos ao sulfato, observou-se que não foram capazes de deteriorar mecanicamente os corpos de prova.

ABSTRACT The scheelite mining, located in Currais Novos, Rio Grande do Norte, has generated large volumes of solid waste that are inadequately destined for environmental systems, causing negative environmental impacts to the municipality. One of the alternatives to destination of these residues is the use in cementitious matrix mortars. However, there are no consistent studies on the technical feasibility of chemical attacks and influence on their durability. In this context, this work aimed to verify whether mortars made with scheelite mining residues in total substitution to conventional aggregate are susceptible to attack by sodium sulfate. The physical, chemical, and mineralogical characterization of the residue was carried out using the following techniques: sieve particle size analysis; determination of unit mass; obtaining the content of powdery material; X-ray fluorescence spectrometry, and X-ray diffraction. In the investigation of the occurrence of sodium sulfate attack was evaluated by the dimensional variation of the specimens when immersed in sodium sulfate solution, according to the methodology of Associação Brasileira de Normas Técnicas NBR 13583. The results of mixture 1:3 indicated that the aggregates from scheelite mining residues showed a reactive behavior to attack by sodium sulfate (expansion greater than 0.06%), as well as an increase in resistance to simple compression by 4.74%. Therefore, although it was found that specimens incorporated with scheelite residues were reactive to sulfate, it was observed that they were not able to mechanically deteriorate the specimens.

Long-term Use of Balanced and Integrated Nutrient Management Improved Soil Aggregation and Carbon Stabilization in a Maize-wheat Cropping Sequence

Sustainable nutrient management practices have the potential to enhance carbon (C) storage capacity of agricultural soils that may help offset increasing atmospheric CO2 concentration. Nutrient management practices on long-term basis differentially influence aggregates and distribution of soil organic C (SOC) present within aggregates, which in time may affect C stabilization. The present study assessed the impact of long-term application of fertilizers for 44 years either alone or integration with farmyard manure (FYM) on bulk density, SOC and C pools, potassium permanganate oxidizable C (KMnO4), aggregate stability and distribution of C fractions within different size aggregate under maize-wheat cropping sequence. The application of 100%NPK+FYM significantly (P< 0.05) improved soil aggregation and mean weight diameter (MWD). The percent of macro-aggregates (MacroA) and meso-aggregates (MesoA) was maximum in 100%NPK+FYM followed by NPK and the minimum in the control treatment. Irrespective of aggregate classes, TOC (g kg-1 aggregate) was maximum in 100%NPK+FYM treatment with an average of 8.42 g kg-1 aggregate as compared to control (5.05 g kg-1 aggregate). If averaged across the treatments, TOC concentration in aggregates followed the order MacroA> MesoA>MicroA. Correspondingly, results for KMnO4-C were similar in different treatments and aggregate classes. Application of FYM with inorganic fertilizers (NPK) or NPK showed a significant increase in all oxidizable organic C fractions particularly recalcitrant C fraction, which reflects the stable nature of OC as compared to very labile and labile C fractions. In general, C present in mineral fraction and large-sized aggregates (MacroA) has higher recalcitrant fractions of SOC as compared to small-sized aggregates (MesoF and MicroF). The study concluded that long-term balanced and integrated nutrient management improved soil aggregation, C distribution within aggregates, and C storage capacity of soils under maize-wheat. Carbon associated with macro aggregate and a mineral fraction has more recalcitrant C fraction compared to meso and micro aggregate fractions.

Aggregates Characterization and Its Associated Organic Carbon in Two Contrasting Lowland Rice Soils of West Bengal

Aims: The present study investigated the effect of lowland rice soils of two regions viz. new alluvial and red-laterite on aggregate characterization and their associated organic carbon (SOC). Study Design: Randomized block design (RBD). Place and Duration of Study: New alluvial soils were collected from Jangipara block of Hooghly, West Bengal and Red-laterite soils were collected from Raghunathpur block of Purulia, West Bengal during 2017-18. Methodology: For each soil types (New alluvial and Red-laterite) five locations were identified and soil samples were collected from three depths i.e. 0-10, 10-20 and 20-30 cm. The aggregate characteristics i.e. water-stable aggregates (WSAs), mean weight diameter (MWD), aggregate stability and aggregate size fractions along with the distribution of carbon in those aggregate size fractions were critically studied. Results: The aggregate size as well as the stability decreased with increasing soil depth from 0 to 30 cm in both soils. New alluvial soils showed higher aggregate stability than red-laterite soils. Mean weight diameter (MWD) values of new alluvial soils were 34, 29 and 87% more than red-laterite soils at 0-10, 10-20 and 20-30 cm depth, respectively. Presence of higher amount of clay and organic matter in new alluvial made the difference in structural coefficient. The surface soil (0-10 cm) had more coarse aggregate (Cmac A >2000μ) fraction, however, microaggregates (<250μ) were dominant in lower depths in both soils. Water stable aggregates (WSA) in surface soils of new alluvial and red-laterite were 57 and 36%, respectively and were decreased with depth. Red-laterite produced higher micro aggregates as compared to new alluvial soils. Coarse macro aggregate fractions (>2000μ) retained maximum amount of soil organic carbon in both soils however, coarse micro aggregate associated carbon (Cmic AC<250μ) was captured in lower depths. New alluvial soils yielded aggregates with higher in diameter and stability coefficient that is due to higher amount of carbon stored in aggregates. Conclusion: The abundance of macro aggregate of New alluvial soils indicates better soil physical quality than Red-laterite soil which was dominated in higher micro aggregates leads to poor in structure and susceptible to water erosion.

Formation of FADD amyloid fiber and its role in immune signaling in Drosophila melanogaster / 生物工程学报

In this research, we studied the formation of Drosophila melanogaster FADD (Fas-associated death domain-containing protein) amyloid fiber and its influence on signal transduction in IMD (Immune deficiency) signaling pathway to better understand the regulation mechanism of Drosophila innate immune signaling pathway, which will provide reference for the immune regulation in other species. First, we purified dFADD protein expressed in Escherichia coli and performed Sulfur flavin T binding and transmission electron microscopy to identify the dFADD amyloid fibers formed in vitro. Then we investigated the formation of dFADD polymers in S2 cells using SDD-AGE and confocal microscope. We also constructed dFADD mutants to find out which domain is essential to fiber formation and its effect on IMD signal transduction. Our results revealed that dFADD could be polymerized to form amyloid fiber polymers in vitro and inside the cells. Formation of fibers relies on DED (Death-effector domain) domain of dFADD, since DED domain-deleted mutant existed as a monomer. Dual luciferase reporter assay showed that intact DED domain was required for the induction of downstream antimicrobial peptides, indicating that fiber formation was the key to IMD signal transduction. Our study revealed the role of dFADD in mediating the cascade between IMD and Dredd in the IMD signaling pathway by forming amyloid fibers, suggesting an evolutionarily conserved regulatory mechanism of innate immune signaling pathway.

Effect of exercise-induced muscle damage on desmin aggregates / 中国组织工程研究

BACKGROUND: Protein degradation is the main reason of exercise induced muscle damage (EIMD). High-intensity exercise may lead to the misfolding of proteins and form aggregates, which is harmful to the ultrastructure of skeleton muscle. OBJECTIVE: To determine the formation and role of protein aggregate (especially the desmin aggregate) in EIMD. METHODS: Sixty-four adult male Sprague-Dawley rats (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., China) were randomly divided into eight groups (control, post-exercise 0, 3, 6, 12, 24, 48, and 72 hours, eight rats in each). A damage model was established by downhill running (-16°, 16 m/min, 90 minutes). All rats were then killed at corresponding time points to take the soleus as specimen. Severity of damage was observed histologically under a transmission electron microscope. Expression of ubiquitin in the insoluble protein of soleus muscle was detected using western blot, and expression of desmin aggregates was observed by immunofluorescence double labeling. All experimental procedures were approved by the Beijing Sport University Institutional Review Board. RESULTS AND CONCLUSION: (1) After a large-load eccentric exercise, the sarcomere of the soleus muscle was widened, the Z-line was broken, and the myofibril was broken and distorted, all of which was most serious at 12 hours after exercise, and recovered completely at 72 hours after exercise. (2) Western blot results indicated that the expression of ubiquitin in the skeletal muscle increased first and then decreased after a large-load eccentric exercise. The expression peaked at 12 hours after exercise and returned to normal at 72 hours after exercise. (3)Overall expression of desmin aggregates significantly raised (P < 0.01) instantly when the EIMD occurred. It reached the peak at 12 hours after exercise, then reduced somewhat (P < 0.01) at 24 and 48 hours, and finally recovered at 72 hours. (4)These findings indicate that desmin aggregation may be one of the causes of EIMD.

Two Cases of Hypersensitivity Reactions Caused by Human Serum Albumin During Therapeutic Plasma Exchange

Iso-oncotic human serum albumin (HSA) is the primary replacement fluid of choice during therapeutic plasma exchange (TPE). Hypersensitivity reactions to HSA are rare, but require proper evaluation and management. In this article, we report two cases of hypersensitivity reactions to 5% HSA during TPE and discuss strategies to address this problem. The first case was a 60-year-old female patient, who was scheduled for TPE for treatment of recurrent focal segmental glomerulosclerosis after ABO-incompatible kidney transplantation. She developed a pruritic rash on her entire body during the first two sessions of TPE using 5% HSA. The third session was conducted using 500 mL normal saline, 1,000 mL 10% pentastarch, and 750 mL 5% HSA, where she eventually developed a pruritic rash when HSA was infused. There were no adverse events during the fourth and fifth session when fresh frozen plasma was used in place of HSA. The second case was a 50-year-old male patient diagnosed with optic neuritis, who was admitted for five sessions of TPE. The patient developed a pruritic rash on his entire body during the first session of TPE using 5% HSA. The patient experienced no adverse events during the following four sessions using fresh frozen plasma. Certain elements contained in HSA, such as albumin aggregates, prekallikrein activator, and caprylate-modified albumin, might be the reason for these hypersensitivity reactions. Careful selection of alternative replacement fluids is important to avoid premature termination of TPE procedures and secure optimal treatment options for patients.

Inclusion complexes and self-assembled cyclodextrin aggregates for increasing the solubility of benzimidazoles

Albendazole and fenbendazole are imidazole derivatives that exhibit broad spectrum activity against parasites, but the low solubility of these drugs considerably reduces their effectiveness. Complexation of albendazole and fenbendazole with cyclodextrins (ß-cyclodextrin and hydroxypropyl-ß-cyclodextrin) in both water and an aqueous solution of polyvinylpyrrolidone (PVP-k30) was studied to determine if it could increase the solubility and dissolution rate of the drugs. In an aqueous solution, ß-cyclodextrin increased the solubility of albendazole from 0.4188 to ~93.47 µg mL-1 (223×), and of fenbendazole from 0.1054 to 45.56 µg mL-1 (432×); hydroxypropyl-ß-cyclodextrin, on the other hand, increased solubility to ~443.06 µg mL-1 (1058×) for albendazole and ~159.36 µg mL-1 (1512×) for fenbendazole. The combination of hydroxypropyl-ß-cyclodextrin and polyvinylpyrrolidone enabled a solubility increase of 1412× (~591.22 µg mL-1) for albendazole and 1373× (~144.66 µg mL-1) for fenbendazole. The dissolution rate of the drugs was significantly increased in binary and ternary systems, with hydroxypropyl-ß-cyclodextrin proving to be more effective. The presence of the water-soluble PVP-k30 increased the dissolution rate and amorphization of the complexes. Analysis of the changes in displacement and the profile of the cyclodextrin bands in the 1H NMR spectra revealed a molecular interaction and pointed to an effective complexation in the drug/cyclodextrin systems. Monomeric forms and nanoclusters of cyclodextrins were observed in the drug/cyclodextrin systems, suggesting that the increase in solubility of the drugs in the presence of cyclodextrins should not be attributed only to the formation of inclusion complexes, but also to the formation of cyclodextrin aggregates

Diet containing seeds of Buchholzia coriacea accelerates healing of acetic acid induced colitis in rats

Objective: To investigate the anti-colitic effects of diet containing seeds of Buchholzia coriacea (B. coriacea) on acetic acid induced ulcerative colitis. Methods: Male Wistar rats (70-100 g) were fed with standard diets (control group) or with same diet containing B. coriacea seeds at 12.5% or 25% for six weeks. At the sixth week, colitis was induced by intra-rectal administration of 1 mL/200 g 6% acetic acid. Animals were sacrificed at days 0 (before induction), 1, 3 and 7 post induction to assess disease severity via evaluation of stool consistency, haematological variables and histomorphometry of colon tissues. Results: A significant increase in body weight was observed in the 12.5% B. coriacea fed group compared to the control. B. coriacea significantly reduced stool consistency and microscopic scores. Histological evaluation revealed significantly decreased inflammatory aggregates in B. coriacea fed groups compared to control after colitis induction. There was a significant decrease in packed cell volume, red blood cell and haemoglobin concentration in the control group at day 1 post colitis induction compared to 12.5% B. coriacea fed groups. Neutrophils and white blood cell were also significantly increased after colitis induction in the control group while these were significantly decreased in the B. coriacea fed groups. Conclusions: The addition of B. coriacea seeds to diets promotes healing of acetic acid induced colitis by suppressing infiltration of inflammatory aggregates and ameliorating anemia.

Diet containing seeds of Buchholzia coriacea accelerates healing of acetic acid induced colitis in rats

Objective:To investigate the anti-colitic effects of diet containing seeds of Buchholzia coriacea (B.coriacea) on acetic acid induced ulcerative colitis.Methods:Male Wistar rats (70-100 g) were fed with standard diets (control group) or with same diet containing B.coriacea seeds at 12.5％ or 25％ for six weeks.At the sixth week,colitis was induced by intra-rectal administration of 1 mL/200 g 6％ acetic acid.Animals were sacrificed at days 0 (before induction),1,3 and 7 post induction to assess disease severity via evaluation of stool consistency,haematological variables and histomorphometry of colon tissues.Results:A significant increase in body weight was observed in the 12.5％ B.coriacea fed group compared to the control.B.coriacea significantly reduced stool consistency and microscopic scores.Histological evaluation revealed significantly decreased inflammatory aggregates in B.coriacea fed groups compared to control after colitis induction.There was a significant decrease in packed cell volume,red blood cell and haemoglobin concentration in the control group at day 1 post colitis induction compared to 12.5％ B.coriacea fed groups.Neutrophils and white blood cell were also significantly increased after colitis induction in the control group while these were significantly decreased in the B.coriacea fed groups.Conclusions:The addition of B.coriacea seeds to diets promotes healing of acetic acid induced colitis by suppressing infiltration of inflammatory aggregates and ameliorating anemia.

Esquistocitos en frotis de sangre periférica como predictor de morbilidad en preeclampsia / Esquistocytes in peripheral blood smear as predictor of morbidity in preeclampsia

Objetivos: determinar la asociación entre alteración orgánica y los hallazgos morfológicos patológicos en frotis de sangre periférica de pacientes con síndrome preeclampsia-eclampsia. Métodos: está enmarcada en un enfoque cuantitativo, descriptivo y transversal, que tiene como muestra a 54 pacientes, de una población de estudio de 137 pacientes que ingresaron con diagnóstico de Preeclampsia en el Hospital Materno Infantil German Urquidi, en periodo comprendido entre mayo y diciembre del 2016. Consistió en realizar una revisión documentada a través de lista de cotejo de las alteraciones clínica-laboratoriales de cada una de las pacientes; así como la visualización microscópica del extendido de frotis de sangre periférica en búsqueda de esquistocitos, equinocitos y agregados plaquetarios, realizado en un laboratorio autorizado por SEDES. Resultados: de 54 pacientes el 85% presento esquistocitos, equinocitos y agregados plaquetarios, considerándose como resultado positivo; el 15% restante no presentó ninguna de estas alteraciones considerándose como resultado negativo. Tomando estos resultados y comparando con las alteraciones clínica-laboratoriales en este grupo de estudio se pudo determinar que existe asociación entre frotis positivo y alteración orgánica; siendo la razón de probabilidad (OR) de 66; traducido en probabilidades es igual a 98.5% de riesgo de alteración orgánica con un frotis de sangre periférico positivo (Tabla 1). Conclusiones: la búsqueda de esquistocitos, equinocitos y agregados plaquetarios en frotis de sangre periferica es un método sencillo de realizar, económico y rápido; que muestra la probabilidad de desarrollar alguna alteración orgánica cuando el frotis de sangre periférica es positivo; anticipándose por lo tanto a la exacerbación de las manifestaciones clínicas y bioquímicas en pacientes con preeclampsia.

Objectives: to determine the association between organic alteration and pathological morphological findings in peripheral blood smears of patients with preeclampsia-eclampsia syndrome. Methods: It is framed in a quantitative, descriptive and transversal approach, which shows 54 patients from a study population of 137 patients admitted with diagnosis of preeclampsia at the Hospital Materno Infantil German Urquidi, between May and December Of 2016. It is consisted of a documented review through a checklist of the clinical-laboratory alterations of each one of the patients; as the microscopic visualization of the spread of peripheral blood smear in search of schistocytes, equinocytes and platelet aggregates, it was performed in a laboratory authorized by SEDES. Results: of 54 patients, 85% presented schistocytes, equinocytes and platelet aggregates, considered as a positive result; the remaining 15% did not present any of these alterations considering as a negative result. Taking these results and comparing with the clinical-laboratory alterations in this study group, it was possible to determine that there is an association between positive smear and organic alteration; The odds ratio (OR) being 66; Translated in probabilities is equal to 98.5% risk of organic alteration with a positive peripheral blood smear (Table 1). Conclusions: The search for schistocytes, equinocytes and platelet aggregates in smears of peripheral blood is a simple, economical and fast method to perform; which shows the probability of developing some organic alteration when the smear of peripheral blood is positive; thus anticipating the exacerbation of clinical and biochemical manifestations in patients with preeclampsia.

Expression of Platelet-neutrophil Aggregates in Patients With ST-segment Elevation Myocardial Infarction / 中国循环杂志

Objective: This work studied platelet-neutrophil aggregates (PNA) expression in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods: Our research included in 2 groups: STEMI group, n=40 patients admitted in our hospital and Control group, n=30 normal subjects. Blood levels of PNA, platelet-monocyte aggregates (PMA) and platelet-lymphocyte aggregates (PlyA) were measured by lfow cytometry at admission and the results were compared between 2 groups. Results: Compared with Control group, STEMI group had similar blood levels of PlyA and PMA,P=0.245 andP=0.312; while increased blood level of PNA,P<0.05. Correlation analysis showed that platelet-leukocyte aggregates (PLA) level was positively related to CK-MB (r=0.4992,P<0.01) and cTnI (r=0.4365,P<0.01). Conclusion: Blood level of PLA, especially PNA was significantly higher in STEMI patients which suggested that increased PNA might be sued as a new index for AMI diagnosis in clinical practice.

Effects of Ginkgo biloba tablet on MPA, CD11 b and MCP-1 levels in patients with unstable angina pectoris / 中国临床药理学与治疗学

AIM:To observe the levels of monocyte-platelet aggregates (MPA) and markers of activated monocytes in patients with unstable angina pectoris (UAP) accepting Ginkgo biloba tablet treatments,and to explore its mechanisms for cardiovascular disease treatments.METHODS:The levels of MPA,CD11b,and MCP-1 were measured in 92 unstable angina pectoris (UAP) and 42 stable angina pectoris (SAP).The UAP patients were randomly assigned into routine treatment group (control group) and combined tablet treatment group (Ginkgo biloba group).The efficacy was assessed,and the levels of MPA,CD11b,and MCP-1 were measured after 28 days of treatment,respectively.RESULTS:The levels of MPA,CD11b,and MCP-1 in UAP group were higher than those in SAP group (P＜0.001).The levels of MPA and CD11b were positively correlated with MCP-1 level (P ＜ 0.01).The total rate of effective Ginkgo biloba tablet treatment was higher than that of non-Ginkgo biloba tablet treatment (P ＜ 0.05).After 28 days of treatments,the levels of MPA,CD11b,and MCP-1 in Ginkgo biloba group were significantly lower than those in control group (P ＜ 0.001).In total effective treatment group,the levels of MPA,CD11b,and MCP-1 were significantly lower after treatment than those before treatment (P ＜ 0.001),and the decreased rates of these markers after treatment were also much higher (P ＜ 0.01).CONCLUSION:There is an obvious efficacy of Ginkgo biloba tablet on unstable angina pectoris by down-regulating the levels of MPA,CD11b and MCP-1.

The level of platelet leukocyte aggregates in patients with acute cerebral infarction and their short term prognosis / 国际检验医学杂志

Objective to investigate the level of platelet leukocyte aggregates in patients with acute cerebral infarction and their short term prognosis.Methods 105 patients with acute cerebral infarction onset within 24 hours were selected continuously,then platelet leukocyte aggregates including neutrophil aggregates (PNA) and platelet monocyte aggregates (PMA) and platelet lymphocyte aggregates (PlyA) were detected by flow cytometry within 24 hours of admission and the incidence of 14 days.modified Rankin Scale(mRS) was performed at 14 days of onset,as a prognostic indicator,and the mRS score was good at 3.The score >3 mRS was divided into poor prognosis.The level of platelet leukocyte aggregates was detected in 50 healthy subjects.Results (1) The platelet leukocyte aggregates in patients with acute cerebral infarction group were significantly higher than that of the healthy group,which was statistically significant (P3 score comparison,the difference of white blood cell aggregates was statistically significant(P<0.05).Conclusion leukocyte aggregates could be used as an index of short-term prognosis in patients with acute cerebral infarction.

Aged Lewis rats exposed to low and moderate doses of rotenone are a good model for studying the process of protein aggregation and its effects upon central nervous system cell physiology / Ratos Lewis idosos expostos a baixa e moderada doses de rotenona são um bom modelo para estudar o processo de agregação proteica e seus efeitos sobre a fisiologia celular do sistema nervoso central

ABSTRACT Cell physiology is impaired before protein aggregation and this may be more relevant than inclusions themselves for neurodegeneration. The present study aimed to characterize an animal model to enable the analysis of the cell biology before and after protein aggregation. Ten-month-old Lewis rats were exposed either to 1 or 2 mg/kg/day of rotenone, delivered subcutaneously through mini-pumps, for one month. Hyperphosphorylated TAU, alpha-synuclein, amyloid-beta peptide and protein carbonylation (indicative of oxidative stress) were evaluated in the hippocampus, substantia nigra and locus coeruleus through immunohistochemistry or western blot. It was found that 2 mg/kg/day rotenone increased amyloid-beta peptide, hyperphosphorylation of TAU and alpha-synuclein. Rotenone at 1mg/kg/day did not alter protein levels. Protein carbonylation remained unchanged. This study demonstrated that aged Lewis rats exposed to a low dose of rotenone is a useful model to study cellular processes before protein aggregation, while the higher dose makes a good model to study the effects of protein inclusions.

RESUMO A fisiologia celular está prejudicada antes da agregação proteica podendo ser mais importante para a neurodegeneração do que as próprias inclusões. Assim, o objetivo deste estudo é caracterizar um modelo animal para analisar os mecanismos e efeitos da agregação proteica. Ratos Lewis com 10 meses de idade foram expostos a rotenona (1 ou 2 mg/kg/dia), administrada subcutaneamente, utilizando minibombas osmóticas. Os níveis de peptídeo beta-amiloide, TAU hiperfosforilada, alfa-sinucleína e proteínas carboniladas (indicativo de estresse oxidativo) foram avaliados por imunohistoquímica e western blot no hipocampo, substância negra e locus coeruleus. Foi demonstrado que 2 mg/kg/dia de rotenona promoveu aumento do peptídeo beta-amiloide, hiperfosforilação da TAU e alfa-sinucleína. Já 1 mg/kg/dia de rotenona não alterou os níveis dessas proteína nessas regiões. As proteínas carboniladas não se alteraram. Foi demonstrado que ratos Lewis idosos expostos a baixas doses de rotenona são modelo de estudo dos processos celulares antes da agregação proteica, enquanto 2 mg/kg/dia de rotenona permite estudos sobre os efeitos da agregação proteica.

Structural regulation by calcium ion in preparing cross-linked enzyme aggregates / 生物工程学报

We studied the effect of calcium ion on particle size and pore structure of cross-linked enzyme aggregates (CLEAs) of glucose oxidase, with activity and stability of the enzyme as evaluation criteria. With calcium ion to prepare CLEA significantly decreased particle sizes of CLEAs whilst the pore structures of CLEAs gradually disappeared with the increase of calcium concentration. When glucose oxidase was precipitated at 0.1 mmol/L Ca²⁺, glucose oxidase in CLEA showed the definitive pore structure. Moreover, glucose oxidase activity in CLEA with Ca²⁺ was 1.69 times higher than that without Ca²⁺. Even at Ca²⁺ as high as 1.0 mmol/L, glucose oxidase activity in CLEA was 42% higher than that of CLEA without Ca²⁺. Furthermore, CLEA prepared with 0.1 mmol/L Ca²⁺ not only exhibited higher substrate conversion and operational stability, but also increased the maximum reaction speed. Therefore, calcium ion improved the performance of glucose oxidase in CLEAs.