Tetradenia riparia leaves, flower buds, and stem essential oils to control of Aedes aegypti larvae

Abstract Tetradenia riparia (Hochst.) Codd (Lamiaceae) is a species native to the African continent and used as an insect repellent. The objective of the study was to evaluate the larvicidal potential of essential oils (EOs) from the leaves, flower buds, and stem of T. riparia, collected in winter against Aedes aegypti larvae. The EOs were extracted by hydrodistillation (3 h) and identified by GC/MS. The EOs were tested against larvae of A. aegypti at concentrations ranging from 12500 to 1.5 µg/mL for 24 h. The insecticide activity was evaluated by probit analysis, and the anticholinesterase activity was determined by bioautographic method. The results of the class projection indicated sesquiterpenes as the majority class, corresponding to 60.66% (leaves), 64.70% (flower buds) and 83.99% (stem), and the bioassays on A. aegypti larvae indicated LC50 of 1590, 675 and 665 µg/mL, respectively. The anticholinesterase activity indicated that the EO of the leaves inhibited the enzyme at a concentration of 780 µg/mL, and those from the flower buds and stem inhibited up to 1560 µg/mL. The results indicated weak activity of essential oils against A. aegypti larvae.

Essential Oil from the Stem Bark of Casuarina equisetifolia Exerts Anti-inflammatory and Anti-nociceptive Activities in Rats

Abstract Herein the chemical constituents and the anti-pain properties of the essential oil from the stem bark of Casuarina equisetifolia L. (Casuarinaceae) grown in Nigeria were evaluated. The essential oil was obtained by hydrodistillation method in an all glass Clevenger-type apparatus, and characterized by gas chromatography (GC-FID) and gas chromatography-mass spectrometry (GC-MS). The hot plate method was used to determine the anti-nociceptive property whereas the anti-inflammatory activity was evaluated by carrageenan-induced and formalin experimental models. The pale-yellow essential oil was obtained in yield of 0.21% (v/w), calculated on a dry weight basis. The main constituents of the essential oil were methyl salicylate (30.4%), a-zingiberene (15.5%), (E)-anethole (9.5%), b-bisabolene (8.6%), b- sesquiphellandrene (6.9%), and ar-curcumene (6.2%). In the anti-nociceptive study, the rate of inhibition increases as the doses of essential oil increases with optimum activity at the 30th and 60th min for all tested doses. The essential oil displayed anti-nociceptive activity independently of reaction time at the highest tested dose (200 mg/kg). The essential oil of C. equisetifolia moderately reduced pain responses in early and late phases of the formalin test. The oil inhibited the paw licking in the neurogenic phase (60-63%) compared to the late phase of the formalin test. The carrageenan- induced oedema model revealed the suppression of inflammatory mediators within the 1st - 3rd h. Thus, C. equisetifolia essential oil displayed both anti-nociceptive and anti-inflammatory activities independent of the dose tested. The anti-inflammatory and anti-nociceptive activities of C. equisetifolia essential oil are herein reported for the first time

Evaluación preliminar de la actividad de Ageratina havanensis contra virus dengue 2 / Preliminary evaluation of the activity of Ageratina havanensis against the dengue 2 virus

. Introducción: El dengue es la enfermedad viral transmitida por mosquitos de más rápida propagación en el mundo. A pesar de la alta incidencia de la enfermedad no existe tratamiento antiviral efectivo ni vacuna aprobada para su uso contra esta infección. La planta Ageratina havanensis ha sido ampliamente utilizada por nuestra cultura popular y en otros países debido a sus diversas propiedades biológicas. Objetivo: Evaluar la actividad antiviral de cuatro extractos provenientes de Ageratina havanensis y dos flavonoides aislados de dicha planta frente al virus dengue 2. Métodos: La pesquisa primaria de actividad antiviral de cada extracto y flavonoide se realizó mediante el ensayo de detección de la productividad vírica, y se evaluó la acción virucida de los extractos etanólico de tallo y butanólico de hoja. Resultados: Todos los extractos inhibieron la replicación viral al evidenciarse disminución del número de placas de lisis, sin embargo, no se evidenció disminución del título infectivo viral en el ensayo virucida. El tratamiento con la sakuranetina mostró valores de inhibición de la productividad viral entre 61-91 por ciento, y para la 7-metoxiaromadendrina se registraron valores de reducción viral entre 64-86 por ciento. Conclusiones: Los extractos y flavonoides aislados de Ageratina havanensis mostraron acción antiviral. Los extractos etanólico de tallo y butanólico de hojas no poseen acción virucida sobre la cepa viral empleada. El presente trabajo constituye el primer informe respecto a la evaluación de extractos y compuestos aislados de A. havanensis frente al virus dengue(AU)

Introduction: Dengue is the fastest spreading mosquito-borne viral disease worldwide. Despite its high incidence, no effective antiviral treatment or vaccine has been approved for use against this infection. The plant Ageratina havanensis has been widely used in our folk culture and in other countries due to its various biological properties. Objective: Evaluate the antiviral activity of four extracts from Ageratina havanensis and two flavonoids isolated from that plant against the dengue 2 virus. Methods: Primary screening of the antiviral activity of each extract and flavonoid was conducted by viral productivity detection assay, and an evaluation was carried out of the virucidal action of the stem ethanolic extract and the leaf butanolic extract. Results: All the extracts inhibited viral replication by a reduction in the number of lysis plates. However, no evidence was found of a reduced viral infective titer in the virucidal assay. Treatment with sakuranetin achieved viral productivity inhibition values of 61 percent-91 percent, whereas 7-methoxyaromadendrin obtained viral reduction values of 64 percent-86 percent Conclusions: The extracts and flavonoids isolated from Ageratina havanensis displayed antiviral activity. The stem ethanolic extract and the leaf butanolic extract do not have a virucidal action against the viral strain used. The paper is the first report about the effect of extracts and compounds isolated from A. havanensis against the dengue virus(AU)

Trasplante de progenitores hematopoyéticos alogénico haploidéntico en aplasia medular, reporte del primer caso en Cuba / Allogenic hematopoietic stem-cell transplantation in marrow aplasia: the first case report in cuba

Introducción: La aplasia medular adquirida grave es una enfermedad hematológica infrecuente caracterizada por una disminución o ausencia de precursores hematopoyéticos en la médula ósea, lo cual se expresa con distintos grados de citopenias. Varios factores, infecciosos o no, pueden incidir en su origen. Su manejo es complejo y puede incluir tratamiento inmunosupresor y trasplante de progenitores hematopoyéticos alogénico. Objetivo: Demostrar la utilidad de la realización del trasplante de progenitores hematopoyéticos alogénico haploidéntico en pacientes con aplasia medular grave. Caso clínico: Paciente masculino de 21 años de edad, con antecedentes de salud, que en octubre del 2018 debutó con íctero, pancitopenia, lesiones purpúrico hemorrágicas en piel y mucosas, en el curso de una hepatitis aguda seronegativa. La biopsia de médula ósea mostró aplasia medular severa. Se inició tratamiento inmunosupresor con globulina antitimocίtica, ciclosporina A y metilprednisolona. Al cabo de los 6 meses mantenía trombocitopenia severa con necesidades transfusionales y en octubre de 2019 se decide realizar trasplante de progenitores hematopoyéticos alogénico con donante haploidéntico y empleando como tratamiento acondicionante globulina antitimocίtica, fludarabina, ciclofosfamida y bajas dosis de irradiación corporal total. En evaluación clίnica de julio de 2020 (dίa + 280 del trasplante) el paciente estaba asintomático y con parámetros hematológicos normales. Conclusiones: Se demostró que el trasplante de progenitores hematopoyéticos alogénico haploidéntico es un proceder realizable y útil en pacientes con aplasia medular grave, lo cual corrobora el beneficio clínico que puede aportar su ejecución en pacientes con esta enfermedad(AU)

Introduction: Acquired severe marrow aplasia is a rare hematological disease characterized by decrease or absence of hematopoietic precursors in bone marrow, which is expressed with different degrees of cytopenias. Several factors, infectious or not, can influence its origin. Its management is complex and may include immunosuppressive treatment and allogeneic hematopoietic stem-cell transplantation. Objective: To demonstrate the usefulness of performing haploidentical allogeneic hematopoietic stem-cell transplantation in patients with severe medullary aplasia. Clinical case: A 21-year-old male patient, with medical history, who first presented, in October 2018, with icterus, pancytopenia, as well as purpuric hemorrhagic lesions on the skin and mucosa, in the course of acute seronegative hepatitis. The bone marrow biopsy showed severe marrow aplasia. Immunosuppressive treatment was started with antithymocytic globulin, cyclosporine A, and methylprednisolone. After six months, he maintained severe thrombocytopenia under transfusion requirements and, in October 2019, the decision was to perform allogeneic hematopoietic stem-cell transplantation with a haploidentical donor and using antithymocyte globulin, fludarabine, cyclophosphamide, and low doses of total body irradiation as conditioning treatment. In the clinical assessment carried out in July 2020 (day +280 after transplantation), the patient was asymptomatic and with normal hematological parameters. Conclusions: Transplantation of haploidentic allogeneic hematopoietic progenitors was shown to be a feasible and useful procedure in patients with severe marrow aplasia, which corroborates the clinical benefit that its execution can bring in patients with this disease(AU)

Influence of Meloidogyne incognita race 1 on the development of clones of Coffea canephora, variety "Jequitibá Incaper 8122" / Influência de Meloidogyne incognita raça 1 no desenvolvimento de clones de Coffea canephora, variedade "Jequitibá Incaper 8122"

Root-knot nematode is one of the most important phytosanitary problems for Conilon coffee, as it reduces productivity and is difficult to handle. We aimed at studying the infectivity and damage caused by M. incognita race 1 in the "Jequitibá Incaper 8122" intermediate maturity coffee variety. The experiment was conducted in a greenhouse, in completely randomized design, with five replicates. The clones composing the variety "Jequitibá Incaper 8122" were inoculated with 2,000 eggs + second-stage juveniles of M. incognita race 1. Uninoculated plants were the control. Evaluations were performed 180 days after inoculation, considering the plant height (H), stem diameter (SD), number of leaves (NOL), leaf area (LA), number of plagiotropic branches (NPB), number of nodes (NN), chlorophyll content (CHLO), shoot dry matter (SDM), root fresh matter (RFM), final population (FNP), and reproduction factor (NRF). The nematode reduced NOL in clones 208 and 209, NRF in clones 201, 203, 207 and 208, NN in clones 203, 207, 208 and 209, CHLO in clones 201, 204, 206, 207 and 209, SDM in clones 201, 203, 204 and 205 and RFM in clones 205 and 207. M. incognita race 1 FNP and NRF were larger in clones 208, 201, 207 and 203. Clone 202 had FNP and NRF equal to zero, being immune to the nematode. Clone 206 presented the lowest NRF value among clones parasitized by M. incognita.(AU)

O nematoide-das-galhas é um dos mais importantes problemas fitossanitários para o cafeeiro conilon, por reduzir a produtividade e ser de difícil manejo. Objetivou-se estudar a infectividade e os danos causados por M. incognita raça 1 na variedade de café conilon de maturação intermediária "Jequitibá Incaper 8122". O experimento foi conduzido em casa de vegetação, em DIC, com cinco repetições. Os clones que compõem a variedade "Jequitibá Incaper 8122" foram inoculados com 2.000 ovos + juvenis de segundo estádio de M. incognita raça 1. Plantas não inoculadas constituíram a testemunha. As avaliações foram realizadas 180 dias após a inoculação, sendo avaliados: altura da planta (ALT), diâmetro do caule (DCA), número de folhas (NFO), área foliar (AFO), número de ramos plagiotrópicos (NRP), número de nós (NN), teor de clorofila (CLO), massa seca da parte aérea (MSA), matéria fresca da raiz (MFR), população final (PFN) e fator de reprodução (FRE). O nematoide reduziu o NFO nos clones 208 e 209, NRP nos clones 201, 203, 207 e 208, NN nos clones 203, 207, 208 e 209, CLO nos clones 201, 204, 206, 207 e 209, MSA nos clones 201, 203, 204 e 205 e MFR nos clones 205 e 207. PFN e FRE de M. incognita raça 1 foram maiores nos clones 208, 201, 207 e 203; o clone 202 teve a PFN e a FRE igual a zero, apresentando-se imune ao nematoide. O clone 206 apresentou o menor valor de FRE entre os clones parasitados por M. incognita.(AU)

Phytochemical and morpho-anatomical study of the vegetative organs of Psychotria fractistipula L. B.Sm., R.M. Klein & Delprete (Rubiaceae)

Psychotria fractistipula L.B.Sm, R.M. Klein & Delprete (Rubiaceae) is found in the existing Atlantic Forest in the South of Brazil, in the Paraná and Santa Catarina States. The members of Rubiaceae are characterized chemically by the presence of alkaloids, some pharmacological properties of which include antifungal, antibacterial, analgesic, antioxidant, antimutagenic, and antiviral activities. In this study, we report the results of anatomical, morphological, and histochemical analyses of the leaves and stems of the Rubiaceae family member, P. fractistipula. The anatomical analysis involved of light microscopy and scanning electron microscopy. Morpho-anatomical and histochemical characterization were made using standard methodology. The isolated compounds were identified by Nuclear Magnetic Resonance spectroscopic analysis. The following structures are highlighted as the distinctive features of the species: presence of petiolate leaves and a petiole base, leaf blade with smooth cuticle on both surfaces, paracytic stomata, and, dorsiventral mesophyll and presence of trichomes on the leaves and stems. Phytosteroids, identified as β-sitosterol and campesterol, were observed in the hexane fraction of the stem. Future studies should focus on the isolation of other fractions of interest and their characterization through specific biological and cytotoxicity assays.

Agrobacterium rhizogenes-mediated transformation of Hypericum sinaicum L. for the development of hairy roots containing hypericin

Hypericum sinaicum L. is an endangered Egyptian medicinal plant of high importance due to the presence of naphthodianthrones (hypericins), which have photodynamic properties and pharmaceutical potential. We sought to assess H. sinaicum ability to develop hairy roots that could be cultured in contained conditions in vitro and used as a source for hypericin production. We used four A. rhizogenes strains differing in their plasmids and chromosomal backgrounds to inoculate excised H. sinaicum root, stem and leaf explants to induce hairy root development. Additionally, inoculum was applied to shoots held in Rockwool cubes supporting their stand after removal of the root system. All explant types were susceptible to A. rhizogenes although stem explants responded more frequently (over 90%) than other explant types. The A4 and A4T A. rhizogenes strains were highly, and equally effective in hairy root induction on 66-72% of explants while the LBA1334 strain was the most effective in transformation of shoots. Sonication applied to explants during inoculation enhanced the frequency of hairy root development, the most effective was 60 s treatment doubling the percentage of explants with hairy roots. However, shoot transformation was the most effective approach as shoots developed hairy roots within 10 days after inoculation. Molecular analyses confirmed that the established hairy root cultures in vitro were indeed obtained due to a horizontal gene transfer from bacteria. These cultures grew fast and the hypericin content in hairy roots was about two fold higher than in H. sinaicum plants as determined by HPLC.

Specification of Bacteriophage Isolated Against Clinical Methicillin-Resistant Staphylococcus Aureus

OBJECTIVES: The emergence of resistant bacteria is being increasingly reported around the world, potentially threatening millions of lives. Amongst resistant bacteria, methicillin-resistant Staphylococcus aureus (MRSA) is the most challenging to treat. This is due to emergent MRSA strains and less effective traditional antibiotic therapies to Staphylococcal infections. The use of bacteriophages (phages) against MRSA is a new, potential alternate therapy. In this study, morphology, genetic and protein structure of lytic phages against MRSA have been analysed. METHODS: Isolation of livestock and sewage bacteriophages were performed using 0.4 μm membrane filters. Plaque assays were used to determine phage quantification by double layer agar method. Pure plaques were then amplified for further characterization. Sulfate-polyacrylamide gel electrophoresis and random amplification of polymorphic DNA were run for protein evaluation, and genotyping respectively. Transmission electron microscope was also used to detect the structure and taxonomic classification of phage visually. RESULTS: Head and tail morphology of bacteriophages against MRSA were identified by transmission electron microscopy and assigned to the Siphoviridae family and the Caudovirales order. CONCLUSION: Bacteriophages are the most abundant microorganism on Earth and coexist with the bacterial population. They can destroy bacterial cells successfully and effectively. They cannot enter mammalian cells which saves the eukaryotic cells from lytic phage activity. In conclusion, phage therapy may have many potential applications in microbiology and human medicine with no side effect on eukaryotic cells.

RNA viruses in trypanosomatid parasites: a historical overview

Viruses of trypanosomatids are now being extensively studied because of their diversity and the roles they play in flagellates' biology. Among the most prominent examples are leishmaniaviruses implicated in pathogenesis of Leishmania parasites. Here, we present a historical overview of this field, starting with early reports of virus-like particles on electron microphotographs, and culminating in detailed molecular descriptions of viruses obtained using modern next generation sequencing-based techniques. Because of their diversity, different life cycle strategies and host specificity, we believe that trypanosomatids are a fertile ground for further explorations to better understand viral evolution, routes of transitions, and molecular mechanisms of adaptation to different hosts.

Avaliação de protetores solares contendo filtros encapsulados/incorporados em matriz de sílica mesoporosa ordenada do tipo SBA-15 / Evaluation of sunscreens containing encapsulated/entrapped UV filters in ordered mesoporous silica type SBA-15

Filtros orgânicos são amplamente utilizados em formulações fotoprotetoras, com habilidade de absorver radiações ultravioleta (UV). Contudo, parte destes compostos possuem limitações, como: fotoinstabilidade, permeação cutânea e fotossensibilização e entre outros. Este trabalho envolveu a síntese de matriz mesoporosa do tipo SBA-15, encapsulação/incorporação de ρ-metoxicinamato de octila (MCO), benzofenona-3 (BZF-3) e avobenzona (AVO) na SBA-15 para aplicação em formulações fotoprotetoras. Fez-se a determinação da eficácia in vitro dos filtros encapsulados/incorporados combinados a ingrediente cosmético; o preparo de bastão fotoprotetor e determinação eficácia estimada; a avaliação do potencial de irritação ocular dos bastões por HET-CAM - Hen's Egg Test - Chorioallantoic Membrane, e a avaliação da permeação/retenção cutânea e perfil de biodistribuição dos filtros. Para a caracterização dos materiais foram empregadas técnicas físico-químicas e analíticas. As medidas de adsorção/dessorção de N2 mostrou que as amostras dos filtros solares encapsulados/incorporados apresentaram diminuição na área superficial e volume de poro (V), indicando que os filtros solares foram encapsulados/incorporados na superfície e nos poros da SBA-15. Os resultados de Espalhamento de raios X a baixo ângulo evidenciaram que os filtros solares não afetaram a estrutura hexagonal da SBA-15. Por TG/DTG e análise elementar foi possível determinar a quantidade de filtros solares na superfície e nos mesoporos da SBA-15. Enquanto, as curvas DSC e DTA revelaram aumento na estabilidade térmica da AVO e BZF-3, quando encapsulados/incorporados. Os resultados da eficácia fotoprotetora in vitro mostraram que a combinação dos três filtros solares encapsulados/incorporados na SBA-15 promoveram aumento de 52% no fator proteção solar (FPS), enquanto que, na formulação fotoprotetora contendo os três filtros encapsulados/incorporados, o aumento foi de 94%. O ensaio de HET-CAM evidenciou que os bastões contendo SBA-15 e os filtros encapsulados/incorporados não foram irritantes. O ensaio de permeação/retenção cutânea mostrou que o processo de encapsulação/incorporação da BZF-3 promoveu diminuição de sua permeação em todos os tempos de exposição. As quantidades permeadas de AVO e MCO ficaram abaixo do limite de quantificação nos tempos 6 e 12 h, no entanto, no tempo de 24 h foi possível quantificá-los. As quantidades dos filtros solares retidas na pele a partir da formulação contendo os filtros solares encapsulados/incorporados na SBA-15 (F4) foram menores (tempos 6 e 12 h) em comparação à formulação contendo os filtros solares não encapsulados (F3). A investigação da biodistribuição dos filtros solares mostrou que a retenção total na pele, como na derme, foi menor na formulação F4 em comparação à F3. O estudo comparativo entre pele suína e a pele humana mostrou que as quantidades de BZF-3 e AVO permeadas e retidas na pele suína foram superiores do que em relação à humana para ambas as formulações (F4 e FR). Pela técnica de biodistribuição, foi possível determinar que os filtros solares oriundos das formulações F3 e referência (FR) apresentaram maior retenção destes compostos na derme do que em outras camadas da pele. Contudo, observou-se que os filtros encapsulados apresentaram taxa reduzida de retenção na derme

Organic Filters are chemical compounds widely used in sunscreens formulations with the ability to absorb ultraviolet radiation (UV). Despite the effectiveness of these compounds in UV radiation protection, disadvantages related to their photo instability, potential skin permeation and photo sensibility pose significant challenges for improving these products. The aim of this work was to synthesize mesoporous matrix SBA-15, encapsulation/entrapping of octyl methoxycinnamate (OMC), benzophenone-3 (BZF-3) and avobenzone (AVO) into SBA-15 for application in photoprotective formulations. It was accessed in vitro photoprotection efficacy and in vitro photostability assay of encapsulated/entrapped UV filters combined with cosmetic ingredient and photoprotective stick formulations; evaluation of the ocular irritation potential of photoprotective stick formulations by in vitro method HET-CAM - Hen's Egg Test - Chorioallantoic Membrane; evaluation the skin permeation/deposition and biodistribution profile of photoprotective stick formulations. The decrease in the surface area and in mesoporous volume (V) observed in the nitrogen adsorption desorption isotherms of encapsulated/entrapped samples indicated that UV filters were efficiently encapsulated/entrapped into SBA-15. Additionally, SAXS results showed that UV filters did not affected the hexagonal structure of the mesoporous material and that these compounds filled the SBA-15 pores. TG/DTG and elemental analysis were efficient tools to confirm the presence and the quantity of UV filters into SBA-15. DTA and DSC curves of encapsulated/entrapped materials showed that the thermal stability of AVO and BZF-3 were increased. On the other hand, DSC curves of encapsulated/entrapped materials demonstrated that thermal stability of OMC was not increase. The in vitro photoprotective efficacy results demonstrated that the combination of the three sunscreens encapsulated/entrapped into SBA-15 increased 52.0% the SPF values, while the stick formulation containing the UV filters encapsulated/entrapped, the increase was 94.0%. Delivery experiments using porcine skin demonstrated that the encapsulation/entrapping process of UV filters resulted the decreased of BZF-3 permeation and deposition in skin (6 and 12 hours). The cutaneous biodistribution profile of UV filters showed that the deposition of these compounds from encapsulated/entrapped stick formulation (F4) was significantly lower than that from UV filters stick formulations (F3) in the total slices of the skin and dermis. The comparative study between porcine skin and human skin demonstrated that the amounts of BZF-3 and AVO permeated and deposited in porcine skin were higher than in human skin for both formulations (F4 and FR - reference formulation). By the biodistribution technique it was possible to determine that the UV filters from the formulations F3 and FR presented higher retention of these compounds in the dermis than in other layers of the skin. On the other hand, it was observed that the encapsulated UV filters presented low retention rate into dermis

Development and evaluation of ultrasound contrast agents with AS1411-conjugated nanoparticles with liquid core / 中南大学学报(医学版)

To prepare AS1411 targeted nano-ultrasonic contrast agent with liquid core, and to evaluate its ability for ultrasonic contrast enhancement and targeting MCF-7 cell in vitro.
 Methods: The modified solvent evaporation, self-synthesized membrane material and perfluorobrominane (PFOB) was used to form nano-ultrasonic contrast agent with PFOB core (nanoparticles, NP); then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS) catalysis was used to connect AS1411 to the surface of NP to prepare NP-AS1411. The transmission electron microscopy was chosen to check the morphology of NP-AS1411. The size, surface charge, encapsulation efficiency, biocompatibility, the contrast grey value and the stability of NP-AS1411 and NP were compared. Whether AS411 was attached to the surface of NP was checked by gel electrophoresis. Fluorescence microscopy and flow cytometry were performed to examine the targeting ability of AS1411.
 Results: NP-AS1411 was a shell-nuclear structure under the electron microscope. Its size was at (245.4±16.5) nm, which was larger than that of NP (P=0.05). There was no significant difference in surface charge and encapsulation efficiency between NP-AS1411 and NP (P>0.05). In the MTT experiment, the cell viability decreased significantly at high concentration of NP-AS411 (25 mg/mL) after incubation for 24 h compared with the control group (0 mg/mL ) (P0.05). The contrast grey value of AS1411-NP was 80.1±9.2 after keeping at room temperature for 24 h, which showed no obviously change comparing with that before the treatment (P>0.05). The size of NP-AS1411didn't change too (P>0.05). The results of gel electrophoresis demonstrated that the AS1411 connecting to the surface of NP was the most when the molar ratio of NP:AS1411 was at 40:1. Flow cytometry analysis confirmed that NP and NP-AS1411 were combined with MCF-7 cells separately but the fluorescence produced by the combination of NP-AS1411 and MCF-7 was more intense.
 Conclusion: The modified solvent evaporation and EDC/NHS catalysis could successfully prepare ultrasound contrast agents with aptamer-conjugated nanoparticles with liquid core. The targeted ultrasonic contrast agents with liquid core possess good ultrasonic contrast enhancement ability in vitro, stability and specificity as well.

Trasplante autológo de células madre hematopoyéticas en pacientes con diagnóstico de neuroblastoma de alto riesgo en la Fundación Valle del Lili, Cali - Colombia / Autologous Haematopoietic Stem-cell Transplantation in Patients with High Risk Neuroblastoma in the Valle del Lili Foundation Hospital, Cali, Colombia

Resumen Objetivo: Describir la experiencia en el manejo de los pacientes con diagnóstico de neuroblastoma de alto riesgo manejados con trasplante autólogo de células madre hematopoyéticas en la Fundación Valle de Lili. Pacientes y métodos: Estudio descriptivo, tipo serie de casos de pacientes con diagnóstico de neuroblastoma de alto riesgo que recibieron trasplante autólogo de células madre hematopoyéticas entre 2001 y 2015. Los desenlaces de este estudio fueron: supervivencia global; supervivencia libre de evento; tiempo de injerto de plaquetas y neutrófilos, e incidencia acumulada de enfermedad veno oclusiva. Se realizó un análisis estadístico descriptivo para todas las variables consideradas en el análisis y para subgrupos seleccionados. El análisis de supervivencia se hizo con el método Kaplan-Meier. Resultados: Entre 2001 y 2015 quince pacientes con diagnóstico de neuroblastoma recibieron trasplante autólogo de células madre hematopoyéticas. La supervivencia globala3y5anos fue del 55% y la supervivencia libre de evento fue del 47%, donde 14 pacientes injertaron neutrófi los entre el día8y19 postrasplante e injerto de plaquetas entre los 9 y 91 días y 2 pacientes desarrollaron enfermedad venooclusiva hepatica como toxicidad a los fármacos quimioterapéuticos de acondicionamiento. Al momento del último seguimiento 10 pacientes permanecen vivos, de los cuales 8 no presentan evidencia clínica y/o paraclínica de la enfermedad. De los 5 pacientes que fallecieron, 2 fue por toxicidad al trasplante en los primeros 100 díasy3por progresión de la enfermedad. Conclusión: El trasplante autólogo de células madre hematopoyéticas es una alternativa facti ble como tratamiento en nuestro medio para pacientes con diagnóstico de neuroblastoma de alto riesgo, el cual ha contribuido a mejorar la supervivencia en este grupo de pacientes.

Abstract Objective: Describe the experience in the management of patients diagnosed with high-risk neuroblastoma treated with autologous haematopoietic stem cell transplantation at the Valle de Lili Foundation Hospital. Patients and Methods: A series of cases of patients with a diagnosis of high-risk neuroblastoma who received an autologous haematopoietic stem cell transplantation between 2001 and 2015. The endpoints of this study were: overall survival, event-free survival, platelet and neutrophil graft time and the cumulative incidence of venous-occlusive disease. A descriptive statistical analysis was performed for all the variables considered in the analysis and for the selected subgroups. Survival analysis was performed using the Kaplan-Meier method. Results: A total of 15 patients diagnosed with high risk neuroblastoma received an autologous haematopoietic stem cell transplantation between 2001 and 2015. Overall survival at 3 and 5 years was 55%, and the event-free survival was 47%. 14/15 patients grafted Neutrophils grafted between day 8 and 19 post-transplant in 14/15 patients, with platelet graft between days 9 and 91 days. Hepatic venous-occlusive disease was observed in 2/15 patients as toxicity to conditioning chemotherapeutic drugs. At the time of the last follow-up, 10/15 patients remained alive, 8 of whom had no clinical and/or para-clinical evidence of the disease. Of the 5/15 patients that died, 2 were due to transplant toxicity in the first 100 days, and 3 due to disease progression. Conclusion: We conclude that autologous haematopoietic stem cell transplantation is a viable alternative as a treatment in our setting for patients with high-risk neuroblastoma, and has contributed to improve survival in this group of patients.

Establishment of a Novel Mouse Model of Coronary Microembolization / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Coronary microembolization (CME) has been frequently seen in acute coronary syndromes and percutaneous coronary intervention. Small animal models are required for further studies of CME related to severe prognosis. This study aimed to explore a new mouse model of CME.</p><p><b>METHODS</b>The mouse model of CME was established by injecting polystyrene microspheres into the left ventricular chamber during 15-s occlusion of the ascending aorta. Based on the average diameter and dosage used, 30 C57BL/6 male mice were randomly divided into five groups (n = 6 in each): 9 μm/500,000, 9 μm/800,000, 17 μm/200,000, 17 μm/500,000, and sham groups. The postoperative survival and performance of the mice were recorded. The mice were sacrificed 3 or 10 days after the surgery. The heart tissues were harvested for hematoxylin and eosin staining and Masson trichrome staining to compare the extent of inflammatory cellular infiltration and fibrin deposition among groups and for scanning transmission electron microscopic examinations to see the ultrastructural changes after CME.</p><p><b>RESULTS</b>Survival analysis demonstrated that the cumulative survival rate of the 17 μm/500,000 group was significantly lower than that of the sham group (0/6 vs. 6/6, P = 0.001). The cumulative survival rate of the 17 μm/200,000 group was lower than those of the sham and 9 μm groups with no statistical difference (cumulative survival rate of the 17 μm/200,000, 9 μm/800,000, 9 μm/500,000, and sham groups was 4/6, 5/6, 6/6, and 6/6, respectively). The pathological alterations were similar between the 9 μm/500,000 and 9 μm/800,000 groups. The extent of inflammatory cellular infiltration and fibrin deposition was more severe in the 17 μm/200,000 group than in the 9 μm/500,000 and 9 μm/800,000 groups 3 and 10 days after the surgery. Scanning transmission electron microscopic examinations revealed platelet aggregation and adhesion, microthrombi formation, and changes in cardiomyocytes.</p><p><b>CONCLUSION</b>The injection of 500,000 polystyrene microspheres at an average diameter of 9 μm is proved to be appropriate for the mouse model of CME based on the general conditions, postoperative survival rates, and pathological changes.</p>

Co-infection by Tritrichomonas foetus and Pentatrichomonas hominis in asymptomatic cats / Co-infecção por Tritrichomonas foetus e Pentatrichomonas hominis em gatos assintomáticos

Tritrichomonas foetus, a parasite well known for its significance as a venereally transmitted pathogen in cattle, has been identified as a cause of chronic large bowel diarrhea in domestic cats in many countries of the world. In Brazil, several studies on the diagnosis of bovine trichomoniasis have been performed, but until now, no study was made regarding feline trichomoniasis. Thus, this is the first study to report the occurrence of T. foetus and Pentatrichomonas hominis in cats using morphological and molecular analysis. Feces from 77 cats were examined, four of which (5.2%) were positive for the presence of parabasalids. Morphological analysis of stained smears revealed piriform trophozoites showing the three anterior flagella, elongated nucleus and axostyle ending abruptly in fillet, characteristic of T. foetus. In scanning and transmission electron microscopy, identification characters similar to those previously reported for T. foetus were observed. The cultures containing trophozoites were submitted for molecular analysis, which resulted positive for T. foetus DNA using specific primers (TFR3 and TFR4), and all samples were positive and subjected to sequencing in which they showed 99.7-100% similarity with another isolate sequencing of T. foetus (JX960422). Although no trophozoite with consistent morphology of P. hominis has been visualized in the samples, differential diagnosis was performed using specific primers for P. hominis (TH3 and TH5) amplicon. In three of the four samples (3.89%) sequencing revealed 100% similarity when compared with another sequence of P. hominis deposited in Genbank (KC623939). Therefore, the present study revealed through the diagnostic techniques employed the simultaneous infection by T. foetus and P. hominis in the feces of cats. However, it was necessary to use more than one technique for the diagnosis of the co-infection...

Tritrichomonas foetus, um parasito bem conhecido por seu significado como um agente patogênico transmitido venereamente em bovinos, também foi identificado como causa de diarreia crónica do intestino grosso em gatos domésticos em muitos países. No Brasil, vários estudos sobre o diagnóstico de tricomonose bovina foram realizados, mas até agora, não há informação disponível em relação à trichomonose felina. Assim, este é o primeiro estudo a relatar a ocorrência de T. foetus e Pentatrichomonas hominis em gatos por meio de análise morfológica e molecular. Fezes de 77 gatos foram examinadas, a partir da qual quatro (5,2%) foram positivas para a presença de parabasalídeos. A análise morfológica de esfregaços corados revelou trophozoitos piriformes com três flagelos anteriores, núcleo alongado e axóstilo cuja projeção termina abruptamente em formato de filete, características estas de identificação morfológica T. foetus. Além disso, microscopia eletrônica de varredura e transmissão, revelaram caracteres morfológicos semelhantes aos descritos na literatura para esta espécie. A análise molecular de culturas utilizando iniciadores específicos para trofozoítos de T. foetus (TFR3 e TFR4), mostrou que as quatro amostras foram positivas para este parasito e osequenciamento dos fragmentos amplificados demonstraram 99,7-100% de similaridade com seqüências depositadas no GeneBank de T. foetus. Nenhum trofozoíto com morfologia consistente com a descrição de P. hominis foi visualizado nas amostras. No entanto, a análise molecular, utilizando iniciadores específicos para esta espécie (TH3 e TH5) detectou que três das quatro amostras (75%) também foram positivas para P. hominis e o sequenciamento de nucleotideos revelou 100% de similaridade dos amplicons quando comparada com o mesmo fragmento de DNA de P. hominis depositado no GenBank. Como tal, o presente estudo relata a coinfecção de gatos com T. foetus e P. hominis e destacou a exigência de uma combinação de métodos...

Cuidado em saúde em adultos com hipertensão arterial autorreferida no Brasil segundo dados da Pesquisa Nacional de Saúde, 2013 / Health care in adults with self-reported hypertension in Brazil according to the National Health Survey, 2013

RESUMO: Objetivo: Descrever indicadores de cuidado em saúde em adultos com hipertensão arterial autorreferida no Brasil, segundo características sociodemográficas. Métodos: Foram utilizados dados da Pesquisa Nacional de Saúde 2013, estudo transversal de base populacional, referentes ao cuidado em saúde com a hipertensão arterial autorreferida quanto ao uso de serviços de saúde. As prevalências e seus intervalos de confiança de 95% (IC95%) foram calculados segundo sexo, idade, raça/cor e escolaridade, representativos para Brasil e grandes regiões. Resultados: A hipertensão arterial foi referida por 21,4% (IC95% 20,8 - 22,0) dos entrevistados, sendo maior em mulheres e em pessoas sem instrução e com ensino fundamental incompleto. Dentre estes, 45,9% (IC95% 44,0 - 47,7) referiram ter recebido assistência médica pela última vez em uma Unidade Básica de Saúde; 81,4% (IC95% 80,1 - 82,7) referiram tomar medicamentos para a hipertensão; e 92,0% (IC95% 91,2 - 92,8) referiram ter realizado todos os exames complementares que foram solicitados. Conclusão: É importante conhecer a cobertura e o acesso aos serviços de saúde para o cuidado aos indivíduos com hipertensão arterial, de modo a avançar na qualidade da assistência prestada e reduzir as desigualdades identificadas.

ABSTRACT: Objective: To describe health care indicators in adults with self-reported hypertension in Brazil, according to socio-demographic characteristics. Methods: Data from the National Health Survey 2013, a cross-sectional population-based study, about health care of self-reported hypertension and health services were used. Prevalence and 95% confidence intervals (95%CI) were calculated for sex, age, race/color skin and schooling levels, representing Brazil and major regions. Results: Hypertension was reported by 21.4% (95%CI 20.8 - 22.0) of respondents, being higher in women and in people without instruction and incomplete middle school. Among these, 45.9% (95%CI 44.0 - 47.7) reported having received medical care for the last time in a basic health unit; 81.4% (95%CI 80.1 - 82.7) reported taking medication for high blood pressure; and 92.0% (95%CI 91.2 - 92.8) reported having taken all requested complementary examinations. Conclusion: It is important to know the coverage and access to health services for the care of patients with hypertension, in order to improve care quality and reduce identified inequalities.

Histochemical study of the oviducal gland and analysis of the sperm storage tubules of Mustelus schmitti Springer, 1939 (Chondrichthyes, Triakidae)

The paired oviducal glands of immature and mature females of Mustelus schmitti were examined macro and microscopically. Findings indicate that these glands possessed the same zonation as in most chondrichthyans from anterior to posterior: club, papillary, baffle and terminal zones. The whole gland is composed by simple tubular glands that connect with transverse grooves all along the organ. The club zone presents a typical indian club shape with a simple columnar and ciliated epithelium including secretory cells PAS (+) and AB (+). The papillary zone is characterized by lamella forming small and long cones in numbers of three. The epithelium of this zone contains ciliated cells with apical nuclei and secretory cells with basal nuclei that stain AB (+)The baffle zone consists of apically flattened lamellae alternating with spinnerets which are small projections disposed by both sides of the plateau. This whole structure is present in number of 8 or 9 units. A simple columnar ciliated epithelium covers the plateau and spinnerets and no AB or PAS staining is observed. The epithelium of the terminal zone is PAS (-) and AB (+), and elongated tubules, that run adjacent to the baffle zone are the site where groups of spermatozoa are clearly observed in the lumen. The epithelium of the sperm storage tubules do not stain with any of the dyes tested. Sperm was also observed in the baffle zone, presumably in its way to the fecundation in the oviduct because it displays no aggregation pattern and was between the folds of the epithelium. By scanning electron microscopy sperm was observed in the club and baffle zones in a gland which belonged to a pregnant female...

Dermatosparaxis in two White Doper lambs in Brazil: case report / Dermatosparaxis em dois cordeiros White Dorper no Brasil: relato de caso

Dermatosparaxis is a genetic disease that affects collagen maturation. This disease is characterized by marked impairment of the resistance of collagen fibers that leads to skin fragility, and it may affect several species. This is the first report of dermatosparaxis in sheep in Brazil. Clinically, two White Dorper lambs had multiple skin lacerations in the neck, dorsum and limbs. Transmission microscopy demonstrated irregular collagen fibers arranged in hieroglyphic shape and scanning electron microscopy demonstrated thin collagen fibrils that were not arranged in bundles as observed in the normal dermis. These findings are consistent with the diagnosis of dermatosparaxis.

A dermatosparaxis é uma doença genética que afeta a maturação do colágeno. Essa doença é caracterizada por redução acentuada da resistência das fibras colágenas que leva à fragilidade da pele. Esse é o primeiro relato de dermatosparaxis em ovinos no Brasil. Clinicamente, dois cordeiros da raça White Dorper tiveram múltiplas lacerações na pele do pescoço, dorso e membros. A microscopia de transmissão demonstrou fibras de colágeno irregularares arranjadas em formato de hieroflifo, e a microscopia de varredura demonstrou fibras finas de colágeno não arranjadas em bandas como na derme do animal normal. Esses achados são consistentes com o diagnóstico de dermatosparaxis.

[Generation of induced pluripotent stem cells from mesenchymal stem cells of human umbilical cord vein]

The aim of this study was to generate induced pluripotent stem cells from human umbilical cord vein mesenchymal stem cells by use of plasmid vector. In this experimental study using type IV collagenase enzyme, we extracted and cultured mesenchymal stem cells obtained from human umbilical cord vein wall.Usingelectroporation method, these cells were transfected with plasmid vector which carried selfrenewal transcription factors of OCT4 and SOX2.After 9 days we observed the induced pluripotent stem cells like colonies. The nature of thesecells were evaluated byimmunocytochemistry and alkalin phosphatase activity of embryonic stem cells. Immunocytochemical analysis showed that these cells express the pluripotency markerssuch as OCT4, SSEA4, TRA1-60, TRA1-81, and also hadalkalin Phosphatase activity. This study revealed that transient expression of self renewal genes of OCT4 and SOX2 could lead to development of induced pluripotent stem cells like colonies from umbilical cord vein mesenchymal stem cells

Comparison of proliferative and multilineage differentiation potential of sheep mesenchymal stem cells derived from bone marrow, liver, and adipose tissue

Despite major progress in our general knowledge related to the application of adult stem cells, finding alternative sources for bone marrow Mesenchymal Stem Cells [MSCs] has remained to be challenged. In this study successful isolation, multilineage differentiation, and proliferation potentials of sheep MSCs derived from bone marrow, adipose tissue, and liver were widely investigated. The primary cell cultures were prepared form tissue samples obtained from sheep 30-35 day fetus. Passage-3 cells were plated either at varying cell densities or different serum concentrations for a week. The Population Doubling Time [PDT], growth curves, and Colony Forming Unit [CFU] of MSCs was determined. The stemness and trilineage differentiation potential of MSCs were analyzed by using molecullar and cytochemical staining approaches. The data was analyzed through one way ANOVA using SigmaStat [ver. 2]. The highest PDT and lowest CFU were observed in adipose tissue group compared with other groups [p<0.001]. Comparing different serum concentrations [5, 10, 15, and 20%], irrespective of cell sources, the highest proliferation rate was achieved in the presence of 20% serum [p<0.001]. Additionally, there was an inverse relation between cell seeding density at culture initiation and proliferation rate, except for L-MSC at 300 cell seeding density. All three sources of fetal sheep MSCs had the identical trilineage differentiation potential. The proliferative capacity of liver and bone marrow derived MSCs were similar at different cell seeding densities except for the higher fold increase in B-MSCs at 2700 cells/cm2 density. Moreover, the adipose tissue derived MSCs had the lowest proliferative indices

validity of preoperative templating in cementless total hip arthroplasty: a pilot study

Preoperative templating plays an important role in orthopaedic surgery for achieving a successful outcome in THA. This includes not only selecting the type and size of implant, but also deciding on the alignment, position, and orientation of these implants, relying on anatomically defined landmarks. The first goal of this study was to evaluate the validity of preoperative templating technique in terms of determining the final implant size and position. The second goal was to investigate the interobserver and Intraobserver reliability of template readings on conventional radiographs. In Aljumhorei hospital, Mosul city a case series; pilot study has been carried for twenty-eight patients had preoperative templating for total hip arthroplasty between 21/10/2009 to 16/5/2012 was carried by the operating surgeon. Preoperative templating by same two orthopedic surgeons carried for 12 patients from the total. Preoperative templating for Seventeen patientsout of the total were studied with the same operating surgeon after 2 years of experiences. 29% compatibility for shell between preoperative templating with intraoperative for the operator, P_ 0.023. While for stem 39%P_ 0.257. Second surgeon 58% compatibility with P_0.564, Stem differences represent 33% [P_ 0.248]. Differences with two years experiences for the operating surgeon indicate 35% compatibility both for shell and the stem with non-significant differences. Preoperative templating appeared to be more accurate for femoral stem measurements than the acetabular templating in determining the accurate sizes. Two years experienced added no more in templating measurements. There were significant differences for interobserver measurements.