Biological Activity of Ganoderma Species (Agaricomycetes) from Sonoran Desert, Mexico.

Ganoderma species have been used in folk medicine against different illnesses and are characterized by producing a diversity of bioactive metabolites (triterpenoids, polysaccharides, flavonoids, and phenols) with numerous medicinal effects (anti-proliferative, antioxidant, anti-inflammatory, and antibacterial). This work aims to evaluate ethanolic extracts of fruiting bodies of Ganoderma oerstedii, G. weberianum, and G. subincrustatum strains from the Sonoran Desert in the anti-proliferative activity by the MTT assay on cancer cell lines; anti-inflammatory effect by quantifying nitric oxide (NO) production; antioxidant activity by DPPH, ABTS, and FRAP assays; total phenolic and flavonoid content by Folin-Ciocalteu and AlCl3 method, respectively; antibacterial activity by the broth microdilution method against Escherichia coli and Staphylococcus aureus. Extracts showed anti-proliferative activity with IC50 < 100 µg/mL on the cancer cell lines MDA-MB-231, A549, and HeLa, except G. subincrustatum extract with an IC50 > 100 µg/mL; anti-proliferative activity was not selective, being affected non-cancerous cell line ARPE-19. Extracts showed significant inhibition of NO release in cells stimulated by LPS, up to 60% with G. subincrustatum and G. oerstedii, and 47% with G. weberianum. All tested assays showed moderate antioxidant potential; the most active was G. lucium (control strain) with IC50 of 69 and 30 µg/mL by DPPH and ABTS respectively; and 271 µg Trolox equivalents/g by FRAP. Total phenols and flavonoids ranged from 38 to 56 mg GAE/g and 0.53 to 0.93 mg QE/g, respectively. A significant correlation was found between the antioxidant activities revealed by DPPH, ABTS, and FRAP with total phenol and flavonoid contents. Antibacterial activity was weak against S. aureus (MIC50 > 10 mg/mL). These results demonstrate that tested Ganoderma mushrooms have medicinal potential such as anti-inflammatory and anti-proliferative.

Neutralization, by a polyspecific antivenom, of the coagulopathy induced by the venom of Bothrops asper: Assessment by standard coagulation tests and rotational thromboelastometry in a murine model.

Venom-induced consumption coagulopathy and thrombocytopenia are common and potentially severe manifestations of viperid snakebite envenoming since they contribute to local and systemic hemorrhage. Therefore, the assessment of the efficacy of antivenoms to neutralize coagulopathic and thrombocytopenic toxins should be part of the preclinical evaluation of these drugs. To evaluate the efficacy of the polyvalent (Crotalinae) antivenom produced in Costa Rica, in this study we have used a mouse model of coagulopathy and thrombocytopenia induced by the venom of Bothrops asper, based on the bolus intravenous (i.v.) injection of venom. When venom and antivenom were incubated before injection, or when antivenom was administered i.v. immediately after venom injection, venom-induced hemostatic alterations were largely abrogated. We also studied the recovery rate of clotting parameters in conditions where antivenom was administered when mice were coagulopathic. Some parameters recovered more rapidly in antivenom-treated mice than in control envenomed animals, but others showed a spontaneous recovery without antivenom. This is due to a rapid clearance of plasma venom levels in these experimental conditions. This implies that models based on the bolus i.v. injection of venom have limitations for assessing the effect of antivenom in the recovery of clotting alterations once coagulopathy has developed. It is suggested that alternative models should be developed based on a slower systemic absorption of venom. Overall, our findings provide a protocol for the preclinical evaluation of antivenoms and demonstrate that the polyvalent antivenom is effective in neutralizing the toxins of B. asper venom responsible for coagulopathy and thrombocytopenia.

Coagulopathy induced by viperid snake venoms in a murine model: Comparison of standard coagulation tests and rotational thromboelastometry.

Viperid snakebite envenoming is often characterized by a venom-induced consumption coagulopathy due to the procoagulant effect of venom components, resulting in the alteration of clotting laboratory tests. There is a growing trend to use rotational thromboelastometry in the assessment of clotting disturbances in a variety of pathologies, although its use in experimental models of envenoming has been limited. An in vivo murine model was implemented to assess the coagulopathy induced by three Central American viperid venoms which have different mechanisms of action on clotting factors, i.e., Bothrops asper, Crotalus simus and Bothriechis lateralis. Venom was injected by the intravenous route and blood samples were collected at 1, 3, 5 and 24 h after envenoming. Coagulopathy was assessed by standard clotting tests and by routine rotational thromboelastometric parameters. In addition, the changes in platelet number were followed. B. asper and C. simus venoms induced coagulopathy and thrombocytopenia 1 h after injection, followed by a slow recovery at 3, 5 and 24 h, although the majority of clotting parameters were still significantly affected by 3 and 5 h, and were corrected by 24 h. In general, a similar time-course of alterations was observed for standard clotting tests and most rotational thromboelastomeric assays. However, some thromboelastometric parameters, especially those related to Fibtem, showed more drastic alterations than standard tests and remained altered even at 24 h in some cases. This is likely related to the low fibrinogen concentration observed at most time intervals. B. lateralis venom did not induce a consumption coagulopathy, although it caused a marked thrombocytopenia.

PLGA nanoparticle preparations by emulsification and nanoprecipitation techniques: effects of formulation parameters.

This study presents the influence of the primary formulation parameters on the formation of poly-dl-lactic-co-glycolic nanoparticles by the emulsification-solvent evaporation, and the nanoprecipitation techniques. In the emulsification-solvent evaporation technique, the polymer and tensoactive concentrations, the organic solvent fraction, and the sonication amplitude effects were analyzed. Similarly, in the nanoprecipitation technique the polymer and tensoactive concentrations, the organic solvent fraction and the injection speed were varied. Additionally, the agitation speed during solvent evaporation, the centrifugation speeds and the use of cryoprotectants in the freeze-drying process were analyzed. Nanoparticles were characterized by dynamic light scattering, laser Doppler electrophoresis, and scanning electron microscopy, and the results were evaluated by statistical analysis. Nanoparticle physicochemical characteristics can be adjusted by varying the formulation parameters to obtain specific sizes and stable nanoparticles. Also, by adjusting these parameters, the nanoparticle preparation processes have the potential to be tuned to yield nanoparticles with specific characteristics while maintaining reproducible results.

Plasmid-DNA lipid and polymeric nanovaccines: a new strategic in vaccines development.

Vaccination is the most effective and least expensive technique used for human diseases prevention and eradication. The need for more vaccine doses and the rapid establishment of facilities for the development of new vaccines are stimulating significate changes in the vaccine industry, which is gradually moving towards cell culture production. One approach is the third generation of vaccines, which are based on the use of plasmid DNA (pDNA) containing transgenes that encode an antigen capable of mimicking intracellular pathogenic infection and triggering both humoral and cellular immune responses. Plasmid DNA vaccination has distinct advantages over other vaccine technologies in terms of safety, ease of fabrication and stability. The effectiveness of pDNA vaccines against viruses, bacteria, parasites and cancer cells has been demonstrated in preclinical and clinical assays. Furthermore, currently there are a few veterinary pDNA vaccines in the market. The application of a simple formulation of naked pDNA as a vaccine is attractive, but a low transfection efficiency is often obtained. The use of nanoparticles to increase transfection efficiency is an approach that has been tested clinically. This review provides a summary of vaccine production, advances and major challenges associated with pDNA lipid and polymeric nanovaccines applications.

Low intensity sonosynthesis of iron carbide@iron oxide core-shell nanoparticles.

Here we demonstrate a simple method for the organic sonosynthesis of stable Iron Carbide@Iron Oxide core-shell nanoparticles (ICIONPs) stabilized by oleic acid surface modification. This robust synthesis route is based on the sonochemistry reaction of organometallic precursor like Fe(CO)5 in octanol using low intensity ultrasonic bath. As obtained, nanoparticles diameter sizes were measured around 6.38 nm ±â€¯1.34 with a hydrodynamic diameter around 25 nm and an estimated polydispersity of 0.27. Core-Shell structure of nanoparticles was confirmed using HR-TEM and XPS characterization tools in which a core made up of iron carbide (Fe3C) and a shell of magnetite (γ-Fe2O3) was found. The overall nanoparticle presented ferromagnetic behavior at 4 K by SQUID. With these characteristics, the ICIONPs can be potentially used in various applications such as theranostic agent due to their properties obtained from the iron oxides and iron carbide phases.

Slow Titration and Delayed Intensification of Basal Insulin Among Patients with Type 2 Diabetes.

BACKGROUND: Clinical inertia in type 2 diabetes mellitus (T2DM) refers to the failure of clinicians to intensify therapy when indicated. Many T2DM patients remain suboptimally controlled after initiating basal insulin. OBJECTIVE: To examine the prevalence of patients treated with basal insulin but in poor glycemic control (hemoglobin A1c [A1c] ≥ 7%) after initiation and subsequent treatment intensification patterns and glycemic outcomes in a real-world setting. METHODS: Adults diagnosed with T2DM newly initiating a basal insulin analog (insulin glargine or detemir) from January 2010 to September 2014 were identified in the QuintilesIMS Real-World Data Adjudicated Claims linked to the QuintilesIMS Real-World Data Electronic Medical Records. Patients were previously naive to insulin and glucagon-like peptide-1 receptor agonists (GLP-1 RAs), were persistent on therapy for ≥ 6 months, and had ≥ 12 months of continuous health plan enrollment after initiation. First treatment intensification (increase in basal insulin dose [of ≥ 10%], addition of bolus insulin, GLP-1 RA, or a new oral antidiabetic drug [OAD]) was assessed among patients in poor glycemic control at 6 months after initiation over the available (minimum ≥ 12-month) follow-up. Subsequent glycemic outcomes and treatment intensification were assessed. Kaplan-Meier (KM) analysis evaluated time-to-treatment intensification and time to A1c goal. RESULTS: Of 427 eligible patients with A1c available at 6 months, 59.3% were male; mean age was 53.9 years; mean follow-up was 29.4 months; and mean dose of the initiated prescription was 29.6 insulin units (U) (median 24U). Six months after initiating basal insulin, 81.0% of patients (n = 346) remained in poor glycemic control, and mean basal insulin dose was 31.0U (median 25U). Most (88.4%; n = 306) of these uncontrolled patients subsequently intensified treatment over the available follow-up. Using KM analysis, these patients intensified treatment in a median of 58 days (range: 17.5 days [GLP-1 RA addition] to 52 days [increase in basal insulin dose]) from the first elevated A1c measurement taken after 6 months, and 72.5% (GLP-1 RA addition) to 91.1% (OAD addition) of patients continued to remain in poor glycemic control at 12 months after intensification. Most patients (66.8%; n = 231/346) first intensified treatment by increasing their basal insulin dose, and mean dose increased to 61.7U (median 38U) at intensification. Six months following basal insulin increase, almost all patients remained on basal insulin therapy and among those with available A1c, 92.1% (140 of 152) were in poor glycemic control. In the subsequent 12 months, only a third (34%) of uncontrolled patients added another antihyperglycemic agent. CONCLUSIONS: The vast majority of patients remained uncontrolled in the 6 months following basal insulin initiation. Basal insulin up-titration was slow and insufficient in the 6 months after initiation, indicating treatment inertia. Subsequently, most patients failed to achieve glycemic targets despite intensification with basal insulin. This finding suggests a substantial unmet need for effective treatment intensification among T2DM patients treated with basal insulin who remain uncontrolled. Improved provider education and guidelines on appropriate intensification are warranted. DISCLOSURES: This study was funded by Novo Nordisk. Mocarski, Guerrero, Langer, and Thorsted are employees and shareholders of Novo Nordisk. Yeaw, Divino, and DeKoven are employed by QuintilesIMS, which received remuneration from Novo Nordisk for work on this study. Study concept and design were contributed by Mocarski, DeKoven, Langer, and Thorsted. Yeaw took the lead in data collection, along with Divino and DeKoven. Data interpretation was performed by Yeaw, Divino, DeKoven, and Guerrero. The manuscript was written by Mocarski and Divino and revised by Guerrero, Langer, and Thorsted, along with Yeaw and DeKoven. Some of the data from this study were presented via poster at the AMCP Annual Meeting in March 2017 and at the 53rd EASD Annual Meeting in September 2017.

Plasmid pVAX1-NH36 purification by membrane and bead perfusion chromatography.

The demand for plasmid DNA (pDNA) has increased in response to the rapid advances in vaccines applications to prevent and treat infectious diseases caused by virus, bacteria or parasites, such as Leishmania species. The immunization protocols require large amounts of supercoiled plasmid DNA (sc-pDNA) challenging the development of efficient and profitable processes for capturing and purified pDNA molecules from large volumes of lysates. A typical bioprocess involves four steps: fermentation, primary recovery, intermediate recovery and final purification. Ion-exchange chromatography is one of the key operations in the purification schemes of pDNA owing the chemical structure of these macromolecules. The goal of this research was to compare the performance of the final purification step of pDNA using ion-exchange chromatography on columns packed with Mustang Q membranes or perfusive beads POROS 50 HQ. The experimental results showed that both matrixes could separate the plasmid pVAX1-NH36 (3936 bp) from impurities in clarified Escherichia coli lysates with an adequate resolution. In addition, a 24- and 21-fold global purification factor was obtained. An 88 and 63% plasmid recuperation was achieved with ion-exchange membranes and perfusion beads, respectively. A better understanding of perfusion-based matrices for the purification of pDNA was developed in this research.

EFFECTS OF LIRAGLUTIDE 3.0 MG ON WEIGHT AND RISK FACTORS IN HISPANIC VERSUS NON-HIPANIC POPULATIONS: SUBGROUP ANALYSIS FROM SCALE RANDOMIZED TRIALS.

OBJECTIVE: Scarce data exist on pharmacotherapy for obesity in Hispanic individuals. This post hoc analysis of pooled data from 4 phase 3a trials compared the efficacy and safety of liraglutide 3.0 mg versus placebo, as adjunct to a reduced-calorie diet and physical activity, in Hispanic versus non-Hispanic subgroups. METHODS: We conducted the double-blind randomized, placebo-controlled trials in adults with a minimum body mass index (BMI) of 27 kg/m2 with at least 1 comorbidity, or a minimum BMI of 30 kg/m2, at clinical research sites worldwide. In this analysis, we investigated possible differences in treatment effects between 534 Hispanics (10.4% of the population) and 4,597 non-Hispanics (89.6%) through statistical tests of interaction between subgroups and treatment. Variables examined included mean and categorical weight change, cardiovascular risk markers, and safety data. RESULTS: Both subgroups achieved clinically significant mean weight loss at end-of-treatment with liraglutide 3.0 mg versus placebo: Hispanics 7.0% versus 1.5%, treatment difference -5.1% (95% CI, -6.2 to -4.0); non-Hispanics 7.5% versus 2.3%, -5.2% (95% CI, -5.5 to -4.8). More individuals in both subgroups lost ≥5%, >10%, and >15% of their baseline weight with liraglutide 3.0 mg than with placebo. Efficacy endpoints generally did not vary with ethnicity (P>.05). Adverse events were comparable between ethnic subgroups, with more gastrointestinal disorders reported with liraglutide 3.0 mg than placebo. CONCLUSION: Efficacy and safety were largely similar between Hispanic and non-Hispanic subgroups. Results support that liraglutide 3.0 mg, used with a reduced-calorie diet and physical activity, can facilitate weight loss in Hispanic individuals. ABBREVIATIONS: A1c = glycated hemoglobin BMI = body mass index CI = confidence interval FPG = fasting plasma glucose GLP-1 = glucagon-like peptide-1 hsCRP = high-sensitivity C-reactive protein SCALE = Satiety and Clinical Adiposity - Liraglutide Evidence in individuals with and without diabetes T2DM = type 2 diabetes mellitus.

Hipotiroidismo congénito, la primera causa de retraso mental prevenible: un desafío para la medicina preventiva / Congenital hypothyroidism, the first cause of preventable mental retardation: a challenge for preventive medicine

El hipotiroidismo congénito es la enfermedad endocrina más frecuente en neonatos y puede ser de manifiesto clínico o subclínico. Es la principalcausa de retraso mental tratable y su pronóstico radica en el tamizajetemprano y la instauración oportuna del tratamiento, por lo cual en Colombia se considera necesaria la implementación de un esquema de tamizaje neonatal con una técnica apropiada y de cobertura nacional, equiparable al de otros países. El objetivo de este artículo es realizar una revisión y análisis de la literatura médica existente sobre el hipotiroidismo congénito centrado en la definición, epidemiología, factores etiológicos, tamizaje, diagnóstico y tratamiento; mediante la búsqueda de artículos en las bases de datos PUBMED, EMBASE, REDALYC, OVID, MEDLINE,DYNAMED y CLINICAL KEY (acceso: mayo 2015). Además se realizó revisiónde los informes del Instituto Nacional de Salud en el año 2013 y 2014, de las guías europeas de la sociedad de endocrinología pediátrica 2014, de laguía mexicana de hipotiroidismo congénito neonatal 2008 y del consensocolombiano para el diagnóstico y manejo de enfermedades tiroideas...

Congenital hypothyroidism is the most common endocrine disease innewborns and can be clinical or subclinical. It is the leading cause oftreatable mental retardation and its prognosis is in the early screening andtimely initiation of treatment; this is the reason why in Colombia theimplementation of a neonatal screening with appropriate nationalcoverage technique, comparable to those in other countries, is considerednecessary. The aim of this article is to review and analyse the literature oncongenital hypothyroidism focused on the definition, epidemiology,etiological factors, screening, diagnosis and treatment; by search inPubMed, EMBASE, REDALYC, OVID, MEDLINE, and CLINICAL KEY Dynameddata (access: May 2015). Besides reviewing the reports of the NationalInstitute of Health in 2013 and 2014, the European guidelines for pediatricendocrinology society of 2014, Mexico’s 2008 guide neonatal congenitalhypothyroidism and Colombian consensus for the diagnosis andmanagement of thyroid diseases...

Segregated growth kinetics of Escherichia coli DH5α-NH36 in exponential-fed perfusion culture for pDNA vaccine production.

The clinical demand of plasmid DNA (pDNA) has been increasing constantly. An exponential-fed perfusion (EFP) culture is a new mode for plasmid production for clinical trials and commercialization. However, the culture conditions may lead to cell filamentation and growth cessation. In this study, the variation of the physiological state and the plasmid contents of Escherichia coli DH5α hosting pVAX1-NH36 in an EFP culture for application as a Leishmaniasis vaccine was investigated. The culture performance was monitored using flow cytometry (FC) and real-time quantitative PCR. The FC studies showed a high viability of cell population and a constant distribution of complexity and size. A high homogeneity of pDNA (>95 % of supercoiled) was obtained, which might be attributed to a better culture environment. The obtained plasmid specific and volumetric yields of 1.8 mg/g dcw and 36.5 mg/L represent typical values for laboratory-scale plasmid production in a defined medium. A segregated kinetic model of the perfusion system was developed and fitted to the experimental data (R(2) > 0.96). A practical conclusion of this work is that a space-time yield analysis of a bioprocess requires a viability evaluation. This new strategy of culture operation might help in the efficient production of pDNA for therapeutic use.

Plasmid DNA primary recovery from E. coli lysates by depth bed microfiltration.

Recently, several studies have been published on the application of plasmid DNA (pDNA) in gene therapy and vaccine production. The bioprocess to obtain pDNA involves the steps of fermentation, primary recovery, secondary recovery and final purification. The pDNA primary recovery, which is the key step to the rest of the process, includes biomass separation, alkaline lysis and clarification of the lysate. In this work, the clarification by depth bed microfiltration of lysates of E. coli DH5α containing the plasmid pVAX1-NH36 was investigated. The studies were conducted using filter capsules with nominal 8.0 µm pore size using fluxes of 0.0027 and 0.004 cm(3)/(cm(2)-s). The results were compared with the conventional clarification by centrifugation. A fiber coating model was used to describe the behavior of the microfiltration system. A 99 % of solids elimination of the lysates was achieved with depth bed filtration method. The removed solids occupied 23 and 43 % (for 4 and 6 cm(3)/min, respectively) of the void volume of the depth bed microfiltration capsule since an early breakthrough curve is characteristic of these processes. The depth bed microfiltration process for removal of solids from the cell lysate showed competitive results compared to clarification by centrifugation.

Dientes supernumerarios en dentición primaria asociados a pólipos palatinos: Reporte de caso / Supernumerary teeth in primary dentition associated to palatal polyps: Case report

Los pólipos son poco frecuentes en niños. En este artículo se presenta un caso clínico de un niño de un año dos meses que acude a la Clínica de Odontopediatría de la DEPeI UNAM con dos pólipos fibroepiteliales palatinos ubicados a ambos lados de la papila incisiva, 10 días posteriores a la excisión quirúrgica se observó la erupción de un diente supernumerario en el paladar, y 25 días después se observó la erupción de un segundo diente supernumerario. Ambos dientes fueron extraídos. El diagnóstico histológico de las lesiones en el paladar fue: fibroma de fibroblastos gigantes; sin embargo, no se encontró evidencia histológica que mostrara alguna posible relación entre la presencia de los pólipos palatinos y los dientes supernumerarios.

Polyps are rare in children. The present article reports the clinical case of a 14 month old male patient brought for treatment to the Pedodontics Clinic of the Graduate School, National School of Dentistry National University of Mexico. He presented two palatal fibro-epithelial polyps, located at both sides of the incisive papilla. 10 days after surgical excision, a supernumerary tooth erupted in the palate. 25 days later, eruption of a second supernumerary tooth was observed. Both teeth were extracted. Histological diagnosis of palatal lesions was giant fibroblast fibroma. Nevertheless, no histological evidence was found to show possible relationship between presence of palatal polyps and supernumerary teeth.

Accuracy and preference assessment of prefilled insulin pen versus vial and syringe with diabetes patients, caregivers, and healthcare professionals.

OBJECTIVES: The primary objective of this study was to investigate the dosing accuracy of the new prefilled FlexTouch insulin pen (FT) in comparison to conventional vial and syringe (V&S) when used by patients (Pts), caregivers (CG) and healthcare professionals (HCPs). METHODS: A total of 120 subjects participated in the trial (40 diabetes patients aged 61 ± 11 [mean ± SD] yrs, 20 caregivers [parents and other relatives], 20 physicians, and 40 nurses/certified diabetes educators). The participants were introduced to the devices in randomized order and were asked to perform injections of 5, 25, 43 and 79 IU doses into laboratory tubes. Dosing accuracy was analyzed by weighing the tubes on a pharmaceutical balance and calculating the mean absolute deviation (MAD) from the intended doses. After completing a device assessment questionnaire, Patient Perception Questionnaire (PPQ), with questions regarding device design and performance, the procedure was repeated for the other device, and the patients were finally asked to complete a device preference questionnaire (DPQ). RESULTS: Dosing accuracy was significantly better for FT when used by any of the cohorts at all doses. (MAD ± SD for FT/V&S; 5 IU: 0.4 ± 0.4/0.6 ± 0.6 IU; 25 IU: 0.3 ± 0.4/0.7 ± 0.9 IU; 43 IU: 0.4 ± 0.4/0.9 ± 1.2 IU; 79 IU: 0.5 ± 0.5/1.7 ± 1.6 IU, p < 0.005 for all doses). Dosing accuracy with FT for all three subgroups was comparable (patients: 0.35-0.59 IU; HCP&CG: 0.29-0.54 IU; n.s.). Dosing accuracy with V&S for all three subgroups was not comparable: HCP and CG performed much better with V&S than patients and delivered the doses with significantly higher accuracy (range of mean MAD; patients: 0.81-2.54 IU; HCP&CG: 0.51-1.30 IU, p < 0.005 at all doses). FT was ranked superior to V&S for all aspects of the PPQ. In the DPQ, 93% of the patients voted for FT (neutral: 5%, V&S: 2%), (CG: 100%/0%/0%; HCPs: 85%/2%/13%; p < 0.001 in all cases). CONCLUSION: FT, compared to V&S, was more accurate at all tested doses and was used with similar accuracy by patients, HCPs, and CGs. Using questionnaires only, and without dexterity assessment, study participants rated FT higher than V&S in every component of the PPQ and the vast majority of them preferred FT. These findings may point to a better alternative for dosing accuracy and improved adherence when using the new prefilled insulin pen compared to V&S for insulin delivery in patients with diabetes.

Healthcare professional and patient perceptions of a new prefilled insulin pen versus vial and syringe.

OBJECTIVE: Usability of a new prefilled insulin pen, FlexTouch® (FT; Novo Nordisk A/S, Bagsvaerd, Denmark), with no push-button extension and low injection force, was compared with vial and syringe (V&S). RESEARCH DESIGN AND METHODS: People with diabetes, and healthcare professionals with diabetes management experience conducted test injections and answered questions on preference, ease of use, confidence, ease of learning and teaching. RESULTS: The study involved 30 needle-naïve patients (naïve to any diabetes injection therapy), 30 V&S-experienced patients, 30 physicians and 30 nurses. In the total population, FT was preferred to V&S for teaching or learning to use (both p < 0.001). Nurses (100 vs. 0%) and physicians (87 vs. 7%) preferred FT to V&S for ease of teaching. V&S-experienced (73 vs. 7%) and needle-naïve patients (83 vs. 7%) preferred FT to V&S for ease of learning. The remainder chose "equally easy/difficult." More participants in each group rated FT "very/fairly easy" for ease of depressing the push-button/plunger (FT vs. V&S: physicians, 93 vs. 80%; nurses, 97 vs. 80%; V&S-experienced patients, 93 vs. 90%; needle-naïve patients, 100 vs. 77%), and injecting three doses. More participants were "very/rather confident" in managing daily injections using FT (FT vs. V&S: physicians, 100 vs. 60%; nurses, 100 vs. 70%; V&S-experienced patients, 93 vs. 90%; needle-naïve patients, 90 vs. 40%). CONCLUSIONS: FT was rated easier to use, learn to use or teach to use than V&S by patients with or without experience of insulin injection with V&S, and by physicians and nurses with diabetes management experience.

Purification of plasmid DNA from Escherichia coli ferments using anion-exchange membrane and hydrophobic chromatography.

A novel downstream bioprocess was developed to obtain purified plasmid DNA (pDNA) from Escherichia coli ferments. The intermediate recovery and purification of the pDNA in cell lysate was conducted using hollow-fiber tangential filtration and frontal anion-exchange membrane and elution hydrophobic chromatographies. The purity of the solutions of pDNA obtained during each process stage was investigated. The results show that the pDNA solution purity increased 30-fold and more than 99% of RNA in the lysate was removed during the process operations. The combination of membrane operations and hydrophobic interaction chromatography resulted in an efficient way to recover pDNA from cell lysates. A better understanding of membrane-based technology for the purification of pDNA from clarified E. coli lysate was developed in this research.

Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography.

A new bioprocess using mainly membrane operations to obtain purified plasmid DNA from Escherechia coli ferments was developed. The intermediate recovery and purification of the plasmid DNA in cell lysate was conducted using hollow-fiber tangential filtration and tandem anion-exchange membrane chromatography. The purity of the solutions of plasmid DNA obtained during each process stage was investigated. The results show that more than 97% of RNA in the lysate was removed during the process operations and that the plasmid DNA solution purity increased 28-fold. One of the main characteristics of the developed process is to avoid the use of large quantities of precipitating agents such as salts or alcohols. A better understanding of membrane-based technology for the purification of plasmid DNA from clarified E. coli lysate was developed in this research. The convenience of anion-exchange membranes, configured in ready-to-use devices can further simplify large-scale plasmid purification strategies.