Effects of Kifunensine on Production and N-Glycosylation Modification of Butyrylcholinesterase in a Transgenic Rice Cell Culture Bioreactor.

The production and N-glycosylation of recombinant human butyrylcholinesterase (BChE), a model highly glycosylated therapeutic protein, in a transgenic rice cell suspension culture treated with kifunensine, a strong α-mannosidase I inhibitor, was studied in a 5 L bioreactor. A media exchange was performed at day 7 of cultivation by removing spent sugar-rich medium (NB+S) and adding fresh sugar-free (NB-S) medium to induce the rice α-amylase 3D (RAmy3D) promoter to produce rice recombinant human BChE (rrBChE). Using a 1.25X-concentrated sugar-free medium together with an 80% reduced working volume during the media exchange led to a total active rrBChE production level of 79 ± 2 µg (g FW)-1 or 7.5 ± 0.4 mg L-1 in the presence of kifunensine, which was 1.5-times higher than our previous bioreactor runs using normal sugar-free (NB-S) media with no kifunensine treatment. Importantly, the amount of secreted active rrBChE in culture medium was enhanced in the presence of kifunensine, comprising 44% of the total active rrBChE at day 5 following induction. Coomassie-stained SDS-PAGE gel and Western blot analyses revealed different electrophoretic migration of purified rrBChE bands with and without kifunensine treatment, which was attributed to different N-glycoforms. N-Glycosylation analysis showed substantially increased oligomannose glycans (Man5/6/7/8) in rrBChE treated with kifunensine compared to controls. However, the mass-transfer limitation of kifunensine was likely the major reason for incomplete inhibition of α-mannosidase I in this bioreactor study.

HRA2pl peptide: a fusion inhibitor for human metapneumovirus produced in tobacco plants by transient transformation.

MAIN CONCLUSION: The HRA2pl peptide expressed by transient transformation in N. tabacum plants is capable of inhibiting the binding of the human metapneumovirus to HEp-2 cells at the fusion stage. Human metapneumovirus (hMPV) is an agent responsible for acute respiratory infections that mainly affects children under 3 years, the elderly and immunocompromised patients. In children younger than 5 years, respiratory tract infections account for 20 % of deaths worldwide. However, there is currently no treatment or vaccine available against hMPV. The production of a safe, efficient and low cost treatment against this virus is a current challenge. Plants provide a system for recombinant protein production that is cost effective and is easier to scale up to an industrial level than other platforms; in addition, the plant tissue may be used as raw food, dried or, alternatively, proteins may be partially or fully purified and administered in aerosol or capsules as dry powder. In this study, we designed a gene expressing an antiviral peptide against hMPV based on the heptad repeat A domain of the F protein of the virus. We produced the recombinant peptide by a viral transient expression system (Magnifection(®)) in Nicotiana tabacum plants. The efficacy of this antiviral peptide was confirmed by in vitro assays in HEp-2 cell line. This is a promising result that can offer a prophylactic approach against hMPV.

Increasing in-person medical interpreter utilization in the NICU through a bundle of interventions.

BACKGROUND: In-person medical interpretation improves communication with patients who have preferred language other than English (PLOE). Multi-dimensional barriers to use of medical interpreters limit their use in the NICU. LOCAL PROBLEM: Medical teams in our NICU were not consistently using in-person medical interpreters, leading to ineffective communication with families with PLOE. METHODS/INTERVENTIONS: Interventions included staff educational sessions and grand rounds regarding equitable language access, distribution of interpreter request cards to families, and allocation of dedicated in-person interpreters for NICU rounds. Interpreter utilization was calculated by total requests per Spanish-speaking person day in the NICU. RESULTS: Interpreter utilization increased five-fold during the intervention period (from 0.2 to 1.0 requests per Spanish-speaking person day). CONCLUSIONS: We substantially increased our unit in-person interpreter utilization through a bundle of multifaceted interventions, many of which were low-cost. NICUs should regard dedicated medical interpreters as a critical part of the care team.

[Current epidemiological overview of the most frequent cancers in Spain. Implications for nursing care]. / Cánceres mas frecuentes en España. Panorama epidemiológico actual.

This article is to describe the epidemiological situation of cancer and prevention strategies, particularly related to the work of nurses (primary prevention and early detection), focusing primarily on lung cancer (LC), breast (BC) and colorectal (CRC). 98,046 cancer deaths occurred in Spain in 2006, which is the leading cause of death in our country. The main action of prevention for the LC is based on smoking cessation, the main risk factor (relative risk (RR) from 10 to 30) beside radon and asbestos exposures. Basically the prevention for the BC is based on early detection, mammography remains the ultimate test because it reduces mortality, and must be done along period between 50 and 70 years old with a two-year interval. CRC mortality is reduced by 16% due to early detection test based on fecal occult blood (TsOH), but it's necessary to insist on changes in lifestyle (diet and exercise) as primary prevention strategies, and in breast cancer too. One of the most effective ways to reduce the impact of cancer today is the prevention, which is carried out mainly by the nurses.

The Aggregation Pheromone of Metamasius spinolae (Coleoptera: Dryophthoridae) Revisited: Less is More.

The weevil Metamasius spinolae (Gyllenhal) is the most important insect pest of cultivated prickly pear in Mexico. A previous work reported that the pheromone of this weevil species was composed by three components. In this study, we reinvestigated the aggregation pheromone of M. spinolae using gas chromatography-electroantennography and gas chromatography-mass spectrometry to locate and identify new potential pheromonal compounds. The behavioral activity of identified compounds was evaluated in laboratory and field trials. Metamasius spinolae males released four compounds: 2-methyl-4-heptanone, 6-methyl-2-hepten-4-one, 2-methyl-4-octanone, and 2-hydroxy-2-methyl-4-heptanone. In the laboratory assays, depending on the concentration, the compounds were attractive, neutral, or repellent to M. spinolae. Field evaluation showed that traps baited with 2-hydroxy-2-methyl-4-heptanone singly or in most of the binary or tertiary blends where this compound was present captured a higher number of M. spinolae compared to live males and the other compounds identified. In conclusion, our results indicate that 2-hydroxy-2-methyl-4-heptanone is the main component of the aggregation pheromone of M. spinolae. We suggest that this compound should be used for developing a monitoring or a mass-trapping system for M. spinolae.

Current advances in the algae-made biopharmaceuticals field.

INTRODUCTION: The biopharmaceuticals industry demands new production platforms to address several challenges; such as cost reduction to make biologics accessible in low-income countries, safety enhancement of the product, development of products administered by noninvasive routes, and expansion of potential biosimilars and biobetters. Microalgae are emerging hosts for biopharmaceuticals production with the potential to meet such requirements. AREAS COVERED: Nowadays successful cases on the production of vaccines, antibodies, antimicrobial peptides, growth factors/cytokines, and hormones in algae have been reported. This review comprises an updated outlook covering protein expression strategies, a compilation of functional biopharmaceuticals produced in algae, and companies investing in this technology. EXPERT OPINION: Key perspectives for the field include optimizing yields, scaling up production and completing preclinical trials. The experience from the field of plant-made biopharmaceuticals is commented as a key reference that will aid in the development of the algae-made biopharmaceuticals field.

What Does Plant-Based Vaccine Technology Offer to the Fight against COVID-19?

The emergence of new pathogenic viral strains is a constant threat to global health, with the new coronavirus strain COVID-19 as the latest example. COVID-19, caused by the SARS-CoV-2 virus has quickly spread around the globe. This pandemic demands rapid development of drugs and vaccines. Plant-based vaccines are a technology with proven viability, which have led to promising results for candidates evaluated at the clinical level, meaning this technology could contribute towards the fight against COVID-19. Herein, a perspective in how plant-based vaccines can be developed against COVID-19 is presented. Injectable vaccines could be generated by using transient expression systems, which offer the highest protein yields and are already adopted at the industrial level to produce VLPs-vaccines and other biopharmaceuticals under GMPC-processes. Stably-transformed plants are another option, but this approach requires more time for the development of antigen-producing lines. Nonetheless, this approach offers the possibility of developing oral vaccines in which the plant cell could act as the antigen delivery agent. Therefore, this is the most attractive approach in terms of cost, easy delivery, and mucosal immunity induction. The development of multiepitope, rationally-designed vaccines is also discussed regarding the experience gained in expression of chimeric immunogenic proteins in plant systems.

Two Cases of SARS-CoV-2 Infection in Pediatric Oncohematologic Patients in Spain.

Since December 2019, severe acute respiratory syndrome coronavirus 2 infection has spread worldwide. We all are concerned about immunocompromised children, especially hematologic and oncologic pediatric patients. We want to share our experience with 2 pediatric cancer patients with severe acute respiratory syndrome coronavirus 2 infection. Both presented mild disease and good outcome. No respiratory symptoms were identified, but both developed diarrhea, one probably secondary to lopinavir/ritonavir. Pediatric cancer patients may have milder disease than adults, but larger studies are needed to make conclusions.

An overview of tolerogenic immunotherapies based on plant-made antigens.

INTRODUCTION: Over the last two decades, genetically engineered plants became attractive and mature platforms for producing vaccines and other relevant biopharmaceuticals. Autoimmune and inflammatory disorders demand the availability of accessible treatments, and one alternative therapy is based on therapeutic vaccines able to downregulate immune responses that favor pathology progression. AREAS COVERED: The current status of plant-made tolerogenic vaccines is presented with emphasis on the candidates under evaluation in test animals. Nowadays, this concept has been assessed in models of food and pollen allergies, autoimmune diabetes, asthma, arthritis, and prevention of blocking antibodies induction against a biopharmaceutical used in replacement therapies. EXPERT OPINION: According to the current evidence generated at the preclinical level, plant-made tolerogenic therapies are a promise to treat several immune-related conditions, and the beginning of clinical trials is envisaged for the next decade. Advantages and limitations for this technology are discussed.

Expression of a Zika virus antigen in microalgae: Towards mucosal vaccine development.

Zika virus (ZIKV) infection has extended rapidly all over the world in the last decades affecting humans of all ages, inducing severe illness such as the autoimmune Guillain-Barré syndrome as well as fetal neurodevelopmental defects. Despite the epidemiological importance of ZIKV, today there are no commercially available drugs or vaccines to combat or prevent this infection. Microalgae are attractive hosts to produce and deliver vaccines, with some candidates under preclinical evaluation. Herein, algae-based expression was assessed for the production of a new vaccine candidate against ZIKV called ZK. The Algevir technology was applied to express an antigenic protein called ZK comprising the B subunit of the heat labile Escherichia coli enterotoxin along with 3 epitopes from the ZIKV envelope glycoprotein. Efficient expression of the ZK antigen was achieved in Schizochytrium sp. with yields of up to 365 µg g-1 microalgae fresh weight. Upon oral administration in mice, the microalgae-made ZK protein elicited significant humoral responses at a higher magnitude to those induced upon subcutaneous immunization. The algae-made ZK vaccine represents a promising candidate to formulate attractive vaccines against ZIKV.

Current developments and prospects on human metapneumovirus vaccines.

INTRODUCTION: Human metapneumovirus (hMPV) has become one of the major pathogens causing acute respiratory infections (ARI) mainly affecting young children, immunocompromised patients, and the elderly. Currently there are no licensed vaccines against this virus. Areas covered: Since the discovery of hMPV in 2001, many groups have focused on developing vaccines against this pathogen. This review presents the outcomes and perspectives derived from preclinical studies performed in cell cultures and animals as well as the only candidate that has reached evaluation in a clinical trial. Limitations of the current vaccine candidates are discussed and perspectives for the development of plant-based vaccines are analyzed. Expert commentary: Several hMPV vaccine candidates are under development with the potential to progress into clinical trials. In parallel, the molecular farming field offers new opportunities to generate innovative vaccines that will offer several advantages in the fight against hMPV.

Plant-based vaccines against respiratory diseases: current status and future prospects.

INTRODUCTION: Respiratory infections have an enormous, worldwide epidemiologic impact on humans and animals. Among the prophylactic measures, vaccination has the potential to neutralize this impact. New technologies for vaccine production and delivery are of importance in this field since they offer the potential to develop new immunization approaches overriding the current limitations that comprise high cost, safety issues, and limited efficacy. Areas covered: In the present review, the state of the art in developing plant-based vaccines against respiratory diseases is presented. The review was based on the analysis of current biomedical literature. Expert commentary: Preclinical and clinical evaluations of several vaccine candidates against influenza, tuberculosis, respiratory syncytial virus, pneumonia, anthrax and asthma are discussed and placed in perspective.

[Clinical implications and prognostic significance of the study on the circadian variation of heart rate variability in patients with severe pulmonary hypertension]. / Implicaciones clínicas y pronósticas del estudio circadiano de la modulación simpático-vagal de la variabilidad de la frecuencia cardiaca en pacientes con hipertensión arterial pulmonar grave.

BACKGROUND: A reduction of heart rate variability (HRV) is currently considered an independent risk factor for morbidity, mortality and severity of severalcardiac disease, however, the dynamic sympathovagal modulation on HRV during 24 hr in primary pulmonary hypertension (PPH) had not been described. METHODS: 24 hr Holter monitoring (HA) were recorded in 32 patients (mean age 34, +/-12, 90% female) with severe primary pulmonary hypertension (mean pulmonary pressure, 90:t:12 mm Hg), and in 34 patients (mean age 36 +/-14, 60% female) with Eisenmenger syndrome (ES) secondary to septal ventricular defect or atent ductus arteriosus. A control group (n=44) paired for age, gender and arterial pulmonary pressure was included. HRV time and spectral parameters (mean, SDNN, SDANN, rMSSD, PNN50, LF, HF and LF/HF ratio) were analyzed during three periods: 24 hr; day (8-22:00), night (23-07:00) and also every hour of recording at 5 min-intervals). After detection of sympatho-vagal balance 15 patients were randomized, Treprostinil (prostaglandin) was administered to 6 patients and subcutaneous placebo to 9. RESULTS: HRV frequency parameters during 24 hr HM were significantly different among groups. LF/HF (day) 5.9:1:12.5:1:1P.001 and LF/HF night) 2.8:tlvs.1.5:l:.8.034. Sympathovagal modulation on 24 hr HRV showed that heart rate circadian rhythm is clearly altered in both PPH and ES, but the sympathetic tone in PPH is higher at l 24 hr. (p < .05), after administering treprostinil a recovery of sympathovagal balance was observed CONCLUSIONS: Autonomic cardiac disturbance is clearly present in PPH and ES. The circadian rhythm of HRV is first lost due to an increase of sympathetic tone. These changes may be markers of autonomic disbalance that favor the development of arrhythmias and sudden death. The sympathovagal balance in PPH could be considered an important risk marker.

Exantema por norovirus / Exanthema by norovirus

Aparte de en gastroenteritis, raramente se ha visto implicado al norovirus en trastornos cutáneos. Presentamos el caso clínico de un lactante de nueve meses de edad que presentó lesiones maculares generalizadas mientras cursaba una gastroenteritis con detección de virus en heces positiva para norovirus

Apart from gastroenteritis, norovirus has been rarely implicated with some cutaneous disorders. We report a clinical case of a 9-month-old boy who developed macular generalized skin eruptions during the course of gastroenteritis in which virus detection was positive for norovirus

Prevalence and characteristics associated with self-reported gall bladder disease in Mexican American elders: results from the Hispanic Established Populations for Epidemiologic Studies in the Elderly (H-EPESE).

BACKGROUND AND AIMS: To identify the prevalence and characteristics of gall bladder disease (GBD) that has been self-reported in Mexican American Elders. METHODS: A prospective survey of a regional probability sample of self-identified Mexican Americans aged 65 and over. The Hispanic Established Population for the Epidemiologic Studies of the Elderly (H-EPESE), a probability sample of non-institutionalized, Mexican Americans, aged 65 and over, residing in Southwestern states of Texas, New Mexico, Colorado, Arizona, and California. In 1993- 1994 (Wave 1), 3050 Mexican Americans, aged 65 and over, were selected at baseline as a weighted probability sample. In 1995-1996 (Wave 2), 2895 remained. Sample weights were used to extrapolate to the estimated 498,176 older Mexican Americans residing in the Southwest United States. Self-reported GBD was collected via in-home interviews. RESULTS: The prevalence of self-reported GBD in Mexican American elders was found to be 18.8% with an average age of 75.05 years. The findings indicate that older Mexican Americans have an increased rate of GBD if they are female, have history of arthritis or hypertension and have more acculturation to the United States. However, the rate decreases when they score poorly on the Mini Mental State Exam. One major limitation was reliance on self-report, as GBD and other co-morbid illnesses may be under-, or overestimated. CONCLUSIONS: Age is not protective in the prevalence of GBD in elder Mexican Americans. Persistent underlying genetics and dietary habits most likely attribute to this consistent high percentage, even in the elderly.