Sustainable access of quality seeds of genetically engineered crops in Eastern Africa - Case study of Bt Cotton.

The genetically engineered bollworm-resistant Bt cotton hybrid varieties offer opportunities for reducing crop losses and enhancing productivity. In Eastern Africa region, Sudan, Ethiopia, and Kenya have approved and released Bt cotton in 2012, in 2018, and in 2019, respectively. The region has potential to grow cotton in over 5 million hectares. For commercial plantings in Ethiopia, Sudan and Kenya, hybrid Bt cotton seeds have been imported from India. Due to the COVID-19 pandemic-induced supply chain disruptions, high shipment costs, bureaucratic procedures for importing seeds, and foreign exchange shortages, farmers have not been able to access Bt cotton seeds. Stakeholders are seeking local production of seeds to provide sustainable access by farmers at affordable cost. Country case studies reveal the importance of enhancing capacity for local seed production and extension advisory services. Revival of the cotton sector needs enhanced public-private partnerships to pave the way for sustainable seeds access in the region.

Cuban Vaccines Abdala and Mambisa against Covid-19

Intro: With the first case of COVID-19 in Cuba on March 11, 2020, the Center for Genetic Engineering and Biotechnology in Havana began an extensive vaccine program. Two vaccines based on RBD recombinant protein were developed, one for systemic administration "Abdala" and one mucosal vaccine "Mambisa". Abdala received the EUA in July 2021 and "Mambisa" completed its clinical development as a booster dose for convalescent subjects. Method(s): Two doses (25 and 50 microg) and two schedules (0-14-28 and 1-28-56 days) were evaluated in phase I clinical trials with volunteers 19 to 54 years old. The phase II and III clinical trials were also double-blind, randomized, and placebo-controlled, and included respectively 660 and 48,000 volunteers from 19 to 80 years. The anti-RBD titers were evaluated using a quantitative ELISA system developed at the Center for Immunoassay, Havana Cuba, and ELECSYS system from Roche. The RBD to ACE2 plate-based binding competitive ELISA was performed to determine the inhibitory activity of the anti-RBD polyclonal sera on the binding of the hFc-ACE2 coated plates. The neutralization antibody titers were detected by a traditional virus microneutralization assay (MN50). Finding(s): The Abdala vaccine reached 92.28% efficacy. The epidemic was frankly under control in Cuba after the vaccine introduction having reached the highest levels of cases and mortality in July 2021 with the dominance of the Delta strain. The peak of the Omicron wave, unlike other countries, did not reach half of the cases of the Delta wave with a significant reduction in mortality. The mucosal vaccine candidate "Mambisa" completed its clinical development as a booster dose for convalescent subjects reaching the trial end-point. Conclusion(s): Vaccine composition based on RBD recombinant antigen alone is sufficient to achieve high vaccine efficacy comparable to mRNA and live vaccine platforms. The vaccine also protects against different viral variants including Delta and Omicron strains.Copyright © 2023

COVID-19, black swans and the anticipation of health disasters: future problems and the future as a problem in public health ethics

The COVID-19 pandemic has recalled the importance of prevention and preparedness for highly disastrous events in community health. Several emerging phenomena pose prospective threats to public health. However, the largely future-oriented character of problems, for instance, such as antibiotic resistance, the impact of climate change on health, or the bioengineering of pathogens generates difficulties of analysis. What are the ethical and epistemological challenges raised by future public health problems? How should the moral problems of potentially catastrophic future scenarios be addressed? This article argues in favour of adopting anticipatory ethical approaches from public health ethics. First, it will be argued that addressing these future problems requires reflection on the future as an ethical and epistemic problem. Second, the characteristics of the emerging anticipatory ethics in the fields of ethics of technology and bioethics will be clarified. Third, the application of foresight and anticipatory methodologies in public health ethics debates will be defended. Finally, some reflections will be offered to strengthen anticipatory normative analyses to prevent and address in advance the adverse effects of future health crises.

SARS-CoV-2 USES ENDOSOMAL MEMBRANES TO BUILD REPLICATION ORGANELLES

Background: The health emergency caused by the COVID-19 pandemic has evidenced that the frequency of spillover episodes of viruses infecting bats to other species, including humans, has significantly increased compared to previous decades. Besides SARS-CoV-2, six other human coronaviruses (NL63, 229E, OC43, HKU1, SARS-CoV and MERS-CoV) emerged in the 20th and 21st century, most likely because of cross-species transmission events from bats. While many of these coronaviruses cause mild respiratory infections, MERS-CoV, SARS-CoV and SARS-CoV-2 can cause severe respiratory distress, particularly in immunocompromised individuals. However, unlike SARS-CoV and MERS-CoV, SARS-CoV-2 is highly contagious, very stable, with many person-to-person transmissions, which can occur even before individuals exhibit any symptoms. While vaccines are readily available, the emergence of new SARS-CoV-2 variants along with the increasing incidence of individuals developing long COVID urge to develop antivirals specific to treat COVID-19. To reach this goal, we need to have a working knowledge of the host-SARS-CoV-2 interactions to identify targets for therapeutic intervention. Method(s): Following that rationale, we focused on understanding how SARSCoV- 2 generates replication organelles (ROs). All coronaviruses need to remodel cellular membranes to create these structures to allow the active replication and transcription of their genome. Due to their relevance for virus replication, disabling RO formation represents a promising strategy to fight SARS-CoV-2. However, the biogenesis mechanism, the origin, and type of these replication organelles are still a major focus of debate. To identify the cellular membranes that SARS-CoV-2 uses to generate ROs we used multiple cell lines and primary cells that were evaluated by fluorescence microscopy, genetic engineering, compounds that specifically inhibit cellular processes, and immunoprecipitation assays to validate protein-protein interactions. We also used RT-qPCR to assess viral genome replication. Result(s): SARS-CoV-2 uses the viral protein NSP6 to remodel endosomal membranes juxtaposed to the ER to generate replication organelles. Specifically, the virus depends on Clathrin, COPB1, and Rab5 for efficient SARSCoV- 2 RNA synthesis. Conclusion(s): Uncovering the origins and mechanism(s) by which SARS-CoV-2 assembles ROs opens new avenues to develop strategies to interfere with RO biogenesis and halt virus replication.

Applications of genetic engineering in COVID-19

We identify major genetic engineering tools and methods developed to date, most targeting the spike protein and virus interaction with host cells. The rapid development of the vaccine and of antigenic and serological tests is the main achievement of one of the darkest pages of human history in the 21st century. This success relies on our capacity to sequence and manipulate genomes. It is also the result of major developments in the understanding of severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle-East respiratory syndrome coronavirus. These viruses were responsible for major outbreaks which foreshadow the coronavirus disease 2019 epidemic. The major role of the SARS-CoV2 spike protein in the host interaction was rapidly confirmed. Subcloning and expression in various vectors enable the development of highly protective antidisease vaccines. A wide range of assays involving recombinant cell lines and vectors allows researchers to search for the next therapies. © 2023 Elsevier Inc. All rights reserved.

American Christianity and the New Eugenics: Consumerism, Human Genetics, and the Challenge to Christian Personhood

Christian leaders and organizations sided overwhelmingly with politicians, scientists, social elites, and various progressive groups to champion the application of "scientific" methods designed to improve the nation's gene pool. Large numbers of Christians believed that genetic improvement of the human species was in keeping with God's command that humankind exercise dominion over creation.2 The unholy association of American Christianity with eugenics laboratories and associations was undermined by 1940 through rec-ognition of Nazi atrocities and realization of the extent to which eugenicists would go in attempts to accelerate and enhance what they saw as "natural" evolutionary processes. The generally favorable disposition of American Christians to the market's moral outcomes, even those that seem to test their reli-gious values, creates a climate in which churches increasingly will be challenged to protect the divine source of human dignity-what German theologian Helmut Thielicke called "alien dignity"-from functional and manipulable views of the person that arise alongside markets for genetic services.3 The entrepreneurial nature of the new "consumer" eugenics also highlights a divide that has formed between American churches and their memberships. Newborn screening and various forms of "reproductive genetics," oncology practices, treatments for auto-immune diseases, and even the response to the COVID-19 pandemic have been boons to this industry, not to mention growth in general interest among Americans eager to learn more about their genealogical and family histories.5 There is even a mail order industry for amateur geneticists who wish to experiment with gene editing in the privacy of their homes.6 It is consumer demand by individuals whose motivations range from the simple desire to experiment with cutting-edge technologies to those who envision possibilities for human "enhancement" to patients who face debil-itating genetic diseases, which fuels fear of a new eugenic age.

Expression of Brucella abortus Omp25 protein in Lactococcus lactis probiotic bacteria

Background and purpose: The sequence of Omp25 is conserved in all Brucella species. The high antigenicity of the product of this gene stimulates the host's immune system. Using engineered probiotic bacteria is an appropriate method for vaccine transport. The aim of this study was to express the Omp25 of the Brucella abortus pathogenic bacterium in Lactococcus lactis probiotic bacterium. Materials and methods: In this experimental study, the required vector was designed and synthesized to include the gene of interest and a signal peptide (pNZ8148-Usp45-Omp25). E. coli strain TOP10F was transformed using the pNZ8148-Usp45-Omp25 expression vector based on induction by nisin. The recombinant plasmid was extracted from the transformed bacteria using a plasmid extraction kit. The L. lactis was transformed by pNZ8148-Usp45-Omp25 vector using electroporation. Evaluation of the expression of Omp25 gene at the RNA level was assessed by reverse transcription method and confirming the presence of recombinant Omp25 protein in the engineered bacteria using SDS-PAGE method. Results: Successful expression of B. abortus Omp25 in L. lactis was verified by RT-PCR. Subsequently, the proteins were separated based on molecular weight using sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). The protein expression analysis showed the expression of Omp25 as a 25 kDa extra band in transformed L. lactis compared to the L. lactis receiving the vector lacking the target gene. Conclusion: This study shows that Omp25 is expressed in L. lactis transformed via pNZ8148-Usp45-Omp25 by electroporation. Transformed L. lactis can be successfully used as a subunit oral vaccine in prevention of Brucellosis.

Editor's Preface

Charles McDaniel, associate professor in the Interdisciplinary Core Program of Baylor University's Honors College, raises such issues in "American Christianity and the New Eugenics: Consumer-ism, Human Genetics, and the Challenge to Christian Personhood." Among Evangelical Christians, after Margaret Mead, perhaps the best-known anthropology major is Billy Graham. In a review and response, Elmer John Thiessen, adjunct professor of phi-losophy at Emmanuel Christian College, reviews Perry L. Glanzer and Na-than F. Alleman's The Outrageous Idea of Christian Teaching.

History of advances in genetic engineering of viruses before COVID-19 pandemic.

Background: On December 31, 2019, the World Health Organization's China Country Office was alerted to cases of pneumonia of unknown cause detected in Wuhan City, Hubei Province of China. Methods: Due to the fact that to date, the question of the origin of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has not been resolved yet, the author analyzed the main advances in the development of genetic engineering of viruses that took place before the onset of the COVID-19 pandemic. Results: The first artificial genetically modified viruses could appear in nature in the mid-1950s. The technique of nucleic acid hybridization was developed by the end-1960s. In the late 1970s, a method called the "reverse genetics" emerged to synthesize ribonucleic acid and deoxyribonucleic acid molecules. In the early 1980-s, it became possible to combine the genes of different viruses and insert the genes of one virus into the genome of another virus. Since that time, the production of vector vaccines began. At present, by modern technologies one can assemble any virus based on the nucleotide sequence available in the virus database or designed by a computer as a virtual model. Conclusion: Scientists around the world are invited to answer the call of Neil Harrison and Jeffrey Sachs of Columbia University, for a thorough and independent investigation into the origin of SARS-CoV-2. Only a full understanding of the origin of the new virus can minimize the likelihood of a similar pandemic in the future.

The History of Ethnic Minority Psychological Associations in the United States

The American Psychological Association (APA, 2021) issued an apology to people of color for their role in "promoting, perpetuating and failing to challenge racism, racial discrimination, and human hierarchy (p.1)." This may be related to the criticisms that American psychology is ahistorical, ignores social realities, (Crenshaw, 1995;Martin-Baro, 1996) and has a Eurocentric bias (Bhatia, 2020). This paper briefly describes historical examples of the oppression of Black, Indigenous and People of Color (BIPOC) in the United States in the health and social sciences fields. It details the context of the historical emergence of ethnic minority psychological associations led by the departure of Black psychologists from APA in the late 1960's. Several other BIPOC groups followed their lead. APA governance then met with the ethnic minority association leaders at the Dulles Conference in 1978. The recommendations of this conference took longer to achieve than what was originally expected yet these demonstrate the diversity, equity and inclusion efforts initiated by ethnic minority psychologists and later supported by funding agencies.

Bioavailability of iron and its potential to improve the immune system and ward off COVID-19: a review

Iron is a mineral that plays an important role, especially to prevent anaemia through the production of red blood cells. Iron also plays a role in physiological processes, such as the activation of enzymes and hormones, as well as increasing the immune system in warding off various viral infections. Therefore, iron bioavailability needs to be considered to take the greatest benefit of iron. This review discussed the factors that can affect the bioavailability of iron, various technologies to increase the bioavailability, and its potential in enhancing the immune system. Iron bioavailability can be increased by fortification, fermentation, the addition of vitamin C, and iron encapsulation. Under conditions of adequate iron intake, iron plays an important role in enhancing the immune system through controlling lymphocytes and T cell proliferation. However, excess iron consumption can be at risk of weakening the host's immune response to viruses. Therefore, the appropriate level of iron intake must be maintained accurately to be used optimally and has the potential to ward off viral infections, including the Sars-CoV-2 virus as the cause of COVID-19.Copyright © 2023, Rynnye Lyan Resources. All rights reserved.

DEVELOPMENT of A MOLECULAR POINT of CARE DETECTION KIT for NEISSERIA GONORRHOEAE

Neisseria gonorrhoeae (NG) is the etiological agent of gonorrhea, one of the four most prevalent sexually transmitted diseases worldwide. The diagnosis of this pathogen is carried out mainly by PCR methodologies, because it is accurate, sensitive and fast. However, it is an expensive technique that is difficult to perform in every health center. Thus, the development of rapid and affordable molecular point-of-care tests (POCT) is very important because it can be applied in isolated places or low complexity centers. Nucleic acid diagnostics generally have three steps: extraction, amplification and visualization. To solve the two last steps, we decided to use an isothermal nucleic acid amplification system coupled to a pH-sensitive dye. In our laboratory (LIGBCM-Laboratory of Genetic Engineering and Cellular and Molecular Biology, of the National University of Quilmes), an isothermal loop-based nucleic acid amplification methodology called Easy Loop Amplification (ELA) was developed. Its application for the diagnosis of both human and veterinary pathogens, showed a great adaptability to different organisms. One of our developed kits was for the detection of SARS CoV-2. In this way, the main objective of this research project is the development of a molecular POCT kit for NG with no need of complex equipment. The development of this kit for NG started with a bioinformatic analysis that allowed the molecular target selection from the genome of NG and afterwards the primer design. With one of the selected target sequences, a positive control was constructed de novo and used for the optimization of the isothermal amplification system. Different reaction parameters were evaluated and optimized (temperature, magnesium concentration, etc.). The specificity and detection limit of ELA were evaluated. The in silico analysis of the primers showed 100% specificity and the detection limit (analyzed by a calibration curve) resulted in less than 1000 molecules. ELA diagnostics of NG showed similar parameters to those found with qPCR techniques. This amplification method could be used with a portable device, allowing the diagnosis in low complexity centers and highlighting its capability to be exploited as a POCT system.

AICTE Sponsored International e-Conference on Pharmacy Practice and Therapeutics, Andhra Pradesh, India, 19-20 August 2020

Pharmacological Targets for Treatment of Covid-19 Infection, Plasma Therapy for Covid-19 Treatment, Safety Measurements for Health Care Providers and Patients at Indian

Covid-19 pandemic and future global catastrophic risks as a challenge for health-care ethics

Purpose>Current covid-19 pandemic challenges health-care ethics. Ones of the most important challenges are medical resources allocation and a duty to treat, often addressed to medical personnel. This paper suggests that there are good reasons to rethink our health-care ethics for future global catastrophic risks. Current pandemic shows how challenging can be an issue of resources allocation even in a relatively small kind of catastrophic event such as covid-19 pandemic. In this paper, the authors show that any future existential bigger catastrophe may require new guidelines for the allocation of medical resources. The idea of assisted dying is considered as a hypothetical scenario.Design/methodology/approach>This is a conceptual work based on conceptual analysis at the intersection of risk studies, health-care ethics and future studies. This study builds the argument on the assumption that the covid-19 pandemic should be treated as a sort of global catastrophic risk. Findings show that there are no such attempts in currently published peer-reviewed academic literature. This is crucial concept for the meta-analysis. This study shows why and how current pandemic can be interpreted in terms of global catastrophic risk even if, literally, covid-19 does not meet all criteria required in the risk studies to be called a global catastrophe.Findings>We can expect an emergence of discriminatory selection policy which will require some actions taken by future patients like, for example, genetic engineering. But even then it is inevitable that there will still be a large number of survivors who require medical assistance, which they have no chance of receiving. This is why this study has considered the concept of assisted dying understood as an official protocol for health-care ethics and resources allocation policy in the case of emergency situations. Possibly more controversial idea discussed in this paper is an idea of assisted dying for those who cannot receive required medical help. Such procedure could be applied in a mass-scale during a global catastrophic event.Research limitations/implications>Philosophers and ethicists should identify and study all possible pros and cons of this discrimination rule. As this study’s findings suggested above, a reliable point of reference is the concept of substantial human enhancement. Human enhancement as such, widely debated, should be studied in that specific context of discrimination of patients in an access to limited medical resources. Last but not least, scientific community should study the concept of assisted dying which could be applied for those survivors who have no chance of obtaining medical care. Such criteria and concepts as cost-benefit analysis, the ethics of quality of life, autonomy of patients and duty of medical personnel should be considered.Practical implications>Politicians and policymakers should prepare protocols for global catastrophes where these discrimination criteria would have to be applied. The same applies to the development of medical robotics aimed at replacing human health-care personnel. We assume that this is important implication for practical policy in healthcare. Our prediction, however plausible, is not a good scenario for humanity. But given this realistic development trajectory, we should do everything possible to prevent the need for the discriminatory rules in medical care described above.Originality/value>This study offers the idea of assisted dying as a health-care policy in emergency situations. The authors expect that next future global catastrophes – looking at the current pandemic only as a mild prelude – will force a radical change in moral values and medical standards. New criteria of selection and discrimination will be perceived as much more exclusivist and unfair than criteria applied today.

Expression of COVID-19 virus N gene in insect cells

In order to obtain COVID-19 nucleocapsid protein with similar function and activity to natural protein and apply it to practical detection. Firstly, according to Bac-to-bac insect expression system and synthetic COVID-19 nucleocapsid protein(N protein)sequence, BamH I and Xba I on pFastBacTMHTB vector were added to upstream and downstream primers respectively. The N gene was amplified by PCR technology, and T-Vector pMD19(simple)vector and pFastBacTMHTB vector were connected successively and recombinant plasmids pMD19-T(simple)-COV19-N and pFastBacTMHTB-COV19-N, and finally construct recombinant bacmid DH10 Bac-pFastBacTMHTB-COV19-N in DH10 Bac cells was expressed in insect cell Sf9. The recombinant protein was obtained and analyzed by SDS-PAGE and WB. The recombinant plasmid pMD19-T(simple)-COV19-N was identified by PCR and double enzyme digestion. The recombinant bacmid DH10 Bac-pFastBacTMHTB-COV19-N was constructed in DH10 Bac cells was identified by PCR and the expected two bands were 2 430,3 690 bp, respectively, which proved that the recombinant bacmid was successfully obtained. The recombinant bacmid was transfected into Sf9 insect cells. At the same time, the recombinant GFP protein control group was established. After 120 h of transfection, the recombinant N protein and recombinant GFP protein were collected and samples were prepared;SDS-PAGE and WB analysis were carried out respectively. HRP-His labeled antibody was used to verify that the transfection was successful, and both recombinant N protein and recombinant GFP protein were successfully expressed in Sf9 cells. The experimental results were consistent with the expectation, and the size of recombinant N protein band was about 46 ku. The eukaryotic expression vector of respiratory coronavirus N gene was successfully constructed and successfully expressed in insect cells, which provides an experimental basis for the establishment of ELISA detection methods and other related research.

Population's perception about vaccines from gene engineering techniques and against COVID-19

The objective of this work was to obtain the perception of the population about: (1) the importance of immunization;(2) knowledge of methodologies (genetic manipulation) used for vaccine production;(3) acceptance of the vaccine against COVID-19. To this end, an online questionnaire produced via Google Forms was made available. This form was disseminated in various virtual environments such as academic and social media. Over six months, 353 responses were obtained. In this survey, responses were obtained from participants residing in 14 states in Brazil. Considering the investigation about vaccines, the results showed that most (90.6%) have already been vaccinated, as well as their families, and understand the importance of immunization. The techniques of genetic manipulation are complex and more widespread in the scientific environment, so it was observed that a large part of the interviewed population does not have an opinion on the subject. The research showed that because of the state of calamity caused by the pandemic, people had urgency in immunization, even with distrust in aspects such as efficiency. Currently, it is clear that vaccination does not prevent infection with the virus, but minimizes the damage caused by the infection. Data from the Ministry of Health show that the number of cases and deaths has reduced by more than 40%, showing once again the effectiveness and necessity of immunization of the population.