CRISPR/Cas9 as an antiviral against Orthopoxviruses using an AAV vector.

A vaccine for smallpox is no longer administered to the general public, and there is no proven, safe treatment specific to poxvirus infections, leaving people susceptible to infections by smallpox and other zoonotic Orthopoxviruses such as monkeypox. Using vaccinia virus (VACV) as a model organism for other Orthopoxviruses, CRISPR-Cas9 technology was used to target three essential genes that are conserved across the genus, including A17L, E3L, and I2L. Three individual single guide RNAs (sgRNAs) were designed per gene to facilitate redundancy in rendering the genes inactive, thereby reducing the reproduction of the virus. The efficacy of the CRISPR targets was tested by transfecting human embryonic kidney (HEK293) cells with plasmids encoding both SaCas9 and an individual sgRNA. This resulted in a reduction of VACV titer by up to 93.19% per target. Following the verification of CRISPR targets, safe and targeted delivery of the VACV CRISPR antivirals was tested using adeno-associated virus (AAV) as a packaging vector for both SaCas9 and sgRNA. Similarly, AAV delivery of the CRISPR antivirals resulted in a reduction of viral titer by up to 92.97% for an individual target. Overall, we have identified highly specific CRISPR targets that significantly reduce VACV titer as well as an appropriate vector for delivering these CRISPR antiviral components to host cells in vitro.

De curandis hominum morbis: An 18th century medical prescription for measles and smallpox in the Nuevo Reino de Granada / De curandis hominum morbis: una receta médica del siglo XVIII para el sarampión y las viruelas en el Nuevo Reino de Granada

A collection of more than one hundred medical recipes from the late 18th century was donated by Presbyter Cipriano Rodríguez Santa María, institutional eponym of the collection at the Historical Archive "Octavio Arizmendi Posada" of the library of the Universidad de La Sabana in Colombia. These texts represent an important historical and medical legacy and they constitute an important basis for understanding the colonial and traditional therapeutics related to various diseases. In this article, we describe one of these recipes for the treatment of smallpox and measles as a contribution to the history of medicine in Colombia.

En el Archivo Histórico de la Biblioteca "Octavio Arizmendi Posada" de la Universidad de La Sabana, se encuentra una colección de más de un centenar de recetas médicas de finales del siglo XVIII donadas por el presbítero Cipriano Rodríguez Santa María, epónimo institucional del archivo. Estos textos son un legado histórico médico y un fundamento para comprender la terapéutica colonial y tradicional de diversas enfermedades. En este artículo, se describen algunas recetas para el tratamiento de la viruela y el sarampión, como aporte a la historia de la medicina en Colombia.

The Role of Brincidofovir in Preparation for a Potential Smallpox Outbreak.

Smallpox (variola) virus is considered a Category A bioterrorism agent due to its ability to spread rapidly and the high morbidity and mortality rates associated with infection. Current recommendations recognize the importance of oral antivirals and call for having at least two smallpox antivirals with different mechanisms of action available in the event of a smallpox outbreak. Multiple antivirals are recommended due in large part to the propensity of viruses to become resistant to antiviral therapy, especially monotherapy. Advances in synthetic biology heighten concerns that a bioterror attack with variola would utilize engineered resistance to antivirals and potentially vaccines. Brincidofovir, an oral antiviral in late stage development, has proven effective against orthopoxviruses in vitro and in vivo, has a different mechanism of action from tecovirimat (the only oral smallpox antiviral currently in the US Strategic National Stockpile), and has a resistance profile that reduces concerns in the scenario of a bioterror attack using genetically engineered smallpox. Given the devastating potential of smallpox as a bioweapon, preparation of a multi-pronged defense that accounts for the most obvious bioengineering possibilities is strategically imperative.

Clinical management of potential bioterrorism-related conditions.

The agents most likely to be used in bioterrorism attacks are reviewed, along with the clinical syndromes they produce and their treatment.

Understanding translational research: a play in four acts.

Translational research (TR) bridges discovery to clinical delivery. All TR also requires the development of an intervention. Classical 'bench to bedside' TR is responsible for many important advances, but cannot account for many others, which begin with clinical observations. My personal involvement in TR has ranged from exploration of long-term mechanical circulatory support devices to amelioration of the progression of Alzheimer's disease to the pharmacologic cure of smallpox. This experience suggests that most TR is opportunistic and inefficient. A strategic approach to TR based on a better understanding of the processes it entails could enhance progress in TR despite its increasing complexity.

[The epidemic of smallpox in Guangdong Province and its prevention and treatment in the Republican period].

During the Republican period, the smallpox broke out in high frequency almost annually in Guangdong. In response to infectious diseases, the Guangdong government established the Health Administration and institutions for infectious disease's prevention and treatment, and prepared smallpox vaccine by themselves. In order to grasp the situation of the epidemic, related institutions collected epidemic data weekly, monthly, and annually with the statistics reported regularly. Meanwhile, infectious disease hospital was established for smallpox patients. Harbor quarantine put smallpox as a key target of inspection. With the joint effort of the government and social organizations, massive "vaccination campaigns" was organized to promote vaccination, in which children, students and young people were the main subjects for inoculations. Prevention knowledge and anti-epidemic concept towards smallpox have been actively publicized and improved by media.

[Treatment and remedies against smallpox outbreaks in Ferrara in the late nineteenth century]. / Cure e rimedi contro le epidemie di vaiolo a Ferrara nella seconda metà del secolo XIX.

Health interventions against smallpox during the two epidemics in the second half of the 19th century are outlined. The 1871 hospital health report and the medical report on smallpox patients treated at the hospital and poorhouse of Ferrara between January 1891 and January 1892, drawn up by Alessandro Bennati, provide both interesting data and insights into the treatments and remedies of the time. The treatment of this illness was - and indeed could be - nothing other than symptomatic, there being no real means to halt the spread of the disease. Rather, other remedies were found by alleviating pain and regaining energy during the various stages of the disease. A close relationship between vaccination and the incidence and gravity of the illness is underlined. When the practice of vaccination started to be widely employed at the end of the century, there were almost no cases of death due to smallpox. The pharmacopoeias of the time, Antonio Campana's Farmacopea ferrarese in particular, proved an essential guide in the analysis of each document.

Biowarfare and bioterrorism.

Bioterrorism is not only a reality of the times in which we live but bioweapons have been used for centuries. Critical care physicians play a major role in the recognition of and response to a bioterrorism attack. Critical care clinicians must be familiar with the diagnosis and management of the most likely bioterrorism agents, and also be adequately prepared to manage a mass casualty situation. This article reviews the epidemiology, diagnosis, and treatment of the most likely agents of biowarfare and bioterrorism.

Secondary and tertiary transmission of vaccinia virus after sexual contact with a smallpox vaccinee--San Diego, California, 2012.

On June 24, 2012, CDC notified Public Health Services, County of San Diego Health and Human Services Agency, of a suspected case of vaccinia virus infection transmitted by sexual contact. The case had been reported to CDC by an infectious disease specialist who had requested vaccinia immune globulin intravenous (VIGIV) (Cangene Corporation, Berwyn, Pennsylvania) for a patient with lesions suspicious for vaccinia. The patient reported two recent sexual contacts: one with a partner who recently had been vaccinated against smallpox and a later encounter with an unvaccinated partner. Infections resulting from secondary transmission of vaccinia virus from the smallpox vaccinee to the patient and subsequent tertiary transmission of the virus from the patient to the unvaccinated partner were confirmed by the County of San Diego Public Health Laboratory. The smallpox vaccine had been administered under the U.S. Department of Defense smallpox vaccination program. The vaccinee did not experience vaccine-associated complications; however, the secondary and tertiary patients were hospitalized and treated with VIGIV. No further transmission was known to have occurred. This report describes the epidemiology and clinical course of the secondary and tertiary cases and efforts to prevent further transmission to contacts.

Cutaneous manifestations of category A bioweapons.

The 2001 anthrax attacks on the United States brought bioterrorism to the forefront of the medical community. Because many bioterrorist agents produce cutaneous disease, dermatologists will likely be first responders during a future attack. Despite this, many dermatologists are not adequately prepared to deal with a bioterrorism attack. It is critical that all first responders be able to recognize symptoms of bioterrorism-related disease and prepared to respond to a bioterrorist attack to minimize threats to public health. This article reviews the diagnosis and treatment of diseases caused by potential biowarfare agents that produce cutaneous manifestations, and also provides information regarding reporting and containment of possible bioterrorism-related diseases.

Development of a multi--electrode electrical stimulation device to improve chronic wound healing.

A new device has been developed for the application of Electrical Stimulation to improve healing of chronic wounds. The device enables the creation of a composite electrode hence matching the electrode(s) to the size and shape of the wound. Up to 49 electrodes in an array can be combined, delivering High Voltage Pulsed Current (HVPC) in the range 60-120 Hz with a pulse duration range from 90-110 mus and the possibility of treating the patient with direct current instead of HVPC. In addition, the software can import the measurements from the ImpediMap device, analysing the electrical impedance of the tissues involved. A test on healthy volunteers did not prove a statistically significant rise in skin temperature, TcPO2 or impedance due to the stimulation, even though a slight reddening of the stimulated site was observed.