Design and Optimization of a Mycoplasma Detection Assay

Mycoplasma are among the smallest free living microorganisms. These bacteria grow slowly, lack a rigid cell wall and are not eliminated by filter sterilization methods used in tissue culture. Mycoplasma infection affects biochemical and genetic aspects of cultured cells, resulting in experimental inconsistency. Therefore, it is necessary to establish routine testing for mycoplasma contamination in tissue culture laboratories. Our goal is to develop a reliable and cost-effective test for mycoplasma in cell culture based on established methods found in literature. We first cloned and sequenced a PCR product from a commercial mycoplasma detection kit. Sequencing revealed the 16s rRNA as the target for mycoplasma detection; we confirmed this target by conducting a literature search. PCR primers were designed using 16s rRNA gene as a target. We set-up reactions and optimized conditions for the real-time PCR assay to detect the target and confirmed amplicon size with agarose gel electrophoresis. We identified that 56oC was the best temperature for the PCR and found that agarose gel electrophoresis was a better detection method because it identified the size to confirm the proper product. The primers we ordered to develop this assay produce the proper band; however, results of several assays have been inconsistent as sometimes a known positive sample fails to amplify. As well, in several PCR reactions the negative showed a signal. The overall reaction needs improvements to have greater reliability and to eliminate all sources of contamination. Research is continuing results are not final

HIV-1 Vpr Causes Synaptodendritic Damage in Neurons

HIV weakens the immune system by infecting and destroying T-cells, leaving the body vulnerable to infection and the development of AIDS. Conventional treatments for HIV, such as combined anti-rectroviral therapy (cART), fail to prevent the development of HIV-associated neurocognitive disorder (HAND). Neurological dysfunction has been directly related to the invasion of HIV in the central nervous system (CNS). HIV produces neurotoxic proteins, such as the Viral Protein R (Vpr), which contribute to HAND. Astrocytes are the most abundant cells in the brain and an important HIV target. We hypothesize that astrocytes expressing Vpr will cause neuronal damage in our co-culture system. Primary astrocytes were transfected with Vpr plasmid or control (pEGFP or mock) using electroporation. Astrocytes were then co-cultured with cortical neurons. At 48 and 72 hours we collected the primary astrocytes to confirm the Vpr expression via western blot analysis. We then measured structural damage in the neurons using immunofluorescence for cytoskeletal (MAP2, f-actin) and synaptic (synaptophysin) damage. Preliminary results showed strong staining of filamentous actin and MAP2 with weak detection of synaptophysin. The positive control for neurotoxicity (2.8µM acrylamide) showed substantial damage to the cellular structure. Results for Vpr expression are pending. After confirming that the immunofluorescence assays are working with our controls, we expect to detect any synaptodendritic damage in the neurons caused by Vpr in our upcoming experiments

Chronic pain, vulnerability and human spirit while living under the umbrella of COVID-19

The purpose of writing this article is to describe what added challenges people like us who are living with chronic pain are experiencing during the COVID-19 pandemic. We explore what this challenging time means to us and how it affects our lives, along with providing insight into our experiences. This is not a research study, but instead an article that shares perspectives from people with lived experience of chronic pain. Our narratives are presented to create an awareness of the plight for people already living with challenging health conditions and how the COVID-19 pandemic has added additional layers of vulnerability. While these stories offer brief accounts of some of the challenges we face, they also provide glimmers of hope that others with similar challenges can look to for inspiration. We also hope that our stories and reflections provide discussion points and perhaps even case studies for clinicians and policymakers who are working to strengthen clinical care and health systems for people with chronic conditions during pandemics such as COVID-19. Sharing our lived experiences with chronic pain during this global crisis may also spark critical conversations among all chronic pain stakeholders to ensure that we could continue to provide excellent care, strong self-management support and optimal health policy-making as this pandemic continues to unfold and for consideration for future health emergencies. Experience Framework This article is associated with the Quality & Clinical Excellence lens of The Beryl Institute Experience Framework. (http://bit.ly/ExperienceFramework) Access other PXJ articles related to this lens. Access other resources related to this lens

Assessing Ozone-Related Health Impacts under a Changing Climate

Climate change may increase the frequency and intensity of ozone episodes in future summers in the United States. However, only recently have models become available that can assess the impact of climate change on O(3) concentrations and health effects at regional and local scales that are relevant to adaptive planning. We developed and applied an integrated modeling framework to assess potential O(3)-related health impacts in future decades under a changing climate. The National Aeronautics and Space Administration–Goddard Institute for Space Studies global climate model at 4° × 5° resolution was linked to the Penn State/National Center for Atmospheric Research Mesoscale Model 5 and the Community Multiscale Air Quality atmospheric chemistry model at 36 km horizontal grid resolution to simulate hourly regional meteorology and O(3) in five summers of the 2050s decade across the 31-county New York metropolitan region. We assessed changes in O(3)-related impacts on summer mortality resulting from climate change alone and with climate change superimposed on changes in O(3) precursor emissions and population growth. Considering climate change alone, there was a median 4.5% increase in O(3)-related acute mortality across the 31 counties. Incorporating O(3) precursor emission increases along with climate change yielded similar results. When population growth was factored into the projections, absolute impacts increased substantially. Counties with the highest percent increases in projected O(3) mortality spread beyond the urban core into less densely populated suburban counties. This modeling framework provides a potentially useful new tool for assessing the health risks of climate change

Epoprostenol (PGI2, Prostacyclin) During High‐Risk Hemodialysis: Preventing Further Bleeding Complications

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/97231/1/j.1552-4604.1988.tb03222.x.pd

Evaluation of the influence of kyphosis and scoliosis on intervertebral disc extrusion in French bulldogs

Although thoracic vertebral malformations with kyphosis and scoliosis are often considered incidental findings on diagnostic imaging studies of screw-tailed brachycephalic breeds, they have been suggested to interfere with spinal biomechanics and intervertebral disc degeneration. It is however unknown if an abnormal spinal curvature also predisposes dogs to develop clinically relevant intervertebral disc herniations. The aim of this study was to evaluate if the occurrence of thoracic vertebral malformations, kyphosis or scoliosis would be associated with a higher prevalence of cervical or thoracolumbar intervertebral disc extrusion in French bulldogs