Revisiting the Genicular Nerve Block: An Up-to-Date Guide Utilizing Ultrasound Guidance and Peripheral Nerve Stimulation - Anatomy Description and Technique Standardization

Background: Over the last decade, several authors have reported that percutaneous peripheral nerve stimulation (PNS) can be used to assist in verifying the position of the procedure needle tip in relation to nerve structures, and that the combined technique using both ultrasound (US) guidance and PNS may serve as a reliable method for confirmation of the correct position of the procedure needle tip. It has also been reported that, when combined with US guidance, PNS may increase the success rate of pain management interventions. Objectives: The aim of this technical report was to standardize an effective and easy to learn illustrated step-by-step technical approach to nerve identification during US-guided genicular nerve blocks, using percutaneous PNS as a verification instrument for procedure needle tip location. Study design: This technical protocol was developed based on the results of the authors' most recent cadaveric study on the innervation of the knee joint capsule. The technique was developed and tested by 4 different interventionists with different levels of expertise in US-guided procedures. Setting: The cadaveric study of the knee joint capsule innervation was performed at the laboratory of the Division of Anatomy of one institution. The technical protocol using US and PNS was later developed at the medical simulation center of a different institution. Methods: A team of anatomists from a division of anatomy of one institution performed the cadaveric study on the innervation of the knee joint capsule. A team of physicians then developed the step-by-step approach to this technical protocol at the medical simulation center of a different institution. Finally, the illustrated step-by-step approach was tested by 4 different interventionists with different levels of expertise in US-guided procedures (1 beginner-level user; 1 intermediate-level user; 2 expert-level users), using a portable percutaneous PNS and 2 different US transducers at 2 different institutions. Results: This technical protocol was successfully developed based on the results of the cadaveric study on the innervation of the knee joint capsule. Additionally, it was later successfully tested by interventionists with various levels of expertise utilizing different US equipment at separate institutions. Limitations: By combining US and nerve stimulation, this protocol requires the availability of both US equipment and necessary equipment for nerve stimulation that must all be made available in the sterile field. Another potential disadvantage is that nerve stimulation controls and the US image screen are generally located on 2 separate display panels, which could cause difficulty with visualization and simultaneous calibration for 2 individual devices. Conclusions: Our illustrated step-by-step technical protocol can be effectively and safely utilized as a reliable method of training, by which physicians with little to moderate US experience can improve their skills in accurately identifying the genicular nerves while performing US-guided examinations with the intent of executing a peripheral nerve block.info:eu-repo/semantics/publishedVersio

Microglia/Astrocytes-Glioblastoma Crosstalk: Crucial Molecular Mechanisms and Microenvironmental Factors

In recent years, the functions of glial cells, namely, astrocytes and microglia, have gained prominence in several diseases of the central nervous system, especially in glioblastoma (GB), the most malignant primary brain tumor that leads to poor clinical outcomes. Studies showed that microglial cells or astrocytes play a critical role in promoting GB growth. Based on the recent findings, the complex network of the interaction between microglial/astrocytes cells and GB may constitute a potential therapeutic target to overcome tumor malignancy. In the present review, we summarize the most important mechanisms and functions of the molecular factors involved in the microglia or astrocytes–GB interactions, which is particularly the alterations that occur in the cell’s extracellular matrix and the cytoskeleton. We overview the cytokines, chemokines, neurotrophic, morphogenic, metabolic factors, and non-coding RNAs actions crucial to these interactions. We have also discussed the most recent studies regarding the mechanisms of transportation and communication between microglial/astrocytes – GB cells, namely through the ABC transporters or by extracellular vesicles. Lastly, we highlight the therapeutic challenges and improvements regarding the crosstalk between these glial cells and GB

Genome of the Avirulent Human-Infective Trypanosome—Trypanosoma rangeli

Background: Trypanosoma rangeli is a hemoflagellate protozoan parasite infecting humans and other wild and domestic mammals across Central and South America. It does not cause human disease, but it can be mistaken for the etiologic agent of Chagas disease, Trypanosoma cruzi. We have sequenced the T. rangeli genome to provide new tools for elucidating the distinct and intriguing biology of this species and the key pathways related to interaction with its arthropod and mammalian hosts.  Methodology/Principal Findings: The T. rangeli haploid genome is ,24 Mb in length, and is the smallest and least repetitive trypanosomatid genome sequenced thus far. This parasite genome has shorter subtelomeric sequences compared to those of T. cruzi and T. brucei; displays intraspecific karyotype variability and lacks minichromosomes. Of the predicted 7,613 protein coding sequences, functional annotations could be determined for 2,415, while 5,043 are hypothetical proteins, some with evidence of protein expression. 7,101 genes (93%) are shared with other trypanosomatids that infect humans. An ortholog of the dcl2 gene involved in the T. brucei RNAi pathway was found in T. rangeli, but the RNAi machinery is non-functional since the other genes in this pathway are pseudogenized. T. rangeli is highly susceptible to oxidative stress, a phenotype that may be explained by a smaller number of anti-oxidant defense enzymes and heatshock proteins.  Conclusions/Significance: Phylogenetic comparison of nuclear and mitochondrial genes indicates that T. rangeli and T. cruzi are equidistant from T. brucei. In addition to revealing new aspects of trypanosome co-evolution within the vertebrate and invertebrate hosts, comparative genomic analysis with pathogenic trypanosomatids provides valuable new information that can be further explored with the aim of developing better diagnostic tools and/or therapeutic targets

Synergistic Interactions between the NS3hel and E Proteins Contribute to the Virulence of Dengue Virus Type 1

Dengue virus constitutes a significant public health problem in tropical regions of the world. Despite the high morbidity and mortality of this infection, no effective antiviral drugs or vaccines are available for the treatment or prevention of dengue infections. The profile of clinical signs associated with dengue infection has changed in recent years with an increase in the number of episodes displaying unusual signs. We use reverse genetics technology to engineer DENV-1 viruses with subsets of mutations previously identified in highly neurovirulent strains to provide insights into the molecular mechanisms underlying dengue neuropathogenesis. We found that single mutations affecting the E and NS3hel proteins, introduced in a different genetic context, had a synergistic effect increasing DENV replication capacity in human and mosquito derived cells in vitro. We also demonstrated correlations between the presence of these mutations and viral replication efficiency, viral loads, the induction of innate immune response genes and pathogenesis in a mouse model. These results should improve our understanding of the DENV-host cell interaction and contribute to the development of effective antiviral strategies