WormBot, an open-source robotics platform for survival and behavior analysis in C. elegans.

Caenorhabditis elegans is a popular organism for aging research owing to its highly conserved molecular pathways, short lifespan, small size, and extensive genetic and reverse genetic resources. Here we describe the WormBot, an open-source robotic image capture platform capable of conducting 144 parallel C. elegans survival and behavioral phenotyping experiments. The WormBot uses standard 12-well tissue culture plates suitable for solid agar media and is built from commercially available robotics hardware. The WormBot is controlled by a web-based interface allowing control and monitoring of experiments from any internet connected device. The standard WormBot hardware features the ability to take both time-lapse bright field images and real-time video micrographs, allowing investigators to measure lifespan, as well as heathspan metrics as worms age. The open-source nature of the hardware and software will allow for users to extend the platform and implement new software and hardware features. This extensibility, coupled with the low cost and simplicity of the system, allows the automation of C. elegans survival analysis even in small laboratory settings with modest budgets.

The Lavender Fertilized Anterior Cruciate Ligament Reconstruction: A Quadriceps Tendon All-Inside Reconstruction Fertilized With Bone Marrow Concentrate, Demineralized Bone Matrix, and Autograft Bone.

The use of biologics in sports medicine is increasing rapidly. Bone marrow concentrate has recently increased in popularity because it includes mesenchymal stem cells which, combined with AlloSync Pure, could lead to better incorporation and healing. The mixture of bone marrow concentrate and Allosync Pure can be used in anterior cruciate ligament reconstruction. We recently expanded on this approach with the addition of saving the host bone normally lost from tunnel reaming, using the GraftNet. After harvesting the autograft bone, we combine it with the AlloSync Pure and bone marrow concentrate. In this Technical Note, we show how this unique biologic composite is obtained and then added back into the tunnels on both the femur and tibia during a quadriceps tendon autograft all-inside anterior cruciate ligament reconstruction.

In vivo measurements reveal a single 5'-intron is sufficient to increase protein expression level in Caenorhabditis elegans.

Introns can increase gene expression levels using a variety of mechanisms collectively referred to as Intron Mediated Enhancement (IME). IME has been measured in cell culture and plant models by quantifying expression of intronless and intron-bearing reporter genes in vitro. We developed hardware and software to implement microfluidic chip-based gene expression quantification in vivo. We altered position, number and sequence of introns in reporter genes controlled by the hsp-90 promoter. Consistent with plant and mammalian studies, we determined a single, natural or synthetic, 5'-intron is sufficient for the full IME effect conferred by three synthetic introns, while a 3'-intron is not. We found coding sequence can affect IME; the same three synthetic introns that increase mcherry protein concentration by approximately 50%, increase mEGFP by 80%. We determined IME effect size is not greatly affected by the stronger vit-2 promoter. Our microfluidic imaging approach should facilitate screens for factors affecting IME and other intron-dependent processes.

Augmentation of Anterior Cruciate Ligament Reconstruction With Bone Marrow Concentrate and a Suture Tape.

The use of biological agents in sports medicine is increasing rapidly. The use of bone marrow concentrate has recently increased in popularity owing to the presence of mesenchymal stem cells. These stem cells combined with Allosync Pure (Arthrex, Naples, FL) could lead to better incorporation and healing. The mixture of bone marrow concentrate and Allosync Pure can be used in anterior cruciate ligament reconstruction, and we have developed a technique to use it in the femoral tunnel and deliver the medium arthroscopically. In this article we present a technique for using suture tape augmentation (InternalBrace; Arthrex) through the interference screw during a bone-patellar tendon-bone anterior cruciate ligament reconstruction.