Health, Medicine, & Biotechnology

A Novel Protocol for Decoating and Permeabilizing Bacterial Spores for Epifluorescent Microscopy

This technique can be used in semiconductor, pharmaceutical, and food processing industries. Based on previously reported procedures for permeabilizing vegetative bacterial cells, and numerous trial-and-error attempts with bacterial endospores, a protocol was developed for effectively permeabilizing bacterial spores, which facilitated the applicability of fluorescent in situ hybridization (FISH) microscopy. Bacterial endospores were first purified from overgrown, sporulated suspensions of B. pumilus SAFR-032. Purified spores at a concentration of ≈10 million spores/mL then underwent proteinase-K treatment, in a solution of 468.5 μL of 100 mM Tris-HCl, 30 μL of 10% SDS, and 1.5 μL of 20 mg/mL proteinase-K for ten minutes at 35 ºC. Spores were then harvested by centrifugation (15,000 g for 15 minutes) and washed twice with sterile phosphate-buffered saline (PBS) solution. This washing process consisted of resuspending the spore pellets in 0.5 mL of PBS, vortexing momentarily, and harvesting again by centrifugation. Treated and washed spore pellets were then resuspended in 0.5 mL of decoating solution, which consisted of 4.8 g urea, 3 mL Milli-Q water, 1 mL 0.5M Tris, 1 mL 1M dithiothreitol (DTT), and 2 mL 10% sodium-dodecylsulfate (SDS), and were incubated at 65 ºC for 15 minutes while being shaken at 165 rpm.

Posted in: Briefs, Medical

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Method and Apparatus for Automated Isolation of Nucleic Acids from Small Cell Samples

Advantages include reduced or eliminated use of toxic reagents and operator-independent extraction. RNA isolation is a ubiquitous need, driven by current emphasis on micro-arrays and miniaturization. With commercial systems requiring 100,000 to 1,000,000 cells for successful isolation, there is a growing need for a small-footprint, easy-to-use device that can harvest nucleic acids from much smaller cell samples (1,000 to 10,000 cells). The process of extraction of RNA from cell cultures is a complex, multi-step one, and requires timed, asynchronous operations with multiple reagents/buffers. An added complexity is the fragility of RNA (subject to degradation) and its reactivity to surface.

Posted in: Briefs, TSP, Medical

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Enabling Microliquid Chromatography by Microbead Packing of Microchannels

The microbead packing is the critical element required in the success of onchip microfabrication of critical microfluidic components for in-situ analysis and detection of chiral amino acids. In order for microliquid chromatography to occur, there must be a stationary phase medium within the microchannel that interacts with the analytes present within flowing fluid. The stationary phase media are the microbeads packed by the process discussed in this work. The purpose of the microliquid chromatography is to provide a lightweight, low-volume, and low-power element to separate amino acids and their chiral partners efficiently to understand better the origin of life.

Posted in: Briefs, Medical

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On-Command Force and Torque Impeding Devices (OC-FTID) Using ERF

This technology is applicable as a rehabilitation or exercise device. Various machines have been developed to address the need for countermeasures of bone and muscle deterioration when humans operate over extended time in space. Even though these machines are in use, each of them has many limitations that need to be addressed in an effort to prepare for human missions to distant bodies in the solar system.

Posted in: Briefs, TSP, Medical

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RECOVIR Software for Identifying Viruses

Most single-stranded RNA (ssRNA) viruses mutate rapidly to generate a large number of strains with highly divergent capsid sequences. Determining the capsid residues or nucleotides that uniquely characterize these strains is critical in understanding the strain diversity of these viruses. RECOVIR (an acronym for “recognize viruses”) software predicts the strains of some ssRNA viruses from their limited sequence data. Novel phylogenetic-treebased databases of protein or nucleic acid residues that uniquely characterize these virus strains are created. Strains of input virus sequences (partial or complete) are predicted through residue-wise comparisons with the databases.

Posted in: Briefs, Medical, Software

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On-Command Force and Torque Impeding Devices (OC-FTID) Using ERF

This technology is applicable as a rehabilitation or exercise device. Various machines have been developed to address the need for countermeasures of bone and muscle deterioration when humans operate over extended time in space. Even though these machines are in use, each of them has many limitations that need to be addressed in an effort to prepare for human missions to distant bodies in the solar system.

Posted in: Briefs, MDB, Briefs, Medical, Rehabilitation & Physical Therapy

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Rapid Detection of Herpes Viruses for Clinical Applications

There are eight herpes viruses that infect humans, causing a wide range of diseases resulting in considerable morbidity and associated costs. Varicella zoster virus (VZV) is a human herpes virus that causes chickenpox in children and shingles in adults. Approximately 1,000,000 new cases of shingles occur each year; post-herpetic neuralgia (PHN) follows shingles in 100,000 to 200,000 people annually. PHN is characterized by debilitating, nearly unbearable pain for weeks, months, and even years. The onset of shingles is characterized by pain, followed by the zoster rash, leading to blisters and severe pain. The problem is that in the early stages, shingles can be difficult to diagnose; chickenpox in adults can be equally difficult to diagnose. As a result, both diseases can be misdiagnosed (false positive/negative).

Posted in: Briefs, Medical, Data Acquisition

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