Quantitative detection of PfHRP2 in saliva of malaria patients in the Philippines.

BACKGROUND: Malaria is a global health priority with a heavy burden of fatality and morbidity. Improvements in field diagnostics are needed to support the agenda for malaria elimination. Saliva has shown significant potential for use in non-invasive diagnostics, but the development of off-the-shelf saliva diagnostic kits requires best practices for sample preparation and quantitative insight on the availability of biomarkers and the dynamics of immunoassay in saliva. This pilot study measured the levels of the PfHRP2 in patient saliva to inform the development of salivary diagnostic tests for malaria. METHODS: Matched samples of blood and saliva were collected between January and May, 2011 from eight patients at Palawan Baptist Hospital in Roxas, Palawan, Philippines. Parasite density was determined from thick-film blood smears. Concentrations of PfHRP2 in saliva of malaria-positive patients were measured using a custom chemiluminescent ELISA in microtitre plates. Sixteen negative-control patients were enrolled at UCLA. A substantive difference between this protocol and previous related studies was that saliva samples were stabilized with protease inhibitors. RESULTS: Of the eight patients with microscopically confirmed P. falciparum malaria, seven tested positive for PfHRP2 in the blood using rapid diagnostic test kits, and all tested positive for PfHRP2 in saliva. All negative-control samples tested negative for salivary PfHRP2. On a binary-decision basis, the ELISA agreed with microscopy with 100 % sensitivity and 100 % specificity. Salivary levels of PfHRP2 ranged from 17 to 1,167 pg/mL in the malaria-positive group. CONCLUSION: Saliva is a promising diagnostic fluid for malaria when protein degradation and matrix effects are mitigated. Systematic quantitation of other malaria biomarkers in saliva would identify those with the best clinical relevance and suitability for off-the-shelf diagnostic kits.

Multielectrode microprobes for deep-brain stimulation fabricated with a customizable 3-D electroplating process.

Although deep-brain stimulation (DBS) can be used to improve some of the severe symptoms of Parkinson's disease (e.g., Bradykinesia, rigidity, and tremors), the mechanisms by which the symptoms are eliminated are not well understood. Moreover, DBS does not prevent neurodegeneration that leads to dementia or death. In order to fully investigate DBS and to optimize its use, a comprehensive long-term stimulation study in an animal model is needed. However, since the brain region that must be stimulated, known as the subthalamic nucleus (STN), is extremely small (500 microm x 500 microm x 1 mm) and deep within the rat brain (10 mm), the stimulating probe must have geometric and mechanical properties that allow accurate positioning in the brain, while minimizing tissue damage. We have designed, fabricated, and tested a novel micromachined probe that is able to accurately stimulate the STN. The probe is designed to minimize damage to the surrounding tissue. The probe shank is coated with gold and the electrode interconnects are insulated with silicon nitride for biocompatibility. The probe has four platinum electrodes to provide a variety of spatially distributed stimuli, and is formed in a novel 3-D plating process that results in a microwire like geometry (i.e., smoothly tapering diameter) with a corresponding mechanically stable shank.

Towards a microcoil for intracranial and intraductal MR microscopy.

Implantable RF-coils have enabled sub-mm resolution magnetic resonance images (MRI) of deep structures. Scaling down the size of RF coils has similarly provided a gain in signal-to-noise ratio in nuclear-magnetic-resonance spectroscopy. By combining both approaches we designed, fabricated, and imaged with an implantable microcoil catheter. While typical implantable catheters use a transverse magnetization, the axial magnetization of the microcoil provides improved sensitivity and allows visualization of the tissue beyond the distal end of the catheter. The microcoil catheter was designed with a diameter of 1 mm for future integration with intracranial devices, and for intraductal use in breast oncology. We modified the NMR-microcoil design to allow implantation of the RF coil, by winding the microcoil on medical-grade silicone tubing and incorporating leads on the catheter to connect circuit components. In order to achieve proper turn spacing, we coated copper wire with 25 microm of biocompatible polymer (Parylene C). Tuning and matching circuitry insured that the impedance of the RF coil was approximately 50 ohm at the operating frequency for 3-T proton MR applications. A duplexer was used to enable use of the microcoil catheter as a transceiver. Experimental verification of the coil design was achieved through ex vivo imaging of neural tissue. As expected, the microcoil catheter provided microscale images with 20-microm in-plane-resolution and 170-microm-thick slices. While 3-T MRI typically provides 1 to 30 voxels per-cubic-millimeter, in this paper we report that the MRI microcoil can provide hundreds, and even thousands of voxels in the same volume.