Microfluidics as an Enabling Technology for Personalized Cancer Therapy

Tailoring patient‐specific treatments for cancer is necessary in order to achieve optimal results but requires new diagnostic approaches at affordable prices. Microfluidics has immense potential to provide solutions for this, as it enables the processing of samples that are not available in large quantities (e.g., cells from patient biopsies), is cost efficient, provides a high level of automation, and allows the set‐up of complex models for cancer studies. In this review, individual solutions in the fields of genetics, circulating tumor cell monitoring, biomarker analysis, phenotypic drug sensitivity tests, and systems providing controlled environments for disease modeling are discussed. An overview on how these early stage achievements can be combined or developed further is showcased, and the required translational steps before microfluidics becomes a routine tool for clinical applications are critically discussed.     

Design and analysis of a high bandwidth disk drive servo systemusing an instrumented suspension

Resonance modes in the suspension of hard disk drives limit the closed-loop bandwidth. The bandwidth of the servo can be increased by active vibration control of the resonance modes. This paper considers the optimal placement of strain gauge sensors on a suspension to observe the vibration states of the suspension. Using a finite-element simulation of an actual suspension, a state-space model is identified for the two normal strains and the shear strain at each finite element. The state-space model includes the dynamics of the three primary resonance modes. A numerical search algorithm is used to determine the sensor location and orientation which maximizes the minimum singular value of the observability grammian. With the strain gauge output signal, a multirate inner loop controller is designed to be used with the existing head-positioning system. Simulations and analysis results suggest that use of an instrumented suspension is a viable candidate method for improved disk drive servo performance     

12 nm tunable WDM source using an integrated laser array

A 12 nm tunable source with up to 15 mW fibre coupled power has been fabricated by integrating four DFB lasers and a booster amplifier to provide a single output. High-yield and low-cost techniques are used, such as quarter-wave-shifted phase-masks for wavelength definition and an integration technique with only two regrowths     

Corneal scarring in the Collaborative Longitudinal Evaluation of Keratoconus (CLEK) Study: baseline prevalence and repeatability of detection

Fibroblast growth factor-16 (FGF-16) is the most recent member of the FGF family to be cloned. Since the biologic activity of rat FGF-16 (rFGF-16) was unknown, and this protein has no apparent signal sequence, we transformed its entire cDNA into Escherichia coli for high-level expression and further characterization of this novel protein. An attempt was made to purify the expressed protein from the supernatant of mechanically lysed cells using sequential cation-exchange chromatography. This resulted in a gradual loss of the protein as precipitate throughout the purification process. In addition to precipitation during purification, sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the partially purified materials showed a cluster of protein bands around 20k to 29k. Sequence analysis of the major bands indicated that two N-terminal truncations had occurred, during E. coli fermentation, purification, or both. The largest truncation resulted in the removal of the 34 N-terminal amino acids, including the initiation codon methionine. We cloned d34 rFGF-16, expressed the gene in E. coli, and developed a purification process for this form. Even with this truncated form, precipitation was a problem. We were largely able to overcome this problem, however, by including EDTA throughout the purification process. We have characterized the structure of purified d34 rFGF-16 extensively using circular dichroism, Fourier transform infrared spectroscopy, and sedimentation velocity analysis. These studies revealed that the protein has a distinct tertiary structure, consists primarily of beta-strands, has a weak tendency to self-associate, and is fairly extended. We then performed biologic assays which showed that d34 rFGF-16 induces oligodendrocyte proliferation in vitro, and induces hepatocellular proliferation and increased liver weight in vivo. In summary, FGF-16, a novel FGF family member, has both unique structural and biological properties.     

Towards automation of color/weave selection in Jacquard design: Model verification

Jacquard woven fabrics are made from colored yarns and different weaves for designing complex pictorial and other patterning effects. The final visualized color effect is the result of assigning weave designs to different areas of the pattern to be created. The current practice in creating Jacquard woven fabric designs is to produce many samples in a trial‐and‐error attempt to match artwork colors. An ability to simulate accurately the appearance of a design prior to manufacture is highly desirable to reduce trial‐and‐error sample production. No automated accurate digital color methodology is yet available to assist designers in matching the patterned woven fabric to the desired artwork. To achieve this, we developed a geometrical model to predict the color contribution of each yarn on the face of the fabric. The geometrical model combined with a Kubelka‐Munk based color mixing model allowed the prediction of the reflectance properties of the final color for a given design. We compared the predicted and experimental values of the reflectance properties for a range of fabrics using the same geometric model with three separate color mixing models. The geometrical model combined with a log‐based color mixing model produced reasonable agreement between predicted and measured ΔE_ab, with an average ΔE_ab of approximately five. © 2009 Wiley Periodicals, Inc. Col Res Appl, 34, 225–232, 2009     

Interface Mutation: an approach for integration testing

The need for test adequacy criteria is widely recognized. Several criteria have been proposed for the assessment of adequacy of tests at the unit level. However, there remains a lack of criteria for the assessment of the adequacy of tests generated during integration testing. We present a mutation based interprocedural criterion, named Interface Mutation (IM), suitable for use during integration testing. A case study to evaluate the proposed criterion is reported. In the study, the UNIX sort utility was seeded with errors and Interface Mutation evaluated by measuring the cost of its application and its error revealing effectiveness. Alternative IM criteria using different sets of Interface Mutation operators were also evaluated. While comparing the error revealing effectiveness of these Interface Mutation-based test sets with same size randomly generated test sets, we observed that in most cases Interface Mutation based test sets are superior. The results suggest that Interface Mutation offers a viable test adequacy criteria for use at the integration level     

Prodrugs for targeting hypoxic tissues: regiospecific elimination of aspirin from reduced indolequinones

We report the cases of three patients with anorexia nervosa (AN) who each recovered rapidly after experiencing a life-threatening episode with severe thrombocytopenia. All three cases were the typical restricting-type of AN, occurring in adolescence. They refused to be admitted to a hospital until their general condition had been severely deteriorated. Their lowest platelet counts were 2.9, 4.6, and 2.3 x 10(4)/mm3, respectively. Apparent hemorrhagic tendencies, such as purpura, gingival and nasal bleeding, and gastrointestinal bleeding were observed. The bone marrow examination showed apparent hypoplasia in two patients. No evidence of disseminated intravascular coagulation or autoantibody to platelets was detected. The platelet counts recovered rapidly by water and nutritional supplementation. The recovery from the AN itself was excellent in all three patients without specific psychotherapy.     

Multiplexed DNA sequencing and diagnostics by hybridization with enriched stable isotope labels

A new DNA diagnostic and sequencing system has been developed that uses time-of-flight resonance ionization mass spectrometry (TOF-RIMS) to provide a rapid method of analyzing stable isotope-labeled oligonucleotides in form 1 sequencing by hybridization (SBH). With form 1, the DNA is immobilized on a nylon membrane and enriched isotope-labeled individual oligonucleotide probes are free to seek out complementary DNAs during hybridization. The major advantage of this new approach is that multiple oligonucleotides can be labeled with different enriched isotopes and can all be simultaneously hybridized to the genosensor matrix. The probes can then be simultaneously detected with TOF-RIMS with high selectivity, sensitivity, and efficiency. By using isotopically enriched tin labels, up to 10 labeled oligonucleotides could be examined in a single hybridization to the DNA matrix. Greater numbers of labels are available if rare earth isotopes are employed. In the present study, matrices containing three different DNAs were prepared and simultaneously hybridized with two different probes under a variety of conditions. The results show that DNAs, immobilized on nylon surfaces, can be specifically hybridized to probes labeled with different enriched in isotopes. Discrimination between complementary and noncomplementary sites of better than 100 was obtained in multiplexed samples. This new SBH method, which employs stable isotopic labels to locate target DNAs and TOF-RIMS to detect the labels, will be a very versatile and extensive multiplexing method.     

Erythema migrans-like rash illness at a camp in North Carolina: a new tick-borne disease?

BACKGROUND: Borrelia burgdorferi, the causative agent of Lyme disease, has never been isolated from a patient thought to have acquired Lyme disease in any southeastern state. OBJECTIVE: To investigate 14 cases of an erythema migrans (EM)-like rash illness that occurred during 2 summers at an outdoor camp in central North Carolina in an effort to determine the etiologic, epidemiological, and clinical aspects of this illness. METHODS: Using active surveillance, we identified cases of clinically diagnosed EM in residents and staff of the camp. We collected clinical and demographic information; history of exposure to ticks; acute and convalescent serum antibodies to B. burgdorferi, Rickettsia rickettsii, and Ehrlichia chaffeensis; and cultures for spirochetes from biopsy specimens of skin lesions. Serum samples from a group of residents and staff who did not develop rashes were tested for the same antibodies. We speciated ticks removed from people and collected from vegetation. RESULTS: We identified 14 cases of EM-like rash illness during the 2 summers. Of the 14 case-patients, 10 had associated mild systemic symptoms and 1 had documented fever. All 14 case-patients had removed attached ticks, and 8 remembered having removed a tick from the site where the rash developed a median of 12 days earlier (range, 2-21 days). One tick removed from the site where a rash later developed was identified as Amblyomma americanum, the Lone Star tick; 97% of ticks collected from vegetation and 95% of ticks removed from people were A. americanum. No spirochetes were isolated from skin biopsy specimens. Paired serum samples from 13 case-patients did not show diagnostic antibody responses to B. burgdorferi or other tick-borne pathogens. CONCLUSIONS: This investigation suggests the existence of a new tick-associated rash illness. We suspect that the disease agent is carried by A. americanum ticks. In the southern United States, EM-like rash illness should no longer be considered definitive evidence of early Lyme disease.