50 research outputs found
Use of streptavidin magnetic beads in single strand conformation polymorphism profiles to detect mutations in rpoB gene of M.tuberculosis
Single strand conformation polymorphism (SSCP) is one of the promising
techniques to identify mutations in short pieces of DNA (Orita et al. 1989). In this
technique, DNA of interest is often amplified by the polymerase chain reaction (PCR)
and then denatured by heat or alkali treatment before electrophoresis on a non
denaturing polyacrylamide gel. Differences in mobility of either of the single strands
compared to the control DNA indicate mutations which affect the secondary structure
and alter the mobility of the DNA. We applied PCR-SSCP for the detection of
mutations in the rifampicin resistance determining region (RRDR) of the rpoB gene of
M. tuberculosis (Telenti et al. 1993a; 1993b). A nested PCR was used to amplify the
RRDR. In the first PCR, 293-bp product was amplified and in the second PCR a 103-
bp of the first PCR product was amplified. However, in our experience using
denaturation by alkali or heating, the denatured PCR product most often reannealed to
form a large proportion of double stranded DNA during the electrophoresis
(Selvakumar et al. 1997a). After visualisation by staining with ethidium bromide or
silver staining, most of the DNA was in the double stranded form, with very little or
no single stranded DNA. The single strands that could be observed often ran close
together, making analysis of any difference in mobility difficult. Therefore an attempt
was made to generate biotinylated PCR product using a biotinylated forward primer
and later the biotinylated strand was separated using sterptavidin magnetic beads. The
separated strands eliminated the problem of strand reannealing during SSCP and were
silver stained to detect the shift in the mobility. Since the nested PCR requires more
time and is more expensive. a biotinylated PCR product was generated in a single PCR
using a biotinylated forward primer and an unbiotinylated reverse primer. This
simplified protocol was applied to clinical isolates in an attempt to detect rifampicin
resistance
AAPT Diagnostic Criteria for Chronic Sickle Cell Disease Pain
Pain in sickle cell disease (SCD) is associated with increased morbidity, mortality, and high health care costs. Although episodic acute pain is the hallmark of this disorder, there is an increasing awareness that chronic pain is part of the pain experience of many older adolescents and adults. A common set of criteria for classifying chronic pain associated with SCD would enhance SCD pain research efforts in epidemiology, pain mechanisms, and clinical trials of pain management interventions, and ultimately improve clinical assessment and management. As part of the collaborative effort between the Analgesic, Anesthetic, and Addiction Clinical Trial Translations Innovations Opportunities and Networks public-private partnership with the U.S. Food and Drug Administration and the American Pain Society, the Analgesic, Anesthetic, and Addiction Clinical Trial Translations Innovations Opportunities and Networks-American Pain Society Pain Taxonomy initiative developed the outline of an optimal diagnostic system for chronic pain conditions. Subsequently, a working group of experts in SCD pain was convened to generate core diagnostic criteria for chronic pain associated with SCD. The working group synthesized available literature to provide evidence for the dimensions of this disease-specific pain taxonomy. A single pain condition labeled chronic SCD pain was derived with 3 modifiers reflecting different clinical features. Future systematic research is needed to evaluate the feasibility, validity, and reliability of these criteria. Perspective: An evidence-based classification system for chronic SCD pain was constructed for the Analgesic, Anesthetic, and Addiction Clinical Trial Translations Innovations Opportunities and Networks-American Pain Society Pain Taxonomy initiative. Applying this taxonomy may improve assessment and management of SCD pain and accelerate research on epidemiology, mechanisms, and treatments for chronic SCD pain
Recommended from our members
Inventory of Freight Transportation in the Southwest. Part II: Motor Carrier Service in the Dallas / Fort Worth Area
See this work in the Center for Transportation Research Library catalog: https://library.ctr.utexas.edu/Presto/catalogid=5734This report provides an inventory of existing motor common carrier freight transportation facilities, services, and practices in the twenty-four Texas and two Oklahoma counties surrounding and including Dallas and Fort Worth. As a manufacturing and distribution hub, Dallas-Fort Worth relies heavily on motor common carrier service to develop its economic potential. For inventory purposes, common carriers have been divided into seven categories: general motor freight, agricultural and other ICC exempt commodities, refrigerated commodities, heavy haulers, bulk commodities, and vehicle haulers. Detailed information for each classification of common carrier includes the number of trucks, estimated number of firms represented, intercity loads, total loads, intercity haulage, and total haulage. The inventory also provides estimates for total volume of general freight, delivery time for truckload and LTL shipments, terminal capacities, load factors, seasonal fluctuations, and the pattern of motor common carrier freight movements.The Department of Transportation, Office of University ResearchCenter for Transportation Researc
Hindlimb kinematics before and after laser fibrotomy in horses with fibrotic myopathy
Reasons for performing study: Fibrotic myopathy can cause incapacitating gait abnormalities. Transection of the fibrotic mass followed by early post operative exercise is the best treatment for fibroticmyopathy. A lasermay be used to transect the fibrotic mass. Assessment of the effectiveness of therapies for fibrotic myopathy has been limited to subjective evaluation. Objectives: To objectively assess gait abnormalities associated with fibrotic myopathy before and after laser fibrotomy followed by early post operative exercise. Methods: Kinematic evaluation of horses with fibrotic myopathy walking and trotting on a treadmill was used to investigate hindfeet trajectories (n = 8) and lameness (n = 5) before and after laser fibrotomy. Hoof flight trajectory length (HFTL), relative protraction length (%PL), maximum hoof height during swing (MXHH), hoof height at end of protraction (HHpro) and retraction (HHret) were measured and differences between fibrotic myopathy affected and nonaffected limbs were calculated. Lameness was quantified by measuring maximum and minimum pelvic height differences between right and left halves of the stride. Results: Before surgery the foot of the fibrotic myopathy affected limb had abnormal trajectories characterised as increased HFTL, MXHH and HHpro and decreased %PL and HHret and the 5 horses objectively evaluated for lamenesswere lame in the fibroticmyopathy affected limb. Immediately after surgery the difference between affected and nonaffected limbs decreased for HFTL, %PL and HHpro. Six to 11weeks after surgery, the HFTL difference increased but was still smaller than before surgery, whichwas interpreted as partial recurrence of the gait abnormality; all horses objectively evaluated for lamenesswere either improved (n = 1) or not lame (n = 4) in the previously affected, operated limb. Conclusions: Fibrotic myopathy affects the foot flight and leads to asymmetric vertical excursion of the pelvis. Laser fibrotomy followed by early post operative exercise can minimise these abnormalities. Potential relevance: Laser fibrotomy combined with early post operative exercise is a viable therapy for fibrotic myopathy.J. Janicek, M. A. F. Lopes, D. A. Wilson, S. Reed and K. G. Keega
Post-mortem brain pathology is related to declining respiratory function in community-dwelling older adults
Damage to brain structures which constitute the distributed neural network that integrates respiratory muscle and pulmonary functions, can impair adequate ventilation and its volitional control. We tested the hypothesis that the level of brain pathology in older adults is associated with declining respiratory function measured during life.1,409 older adults had annual testing with spirometry and respiratory muscle strength based on maximal inspiratory and maximal expiratory pressures. Those who died underwent structured brain autopsy. On average, during 5 years of follow-up, spirometry and respiratory muscle strength showed progressive decline which was moderately correlated (ρ=0.57, p<0.001). Among decedents (N=447), indices of brain neuropathologies showed differential associations with declining spirometry and respiratory muscle strength. Nigral neuronal loss was associated with the person-specific decline in spirometry (Estimate, -0.016 unit/year, S.E. 0.006, p=0.009) and reduction of the slope variance was equal to 4%. By contrast, Alzheimer’s disease (AD) pathology (Estimate, -0.030 unit/year, S.E. 0.009, p<0.001) and macroscopic infarcts (-0.033 unit/year, S.E., 0.011, p=0.003) were associated with the person-specific decline in respiratory muscle strength and reduction of the slope variance was equal to 7%. These results suggest that brain pathology is associated with the rate of declining respiratory function in older adults
Nanoparticle Based Surface-Enhanced Raman Spectroscopy
Talley CE, Huser T, Hollars CW, Jusinski L, Laurence T, Lane SM. Nanoparticle Based Surface-Enhanced Raman Spectroscopy. In: Wison BC, Tuchin VV, Tanev S, eds. Advances in Biophotonics. NATO Science Series, Amsterdam: IOS Press. Vol 369. 2005: 182-195