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Biomedical subjects

J W Berger

Publications and source records attributed to J W Berger.

28 records · Page 2Linked to original sources

Temperature measurements during phacoemulsification and erbium:YAG laser phacoablation in model systems.

PURPOSE: To compare the potential for thermal injury to ocular structure resulting from phacoemulsification ultrasound energy and erbium:YAG (Er:YAG) laser output. SETTING: Morse Laser Laboratory, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston. METHODS: Ultrasonic phacoemulsification energy and ER:YAG laser output (10 J/cm2, 10 Hz, 0.3 watts) sufficient for lens removal were applied to model systems and human cadaver eyes. Temperatures were measured with ultrafine thermocouples interfaced to a microcomputer data acquisition system. RESULTS: Although greater than 95% of energy from laser output is converted to thermal energy, temperature rise in model systems and cadaver eyes was 10 to 15 times greater after pulsed application of ultrasound energy than after Er:YAG laser application. At 100% power, approximately 4 watts of ultrasound power is converted to heat. Temperature rise following both laser and ultrasound applications decreased with irrigation in cadaver eyes and increasing volume in a model system. With continuous irrigation (20 cc/min), the temperature rise at 2 minutes measured at the corneal endothelial surface, within the corneal stroma, and in the anterior chamber angle of cadaver eyes was approximately 0.5 degrees Celsius (degrees C) after laser application and 7.0 degrees C after ultrasound application. Without irrigation, temperatures rose 2.5 degrees C after laser application and 35.0 degrees C after ultrasound application. CONCLUSION: At operating parameters sufficient for lens removal, the Er:YAG laser imparted less thermal energy to whole eyes and model systems than ultrasonic phacoemulsification.

Anterior Chamber↗

The anaerobic photolysis of lens alpha-crystallin: evidence for triplet state mediated photodamage.

Anaerobic solutions of lens alpha-crystallin were subjected to near-UV (greater than 295 nm) irradiation, and the photoproducts were analyzed by fluorescence and room-temperature phosphorescence spectroscopy. The principal photoproduct was excited maximally at 340 nm, fluoresced maximally at 430 nm, and phosphoresced with an emission maximum at 510 nm. The phosphorescence intensity decay of this species was well fit by a sum of two exponentials with lifetimes of 9.2 ms (78%) and 61 ms (22%); this report is the first demonstration of a long-lived triplet state associated with a protein photolysis product. As reported previously, 3trp* is also long-lived in deoxygenated alpha-crystallin solution at room-temperature (Berger and Vanderkooi, 1989, Biochemistry 28, 5501-5508), hence both tryptophan and photoproduct triplet states are good candidates to mediate photodamage. Photolysis experiments in the presence of agents known to alter the tryptophan triplet yield provide evidence for the importance of triplet-state-mediated photodamage of lens crystallins in anaerobic solution. In 30 mM acrylamide where 3trp*, but not 1trp*, is efficiently quenched, anaerobic solutions exhibited marked resistance to protein photodamage, whereas the photoprotection in aerobic solution was minimal. In D2O, where photoionization is suppressed but triplet states are longer-lived, photodamage was accelerated in anaerobic solution but reduced in aerobic solutions. Finally, the anaerobic photodestruction rate was increased in 500 mM Cs+ solution where the triplet yield is increased by a heavy atom effect.

Anaerobiosis↗

Characterization of lens alpha-crystallin tryptophan microenvironments by room temperature phosphorescence spectroscopy.

Room temperature phosphorescence techniques were used to study the structural and dynamic features of the tryptophan residues in bovine alpha-crystallin. Upon excitation at 290 nm, the characteristic signature of tryptophan phosphorescence was observed with an emission maximum at 442 +/- 2 nm. The phosphorescence intensity decay was biphasic with lifetimes of 5.4 ms (71%) and 42 ms (29%). Phosphorescence quenching measurements strongly suggest that each component corresponds to one class of tryptophans with the more buried residues having the longer emission lifetime. Three small-molecule quenchers were surveyed, and in order of increasing quenching efficiency: iodide less than nitrite less than acrylamide. A heavy-atom effect was observed in iodide solutions, and an upper limit of 5% was placed on the quantum yield of triplet formation in iodide-free solutions, while the phosphorescence quantum yield was estimated to be approximately 3.2 x 10(-4). The temperature dependence of the phosphorescence lifetime was measured between 5 and 40 degrees C. Arrhenius plots exhibited discontinuities at 26 and 29 degrees C for the short- and long-lived components, respectively, corresponding to abrupt transitions in segmental flexibility. Denaturation studies revealed conformational transitions between 1 and 2 M guanidine hydrochloride, and 4 and 6 M urea. Long-lived phosphorescence lifetimes of 3 and 7 ms were measured in 6 M guanidine hydrochloride and 8 M urea, respectively, suggesting that some structural features are preserved even at very high concentrations of denaturant. Our studies demonstrate the sensitivity of room temperature phosphorescence spectroscopy to the structure of alpha-crystallin, and the applicability of this technique for monitoring conformational changes in lens crystallin proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrylamide↗

Phosphorescence measurements of calf gamma-II, III, and IV crystallins at 77 and 293 K.

Structural and dynamical features of bovine gamma-crystallin tryptophan residues were investigated by phosphorescence measurements at 77 and 293 K. The low temperature phosphorescence spectra and lifetimes of calf gamma-II, III, and IV crystallins did not reflect heterogeneity among the gamma-crystallins. The 0-0 bands were all at 414 +/- 1 nm and the emission lifetimes were all single-exponential with lifetimes of 5.1, 5.3 and 5.3 +/- 0.3 sec, respectively. In contrast, phosphorescence measurements at room temperature were sensitive to subtle differences in exposure, accessibility, and flexibility of gamma-crystallin tryptophan residues. Thorough deoxygenation allowed for measurement of the normally-quenched room-temperature phosphorescence, and we report the first native phosphorescence measurements of lens crystallins at ambient temperature. The emission maxima for gamma-II, III and IV were 446, 442, and 440 +/- 2 nm, respectively. The intensity decay curves were all non-single exponential, and the decays were fit to a sum of two exponentials with lifetimes of 9.1 and 93 msec (gamma-II), 11 and 75 msec (gamma-III), and 4.2 and 68 msec (gamma-IV), +/- 10%. The components of the gamma-II emission were assigned to the four tryptophans based on X-ray structural information. Quantum yields of the phosphorescence emission were in the ratio of 20:7:1 for gamma-II, III and IV, and comparison of lifetimes and quantum yields suggests that tryptophan rigidity increases in the order gamma-IV less than III less than II. Acrylamide quenching constants for the long-lived components of gamma-II and III were roughly equal, while the short-lived tryptophans of gamma-III were an order of magnitude more accessible than those of gamma-II. The wide range of phosphorescence lifetimes and quenching constants allowed for discrimination of distinct contributions to the phosphorescence emission, and we suggest that room-temperature phosphorescence measurements will be an effective tool for studying conformational changes of lens crystallins.

Animals↗

Brownian dynamics simulations of intramolecular energy transfer.

A novel technique for modelling intramolecular energy transfer is presented. Brownian dynamics calculations are used to compute the trajectories of donor and acceptor species, and the instantaneous orientation factor is calculated during each temporal iteration. In this work, several model systems are considered. Trajectories were computed for energy transfer between a flexible donor and a rigidly fixed acceptor. We have considered configurations where the donor is, (1) tethered to a fixed point in space, but free to diffuse rotationally, and (2) constrained to wobble in a cone. The luminescence decay of the donor is 'measured', and a non-single-exponential decay is observed for configurations of efficient energy transfer. Luminescence anisotropy measurements of constrained and unconstrained donors reflect the contribution of both energy transfer and rotational diffusion to the shape of the anisotropy decay curve.

Chemical Phenomena↗

Regulatory issues in pain management.

Undertreatment of pain continues to be a serious public health problem. The use of opioid analgesics is considered essential for chronic pain management and palliative care. Pharmacists are encouraged to become involved in initiatives to improve pain management.

Analgesics, Opioid↗