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J Orenstein

Publications and source records attributed to J Orenstein.

41 records · Page 3Linked to original sources

Passive stiffness of isolated cardiac and skeletal myocytes in the hamster.

Single cardiac myocytes and skeletal myocyte fragments, devoid of interstitial collagen but with intact glycocalyx, were prepared by mechanical disaggregation of hamster ventricular myocardium and caudal gracilis muscle, respectively. Passive stiffness was studied by examining the sarcomere length-tension relationship over the approximate Eulerian stress range of 0-20 mN/mm2 for cardiac myocytes and 0-120 mN/mm2 for skeletal myocytes. Creep and stress-relaxation became apparent only when cells were stretched to sarcomere lengths close to, or exceeding, 2.2 micron for the cardiac myocytes, and 2.7 micron for the skeletal myocytes. Stress-relaxation and creep occurred simultaneously, suggesting that the sarcomere is at least one of the structural components responsible for viscoelasticity. The differential strain stiffness constant was calculated from the regression of natural stress [Ln(mN/mm2)] against differential strain [(L-Lo)/Lo] and found to be 7.48 +/- 1.73 for the ventricular myocytes and 5.77 +/- 0.87 for the skeletal myocyte fragments. The natural strain stiffness constant was obtained from the regression of natural stress against natural strain [Ln(L/Lo)]. The natural strain stiffness constant was 30-50% higher than the differential strain constant. The high correlation coefficients obtained for both regressions indicate that the length-tension relationships for these isolated cardiac and skeletal myocytes can be very closely fitted to the single exponential function, sigma = C X exp[K(epsilon)]. The length-tension curves obtained for the skeletal myocyte fragments are qualitatively and quantitatively similar to those obtained by others with intact skeletal muscle. The cardiac myocyte length-tension curves are qualitatively, but not quantitatively, similar to those obtained with cardiac muscle. Isolated ventricular myocytes are stiffer than similarly isolated skeletal myocytes. These findings suggest that cellular structures contribute to myocardial stiffness in the hamster.

Animals↗

Morphologic changes in human immunodeficiency virus type 1 virions secondary to intravirion reverse transcription: evidence indicating that reverse transcription may not take place within the intact viral core.

INTRODUCTION: In the past, retroviral endogenous reverse transcription (ERT) was considered an artificial process, secondary to permeabilization of the viral envelope by detergents or amphipathic peptides. However, recently we have demonstrated that ERT may occur in a variety of lentiviruses without detergent treatment and may lead to increased infectivity of lentivirions in initially quiescent T lymphocytes and nonproliferating cells, such as macrophages. As full-length reverse transcripts could be synthesized within lentiviral particles, it is worth evaluating the potential alterations in lentiviral morphology due to the stimulation of intravirion reverse transcription. METHODS: Using quantitative DNA-polymerase chain reaction (PCR) and transmission electron microscopy (TEM), we characterized critical alterations in human immunodeficiency virus type 1 (HIV-1) virions after stimulation of intravirion reverse transcription. RESULTS: Intravirion reverse transcription in HIV-1 virions was stimulated using deoxyribonucleoside triphosphates (dNTPs) and physiologic polyamines. Our studies indicated that HIV-1 virions, in which intravirion reverse transcription was stimulated, showed dissolution of the p24-shelled viral core and absence of the core-envelope linkage (CEL) region by TEM. These changes in the structure of the core correlate with the in vitro alterations in virion infectivity on primary cells. CONCLUSIONS: Stimulation of intravirion HIV-1 reverse transcription leads to morphologic changes in the viral particles that suggest changes in the compact viral core, which is consistent with active reverse transcription before infection of target cells. Further, via this unique approach, we suggest that intravirion or intracellular reverse transcription of HIV-1 is unlikely to take place within intact viral cores made up of p24-containing outer shells. As such, these results suggest a new approach to further dissect the intravirion or intracellular reverse transcription machinery of lentiviruses.

DNA, Viral↗