Dataset Information

Massively Parallelized Molecular Force Manipulation with On-Demand Thermal and Optical Control.

ABSTRACT: In single-molecule force spectroscopy (SMFS), a tethered molecule is stretched using a specialized instrument to study how macromolecules extend under force. One problem in SMFS is the serial and slow nature of the measurements, performed one molecule at a time. To address this long-standing challenge, we report on the origami polymer force clamp (OPFC) which enables parallelized manipulation of the mechanical forces experienced by molecules without the need for dedicated SMFS instruments or surface tethering. The OPFC positions target molecules between a rigid nanoscale DNA origami beam and a responsive polymer particle that shrinks on demand. As a proof-of-concept, we record the steady state and time-resolved mechanical unfolding dynamics of DNA hairpins using the fluorescence signal from ensembles of molecules and confirm our conclusion using modeling.

SUBMITTER: Su H

PROVIDER: S-EPMC9066152 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature

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Massively Parallelized Molecular Force Manipulation with On-Demand Thermal and Optical Control.

Su Hanquan H Brockman Joshua M JM Duan Yuxin Y Sen Navoneel N Chhabra Hemani H Bazrafshan Alisina A Blanchard Aaron T AT Meyer Travis T Andrews Brooke B Doye Jonathan P K JPK Ke Yonggang Y Dyer R Brian RB Salaita Khalid K

Journal of the American Chemical Society 20211111 46

In single-molecule force spectroscopy (SMFS), a tethered molecule is stretched using a specialized instrument to study how macromolecules extend under force. One problem in SMFS is the serial and slow nature of the measurements, performed one molecule at a time. To address this long-standing challenge, we report on the origami polymer force clamp (OPFC) which enables parallelized manipulation of the mechanical forces experienced by molecules without the need for dedicated SMFS instruments or sur ...[more]

PMID: 34762807

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Project description:BackgroundThe aim of the current study was to evaluate the effect of different thermal manipulation (TM) protocols during embryogenesis on thermotolerance acquisition parameters during subsequent thermal challenge (TC) at posthatch day 28. A total of 1500 fertile chicken eggs were divided randomly into five treatments (300 eggs each): control was maintained at 37.8 °C and 56 % relative humidity (RH) whereas, TM1, TM2, TM3 and TM4 were subjected to 38.5, 39, 39.5 and 40 °C for 18 h and 65 % RH daily during embryonic days ED 12-18. Hatched chicks from each treatment group allocated randomly into two sub-treatment groups (thermo-neutral, naïve (TN) and thermal challenge (TC). At day 28 of age, chicks subjected to TC by adjusting room temperature to 42 °C for 6 h while naïve chicks kept under regular conditions (22 ± 1 °C and 50-60 % RH). Chick's response to TC evaluated by determination of plasma T3, T4, corticosterone, total proteins, albumin, selected enzymes and some electrolytes at the beginning (0 h) and after 1, 3 and 5 h of TC in TM and TN chicks. Furthermore, pectoral and thigh muscles mRNA expression of Atrogin-1, CK, avUCP, DIO3, DIO2 were evaluated in TC and TN sub-treatment groups.ResultsTM induced a significant reduction in free T3 and elevation in total proteins and albumin in plasma with significant down-regulation of Atrogin-1 and DIO2 and significant up-regulation of DIO3 mRNA expression in muscle of TM chicks compare to control. During TC at day 28, decrease in the concentrations of plasma free T3, total proteins and albumin with increase in T4 have been detected in control and TM chicks. TC induced up-regulation of Atrogin-1 and DIO3 with down-regulation of DIO2 gene expression in muscles of all TC chicks.ConclusionThe present study indicated that, TM improved thermotolerance acquisition by decreasing basal metabolic rate and muscle injury during thermal stress. Basal metabolic rate decreased via reduction of plasma T3 concentration with up and down regulation of expression of DIO3 and DIO2, respectively in muscles. Muscle injury protected by stimulation of protein biosynthesis and down-regulation of Atrogin-1 expression.

Project description:The first third of incubation is critical for embryonic development, and environmental changes during this phase can affect the physiology and survival of the embryos. We evaluated the effects of low (LT), control (CT), and high (HT) temperatures during the first 5 days of incubation on ventilation ( V. E ), body temperature (Tb), oxygen consumption ( V. O2), respiratory equivalent ( V. E / V. O2), and brain monoamines on 3-days-old (3d) and 14-days-old (14d) male and female chickens. The body mass of LT animals of both ages and sexes was higher compared to HT and CT animals (except for 3d males). The heart mass of 14d HT animals was higher than that of CT animals. Thermal manipulation did not affect V. E , V. O2 or V. E / V. O2 of 3d animals in normoxia, except for 3d LT males V. E , which was lower than CT. Regarding 14d animals, the HT females showed a decrease in V. E and V. O2 compared to CT and LT groups, while the HT males displayed a lower V. O2 compared to CT males, but no changes in V. E / V. O2. Both sexes of 14d HT chickens presented a greater Tb compared to CT animals. Thermal manipulations increased the dopamine turnover in the brainstem of 3d females. No differences were observed in ventilatory and metabolic parameters in the 3d animals of either sexes, and 14d males under 7% CO2. The hypercapnic hyperventilation was attenuated in the 14d HT females due to changes in V. O2, without alterations in V. E . The 14d LT males showed a lower V. E , during hypercapnia, compared to CT, without changes in V. O2, resulting in an attenuation in V. E / V. O2. During hypoxia, 3d LT females showed an attenuated hyperventilation, modulated by a higher V. O2. In 14d LT and HT females, the increase in V. E was greater and the hypometabolic response was attenuated, compared to CT females, which resulted in no change in the V. E / V. O2. In conclusion, thermal manipulations affect hypercapnia-induced hyperventilation more so than hypoxic challenge, and at both ages, females are more affected by thermal manipulation than males.

Dataset Information

Massively Parallelized Molecular Force Manipulation with On-Demand Thermal and Optical Control.

Publications

Massively Parallelized Molecular Force Manipulation with On-Demand Thermal and Optical Control.

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