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The possibility of artificial induction of a torpid state in animals that do not naturally do so, as well as in humans, offers a great potential in biomedicine and in human spaceflight. However, the mechanisms of action that provide a coordinated and concomitant downregulation with a safe recovery from this state are poorly understood. In our previous study, we demonstrated that the metabolic rate of mice can be reduced by nearly 94% and can remain stable under hypothermic conditions for a prolonged period of up to 11 h. The present study was carried out in order to test the limitations and identify potential factors that can enable the safe and reversible arousal of non-hibernating mice from deep artificially-induced torpor to an active state. Results demonstrate that the energy budget may be a limiting factor for the prolongation and safe recovery from the hypometabolic state. While the continuation of torpor may be possible for additional hours, we found that a reduction of 40% or more in the plasma glucose level increases the risk of heart fibrillation, which results in death during arousal. Therefore, the plasma glucose level could be a component of the criteria indicating the reversibility of torpor. Another important factor complementing the energetic necessity that may limit the duration of torpor in mice is a gradual reduction in body mass during torpor. Under the conditions of our experiment, body mass declines by nearly 15% after 16 h from the initiation of torpor and may continue to decline if the mice are allowed to remain in torpor longer. Extrapolation of this data suggests that there may be a critical mass relating to animal mortality and thus limiting the duration of torpor. Control and maintenance of the body mass and glucose level in a torpid animal may extend the longevity of torpor and mitigate the risk of cardiac failure during rewarming to the metabolically active state. The cardiac complications that occur during arousal from torpor in many cases could be mitigated and even avoided by applying appropriate temperature-arising kinetics and providing a sufficient dynamic range to maintain cardiac output.

The effect of UV- and 137Cs gamma radiation on the structural and chemical integrity of human hair was studied to determine the feasibility of using human hair as a non-invasive biomarker of radiation exposure to ionized gamma- and non-ionized UV-radiation. Steady state tryptophan (Trp) fluorescence and chemical analytical methods were used to evaluate the molecular integrity of Trp fluorophores and SH-groups in hair proteins and to assess the radiation induced damage quantitatively. It was found that human hair fibers were progressively damaged by exposure to both UV- and ionized gamma radiation. Damage to the hair was evidenced by a decrease in the fluorescence intensity as a result of observed depletion of the amino acid tryptophan as well as significant reduction in a number of free SH-groups in hair proteins. Hair damage was dose-dependent for exposures between 0 and 10.0 Gy and 0 - 20 J/cm2 of UV-radiation. Additional results demonstrate that hair-fibers exposed to gamma rays, with much higher quantum energy than UV, undergo a smaller extent of changes in Trp fluorescence than when exposed to lower or equal energy of UV-irradiation. The stable Trp fluorophore appears to be extremely sensitive to UV-radiation in contrast to the ionized gamma radiation whose damage is originated from the reaction of free radicals and direct deposition of energy. We conclude that fluorescence spectroscopy represents a useful tool in the quantitative evaluation of the radiation exposure and could also be used for the rapid and non-invasive assessment of radiation dose i.e. biodosimeter. The approach is simple, non-invasive and appears to have considerable potential that enables quantitative evaluation of radiation dose exposure in a single hair fiber.

One of the major problems associated with the therapeutic application of β-lactam antibiotics is their significant instability [1]-[3]. Analogs from that group are easily degraded in aqueous solutions and in solid state, which happens predominantly as a result of β-lactam ring chemical degradation. Because of this, the chemical integrity of β-lactam antibiotics must be closely monitored to meet FDA requirements. The original iodometric method based on visual detection of the end products of beta-lactamase hydrolysis that has been used to test beta-lactamase activity [4] was modified to quantitatively assess the amount of degraded product in amoxicillin. A UV-visible spectrophotometric method has been used, focusing on the ability of the products from the degradation of β-lactam analogs to absorb iodine from bulk solution, resulting in a significant reduction in the absorption spectra. Amoxicillin trihydrate tablets with different expiration dates and amoxicillin samples pre-loaded with known amounts of degraded amoxicillin have been used to generate a calibration curve and quantify the presence of degraded materials. Results show that the changes in the absorption spectra of iodine reagent directly correlate with the fraction of degraded β-lactam antibiotics, such as amoxicillin, in commercial products. The proposed assay might be as close as one step from simpler and cheaper methods of quantitative analysis of degraded β-lactam analogs, with estimated sensitivity in the range of 1% - 2%, which is better than the current FDA requirements.

The effect of UV- and 60Co gamma radiations on the structural and chemical integrity of human hair has been studied to determine the feasibility of using human hair as a non-invasive biomarker of radiation exposure to ionized and non-ionized radiation. The goal was to identify the most sensitive molecular parameter associated with radiation-induced damage and to evaluate effects of dose range from 0.5Gy to 10Gy. Steady state tryptophane fluorescence and chemical analytical methods were utilized to quantitatively evaluate molecular integrity of Trp fluorophor and SH-groups in hair proteins to asses the radiation induced damage.The study found that human hair fibers were progressively damaged by exposure to both UV- and ionized gamma radiation up to 10Gy. Damage to the hair was evidenced by a decrease in the fluorescence intensity due to depletion of the amino acid tryptophan as well as significant reduction in a number of free SH- groups.The results show that hair-fibrils exposed to gamma rays of higher quantum energy than UV, undergo much smaller extent of changes in Trp fluorescence and SH-group modification, then when exposed to equal energy of UV-irradiation. The changes appear faster under the effect of UV than by gamma-irradiation and their intensity and character depend on the dose. The stable Trp fluorophore is extremely sensitive to UV-B and UV-C radiation in contrast to the ionized gamma radiation, which causes damage from the reaction of free radicals and direct deposition of energy.We conclude that fluorescence intensity of the hair is a sensitive parameter for the quantitative evaluation of the radiation exposure.