Relationship between quantity of IFNT estimated by IFN-stimulated gene expression in peripheral blood mononuclear cells and bovine embryonic mortality after AI or ET

<p>Abstract</p> <p>Background</p> <p>Interferon tau (IFNT), which is secreted into the uterine cavity during the maternal recognition period (MRP), is a key factor for establishment of pregnancy. The present study aims to clarify the relationship between the ability of a bovine conceptus to produce IFNT during the MRP and the conceptus's ability to establish pregnancy.</p> <p>Methods</p> <p>In the first experiment, IFNT (0, 500, or 1000 micrograms) was administered into the uterine horn ipsilateral to the CL 16 or 17 d after standing estrus, and mRNA levels of IFN-stimulated gene 15-kDa protein (<it>ISG15</it>) and <it>Mx2 </it>in peripheral blood mononuclear cells (PBMCs) were determined. In the second experiment, we investigated <it>ISG15 </it>mRNA expression in PBMCs during the MRP in cattle after either artificial insemination (AI) or embryo transfer (ET).</p> <p>Results</p> <p>Intrauterine administration of IFNT stimulated <it>ISG15 </it>and <it>Mx2 </it>gene expressions in PBMCs in cattle, and there was a positive correlation between the expressions of peripheral markers and the quantity of IFNT administered. In pregnant and normal interestrous interval (< 25 d) cattle (nIEI cattle), expression levels of the <it>ISG15 </it>gene showed similar patterns after AI and ET, and <it>ISG15 </it>mRNA expression was increased in pregnant cattle but unchanged in nIEI cattle. In contrast, <it>ISG15 </it>gene expression in extended interestrous interval (greater than or equal to 25 d) cattle (eIEI cattle) differed after ET compared with AI. In eIEI cattle after ET, <it>ISG15 </it>gene expression increased, such that the value on day 18 was intermediate between those of pregnant and nIEI cattle. In eIEI cattle after AI, <it>ISG15 </it>gene expression did not increase throughout the observation period.</p> <p>Conclusions</p> <p>The results of the current study indicate that the quantity of conceptus-derived IFNT can be estimated by measuring <it>ISG15 </it>mRNA levels in PBMCs from cattle. Using this approach, we demonstrate that <it>ISG15 </it>gene expression during the MRP in eIEI cattle differed after ET compared with AI. In addition, the modest increase in <it>ISG15 </it>gene expression in eIEI cattle after ET suggests that late embryo losses were due to delayed or insufficient growth of the conceptus during the MRP in cattle.</p

Renaturation of the mature subtilisin BPN' immobilized on agarose beads

AbstractWe report here another example of renaturation of subtilisin BPN′(Sbtl) by using an immobilized preparation instead of applying a digestible mutant of Streptomyces subtilisin inhibitor (SSI), a proteinaceous inhibitor of Sbtl [M. Matsubara et al. (1994) FEBS Letters 342, 193–196]. The mature Sbtl was immobilized on agarose beads employing the amino group of the protein. After thorough washing, the immobilized Sbtl was subjected to denaturation in 6 M guanidine hydrochloride (GdnHCl) at pH 2.4 for 4 h, followed by renaturation in 2 M potassium acetate at pH 6.5 for 24 h. This denaturation/renaturation cycle was repeated five times. The recovered activity of the renatured immobilized Sbtl settled at a constant level after the third denaturation/renaturation cycle, demonstrating that almost 100% renaturation was attained by use of the immobilized Sbtl. This immobilized Sbtl preparation could well be utilized for the mechanistic study of protein folding. We then found that 2 M potassium acetate was superior to 2 M potassium chloride as a refolding medium and that the ability of SSI to induce the correct shape of the mature Sbtl was lacking in several refolding media in both thermodynamic and kinetic criteria. Thus the main cause for the increase of refolding yield of Sbtl by coexistence of SSI was prevention of the autolysis of Sbtl

Development of Safety Measures of Bicycle Trafflc by Observation wffh Deep-Leamlng, Drive Recorder Data, Probe Blcycle wlth LIDAR, and Connected Simulators

This research outlines the development of evaluating safety measures for bicycle traffic using state-of-the-art technology, which was started since 2020 as a four-year project. The project is funded by the Commission on Advanced Road Technology in the Ministry of Land, Infrastructure, Transport and Tourism(MLIT). While Japan has a high bicycle modal share of 12% (2010), bicycle-related fatalities are relatively high among other countries in the IRTAD database (2019). Under these circumstances, since 2007, various measures for bicycle traffic measures have been implemented to improve the safe bicycle traffic environment, including the revision of the Road Traffic Act and the formulation of a national plan to promote bicycle use. However, serious accidents involving bicycles are remained in some specific cases. According to the government's traffic accident analysis results (2019), right-hook crash at signalized intersections are one of the most serious types of collision involving bicycles, along with accidents at unsignalized intersections involving vehicles turning left, rear-end collisions, and single vehicle accidents due to off-road deviation. In particular, proactive safety measures are required at signalized intersections along arterial roads, where electric personal mobility vehicles traveling at speeds of up to 20 km/h are expected to share with bicycles in the future. In order to evaluate safety measures for bicycle-vehicle crashes, this project set the following goals. 1) Identify factors influencing near-miss incidents and collisions through analysis of drive recorder data and accident statistical data. 2) Detailed analysis of traffic conditions from the cyclist's perspective using a probe bicycle equipped with a LiDAR sensor. 3) Development of an experimental environment using a connected simulator for evaluation of cooperative driving behavior. 4) Clarification of experimental conditions to evaluate different scenarios and conditions with and without intervention. 5) Proposal of effective interventions to improve crash cases based on experiments

Stigmatic Microscopy Enables Low-Cost, 3D, Microscale Particle Imaging Velocimetry in Rehydrating Aqueous Two-Phase Systems

This paper describes the construction of a novel stigmatic microscope and image analysis algorithm to simultaneously analyze convective mixing both inside and outside of rehydrating μL-scale aqueous two-phase system (ATPS) droplets. Stigmatic microscopy is inexpensive and advantageous because it modifies the point-spread function of fluorescent particles to enable measurement of their 3D positions from single 2D images, without needing to take slices. In one application of the technique, the convection patterns captured clarify how different ATPS formulations succeed or fail to exclude cells for patterning. Particle flow traces reveal speed and directionality of circulation, indicating temporary eddies at the outer edge of the rehydrating droplet. In another application, the speed of circulation during rehydration was analyzed for different ATPS formulations and the results used to develop a new fast ELISA procedure. While this paper focuses on ATPS rehydration, the microscope and algorithm should be applicable to a broad range of microfluidic flows where microscale 3D convection is important

Nutrition Support for Head and Neck Squamous Cell Carcinoma Patients Treated with Chemoradiotherapy: How Often and How Long?

Background. Oral intake of many patients with locally advanced head and neck cancer (LAHNC) decrease during chemoradiotherapy (CRT). Although prophylactic percutaneous endoscopic gastrostomy (PEG) is recommended, not a few patients complete CRT without using PEG tube. Patients and Methods. The subjects were patients with LAHNC who received CRT. We retrospectively investigated the incidence and duration of nutritional support during and after CRT, and predicting factors of nutritional support. For patients who required nutritional support, we also checked the day of initiation and the duration of nutritional support. Results. Of 53 patients, 29 patients (55%) required nutritional support during and/or after CRT. While no clear relation between requirement of nutritional support and variables including age, T stage, N stage, clinical stage and chemotherapy regimen, there could be some relationships between tumor primary sites and the requirement and duration of nutritional support. 17 (77%) of 22 patients with oropharynx cancer(OP) required nutritional support and prolonged for 4.4 months, and 11 (46%) of 24 patients with hypopharynx cancer(HP) required nutritional support and prolonged for 21.9 months. Conclusion. Nutritional support is indicated many HNC patients treated with CRT and primary sites may have some relation to its indication and duration

Induced-fit expansion and contraction of a self-assembled nanocube finely responding to neutral and anionic guests

Induced-fit or conformational selection is of profound significance in biological regulation. Biological receptors alter their conformation to respond to the shape and electrostatic surfaces of guest molecules. Here we report a water-soluble artificial molecular host that can sensitively respond to the size, shape, and charged state of guest molecules. The molecular host, i.e. nanocube, is an assembled structure consisting of six gear-shaped amphiphiles (GSAs). This nanocube can expand or contract its size upon the encapsulation of neutral and anionic guest molecules with a volume ranging from 74 to 535 Å3 by induced-fit. The responding property of this nanocube, reminiscent of a feature of biological molecules, arises from the fact that the GSAs in the nanocubes are connected to each other only through the hydrophobic effect and very weak intermolecular interactions such as van der Waals and cation-π interactions

Development of early neutropenic fever, with or without bacterial infection, is still a significant complication after reduced-intensity stem cell transplantation

AbstractLittle information is available on the clinical characteristics of infectious complications that occur in the early period after reduced-intensity stem cell transplantation (RIST). We retrospectively investigated the clinical features of neutropenic fever and infectious episodes within 30 days after RIST in 76 patients who had received fluoroquinolones as part of their antibacterial prophylaxis. Preparative regimens included cladribine 0.66 mg/kg or fludarabine 180 mg/m2 plus busulfan 8 mg/kg. All but 1 patient survived 30 days after transplantation, and 75 patients (99%) became neutropenic within a median duration of 9 days. Neutropenic fever was observed in 29 patients (38%), and bacterial infection was confirmed in 15 (20%) of these, including bacteremia (n = 13), bacteremia plus pneumonia (n = 1), and urinary tract infection (n = 1). The causative organisms were gram-positive (n = 9) and gram-negative organisms (n = 7), with a mortality rate of 6%. Neither viral nor fungal infection was documented. Multivariate analysis showed that the presence of neutropenia at the initiation of preparative regimens was an independent risk factor for subsequent documented bacterial infections (P = .026; 95% confidence interval, 1.25–35.1). We conclude that neutropenic fever and bacteremia remain common complications in RIST