The influence of the blood handling process on the measurement of circulating TGF-β1

In order to evaluate the impact of blood sample handling processes on circulating TGF-β1 levels, blood specimens were obtained from 13 healthy volunteers using different handling processes (kept at room temperature (RT) or on ice before centrifugation, using different centrifugal forces). TGF-β1 levels were measured using an enzyme-linked immunosorbent assay. A paired-T test was used for statistical analysis. The TGF-β1 level in on-ice serum was significantly lower than that in room-temperature serum (P<0.001), and both were significantly higher than that found in on-ice plasma (P<0.001). Compared with on-ice plasma samples, the longer the samples were kept at RT, the higher the levels of TGF-β1 in plasma (P=0.268, 0.040, and 0.0015 for 5 min, 30 min, and 60 min in RT, respectively). Compared with plasma centrifuged at 2,500×g for 30?min, the TGF-β1 levels were much lower than those found in plasma centrifuged at 1,200×g for 10?min (P=0.003); and a double centrifugation before TGF-β1 detection, significantly decreased the level (P<0.001). It is suggested that the optimal sampling conditions for the detection of TGF-β1 should be plasma prepared on ice and spun down at a higher centrifugal force.     

Influence of Salt Intake on Association of Blood Uric Acid with Hypertension and Related Cardiovascular Risk

BackgroundA relationship of blood uric acid (UA) with hypertension and cardiovascular risk is under debate thus salt intake is hypothesized to contribute to such associations.MethodsIn this cross-sectional study, stratified cluster random sampling elicited a sample of 1805 Kazakhs with 92.4% compliance. Hypertension and moderate-or-high total cardiovascular risk (mTCR) were defined according to guidelines. Sodium intake was assessed by urinary sodium excretion. Prevalence ratios (PRs) were used to express associations of UA with hypertension and mTCR.ResultsIn the highest tertile of sodium intake in women, the adjusted PRs (95% confidence intervals) of low to high quartiles compared with the lowest quartile of UA, were 1.22(0.78–1.91), 1.18(0.75–1.85), and 1.65(1.09–2.51) for hypertension and 1.19(0.74–1.90), 1.39(0.91–2.11), and 1.65(1.10–2.47) for mTCR (P for trend <0.05). However, these findings were not shown for other sodium intake levels. There were similar results in men. PRs markedly increased with a concomitant increase in UA and sodium intake and there was a significant interaction (P = 0.010) for mTCR with PRs of 1.69(1.10–2.60) for men and 3.70(2.09–6.52) for women in those with the highest compared with the lowest quartile of UA and tertile of sodium intake. Similar findings were shown for hypertension.ConclusionsThis study implied that a high salt intake may enhance the associations of UA with hypertension and cardiovascular risk.     

Assessing hemorrhagic shock: Feasibility of using an ultracompact photoacoustic microscope

Hemorrhagic shock, as an important clinical issue, is regarding as a critical disease with a high mortality rate. Unfortunately, existing clinical technologies are inaccessible to assess the hemorrhagic shock via hemodynamics in microcirculation. Here, we propose an ultracompact photoacoustic microscope to assess hemorrhagic shock using a rat model and demonstrate its clinical feasibility by visualizing buccal microcirculation of healthy volunteers. Both functional and morphological features of the microvascular network including concentration of total hemoglobin (C_HbT), number of blood vessels (VN), small vascular density (SVD) and vascular diameter (VD) were derived to assess the microvascular hemodynamics of different organs. Animal studies show the feasibility of the proposed tool to assess and stage the hemorrhagic shock via microcirculation. in vivo oral imaging of healthy volunteers indicates the translational possibility of this technique for clinical evaluation of hemorrhagic shock.      

Directional freezing as an alternative method for cryopreserving rhesus macaque (Macaca mulatta) sperm

The objective was to develop a freezing protocol using a directional freezing (DF) technique for cryopreservation of rhesus macaque sperm and achieve a survival rate comparable to that achieved with a conventional freezing (CF) technique. Rhesus macaque sperm frozen with a DF technique, with cooling rates of 12 or 16 °C/min, had higher post-thaw motility (P < 0.05) than those cooled at 7 °C/min (59.3, 61.1, and 50.3%, respectively). Furthermore, sperm cryopreserved with 5% glycerol and a DF technique had similar frozen-thawed sperm motility to those cryopreserved by a CF technique (63.7 vs. 53.9%, P > 0.05). The function of sperm cryopreserved at the optimized cooling rate using a DF technique was evaluated by in vitro fertilization of oocytes collected from gonadotropin-stimulated rhesus macaques. Of the 38 mature oocytes collected, 78.9% were fertilized and 71.1, 47.4, and 42.1% of the oocytes developed to the 2-cell, morulae, and blastocyst stages, respectively. In conclusion, rhesus macaque sperm was effectively cryopreserved using a DF technique, providing a new and effective method for genetic preservation in this important species.     

Alpha-tocopherol is essential for acquired chill-light tolerance in the cyanobacterium Synechocystis sp. strain PCC 6803

Unlike Escherichia coli, the cyanobacterium Synechocystis sp. strain PCC 6803 is insensitive to chill (5°C) in the dark but rapidly losses viability when exposed to chill in the light (100 μmol photons m⁻² s⁻¹). Preconditioning at a low temperature (15°C) greatly enhances the chill-light tolerance of Synechocystis sp. strain PCC 6803. This phenomenon is called acquired chill-light tolerance (ACLT). Preconditioned wild-type cells maintained a substantially higher level of α-tocopherol after exposure to chill-light stress. Mutants unable to synthesize α-tocopherol, such as slr1736, slr1737, slr0089, and slr0090 mutants, almost completely lost ACLT. When exposed to chill without light, these mutants showed no or a slight difference from the wild type. When complemented, the slr0089 mutant regained its ACLT. Copper-regulated expression of slr0090 from P_petE controlled the level of α-tocopherol and ACLT. We conclude that α-tocopherol is essential for ACLT of Synechocystis sp. strain PCC 6803. The role of α-tocopherol in ACLT may be based largely on a nonantioxidant activity that is not possessed by other tocopherols or pathway intermediates.     

Vitrification of Yunnan Yellow Cattle oocytes: work in progress

The objective of this study was to provide a simple cryopreservation method for oocytes from Yunnan Yellow Cattle and facilitate preservation efforts in this native Chinese breed, which is threatened by agricultural modernization. Cumulus-oocyte complexes (COCs) were collected from slaughterhouse ovaries and matured in vitro for 22-24 h, then selected for cryopreservation. Vitrification in open pulled straws (OPS) or in microdrops on a cooled metal surface (solid surface vitrification, SSV) was compared. The OPS vitrification solution consisted of 20% ethylene glycol (EG) and 20% DMSO. The SSV solution was a mixture of 35% EG, 5% polyvinyl-pyrrolidon (PVP) and 0.4 M trehalose. Vitrified and warmed oocytes were either fertilized in vitro or parthenogenetically activated. The rates of cleavage and development to blastocysts of fertilized oocytes following OPS versus SSV were not statistically different (38.3 and 12.5% versus 35.8 and 6.0%, respectively). The corresponding rates of parthenogenetic development to blastocysts were also not different (8.2 versus 3.5%, respectively). Development to blastocysts of non-vitrified controls following fertilization was significantly higher than that of the vitrified oocytes (22.6%, P < 0.05). These results demonstrate for the first time, that although both OPS and SSV procedures reduced embryonic development, Yunnan Yellow Cattle oocytes are capable of developing to blastocysts following cryopreservation.     

Synthesis and biological evaluation of dithiocarbamate esters of parthenolide as potential anti-acute myelogenous leukaemia agents

A series of dithiocarbamate esters of parthenolide (PTL) was designed, synthesised, and evaluated for their anti- acute myelogenous leukaemia (AML) activities. The most promising compound 7l showed greatly improved potency against AML progenitor cell line KG1a with IC₅₀ value of 0.7?μM, and the efficacy was 8.7-folds comparing to that of PTL (IC₅₀?=?6.1?μM). Compound 7l induced apoptosis of total primary human AML cells and leukaemia stem cell (LSCs) of primary AML cells while sparing normal cells. Furthermore, 7l suppressed the colony formation of primary human leukaemia cells. Moreover, compound 12, the salt form of 7l, prolonged the lifespan of mice in two patient-derived xenograft models and had no observable toxicity. The preliminary molecular mechanism study revealed that 7l-mediated apoptosis is associated with mitogen-activated protein kinase signal pathway. On the basis of these investigations, we propose that 12 might be a promising drug candidate for ultimate discovery of anti-LSCs drug.     

Identification of Regulatory Sequences and Expression Analysis of OmpR Gene Under Different Stress Conditions in the Antarctic Bacterium Psychrobacter sp. G

An OmpR gene, named OmpR503, was cloned from the Antarctic psychrotrophic bacterium Psychrobacter sp. G according to its genomic draft. The deduced amino acid sequences of OmpR503 were highly conserved with other known protein members of OmpR family. qRT-PCR analysis showed that the expression of OmpR503 gene was significantly enhanced by high salinity (90, 120). The expression of OmpR503 gene was also significantly increased at low temperature (0, 10 °C), whereas depressed at high temperature (30 °C). When the strain was subjected to combined stress (0 °C with a salinity of 90), the expression of OmpR503 gene was increased significantly, which was up to 3.0-fold. In Antarctica, freezing tolerance of psychrotrophic bacteria is often accompanied by tolerance to osmotic stress caused by a lack of free water, thus the cold inducibility of OmpR503 gene might help the strain adapt to the harsh environment more efficiently.