Control of acetylcholine receptor mobility and distribution in cultured muscle membranes. A fluorescence study

The molecular control of the distribution and motion of acetylcholine receptors in the plasma membrane of developing rat myotubes in primary cell culture was investigated by fluorescence techniques. Acetylcholine receptors were marked with tetramethylrhodamine-labeled α-bungarotoxin and lateral molecular motion in the membrane was measured by the fluorescence photobleaching recovery technique. Three types of experiments are discussed: (I) The effect of enzymatic cleavages, drugs, cross-linkers, and physiological alterations on the lateral motion of acetylcholine receptors and on the characteristic distribution of acetylcholine receptors into patch and diffuse areas. (II) Observation of the distribution and/or motion of fluorescence-labeled concanavalin A receptors, lipid probes, cell surface protein, and stained cholinesterase in acetylcholine receptor patch and diffuse areas. (III) The effect of a protein synthesis inhibitor and electrical stimulation on membrane incorporation of new acetylcholine receptors.Some of the main conclusions are: (a) acetylcholine receptor lateral motion is inhibited by concanavalin A plant lectin and by anti-α-bungarotoxin antibody, but marginally enhanced by treatment with a local anesthetic; (b) patches are stabilized by an immobile cellular structure consisting of molecules other than the acetylcholine receptors themselves; (c) this structure is highly selective for acetylcholine receptors and not for other cell membrane components; (d) acetylcholine receptor patch integrity and diffuse area motion are independent of direct metabolic energy requirements and are sensitive to electrical excitation of myotube; (e) lipid molecules can move laterally in both acetylcholine receptor patches and diffuse areas; and (f) acetylcholine receptor lateral motion in diffuse areas and immobility in patch areas are not altered by specific agents which are known to affect extrinsic cell surface proteins, or cytoplasmic microfilaments and microtubules.     

Incidence of thyroid diseases in Zhejiang Province,China, after 15 years of salt iodization

Thyroid diseases(TD) can be induced by either deficient or excessive iodine intake. Universal Salt Iodization(USI) program has been implemented in China since 1995, to prevent iodine deficiency disorders (IDD). To evaluate the current conditions of TD and the role of USI, a multi-stage stratified random sampling scheme was used to perform a cross-sectional survey on the incidence of TD among participants in 6600 households in Zhejiang Province, a coastal area in China. Iodine nutrition status of the population was assessed by dietary iodine intake recall and urinary iodine concentration (UIC) of the participants, and TD were diagnosed by thyroid ultrasonography for 15122 participants and for 5873 participants by serum criteria for thyroid function(fT3, fT4, TSH, TRAb, TgAb, TPOAb; see Introduction for abbreviations). The median UIC of the surveyed population was 163 μg iodine/L. From the participants 23.2% had UIC < 100 μg/L which is moderately iodine-deficient according to WHO classification. Diffuse goiter was present in 2.3% of the population and thyroid nodule in 20.9%. The incidence of hyperthyroidism, subclinical hyperthyroidism, hypothyroidism, subclinical hypothyroidism, Graves’ disease and chronic lymphocytic thyroiditis was 0.5%, 0.6%, 0.6%, 7.8%, 0.2% and 0.3%, respectively. The proportion of several TD for participants with non-iodized salt intake was higher than that for participants with iodized salt intake.     

Scale‐down simulators for mammalian cell culture as tools to access the impact of inhomogeneities occurring in large‐scale bioreactors

During the scale‐up of a bioprocess, not all characteristics of the process can be kept constant throughout the different scales. This typically results in increased mixing times with increasing reactor volumes. The poor mixing leads in turn to the formation of concentration gradients throughout the reactor and exposes cells to varying external conditions based on their location in the bioreactor. This can affect process performance and complicate process scale‐up. Scale‐down simulators, which aim at replicating the large‐scale environment, expose the cells to changing environmental conditions. This has the potential to reveal adaptation mechanisms, which cells are using to adjust to rapidly fluctuating environmental conditions and can identify possible root causes for difficulties maintaining similar process performance at different scales. This understanding is of utmost importance in process validation. Additionally, these simulators also have the potential to be used for selecting cells, which are most robust when encountering changing extracellular conditions. The aim of this review is to summarize recent work in this interesting and promising area with the focus on mammalian bioprocesses, since microbial processes have been extensively reviewed.     

Development,characterization, and application of a 2‐Compartment system to investigate the impact of pH inhomogeneities in large‐scale CHO‐based processes

Large‐scale bioreactors for the production of monoclonal antibodies reach volumes of up to 25 000 L. With increasing bioreactor size, mixing is however affected negatively, resulting in the formation of gradients throughout the reactor. These gradients can adversely affect process performance at large scale. Since mammalian cells are sensitive to changes in pH, this study investigated the effects of pH gradients on process performance. A 2‐Compartment System was established for this purpose to expose only a fraction of the cell population to pH excursions and thereby mimicking a large‐scale bioreactor. Cells were exposed to repeated pH amplitudes of 0.4 units (pH 7.3), which resulted in decreased viable cell counts, as well as the inhibition of the lactate metabolic shift. These effects were furthermore accompanied by increased absolute lactate levels. Continuous assessment of molecular attributes of the expressed target protein revealed that subunit assembly or N‐glycosylation patterns were only slightly influenced by the pH excursions. The exposure of more cells to the same pH amplitudes further impaired process performance, indicating this is an important factor, which influences the impact of pH inhomogeneity. This knowledge can aid in the design of pH control strategies to minimize the effects of pH inhomogeneity in large‐scale bioreactors.     

Membrane‐Free Zn/MnO2 Flow Battery for Large‐Scale Energy Storage

The traditional Zn/MnO₂ battery has attracted great interest due to its low cost, high safety, high output voltage, and environmental friendliness. However, it remains a big challenge to achieve long‐term stability, mainly owing to the poor reversibility of the cathode reaction. Different from previous studies where the cathode redox reaction of MnO₂/MnOOH is in solid state with limited reversibility, here a new aqueous rechargeable Zn/MnO₂ flow battery is constructed with dissolution–precipitation reactions in both cathodes (Mn²⁺/MnO₂) and anodes (Zn²⁺/Zn), which allow mixing of anolyte and catholyte into only one electrolyte and remove the requirement for an ion selective membrane for cost reduction. Impressively, this new battery exhibits a high discharge voltage of ≈1.78 V, good rate capability (10C discharge), and excellent cycling stability (1000 cycles without decay) at the areal capacity ranging from 0.5 to 2 mAh cm^‐2. More importantly, this battery can be readily enlarged to a bench scale flow cell of 1.2 Ah with good capacity retention of 89.7% at the 500th cycle, displaying great potential for large‐scale energy storage.     

2D Materials for Large‐Area Flexible Thermoelectric Devices

The rapid development of the concept of the “Internet of Things (IoT)” requires wearable devices with maintenance‐free batteries, and thermoelectric energy conversion based on large‐area flexible materials has attracted much attention. Among large‐area flexible materials, 2D materials, such as graphene and related materials, are promising for thermoelectric applications due to their excellent transport properties and large power factors. In this Review, both single‐crystalline and polycrystalline 2D materials are surveyed using the experimental reports on thermoelectric devices of graphene, black phosphorus, transition metal dichalcogenides, and other 2D materials. In particular, their carrier‐density dependent thermoelectric properties and power factors maximized by Fermi level tuning techniques are focused. The comparison of the relevant performances between 2D materials and commonly used thermoelectric materials reveals the significantly enhanced power factors in 2D materials. Moreover, the current progress in thermoelectric module applications using large‐area 2D material thin films is summarized, which consequently offers great potential for the use of 2D materials in large‐area flexible thermoelectric device applications. Finally, important remaining issues and future perspectives, such as preparation methods, thermal transports, device designs, and promising effects in 2D materials, are discussed.     

Structural and immunogenomic insights into B-cell receptor activation

B cells express B-cell receptors(BCRs) which recognize antigen to trigger signaling cascades for B-cell activation and subsequent antibody production. BCR activation has a crucial influence on B-cell fate. How BCR is activated upon encountering antigen remains to be solved, although tremendous progresses have been achieved in the past few years. Here, we summarize the models that have been proposed to explain BCR activation, including the cross-linking model, the conformation-induced oligomerization model, the dissociation activation model, and the conformational change model. Especially, we elucidate the partially resolved structures of antibodies and/or BCRs by far and discusse how these current structural and further immunogenomic messages and more importantly the future studies may shed light on the explanation of BCR activation and the relevant diseases in the case of dysregulation.     

Effect of litter size,milk replacer and housing on production results of hyper-prolific sows

The modern hyper-prolific sow gives birth to more piglets than she has functional teats (in the following called supernumerary piglets). The aim of the present study was (1) to investigate the production consequences of hyper-prolific sows rearing supernumerary piglets equal to the mean live-born litter size, and (2) investigate whether potential negative effects on survival and growth could be alleviated by providing access to milk replacer and/or providing easier access to the udder (by loose housing). At day 1 (D1) postpartum (pp), 93 litters were standardised to 14 or 17 piglets (LS14/LS17) after which no piglets were moved between sows leading to decreased litter size if piglets died. Litters were provided with or without milk replacer in milk cups (+MILK/−MILK), and sows were either crated or loose housed (CRATE/LOOSE) in a 2 × 2 × 2 factorial design. Piglet mortality was higher in LS17 compared to LS14 (P < 0.01; OR = 2.0), higher in −MILK compared to +MILK (P = 0.01; OR = 1.2) and higher in LOOSE compared to CRATE (P = 0.02; OR = 1.8). This study showed that sow rearing of supernumerary piglets while supplying with milk replacer can increase piglet survival. It also showed that early mortality before piglets learned to drink milk replacer posed a challenge using this automatic milk replacer system. An interaction between access to milk replacer and the standardised litter size D1 affected litter weight (P < 0.01) and piglet weight day 28 (D28) (P = 0.03). The highest litter weight D28 was found in LS17 +MILK (P < 0.01) but with a lower individual piglet weight than in LS14 −MILK. Piglet weight D28 was higher in LS14 −MILK compared to LS17 regardless of access to milk replacer. Heterogeneity in piglet weight within litters D28 was larger in LS17 (P = 0.03) but could be reduced with +MILK in CRATE (P < 0.01). No effects were found on sow weight loss and feed intake (P > 0.05). In conclusion, the results showed that sows cannot rear the supernumerary piglets without further management interventions to reduce mortality. Supplying supernumerary piglets equal to the mean live-born litter size of hyper-prolific sows with milk replacer can from results of this study be an alternative strategy to the use of nurse sows.