Hydrogen Bond-Induced Activation of Photocatalytic and Piezophotocatalytic Properties in Calcium Nitrate Doped Electrospun PVDF Fibers.

In this study, polyvinylidene fluoride (PVDF) fibers doped with hydrated calcium nitrate were prepared using electrospinning. The samples were analyzed using scanning electron microscopy (SEM), X-ray diffraction (XRD), optical spectroscopy, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared (FTIR), Raman, and photoluminescence (PL) spectroscopy. The results are complementary and confirm the presence of chemical hydrogen bonding between the polymer and the dopant. Additionally, there was a significant increase in the proportion of the electroactive polar beta phase from 72 to 86%. It was shown that hydrogen bonds acted as a transport pathway for electron capture by the conjugated salt, leading to more than a three-fold quenching of photoluminescence. Furthermore, the optical bandgap of the composite material narrowed to the range of visible light energies. For the first time, it the addition of the salt reduced the energy of the PVDF exciton by a factor of 17.3, initiating photocatalytic activity. The calcium nitrate-doped PVDF exhibited high photocatalytic activity in the degradation of methylene blue (MB) under both UV and visible light (89 and 44%, respectively). The reaction rate increased by a factor of 2.4 under UV and 3.3 under visible light during piezophotocatalysis. The catalysis experiments proved the efficiency of the membrane design and mechanisms of catalysis are suggested. This study offers insight into the nature of chemical bonds in piezopolymer composites and potential opportunities for their use.

3D bioprinting of in situ vascularized tissue engineered bone for repairing large segmental bone defects.

Large bone defects remain an unsolved clinical challenge because of the lack of effective vascularization in newly formed bone tissue. 3D bioprinting is a fabrication technology with the potential to create vascularized bone grafts with biological activity for repairing bone defects. In this study, vascular endothelial cells laden with thermosensitive bio-ink were bioprinted in situ on the inner surfaces of interconnected tubular channels of bone mesenchymal stem cell-laden 3D-bioprinted scaffolds. Endothelial cells exhibited a more uniform distribution and greater seeding efficiency throughout the channels. In vitro, the in situ bioprinted endothelial cells can form a vascular network through proliferation and migration. The in situ vascularized tissue-engineered bone also resulted in a coupling effect between angiogenesis and osteogenesis. Moreover, RNA sequencing analysis revealed that the expression of genes related to osteogenesis and angiogenesis is upregulated in biological processes. The in vivo 3D-bioprinted in situ vascularized scaffolds exhibited excellent performance in promoting new bone formation in rat calvarial critical-sized defect models. Consequently, in situ vascularized tissue-engineered bones constructed using 3D bioprinting technology have a potential of being used as bone grafts for repairing large bone defects, with a possible clinical application in the future.

An action recognition method for manual acupuncture techniques using a tactile array finger cot.

The current measurement systems for the physical parameters (rotation frequency, and amplitude) of Traditional Chinese Medicine (TCM) manual acupuncture tend to cause disturbance and inconvenience in clinical application and do not accurately capture the tactile signals from the physician's finger during manual acupuncture operations. In addition, the literature rarely discusses classification of the four basic manual acupuncture techniques (reinforcing by twirling and rotating (RFTR), reducing by twirling and rotating (RDTR), reinforcing by lifting and thrusting (RFLT), and reducing by lifting and thrusting (RDLT)). To address this problem, we developed a multi-PVDF film-based tactile array finger cot to collect piezoelectric signals from the acupuncturist's finger-needle contact during manual acupuncture operations. In order to recognize the four typical TCM manual acupuncture techniques, we developed a method to capture piezoelectric signals in related "windows" and subsequently extract features to model acupuncture techniques. Next, we created an ensemble learning-based action classifier for manual acupuncture technique recognition. Finally, the proposed classifier was employed to recognize the four types of manual acupuncture techniques performed by 15 TCM physicians based on the piezoelectric signals collected using the tactile array finger cot. Among all the approaches, our proposed feature-based CatBoost ensemble learning model achieved the highest validation accuracy of 99.63% and the highest test accuracy of 92.45%. Moreover, we provide the efficiency and limitations of using this action recognition method.

Enhanced Energy Storage Performance of PVDF-Based Composites Using BN@PDA Sheets and Titania Nanosheets.

With the rapid development of modern electrical and electronic applications, the demand for high-performance film capacitors is becoming increasingly urgent. The energy density of a capacitor is dependent on permittivity and breakdown strength. However, the development of polymer-based composites with both high permittivity (εr) and breakdown strength (Eb) remains a huge challenge. In this work, a strategy of doping synergistic dual-fillers with complementary functionalities into polymer is demonstrated, by which high εr and Eb are obtained simultaneously. Small-sized titania nanosheets (STNSs) with high εr and high-insulating boron nitride sheets coated with polydopamine on the surface (BN@PDA) were introduced into poly(vinylidene fluoride) (PVDF) to prepare a ternary composite. Remarkably, a PVDF-based composite with 1 wt% BN@PDA and 0.5 wt% STNSs (1 wt% PVDF/BN@PDA-STNSs) shows an excellent energy storage performance, including a high εr of ~13.9 at 1 Hz, a superior Eb of ~440 kV/mm, and a high discharged energy density Ue of ~12.1 J/cm3. Moreover, the simulation results confirm that BN@PDA sheets improve breakdown strength and STNSs boost polarization, which is consistent with the experimental results. This contribution provides a new design paradigm for energy storage dielectrics.

A novel phenolic formulation for treating hepatic and peripheral insulin resistance by regulating GLUT4-mediated glucose uptake.

Chronic insulin resistance suppresses muscle and liver response to insulin, which is partially due to impaired vesicle trafficking. We report here that a formula consisting of resveratrol, ferulic acid and epigallocatechin-3-O-gallate is more effective in ameliorating muscle and hepatic insulin resistance than the anti-diabetic drugs, metformin and AICAR. The formula enhanced glucose transporter-4 (GLUT4) translocation to the plasma membrane in the insulin-resistant muscle cells by regulating both insulin-independent (calcium and AMPK) and insulin-dependent (PI3K) signaling molecules. Particularly, it regulated the subcellular location of GLUT4 through endosomes to increase glucose uptake under insulin-resistant condition. Meanwhile, this phytochemicals combination increased glycogen synthesis and decreased glucose production in the insulin-resistant liver cells. On the other hand, this formula also showed anti-diabetic potential by the reduction of lipid content in the myotubes, hepatocytes, and adipocytes. This study demonstrated that the three phenolic compounds in the formula could work in distinct mechanisms and enhance both insulin-dependent and independent vesicles trafficking and glucose transport mechanisms to improve carbohydrate and lipid metabolism.

PVDF/KNO3 Composite Sub-Microfibers Produced by Solution Blow Spinning as a Hydrophobic Matrix for Fertilizer Delivery System.

Nutrient supplementation is a common practice in agriculture to increase crop productivity in the field. This supplementation is usually excessive, causing nutrient leaching in periods of rainfall leading to environmental problems. To overcome such issues, many studies have been devoted to developing polymeric matrices for the controlled and continuous release of nutrients, reducing losses, and keeping plants nourished for as long as possible. However, the release mechanism of these matrices is based on water diffusion. They start immediately for swellable polymeric matrices, which is not interesting and also may cause some waste, because the plant only needs nutrition only after the germination process. Here, as proof of concept, we tested a hydrophobic polymeric matrix based on sub-microfibers mats, produced by solution blow spinning, filled with potassium nitrate (KNO3) for the controlled release of nutrients to plants. In this work, we used the polyvinylidene fluoride (PVDF) polymer to produce composite nanofibers containing pure potassium nitrate in the proportion of 10% weight. PVDF/KNO sub-microfibers mats were obtained with 370 nm average diameter and high occurrence of beads. We performed a release test using PVDF/KNO3 mats in a water bath. The release kinetic tests showed an anomalous delivery mechanism, but the composite polymeric fibrous mat showed itself to be a promising alternative to delay the nutrient delivery for the plants.

MAP30 protein from Momordica charantia is therapeutic and has synergic activity with cisplatin against ovarian cancer in vivo by altering metabolism and inducing ferroptosis.

Increasing evidence shows that Traditional Chinese Medicine (TCM) has an obvious appeal for cancer treatment, but there is still a lack of scientific investigation of its underlying molecular mechanisms. Bitter melon or bitter gourd (Momordica charantia) is an edible fruit that is commonly consumed, and it is used to cure different diseases in various ancient folk medical practices. We report that a bioactive protein, MAP30, isolated from bitter melon seeds exhibited potent anticancer and anti-chemoresistant effects on ovarian cancer cells. Functional studies revealed that MAP30 inhibited cancer cell migration, cell invasion, and cell proliferation in various ovarian cancer cells but not normal immortalized ovarian epithelial cells. When administered with cisplatin, MAP30 produced a synergistic effect on cisplatin-induced cell cytotoxicity in ovarian cancer cells. When low doses of cisplatin and MAP30 were co-injected intraperitoneally, a remarkable reduction of tumor dissemination and tumor growth was observed in an ovarian cancer ascites mouse model. Notably, blood tests confirmed that MAP30 did not cause any adverse effects on liver and kidney functions in the treated mice. MAP30 activated AMP-activated protein kinase (AMPK) signaling via CaMKKß and induced cell cycle arrest in the S-phase. MAP30 modulated cell metabolism of ovarian cancer cells via suppression of GLUT-1/-3-mediated glucose uptake, adipogenesis, and lipid droplet formation in tumor development and progression. MAP30 also induced an increase in intracellular Ca2+ ion concentration, which triggered ROS-mediated cancer cell death via apoptosis and ferroptosis. Collectively, these findings suggest that natural MAP30 is a non-toxic supplement that may enhance chemotherapeutic outcomes and benefit ovarian cancer patients with peritoneal metastases.

Inhibition of NADPH oxidase alleviates germ cell apoptosis and ER stress during testicular ischemia reperfusion injury.

Testicular torsion and detorsion (TTD) is a serious urological condition affecting young males that is underlined by an ischemia reperfusion injury (tIRI) to the testis as the pathophysiological mechanism. During tIRI, uncontrolled production of oxygen reactive species (ROS) causes DNA damage leading to germ cell apoptosis (GCA). The aim of the study is to explore whether inhibition of NADPH oxidase (NOX), a major source of intracellular ROS, will prevent tIRI-induced GCA and its association with endoplasmic reticulum (ER) stress. Sprague-Dawley rats (n = 36) were divided into three groups: sham, tIRI only and tIRI treated with apocynin (a NOX inhibitor). Rats undergoing tIRI endured an ischemic injury for 1 h followed by 4 h of reperfusion. Spermatogenic damage was evaluated histologically, while cellular damages were assessed using real time PCR, immunofluorescence staining, Western blot and biochemical assays. Disrupted spermatogenesis was associated with increased lipid and protein peroxidation and decreased antioxidant activity of the enzyme superoxide dismutase (SOD) as a result of tIRI. In addition, increased DNA double strand breaks and formation of 8-OHdG adducts associated with increased phosphorylation of the DNA damage response (DDR) protein H2AX. The ASK1/JNK apoptosis signaling pathway was also activated in response to tIRI. Finally, increased immuno-expression of the unfolded protein response (UPR) downstream targets: GRP78, eIF2-α1, CHOP and caspase 12 supported the presence of ER stress. Inhibition of NOX by apocynin protected against tIRI-induced GCA and ER stress. In conclusion, NOX inhibition minimized tIRI-induced intracellular oxidative damages leading to GCA and ER stress.

Insight into the magnetic lime coagulation-membrane distillation process for desulfurization wastewater treatment: From pollutant removal feature to membrane fouling.

The high suspended solid (SS) and salts were main issues for flue gas desulfurization wastewater (FGDW). A magnetic lime coagulation (MLC)-membrane distillation (MD) integrated process was firstly applied with a self-made poly (vinylidene fluoride-co-chlorotrifluoroethylene) (PVDF-CTFE) membrane and the pollutants remove feature and membrane fouling were discussed. The SS was nearly 100 % removed and magnetic seed significantly accelerate the settleability. The flux was 43.00 kg/m2 h with a salt rejection >99 %. It was higher than 13 kg/m2 h in the first 125 h during the 18d continuous test, and the rejection for all cations, anions, total organic carbon (TOC) and total inorganic carbon (TIC) were higher than 99.95 %, 99.00 %, 98.81 %, and 99.65 %, respectively. Humic substances and tryptophan with 100-5000 Da were main dissolved organic matter (DOM), which were significantly removed. However, membrane fouling and wetting happened after 150 h. Scaling was the main foulants, while the organic fouling and biofouling were also detected. A new "bricklaying model" was induced to depict the formation of foulant layer, the colloids, organic matters (OMs) and microbe communities act as the "concrete", while the inorganic crystals (magnesium and calcium oxysulphides) were the "bricks". This contribution offers a new method for FGDW treatment and the membrane fouling mechanism of MD process.

Nano CuAl2O4 spinel mineral as a novel antibacterial agent for PVDF membrane modification with minimized copper leachability.

The employment of copper-containing antibacterial agents for membrane modification can constrain undesirable bacterial adhesion and growth in an effective and economical way. However, copper ion may be leached out and cause further toxicity after applying those materials for membrane processes. Therefore, in this study, nano CuAl2O4 spinel was synthesized as a novel copper-containing material which was expected to have good antibacterial activity and simultaneously stabilize copper ions by its intrinsic structure. The obtained nano spinel was applied for membrane modification via both doping and coating methods. Results show that the addition of nano CuAl2O4 spinel during doping process can change polyvinylidene fluoride (PVDF) membrane properties (crystallization, tensile strength, porosity, pore size distribution, permeate flux and bovine serum albumin (BSA) rejection) obviously, but no obvious change was observed for the coated membrane except increased hydrophilicity and permeate flux. Most importantly, the amount of bacteria attachment was found to reduce significantly (˜68%) on the membrane coated with nano spinel, which indicates the successful application of nano CuAl2O4 spinel as a novel antibacterial agent for membrane modification without extra concern of copper toxicity.

Salidroside stimulates the Sirt1/PGC-1α axis and ameliorates diabetic nephropathy in mice.

BACKGROUND: Salidroside, an active component from Traditional Chinese Medicine Rhodiola rosea L., has various pharmacological functions including anti-inflammatory, anti-cancer and anti-oxidative properties. However, whether salidroside plays a beneficial role in diabetic nephropathy is still unclear. PURPOSE: The objective of this work was to investigate the potential roles of salidroside against diabetic nephropathy and the underlying molecular mechanisms. METHODS: Streptozocin was given to obese mice to generate diabetic nephropathy animal model. Salidroside was administered to these mice and proteinuria, podocyte integrity, renal morphology and fibrosis, mitochondrial biogenesis were examined. RESULTS: Our results showed that salidroside treatment greatly attenuates diabetic nephropathy as evidenced by decreased urinary albumin, blood urea nitrogen and serum creatinine. Morphological analysis indicated that salidroside improves renal structures in diabetic nephropathy. The decreases in nephrin and podocin expression were markedly reversed by salidroside. Moreover, kidney fibrosis in diabetic nephropathy mice was largely prevented by salidroside. Mechanistically, in salidroside-treated mice, the mitochondrial DNA copy and electron transport chain proteins were significantly enhanced. Meanwhile, the reduced Sirt1 and PGC-1α expression in diabetic nephropathy was almost completely counteracted in the presence of salidroside. CONCLUSIONS: Our data showed that salidroside plays a beneficial role against diabetic nephropathy in mice, which probably via Sirt1/PGC-1α mediated mitochondrial biogenesis.

PVDF piezoelectric neural conduit incorporated pre-differentiated adipose-derived stem cells may accelerate the repair of peripheral nerve injury.

Peripheral nerve injury is a common trauma disease which often results in sensory and motor dysfunction. However, the surgical repair for peripheral nerve injury, especially for large segmental defects, is not satisfactory. Growing evidences suggest that artificial neural conduit combined with stem cells is potential tissue engineering remediation method for peripheral nerve injury. But, selections of biomaterials and stem cells are still being debated. Based on the findings from previous studies, we hypothesize that PVDF piezoelectric neural conduit incorporated Schwann-like cells which pre-differentiated from adipose-derived stem cells may substantially promote the repair of peripheral nerve injury. This novel technique may help clinical surgeons cure the seriously injured patients better and point out a new direction for neural tissue engineering researchers select the suitable biomaterials and seed cells.

Flow injection spectrophotometric determination of V(V) involving on-line separation using a poly(vinylidene fluoride-co-hexafluoropropylene)-based polymer inclusion membrane.

A poly(vinylidene fluoride-co-hexafluoropropylene)-based polymer inclusion membrane (PIM) using Cyphos® IL 101 (i.e. trihexyl(tetradecyl)phosphonium chloride) as the carrier and 2-nitrophenyl octyl ether as a plasticizer in a mass ratio of 55/35/10 was employed for the on-line extractive separation of V(V) prior to its spectrophotometric determination in a flow injection analysis (FIA) system using xylenol orange as the colorimetric reagent. The selectivity of the membrane allowed the determination of V(V) in sulfate solutions in the presence of a variety of cations and anions. The interference of molybdenum(VI) was eliminated by off-line extraction using the same PIM. A univariate sequential optimization of the newly developed FIA system was conducted and under optimal conditions the system is characterized by a linear concentration range of 0.5-8.0mgL-1, detection limit of 0.08mgL-1 and sample throughput of 4h-1. The relative standard deviation at the 3mgL-1 level of V(V) was 2.9% based on 8 replicate determinations. The membrane was stable, which was reflected by the standard deviation value for determinations over three consecutive days (24 determinations of 3mgL-1 V(V)) of 3.6%. The newly developed FIA system was applied to the determination of V(V) in water and dietary supplements samples and a good agreement with inductively coupled plasma optical emission spectrometry was observed.

A maternal high-fat, high-sucrose diet alters insulin sensitivity and expression of insulin signalling and lipid metabolism genes and proteins in male rat offspring: effect of folic acid supplementation.

A maternal high-fat, high-sucrose (HFS) diet alters offspring glucose and lipid homoeostasis through unknown mechanisms and may be modulated by folic acid. We investigated the effect of a maternal HFS diet on glucose homoeostasis, expression of genes and proteins associated with insulin signalling and lipid metabolism and the effect of prenatal folic acid supplementation (HFS/F) in male rat offspring. Pregnant Sprague-Dawley rats were randomly fed control (CON), HFS or HFS/F diets. Offspring were weaned on CON; at postnatal day 70, fasting plasma insulin and glucose and liver and skeletal muscle gene and protein expression were measured. Treatment effects were assessed by one-way ANOVA. Maternal HFS diet induced higher fasting glucose in offspring v. HFS/F (P=0·027) and down-regulation (P<0·05) of genes coding for v-Akt murine thymoma viral oncogene homolog 2, resistin and v-Raf-1 murine leukaemia viral oncogene homolog 1 (Raf1) in offspring skeletal muscle and acetyl-CoA carboxylase (Acaca), fatty acid synthase and phosphatidylinositol-4,5-biphosphate 3-kinase, catalytic subunit ß in offspring liver. Skeletal muscle neuropeptide Y and hepatic Kruppel-like factor 10 were up-regulated in HFS v. CON offspring (P<0·05). Compared with CON, Acaca and Raf1 protein expression levels were significantly lower in HFS offspring. Maternal HFS induced higher homoeostasis model of assessment index of insulin resistance v. CON (P=0·030) and HFS/F was associated with higher insulin (P=0·016) and lower glucose (P=0·025). Maternal HFS diet alters offspring insulin sensitivity and de novo hepatic lipogenesis via altered gene and protein expression, which appears to be potentiated by folate supplementation.

Tissue specific effects of feeds supplemented with grape pomace or olive oil mill wastewater on detoxification enzymes in sheep.

The aim of the present study was to investigate the effects of livestock feed supplemented with grape pomace (GP) or olive oil mill wastewater (OMW) byproducts on the enzymatic activity and protein expression of antioxidants enzymes, in liver and spleen tissue of sheep. Thus, 36 male sheep of Chios breed were divided into 3 homogeneous groups, control group (n = 12), GP group (n = 12) and OMW group (n = 12), receiving standard or experimental feed. Liver and spleen tissues were collected at 42 and 70 days post-birth. The enzymatic activity of superoxide dismutase (SOD) and glutathione-s-transferase (GST) and also the protein expression of γ-synthase glutamyl custeine (γ-GCS) were determined in these tissues. The results showed GP group exhibited increased enzymatic activity of GST and protein expression of γ-GCS in liver compared to control group. In GP group's spleen, GST activity was increased compared to control but γ-GCS expression was not affected. In OMW group's liver, GST activity was increased and γ-GCS expression was reduced compared to control. In OMW group's spleen, GST activity was increased but GCS expression was not affected. SOD activity was not affected in both tissues either in GP or OMW group.

Adhesion prevention in ventral hernia repair: an experimental study comparing three lightweight porous meshes recommended for intraperitoneal use.

BACKGROUND: In ventral hernia repair, when prosthetic material is placed intraperitoneally, it may lead to an inflammatory reaction resulting in adhesions between the mesh and abdominal viscera. Several meshes have been developed to minimize this process. In this experimental study, the ability of different combined meshes to attenuate the adhesion formation was examined. METHODS: Three commercially available lightweight porous combined meshes were placed intraperitoneally to repair an abdominal wall defect in rats: DynaMesh-IPOM (PVDF + PP), TiMesh (titanium-coated filament PP) and C-QUR/FX (omega-3 fatty acid-coated filament PP). The DynaMesh-CICAT (PVDF) was implanted in the control group. Adhesion formation was macroscopically evaluated and scored after 7 and 21 days. RESULTS: All animals except two presented intra-abdominal adhesions. None of the meshes examined in the study demonstrated to prevent adhesions. C-QUR/FX reduced adhesion formation at 7 days' follow-up compared with all other meshes but by 21 days this effect was diminished. Between 7 and 21 days adhesion extension significantly decreased for TiMesh. TAS did not show significant modifications between 7 and 21 days' follow-up for each mesh. CONCLUSIONS: The combined porous meshes tested in the present study demonstrated to reduce but not to prevent the adhesion formation, even if with some differences. Combined porous meshes could be chosen instead of simple meshes for retro-rectus preperitoneal prosthetic ventral hernia repair.

Effect of ultrahigh diluted homeopathic medicines on the electrical properties of PVDF-HFP

In an effort to improve the electrical properties of the electroactive Poly(vinylidene fluoridehexafluoropropylene) (PVdF-HFP), we introduced a novel and simple approach to synthesize PVDFHFP composite films by incorporating ultrahigh dilutions of two homeopathic medicines Ferrum metallicum (FM) and Zincum oxidatum (ZO) in different potencies. The homeo-PVDF-composite films (HPCF) were synthesized by simple solution casting technique. XRD, FESEM, FTIR studies were performed to check the presence of nanoparticles in the film. The electrical properties of the HPCF samples get enhanced significantly due to the incorporation of the medicines and the effect increases with the increase in potency of the medicines. (AU)

Effect of ultrahigh diluted homeopathic medicines on the electrical properties of PVDF-HFP

In an effort to improve the electrical properties of the electroactive Poly(vinylidene fluoride-hexafluoropropylene) (PVdF-HFP), we introduced a novel and simple approach to synthesize PVDFHFP composite films by incorporating ultrahigh dilutions of two homeopathic medicines Ferrum metallicum (FM) and Zincum oxidatum (ZO) in different potencies. The homeo-PVDF-composite films (HPCF) were synthesized by simple solution casting technique. XRD, FESEM, FTIR studies were performed to check the presence of nanoparticles in the film. The electrical properties of the HPCF samples get enhanced significantly due to the incorporation of the medicines and the effect increases with the increase in potency of the medicines.

Preparation and characterization of Pd/Fe bimetallic nanoparticles immobilized on Al2O3/PVDF membrane: Parameter optimization and dechlorination of dichloroacetic acid.

Using a liquid-solid phase inversion method, a hybrid matrix poly(vinylidene fluoride) (PVDF) membrane was prepared with alumina (Al2O3) nanoparticle addition. Pd/Fe nanoparticles (NPs) were successfully immobilized on the Al2O3/PVDF membrane, which was characterized by Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The micrographs showed that the Pd/Fe NPs were dispersed homogeneously. Several important experimental parameters were optimized, including the mechanical properties, contact angle and surface area of Al2O3/PVDF composite membranes with different Al2O3 contents. At the same time, the ferrous ion concentration and the effect of hydrophilization were studied. The results showed that the modified Al2O3/PVDF membrane functioned well as a support. The Al2O3/PVDF membrane with immobilized Pd/Fe NPs exhibited high efficiency in terms of dichloroacetic acid (DCAA) dechlorination. Additionally, a reaction pathway for DCAA dechlorination by Pd/Fe NPs immobilized on the Al2O3/PVDF membrane system was proposed.

Translation attenuation via 3' terminal codon usage in bovine csn1s2 is responsible for the difference in αs2- and ß-casein profile in milk.

The rate of secretion of αs2-casein into bovine milk is approximately 25% of that of ß-casein, yet mammary expression of their respective mRNA transcripts (csn1s2 and csn2) is not different. Our objective was to identify molecular mechanisms that explain the difference in translation efficiency between csn1s2 and csn2. Cell-free translational efficiency of csn2 was 5 times that of csn1s2. Transcripts of csn1s2 distributed into heavier polysomes than csn2 transcripts, indicating an attenuation of elongation and/or termination. Stimulatory and inhibitory effects of the 5' and 3' UTRs on translational efficiency were different with luciferase and casein sequences in the coding regions. Substituting the 5' and 3' UTRs from csn2 into csn1s2 did not improve csn1s2 translation, implicating the coding region itself in the translation difference. Deletion of a 28-codon fragment from the 3' terminus of the csn1s2 coding region, which displays codons with low correlations to cell fitness, increased translation to a par with csn2. We conclude that the usage of the last 28 codons of csn1s2 is the main regulatory element that attenuates its expression and is responsible for the differential translational expression of csn1s2 and csn2.