Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells.

Mammalian cell lines are frequently used as the preferred host cells for producing recombinant therapeutic proteins (RTPs) having post-translational modified modification similar to those observed in proteins produced by human cells. Nowadays, most RTPs approved for marketing are produced in Chinese hamster ovary (CHO) cells. Recombinant therapeutic antibodies are among the most important and promising RTPs for biomedical applications. One of the issues that occurs during development of RTPs is their degradation, which caused by a variety of factors and reducing quality of RTPs. RTP degradation is especially concerning as they could result in reduced biological functions (antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity) and generate potentially immunogenic species. Therefore, the mechanisms underlying RTP degradation and strategies for avoiding degradation have regained an interest from academia and industry. In this review, we outline recent progress in this field, with a focus on factors that cause degradation during RTP production and the development of strategies for overcoming RTP degradation. KEY POINTS: • The recombinant therapeutic protein degradation in CHO cell systems is reviewed. • Enzymatic factors and non-enzymatic methods influence recombinant therapeutic protein degradation. • Reducing the degradation can improve the quality of recombinant therapeutic proteins.

[Research Progress in the Association Between Sarcopenic Obesity and Cancer].

As the global prevalence of obesity and the elderly population continues to increase,the incidence of sarcopenic obesity is also on the rise and becoming a global public health concern.Sarcopenic obesity not only increases the incidence of cancer,but is also associated with poor clinical outcomes in various cancers,such as surgical complications,increased risk of death,and possibly even an impact on chemotherapy as well.Therefore,sarcopenic obesity is emerging as an important indicator of prognosis in cancer patients.However,there are limited relevant studies on the association between sarcopenic obesity and cancer in China.This article reviews the definition and diagnosis of sarcopenic obesity,the clinical correlation between sarcopenic obesity and cancer,and the potential mechanisms,with a view to providing a reference for future clinical practice in China.

Identification and expression analysis of invertase family genes during grape (Vitis vinifera L.) berry development under CPPU and GA treatment.

Sugar is crucial for grape berry, whether used for fresh food or wine. However, berry enlargement treatment with forchlorfenuron (N-(2-chloro4-pyridyl)-N'-phenylurea) (CPPU, a synthetic cytokinin) and gibberellin (GA) always had adverse effects on sugar accumulation in some grape varieties, especially CPPU. Therefore exploring the molecular mechanisms behind these adverse effects could provide a foundation for improving or developing technology to mitigate the effects of CPPU/GA treatments for grape growers. In the present study, invertase (INV) family, the key gene controlling sugar accumulation, was identified and characterized on the latest annotated grape genome. Their express pattern, as well as invertase activity and sugar content, were analyzed during grape berry development under CPPU and GA3 treatment to explore the potential role of INV members under berry enlargement treatment in grapes. Eighteen INV genes were identified and divided into two sub-families: 10 neutral INV genes (Vv-A/N-INV1-10) and 8 acid INV genes containing 5 CWINV (VvCWINV1-5) and 3 VIN (VvVIN1-3). At the early development stage, both CPPU and GA3 treatment decreased the hexose level in berries of 'Pinot Noir' grape, whereas the activity of three types inverstase (soluble acid INV, insoluble acid INV, and neutral INV) increased. Correspondingly, most of INV members were up-regulated by GA3 /CPPU application at least one sampling time point during early berry development, including VvCWINV1, 2, 3, 4, 5, VvVIN1, 2, 3 and Vv-A/N-INV1, 2, 5, 6, 7, 8, 10. At maturity, the sugar content in CPPU-treated berries is still lower than that in the control. Soluble acid INV and neutral INV, rather than insoluble acid INV, presented lower activity in CPPU-treated berries. Meanwhile, several corresponding genes, such as VvVIN2 and Vv-A/N-INV2, 8, 10 in ripening berries were obviously down-regulated by CPPU treatment. These results suggested that most of INV members could be triggered by berry enlargement treatment during early berry development, whereas VvVINs and Vv-A/N-INVs, but not VvCWINVs, could be the limiting factor resulting in decreased sugar accumulation in CPPU-treated berries at maturity. In conclusion, this study identified the INV family on the latest annotated grape genome and selected several potential members involving in the limit of CPPU on final sugar accumulation in grape berry. These results provide candidate genes for further study of the molecular regulation of CPPU and GA on sugar accumulation in grape.

Partially coherent twisted vector vortex beam enabling manipulation of high-dimensional classical entanglement.

In this paper, we present a novel form of a partially coherent beam characterized by classical entanglement in higher dimensions. We coin the term "twisted vector vortex (TVV) beam" to describe this phenomenon. Similar to multi-partite quantum entangled states in higher dimensions, the partially coherent twisted vector vortex beam possesses distinct properties such as non-uniform polarization, vortex phase, and twist phase. Through experiments, we offer empirical evidence for these three degrees-of-freedom in the light field. The results demonstrate that the state of the light is inseparable in terms of polarization and orbital angular momentum (OAM) modes. Additionally, the twist phase introduces an additional dimension in controlling the vector vortex beam. This research reveals the possibility of new controlling dimensions in classical entanglement through the chirality of coherence within partially coherent light. Consequently, this opens up new avenues for the utilization of partially coherent light in both classical and quantum domains.

Generating a hollow twisted correlated beam using correlated perturbations.

In this study, a twisted correlated optical beam with a dark hollow center in its average intensity is synthesized by correlated correlation perturbation and incoherent mode superposition. This new hollow beam has a topological charge (TC) mode with a zero value compared with a coherence vortex that has a TC mode with a nonzero value. We transform the twisted correlated beam from solid centered to dark hollow centered by constructing a correlation between the twist factor and the spot structure parameter. Theoretical and experimental results show that twist correlation makes the random optical beam an asymmetric orbital angular momentum spectral distribution and a tunable intensity center. Controlling the correlation parameters can make the focal spot of the twisted beam a dark core when the dominant mode of the TC is still zero. The new nontrivial beams and their proposed generation method provide important technical preparations for the optical particle manipulation with low coherence environment.

Progress in fed-batch culture for recombinant protein production in CHO cells.

Nearly 80% of the approved human therapeutic antibodies are produced by Chinese Hamster Ovary (CHO) cells. To achieve better cell growth and high-yield recombinant protein, fed-batch culture is typically used for recombinant protein production in CHO cells. According to the demand of nutrients consumption, feed medium containing multiple components in cell culture can affect the characteristics of cell growth and improve the yield and quality of recombinant protein. Fed-batch optimization should have a connection with comprehensive factors such as culture environmental parameters, feed composition, and feeding strategy. At present, process intensification (PI) is explored to maintain production flexible and meet forthcoming demands of biotherapeutics process. Here, CHO cell culture, feed composition in fed-batch culture, fed-batch culture environmental parameters, feeding strategies, metabolic byproducts in fed-batch culture, chemostat cultivation, and the intensified fed-batch are reviewed. KEY POINTS: • Fed-batch culture in CHO cells is reviewed. • Fed-batch has become a common technology for recombinant protein production. • Fed batch culture promotes recombinant protein production in CHO cells.

Control of orbital angular momentum of optical vortex beams with complex wandering perturbations.

This work investigates how independent perturbations and cross-correlation perturbations affect optical vortex beams. Theoretical and experimental results show that both perturbations cause the intensity, average orbital angular momentum (OAM), and the OAM spectrum of the vortex beam to vary periodically with the perturbation direction, but with different periods. When the beam is subjected to independent perturbations, the average OAM changes periodically with θ in every π/2; when the beam is subjected to cross-correlation perturbations, the average OAM varies with θ in every π. The results of this work provide a method to control the OAM and regulate low-coherence vortex beams in turbulent environments.

Ultra-high sensitivity strain sensor based on biconical fiber with a bulge air-bubble.

An ultra-high sensitivity Fabry-Perot interferometer (FPI) strain sensor is proposed and experimentally demonstrated. It is composed of a biconical fiber with a bulge air-bubble at its waist. Because of the good stress concentration capability of the tapered fiber, the bulge air-bubble is easily deformed when it is stressed, resulting in an ultra-high strain sensitivity of 101.7 pm/µÉ›, which is the highest strain sensitivity among the direct wavelength interrogation-based strain sensors reported so far, to the best of our knowledge. Moreover, the proposed sensor has a low temperature sensitivity of ∼1pm/∘C; thus an extreme low temperature cross sensitivity of less than 10 nɛ/°C can be achieved.

Progress of cationic gene delivery reagents for non-viral vector.

Gene delivery systems play a vital role in gene therapy and recombinant protein production. The advantages of using gene delivery reagents for non-viral vector include the capacity to accommodate a large packaging load and their low or absent immunogenicity. Furthermore, they are easy to produce at a large scale and preserve. Gene delivery reagents for non-viral vector are commonly used for transfecting a variety of cells and tissues. It is mainly composed of liposomes and non-liposome cationic polymers. According to the different head structures used, the non-viral cationic transfection reagents include a quaternary ammonium salt, amine, amino acid or polypeptide, guanidine salt, and a heterocyclic ring. This article summarizes these approaches and developments of types and components of transfection reagents and optimization of gene delivery. The optimization of mammalian cell transient recombinant protein expression system and cationic reagents for clinical or clinical trials are also discussed.

[Selection of q RT-PCR reference genes for Amomum tsaoko seeds during dormancy release].

Freshly collected seeds of Amomum tsaoko demonstrate obvious dormancy. Therefore, the selection of stable reference genes during seed dormancy release is very important for the subsequent functional research of related genes. In this study, ten commonly used reference genes(GAPDH, 40S, actin, tubulin, EIF4A-9, EIF2α, UBC, UBCE2, 60S, and UBQ) were selected as candidates for quantitative Real-time polymerase chain reaction(qRT-PCR) of the embryo samples of A. tsaoko at different dormancy release stages. Three kinds of software(BestKeeper, geNorm, and Normfinder) and the Delta CT method were used to evaluate the expression stability of the candidate reference genes, and the RefFinder online tool was employed to integrate the results and generate a comprehensive ranking. The results showed that the expression levels of the ten candidate reference genes differed greatly in different embryo samples. GAPDH and UBC had high expression levels, as manifested by the small Ct values. GeNorm identified 40S and UBCE2 as the most stable genes. NormFinder ranked EIF2α as the most stable gene and UBC as the least stable gene. UBCE2 was found to be the most stable gene and actin the least stable one by BestKeeper. Delta CT analysis suggested that the expression of 40S was most stable. UBCE2 was recommended as the most stably expressed gene by RefFinder. Thus, UBCE2 is the ideal reference gene for qRT-PCR analysis of A. tsaoko seeds at different dormancy release stages. The results may lay a foundation for analyzing the expression of related genes during seed dormancy release of A. tsaoko.

Cardiovascular Risk and Safety Evaluation of a Dual Peroxisome Proliferator-Activated Receptor-Alpha/Gamma Agonist, Aleglitazar, in Patients With Type 2 Diabetes: A Meta-analysis.

This study evaluates the cardiovascular risk and safety of a dual peroxisome proliferator-activated receptor alpha and gamma (PPARα&γ), aleglitazar, for the management of type 2 diabetes mellitus. Studies were identified after a literature search in electronic databases and included in the meta-analysis according to eligibility criteria. Meta-analyses of mean differences in the changes from the baseline or odds ratios of selected indices between the aleglitazar- and the placebo/comparator-treated participants were performed. Seven studies {11,832 individuals; age 59.3 years [95% confidence interval (CI) 56.4-61.9]; body mass index 30.8 kg/m [95% CI 30.1-31.7]; sex, 54% males [44-64]} were included. In comparison with the placebo or pioglitazone, the aleglitazar treatment significantly improved %HbA1c, high-density lipoprotein-cholesterol (HDL-chol), and triglycerides. Aleglitazar also significantly decreased fasting plasma glucose and apolipoprotein B compared with the placebo. However, compared with the placebo or pioglitazone, aleglitazar significantly increased serum creatinine levels and significantly decreased the estimated glomerular filtration rate. In addition, the aleglitazar treatment was associated with a significantly increased body weight. Incidence of hypoglycemia, gastrointestinal hemorrhage, bone fractures, heart failure, cardiovascular death, and malignancy was higher in the aleglitazar group. Despite efficacy in glycemic and lipidic control, the aleglitazar treatment was associated with a poor safety profile.

Effects of different 2A peptides on transgene expression mediated by tricistronic vectors in transfected CHO cells.

Multicistronic vectors can increase transgene expression and decrease the imbalance of gene expression in the Chinese hamster ovary (CHO) cell expression system. Small, self-cleaving 2A peptides have a high cleavage efficiency and are essential for constructing high-expression multicistronic vectors. In this study, we investigated the effects of two different 2A peptides on transgene expression in CHO cells via their mediating action on tricistronic vectors. The enhanced green fluorescent protein (eGFP) and red fluorescent protein (RFP) genes were linked by the porcine teschovirus-1 (P2A) and Thosea asigna virus (T2A) peptides in a multicistronic vector. We transfected CHO cells with these vectors and screened for the presence of blasticidin-resistant colonies. Flow cytometry and real-time quantitative PCR (qPCR) were used to detect the expression levels of eGFP and RFP and the copy numbers of stably transfected cells. The results showed that P2A could enhance eGFP and RFP expression by 1.48- and 1.47-fold, respectively, compared to T2A. The expression levels of the genes were not proportional to their copy numbers. In conclusion, we found that P2A can effectively drive transgene expression in CHO cells and a potent 2A peptide can be used for recombinant protein production in the CHO cell system.

Albendazole solid dispersions prepared using PEG6000 and Poloxamer188: formulation, characterization and in vivo evaluation.

This study aimed to optimize the preparation process of albendazole (ABZ) solid dispersion (SD) and enhance its dissolution rate and oral bioavailability in dogs. The ABZ-SD formulations were prepared by a fusion method with ABZ and polyethylene glycol 6000 (PEG 6000), poloxamer 188 (P 188) polymers at various weight ratios or the combination of PEG 6000&P 188. The characterizations of the optimal formulations were performed by scanning electron microscopy (SEM), powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FTIR), in vitro dissolution test and molecular docking. The in vivo pharmacokinetic study was conducted in beagle dogs. As a result, ABZ solid dispersion based on PEG 6000&P 188 (1:2) was successfully prepared. The ABZ-SD formulation could significantly improve the apparent solubility and dissolution rate of ABZ compared with commercial tablets. Furthermore, the water solubility of ABZ-SD was improved mainly based on hydrogen bond association. Besides, at an oral dosage of 15 mg/kg ABZ, the SDs had higher Cmax values and areas under the curve (AUCs) compared to those of commercial ABZ tablets. Preparation of ABZ-loaded SDs by PEG 6000&P 188 is a promising strategy to improve the oral bioavailability of ABZ.

Deciphering an Abnormal Layered-Tunnel Heterostructure Induced by Chemical Substitution for the Sodium Oxide Cathode.

Demands for large-scale energy storage systems have driven the development of layered transition-metal oxide cathodes for room-temperature rechargeable sodium ion batteries (SIBs). Now, an abnormal layered-tunnel heterostructure Na0.44 Co0.1 Mn0.9 O2 cathode material induced by chemical element substitution is reported. By virtue of beneficial synergistic effects, this layered-tunnel electrode shows outstanding electrochemical performance in sodium half-cell system and excellent compatibility with hard carbon anode in sodium full-cell system. The underlying formation process, charge compensation mechanism, phase transition, and sodium-ion storage electrochemistry are clearly articulated and confirmed through combined analyses of in situ high-energy X-ray diffraction and ex situ X-ray absorption spectroscopy as well as operando X-ray diffraction. This crystal structure engineering regulation strategy offers a future outlook into advanced cathode materials for SIBs.

Vernier effect of two cascaded in-fiber Mach-Zehnder interferometers based on a spherical-shaped structure.

The Vernier effect of two cascaded in-fiber Mach-Zehnder interferometers (MZIs) based on a spherical-shaped structure has been investigated. The envelope based on the Vernier effect is actually formed by a frequency component of the superimposed spectrum, and the frequency value is determined by the subtraction between the optical path differences of two cascaded MZIs. A method based on band-pass filtering is put forward to extract the envelope efficiently; strain and curvature measurements are carried out to verify the validity of the method. The results show that the strain and curvature sensitivities are enhanced to -8.47 pm/µÎµ and -33.70 nm/m-1 with magnification factors of 5.4 and -5.4, respectively. The detection limit of the sensors with the Vernier effect is also discussed.

Extensive sequence divergence between the reference genomes of two elite indica rice varieties Zhenshan 97 and Minghui 63.

Asian cultivated rice consists of two subspecies: Oryza sativa subsp. indica and O. sativa subsp. japonica Despite the fact that indica rice accounts for over 70% of total rice production worldwide and is genetically much more diverse, a high-quality reference genome for indica rice has yet to be published. We conducted map-based sequencing of two indica rice lines, Zhenshan 97 (ZS97) and Minghui 63 (MH63), which represent the two major varietal groups of the indica subspecies and are the parents of an elite Chinese hybrid. The genome sequences were assembled into 237 (ZS97) and 181 (MH63) contigs, with an accuracy >99.99%, and covered 90.6% and 93.2% of their estimated genome sizes. Comparative analyses of these two indica genomes uncovered surprising structural differences, especially with respect to inversions, translocations, presence/absence variations, and segmental duplications. Approximately 42% of nontransposable element related genes were identical between the two genomes. Transcriptome analysis of three tissues showed that 1,059-2,217 more genes were expressed in the hybrid than in the parents and that the expressed genes in the hybrid were much more diverse due to their divergence between the parental genomes. The public availability of two high-quality reference genomes for the indica subspecies of rice will have large-ranging implications for plant biology and crop genetic improvement.

Saturable absorber based on a single mode fiber - graded index fiber - single mode fiber structure with inner micro-cavity.

An Er-doped mode-locked fiber laser with a saturable absorber based on single mode - graded index multimode - single mode fiber (SMF-GIMF-SMF) with inner micro-cavity is demonstrated. The modulation depth of the saturable absorber was measured to be 1.9% when the SMF-GIMF-SMF structure is bent to a certain state. Such a simple saturable absorber enables the mode-locking operation in a ring Er-doped fiber laser and ultrafast pulses with pulse energy of 0.026 nJ and pulse width of 528 fs at the fundamental repetition rate of 14.34 MHz can be generated. In addition, the harmonic mode-locking operation can also be achieved.

Stretched graded-index multimode optical fiber as a saturable absorber for erbium-doped fiber laser mode locking.

A novel mode-locking method based on the nonlinear multimode interference in the stretched graded-index multimode optical fiber (GIMF) is proposed in this Letter. The simple device geometry, where the light is coupled in and out of the stretched GIMF via single-mode fibers, is demonstrated to exhibit the temporal intensity discrimination required for mode locking. The nonlinear saturable absorber (SA) characteristics of the device are controllable by simply adjusting the strength of the stretching applied. The modulation depth of the device, which consists of ∼23.5 cm GIMF, is tuned from 10.37% to 22.27%. Such a simple SA enables the wavelength-switchable mode-locking operation in a ring Er-doped fiber laser, and ultrafast pulses with a pulse width of 506 fs at 1572.5 nm and 416 fs at 1591.4 nm were generated. The versatility and simplicity of the SA device, together with the possibility of scaling the pulse energy, make it highly attractive in ultrafast photonics.

Spatial decorrelation of wind noise with porous microphone windscreens.

This paper explores the wind noise reduction mechanism of porous microphone windscreens by investigating the spatial correlation of wind noise. First, the spatial structure of the wind noise signal is studied by simulating the magnitude squared coherence of the pressure measured with two microphones at various separation distances, and it is found that the coherence of the two signals decreases with the separation distance and the wind noise is spatially correlated only within a certain distance less than the turbulence wavelength. Then, the wind noise reduction of the porous microphone windscreen is investigated, and the porous windscreen is found to be the most effective in attenuating wind noise in a certain frequency range, where the windscreen diameter is approximately 2 to 4 times the turbulence wavelengths (2 < D0/ξ < 4), regardless of the wind speed and windscreen diameter. The spatial coherence between the wind noise outside and inside a porous microphone windscreen is compared with that without the windscreen, and the coherence is found to decrease significantly when the windscreen diameter is approximately 2 to 4 times the turbulence wavelengths, corresponding to the most effective wind noise reduction frequency range of the windscreen. Experimental results with a fan are presented to support the simulations. It is concluded that the wind noise reduction mechanism of porous microphone windscreens is related to the spatial decorrelation effect on the wind noise signals provided by the porous material and structure.

PDMS-coated fiber volatile organic compounds sensors.

The functionality of poly(dimethylsiloxane) (PDMS)-based interferometric fiber sensors for volatile organic compounds (VOCs) detection is investigated and experimentally demonstrated. Two interferometric configurations are considered in this work, namely Fabry-Perot (FP) and Sagnac interferometers (SI). Both sensors are functionalized with a thin layer of VOC-sensitive polymer: PDMS, whose degree of swelling varies as a function of VOC concentrations. This swelling effect will result in an optical path length and birefringence modulation for FP and SI sensors, respectively. In this paper, the two common VOCs, ethanol and 2-propanol, were detected by the proposed sensor and the inverse matrix method was used to differentiate the VOC in gas mixture.