Exploring the effects of resin particle sizes on enhancing antibody binding capacity of a hybrid biomimetic ligand.

Particle size is a critical parameter of chromatographic resins that significantly affects protein separation. In this study, effects of resin particle sizes (31.26 µm, 59.85 µm and 85.22 µm named Aga-31, Aga-60 and Aga-85, respectively) on antibody adsorption capacity and separation performance of a hybrid biomimetic ligand were evaluated. Their performance was investigated through static adsorption and breakthrough assays to quantify static and dynamic binding capacity (Qmax and DBC). The static adsorption results revealed that the Qmax for hIgG was 152 mg/g resin with Aga-31, 151 mg/g resin with Aga-60, and 125 mg/g resin with Aga-85. Moreover, the DBC at 10% breakthrough for hIgG with a residence time of 2 min was determined to be 49.4 mg/mL for Aga-31, 45.9 mg/mL for Aga-60, and 38.9 mg/mL for Aga-85. The resins with smaller particle sizes exhibited significantly higher capacity compared to typical commercial agarose resins and a Protein A resin (MabSelect SuRe). Furthermore, the Aga-31 resin with the hybrid biomimetic ligand demonstrated exceptional performance in terms of IgG purity (>98%) and recovery (>96%) after undergoing 20 separation cycles from CHO cell supernatant. These findings are helpful in further chromatographic resin design for the industrial application of antibody separation and purification.

Maresin 1 Activates LGR6 to Alleviate Neuroinflammation via the CREB/JMJD3/IRF4 Pathway in a Rat Model of Subarachnoid Hemorrhage.

Neuroinflammation is an early event of brain injury after subarachnoid hemorrhage (SAH). Whether the macrophage mediators in resolving inflammation 1 (MaR1) is involved in SAH pathogenesis is unknown. In this study, 205 male Sprague-Dawley rats were subjected to SAH via endovascular perforation in the experimental and control groups. MaR1 was dosed intranasally at 1 h after SAH, with LGR6 siRNA and KG-501, GSK-J4 administered to determine the signaling pathway. Neurobehavioral, histological and biochemical data were obtained from the animal groups with designated treatments. The results showed: (i) The leucine-rich repeat containing G protein-coupled receptor 6 (LGR6) was decreased after SAH and reached to the lowest level at 24 h after SAH. Jumonji d3 (JMJD3) protein levels tended to increase and peaked at 24 h after SAH. LGR6 and JMJD3 expression were co-localized with microglia. (ii) MaR1 administration mitigated short-term neurological deficits, brain edema and long-term neurobehavioral performance after SAH, and attenuated microglial activation and neutrophil infiltration. (iii) Knockdown of LGR6, inhibition of CREB phosphorylation or JMJD3 activity abolished the anti-neuroinflammatory effect of MaR1 on the expression of CREB, CBP, JMJD3, IRF4, IRF5, IL-1ß, IL-6 and IL-10, thus prevented microglial activation and neutrophil infiltration. Together, the results show that MaR1 can activate LGR6 and affect CREB/JMJD3/IRF4 signaling to attenuate neuroinflammation after SAH, pointing to a potential pharmacological utility in this disorder.

Synthesis and Bioevaluation of 2-Styrylquinoxaline Derivatives as Tau-PET Tracers.

This study focused on designing and evaluating Tau-PET tracers for noninvasive positron emission computed tomography (PET) imaging of neurofibrillary tangles (NFTs), a hallmark pathology of Alzheimer's disease (AD). The tracers were synthesized with a 2-styrylquinoxaline scaffold and varying lengths of FPEG chains. The compound [18F]15, which had two ethoxy units, showed high affinity for recombinant K18-Tau aggregates (Ki = 41.48 nM) and the highest selectivity versus Aß1-42 aggregates (8.83-fold). In vitro autoradiography and fluorescent staining profiles further validated the binding of [18F]15 or 15 toward NFTs in brain sections from AD patients and Tau-transgenic mice. In normal ICR mice, [18F]15 exhibited an ideal initial brain uptake (11.21% ID/g at 2 min) and moderate washout ratio (2.29), and micro-PET studies in rats confirmed its ability to penetrate the blood-brain barrier with the peak SUV value of 1.94 in the cortex. These results suggest that [18F]15 has the potential to be developed into a useful Tau-PET tracer for early AD diagnosis and evaluation of anti-Tau therapeutics.

Hydroxyethyl-Modified Cycloheptatriene-BODIPY Derivatives as Specific Tau Imaging Probes.

Near-infrared fluorescence (NIRF) imaging as an exquisite sensitive, high spatial-resolution, and real-time tool plays an important role in visualizing pathologies in the brain. In this study, we designed and synthesized a series of NIR probes of hydroxyethyl cycloheptatriene-BODIPY derivatives that have demonstrated strong binding specificity to native neurofibrillary tangles (NFTs) in Alzheimer's disease (AD) brain sections. The improved hydrophilicity of TNIR7-9 and TNIR7-11 resulted in faster clearance rates from healthy brains (4.2 and 10.9, respectively) compared to previously reported compounds. Furthermore, TNIR7-13, which features a fluorinated modification, exhibited a high binding affinity to Tau aggregates (Kd = 11.8 nM) and held promise for future PET studies.

METTL14 promotes fibroblast-like synoviocytes activation via the LASP1/SRC/AKT axis in rheumatoid arthritis.

The objective of this study is to explore the specific roles of a crucial N6-methyladenosine (m6A) methyltransferase, methyltransferase-like 14 (METTL14), in fibroblast-like synoviocytes (FLSs) activation of rheumatoid arthritis (RA). RA rat model was induced by administering intraperitoneally collagen antibody alcohol. Primary fibroblast-like synoviocytes (FLSs) were isolated from joint synovium tissues in rats. shRNA transfection tools were used to downregulate METTL14 expression in vivo and vitro. The injury of joint synovium was shown by hematoxylin and eosin (HE) staining. The cell apoptosis of FLSs was determined by flow cytometry. The levels of IL-6, IL-18, and C-X-C motif chemokine ligand (CXCL)10 in serum and culture supernatants were measured by ELISA kits. The expressions of LIM and SH3 domain protein 1 (LASP1), p-SRC/SRC, and p-AKT/AKT in FLSs and joint synovium tissues were determined by Western blots. The expression of METTL14 was greatly induced in the synovium tissues of RA rats compared with normal control rats. Compared with sh-NC-treated FLSs, METTL14 knockdown significantly increased cell apoptosis, inhibited cell migration and invasion, and suppressed the production of IL-6, IL-18, and CXCL10 induced by TNF-α. METTL14 silencing suppresses the expression of LASP1 and the activation of Src/AKT axis induced by TNF-α in FLSs. METTL14 improves the mRNA stability of LASP1 through m6A modification. In contrast, these were reversed by LASP1 overexpression. Moreover, METTL14 silencing clearly alleviates FLSs activation and inflammation in a RA rat model. These results suggested that METTL14 promotes FLSs activation and related inflammatory response via the LASP1/SRC/AKT signaling pathway and identified METTL14 as a potential target for treating RA.

Doublecortin-Expressing Neurons in Human Cerebral Cortex Layer II and Amygdala from Infancy to 100 Years Old.

A cohort of morphologically heterogenous doublecortin immunoreactive (DCX +) "immature neurons" has been identified in the cerebral cortex largely around layer II and the amygdala largely in the paralaminar nucleus (PLN) among various mammals. To gain a wide spatiotemporal view on these neurons in humans, we examined layer II and amygdalar DCX + neurons in the brains of infants to 100-year-old individuals. Layer II DCX + neurons occurred throughout the cerebrum in the infants/toddlers, mainly in the temporal lobe in the adolescents and adults, and only in the temporal cortex surrounding the amygdala in the elderly. Amygdalar DCX + neurons occurred in all age groups, localized primarily to the PLN, and reduced in number with age. The small-sized DCX + neurons were unipolar or bipolar, and formed migratory chains extending tangentially, obliquely, and inwardly in layers I-III in the cortex, and from the PLN to other nuclei in the amygdala. Morphologically mature-looking neurons had a relatively larger soma and weaker DCX reactivity. In contrast to the above, DCX + neurons in the hippocampal dentate gyrus were only detected in the infant cases in parallelly processed cerebral sections. The present study reveals a broader regional distribution of the cortical layer II DCX + neurons than previously documented in human cerebrum, especially during childhood and adolescence, while both layer II and amygdalar DCX + neurons persist in the temporal lobe lifelong. Layer II and amygdalar DCX + neurons may serve as an essential immature neuronal system to support functional network plasticity in human cerebrum in an age/region-dependent manner.

Bicalutamide, an androgen receptor antagonist, effectively alleviate allergic rhinitis via suppression of PI3K-PKB activity.

OBJECTIVES: To investigate the therapeutic effect of Bicalutamide, an androgen receptor antagonist on the onset and development of allergic rhinitis in an animal model. METHODS: 40 male BALB/c mice were randomly divided into five groups (eight mice per group). Aluminum hydroxide powder was used as an adjuvant, combined with Ovalbumin (OVA) to establish the mouse model of allergic rhinitis via ultrasonic nebulization of OVA to stimulate the nasal cavity. Mice in Bica#1 group were intraperitoneally injected with 0.02 mg Bicalutamide/0.5 ml of normal saline daily for 7 consecutive days; mice in Bica#2 group were administered 0.02 mg Bicalutamide/0.5 ml of normal saline via intraperitoneal injection for 5 consecutive days, and then the same amount of normal saline was injected intraperitoneally for 2 consecutive days. Enzyme-linked immunosorbent assay was adopted to detect the serological levels of IgE, IL-4, and IL-6 production. Eosinophil infiltration was observed under microscope after hematoxylin and eosin staining of nasal mucosa. Quantitative PCR and Western blot were employed for determination of histamine receptors mRNA expression and PI3K/PKB associated protein levels, respectively. RESULTS: Histological analysis shown that allergic lesion was induced after OVA sensitization. Intraperitoneal injection with 0.02 mg Bicalutamide daily for 7 consecutive days significantly reduced the allergic lesion; however, mice injected with the same amount of normal saline at the same time demonstrated no allergic rhinitis symptoms. In addition, there was a significant reduction in eosinophils number in Bicalutamide treated mice (n = 8) compared to the OVA group (n = 8) (OVA: 19.6 ± 5.3 vs. Bica#1: 7.7 ± 0.8 vs. Bica#2: 9.4 ± 1.2, both p < 0.01). Furthermore, ELISA results revealed that the serological levels of IgE (OVA: 17.3 ± 1.7 µg/ml vs. Bica#1: 9.2 ± 0.6 vs. Bica#2: 10.4 ± 2.3, both p < 0.05), IL-4 (OVA: 164.3 ± 5.1 pg/ml vs. Bica#1: 110.2 ± 3.1 vs. Bica#2: 115.3 ± 4.1, both p < 0.05) and IL-6 (OVA: 167.3 ± 3.7 pg/ml vs. Bica#1: 117.5 ± 6.5 vs. Bica#2: 114.8 ± 2.4, both p < 0.05) were significantly decreased after two different dosage of Bicalutamide treatment. Similarly, histamine receptors in mast cells were significantly reduced after two different dosage of Bicalutamide treatment. More importantly, p-PKB protein was notably reduced after two different dosage of Bicalutamide treatment compared to the OVA group, mTOR protein levels were also down regulated after two different dosage of Bicalutamide treatment. CONCLUSIONS: Our data demonstrated that androgen receptor antagonist Bicalutamide can significantly alleviate allergic rhinitis lesion in the animal model. PI3K/PKB activity in mast cells was suppressed after Bicalutamide injection. Our results provide important implication in allergic rhinitis prevention and treatment.

Effects of Drought and Host on the Growth of Santalum album Seedlings in Pot Culture.

Santalum album is a semi parasitic plant and its growth is often restricted due to a lack of a host or water during plantation establishment. In this study, the effects of water and the host on the growth of S. album seedlings were studied in pot culture. The results showed that the net photosynthetic rate and height of S. album seedlings decreased significantly under drought stress. Compared with the seedlings of S. album grown without a host, the host could significantly increase the growth of S. album seedlings. The contents of soluble sugar and proline in S. album leaves increased significantly under drought stress. Drought stress resulted in a significant accumulation of malondialdehyde, increments of antioxidant enzymes activity, and non-enzymatic antioxidant substances. Antioxidant capacity was stronger and malondialdehyde content was lower in the seedling leaves of S. album with a host than in the seedlings without a host. RNA-seq was used to analyze the transcription expression profiles of S. album leaves and the results were consistent with the physiological data. These results indicate that the host can promote the seedling growth of S. album and it can increase the antioxidant capacity and osmotic adjustment substance content of the seedlings of S. album, alleviating the damage caused by drought.

Induction of heartwood formation in young Indian sandalwood (Santalum album L.) by gas elicitors.

Induction of heartwood formation in 6-year-old Indian sandalwood (Santalum album L.) trees by treatment with carbon dioxide, ethylene, nitrogen, and wounding was investigated. All treatments induced fragrant heartwood formation upward and downward from the drill hole. The amount of heartwood formed above and below the drill hole depended on the treatment in the order nitrogen>carbon dioxide>ethylene>wounding, whereas the radial extension proportion was, in order, nitrogen>carbon dioxide>ethylene=wounding. Based on the chemical analysis (GC-MS) and evaluation of the essential oil quality and heartwood properties, heartwood induced by carbon dioxide showed the maximum similarities to naturally formed heartwood, which included the same color, similar chemical composition, reasonable oil content, and quality essential oil, whereas ethylene, nitrogen, and wounding treatment showed fewer similarities to natural heartwood. The results suggest that carbon dioxide is a promising candidate gas elicitor for inducing heartwood formation in young S. album.

Intraneuronal sortilin aggregation relative to granulovacuolar degeneration, tau pathogenesis and sorfra plaque formation in human hippocampal formation.

Extracellular ß-amyloid (Aß) deposition and intraneuronal phosphorylated-tau (pTau) accumulation are the hallmark lesions of Alzheimer's disease (AD). Recently, "sorfra" plaques, named for the extracellular deposition of sortilin c-terminal fragments, are reported as a new AD-related proteopathy, which develop in the human cerebrum resembling the spatiotemporal trajectory of tauopathy. Here, we identified intraneuronal sortilin aggregation as a change related to the development of granulovacuolar degeneration (GVD), tauopathy, and sorfra plaques in the human hippocampal formation. Intraneuronal sortilin aggregation occurred as cytoplasmic inclusions among the pyramidal neurons, co-labeled by antibodies to the extracellular domain and intracellular C-terminal of sortilin. They existed infrequently in the brains of adults, while their density as quantified in the subiculum/CA1 areas increased in the brains from elderly lacking Aß/pTau, with pTau (i.e., primary age-related tauopathy, PART cases), and with Aß/pTau (probably/definitive AD, pAD/AD cases) pathologies. In PART and pAD/AD cases, the intraneuronal sortilin aggregates colocalized partially with various GVD markers including casein kinase 1 delta (Ck1δ) and charged multivesicular body protein 2B (CHMP2B). Single-cell densitometry established an inverse correlation between sortilin immunoreactivity and that of Ck1δ, CHMP2B, p62, and pTau among pyramidal neurons. In pAD/AD cases, the sortilin aggregates were reduced in density as moving from the subiculum to CA subregions, wherein sorfra plaques became fewer and absent. Taken together, we consider intraneuronal sortilin aggregation an aging/stress-related change implicating protein sorting deficit, which can activate protein clearance responses including via enhanced phosphorylation and hydrolysis, thereby promoting GVD, sorfra, and Tau pathogenesis, and ultimately, neuronal destruction and death.

One kind of challenging tetrapeptide biomimetic chromatographic resin for antibody separation.

Short peptide biomimetic chromatography technology as a developing protein separation technology has huge potential for antibody purification. In this study, four tetrapeptide ligands (Ac-FYKH, Ac-YEHF, Ac-YFLH and Ac-FYHI) with high potential binding ability to antibody were selected for the optimal ligand to antibody purification. The results showed that Ac-YEHF-4FF resin had higher binding capacity and selectivity for hIgG among the four resins. And at pH 7.0 and 0.3 ml/min, the highest Q10%-hIgG of Ac-YEHF-4FF resin was 26.2 mg/ml resin while its Q10%-BSA was just 2.2 mg/ml resin. Further, Ac-YEHF-4FF resin was used to purify protein mixtures. By binding at pH 7.0 and being eluted at pH 5.0 and pH 4.0, Ac-YEHF-4FF resin was well used to separate hIgG from BSA containing feedstock, HSA containing feedstock and human serum with the purity and yield both more than 95 %. And the screened resin could also separate mAb from CHO cell culture supernatant with purity 94.3 % and yield 97.5 %. The adsorption and separation results of Ac-YEHF-4FF resin indicated that the goal of getting the efficacy of critical residues from protein A to biomimetic its structure and function could be achieved, which had great significance to the establishment and improvement of tetrapeptide biomimetic chromatography, and also provided a new method for the field of antibody separation and purification.

Model-based evaluation and model-free strategy for process development of three-column periodic counter-current chromatography.

Multi-column counter-current chromatography is an advanced technology used for continuous capture processes to improve process productivity, resin capacity utilization and product consistency. However, process development is difficult due to process complexity. In this work, some general and convenient guidances for three-column periodic counter-current chromatography (3C-PCC) were developed. Boundaries and distributions of operating windows of 3C-PCC processes were clarified by model-based predictions. Interactive effects of feed concentration (c0), resin properties (qmax and De), recovery and regeneration times (tRR) were evaluated over a wide range for maximum productivity (Pmax). Furthermore, variation of Pmax was analyzed considering the constraint factors (capacity utilization target and flow rate limitation). The plateau value of Pmax was determined by qmax and tRR. The operating conditions for Pmax were controlled by qmax, tRR and c0 interactively, and a critical concentration existed to judge whether the operating conditions of Pmax under constraints. Based on the comprehensive understanding on 3C-PCC processes, a model-free strategy was proposed for process development. The optimal operating conditions could be determined based on a set of breakthrough curves, which was used to optimize process performance and screen resins. The approach proposed was validated using monoclonal antibody (mAb) capture with a 3C-PCC system under various mAb and feed concentrations. The results demonstrated that a thorough model-based process understanding on multi-column counter-current chromatography is important and could improve process development and establish a model-free strategy for more convenient applications.

Adaptation of the Invasive Plant (Sphagneticola trilobata L. Pruski) to a High Cadmium Environment by Hybridizing With Native Relatives.

Invasive species can evolve rapidly in the invasion areas to adapt to new habitats. Sphagneticola trilobata L. Pruski, an invasive species, was studied for its tolerance to cadmium (Cd) in the soil and compared with its natural hybrid. From the perspective of photosynthetic physiology, antioxidant characteristics, and leaf hormone levels, the differences between the leaves of the two species before and after Cd treatment were compared. The results showed that the hybrid had stronger tolerance to Cd stress than invasive species. After Cd stress, the indexes of gas-exchange [net photosynthetic rate (Pn), intercellular CO2 concentration (Ci), stomatal conductance (Gs), and transpiration rate (Tr)] of the hybrid was higher than invasive species, while the content of non-enzymatic antioxidants (flavonoids and total phenols) and antioxidant enzyme activities [peroxidase (POD) and superoxide dismutase (SOD)] was lower in hybrid than in invasive species. The changes in the content of plant hormones [auxin (IAA) and abscisic acid (ABA)] under Cd stress showed that hybrid can still maintain growth and prevent leaf senescence. Furthermore, the differences in gene expression between hybrid and invasive species in photosynthetic physiology, the antioxidant capacity of leaves, and endogenous hormone (IAA and ABA) synthesis pathway also showed that hybrid has stronger Cd tolerance than invasive species. This suggests that invasive species will realize the invasion through hybridization with the native relatives to overcome the stress from environmental factors. The study implied that hybridization between invasive species and native relatives is an important way for invasive species to spread in a wider and new environment that invasive species have not experienced in the area of origin.

Study on antibody adsorption and elution performance of carboxyl and hydrophobic groups on mixed-mode ligands.

Molecular interactions between ligands and target biomolecules are crucial in the development of chromatographic techniques for the separation and purification of biotherapeutics. In this study, the role of functional moieties on a mixed-mode ligand (phenylalanine-tyrosine-glutamate-5-aminobenzimidazole) for human immunoglobulin G purification was investigated and a detailed mechanism was discussed. A similar ligand with glutamic acid substituted by glutamine (phenylalanine-tyrosine-glutamine-5-aminobenzimidazole) together with other resins including a commercial resin (CM Bestarose Fast Flow), phenylalanine-tyrosine-glutamate, glutamate-5-aminobenzimidazole, and 5-aminobenzimidazole resins were prepared for comparison. Molecular dynamics simulation and experimental studies were used to analyze the difference between these ligands. The results showed that the carboxyl group of phenylalanine-tyrosine-glutamate-5-aminobenzimidazole contributed 70% of the electrostatic interaction during human immunoglobulin G binding, and 5-aminobenzimidazole provided electrostatic repulsion for desorption, which showed low selectivity and binding capacities at pH 4.0 (dynamic binding capacities at 10% breakthrough of human immunoglobulin G = 1.0 mg/ml resin, dynamic binding capacities at 10% breakthrough of human serum albumin = 1.2 mg/ml resin) when used as an individual resin ligand. The results showed in this study demonstrated that it is possible to achieve optimal antibody separation and purification through reasonable ligand design by understanding the performance of key functional moieties in binding and elution processes.

Tetrandrine Prevents Neomycin-Induced Ototoxicity by Promoting Steroid Biosynthesis.

Aminoglycoside antibiotics are widely used for the treatment of serious acute infections, life-threatening sepsis, and tuberculosis, but all aminoglycosides cause side effects, especially irreversible ototoxicity. The mechanisms underlying the ototoxicity of aminoglycosides need further investigation, and there are no effective drugs in the clinic. Here we showed that tetrandrine (TET), a bioactive bisbenzylisoquinoline alkaloid derived from Stephania tetrandra, ameliorated neomycin-induced cochlear hair cell injury. In both in vitro and in vivo experiments we found that TET administration significantly improved auditory function and reduced hair cell damage after neomycin exposure. In addition, we observed that TET could significantly decrease oxidative stress and apoptosis in hair cells after neomycin exposure. Finally, RNA-seq analysis suggested that TET protected against neomycin-induced ototoxicity mainly by promoting steroid biosynthesis. Collectively, our results provide pharmacological evidence showing that TET may be a promising agent in preventing aminoglycosides-induced ototoxicity.

Decompressive craniectomy can improve the recovery of neurological function, daily living ability and life quality of patients with intracerebral hemorrhage after surgery.

OBJECTIVE: To determine the effect of decompressive craniectomy (DC) on the recovery of neurological function, daily living ability and life quality of patients with intracerebral hemorrhage (ICH) after surgery. METHODS: Totally 290 patients with ICH admitted to our hospital from January 2018 to June 2020 were retrospectively enrolled and assigned to two groups according to different surgical methods. Among them, 138 patients who received craniotomy evacuation of hematoma (CEH) only were assigned to a control group (Con group), while the other 152 who received CEH combined with DC to a research group (Res group). The two groups were compared in the total effective rate, hematoma clearance rate, and complication rate. Additionally, the ICP and MMP-9 levels after surgery, National Institutes of Health Stroke Scale (NIHSS), activities of daily living (ADL), Fugl-Meyer Assessment of motor function (FMA), Glasgow outcome scale (GOS), Glasgow coma scale (GCS), and MOS 36-Item Short-Form Health Survey (SF-36) scores before and after surgery were also compared between the two groups. RESULTS: After treatment, the Res group showed a notably higher total effective rate, hematoma clearance rate, and a notably lower complication rate than the Con group. On postoperative day 3 and 7, the Res group showed notably lower ICP than the Con group, and on postoperative day 7, the Res group showed a notably lower MMP-9 level as compared with the Con group. Additionally, 6 months after the surgery, the Res group got notably lower NIHSS scores and higher ADL, GOS, and SF-36 scores as compared with the Con group, and at 1 month after surgery, the Res group got notably higher FMA scores as compared to the Con group. Moreover, on postoperative day 7, the Res group got notably higher GCS scores than the Con group. CONCLUSION: DC can improve the recovery of neurological function, daily living ability and life quality of patients with ICH after surgery.

Responses of Sphagneticola trilobata, Sphagneticola calendulacea and Their Hybrid to Drought Stress.

Sphagneticola trilobata is an invasive plant in South China. A hybrid between S. trilobata and Sphagneticola calendulacea (a native related species) has also been found in South China. The drought resistance of S. calendulacea, S. trilobata and their hybrid was studied in this paper. Under drought stress, the leaves of S. trilobata synthesized more abscisic acid (ABA) than those of the other species to reduce stomatal opening and water loss. The activities of antioxidant enzymes were the highest in S. trilobata and the lowest in S. calendulacea. The leaves of S. calendulacea suffered the most serious damage, and their maximum photochemical efficiency was the lowest. RNA-sequencing ware used to analyze the expression levels of genes in ABA, antioxidant enzyme, sugar and proline synthesis and photosynthesis pathways. Further real-time PCR detection verified the RNA-sequence results, and the results were in accordance with the physiological data. The results showed that S. trilobata was the most drought tolerant, and the drought tolerance of the hybrid did not show heterosis but was higher than S. calendulacea. Therefore, compared with S. trilobata and the hybrid, the population number and distribution of S. calendulacea may be less in arid areas.

Expression of the Excitatory Postsynaptic Scaffolding Protein, Shank3, in Human Brain: Effect of Age and Alzheimer's Disease.

Shank3 is a postsynaptic scaffolding protein of excitatory synapses. Mutations or variations of SHANK3 are associated with various psychiatric and neurological disorders. We set to determine its normal expression pattern in the human brain, and its change, if any, with age and Alzheimer's disease (AD)-type ß-amyloid (Aß) and Tau pathogenesis. In general, Shank3 immunoreactivity (IR) exhibited largely a neuropil pattern with differential laminar/regional distribution across brain regions. In youth and adults, subsets of pyramidal/multipolar neurons in the cerebrum, striatum, and thalamus showed moderate IR, while some large-sized neurons in the brainstem and the granule cells in the cerebellar cortex exhibited light IR. In double immunofluorescence, Shank3 IR occurred at the sublemmal regions in neuronal somata and large dendrites, apposing to synaptophysin-labeled presynaptic terminals. In aged cases, immunolabeled neuronal somata were reduced, with disrupted neuropil labeling seen in the molecular layer of the dentate gyrus in AD cases. In immunoblot, levels of Shank3 protein were positively correlated with that of the postsynaptic density protein 95 (PSD95) among different brain regions. Levels of Shank3, PSD95, and synaptophysin immunoblotted in the prefrontal, precentral, and cerebellar cortical lysates were reduced in the aged and AD relative to youth and adult groups. Taken together, the differential Shank3 expression among brain structures/regions indicates the varied local density of the excitatory synapses. The enriched Shank3 expression in the forebrain subregions appears inconsistent with a role of this protein in the modulation of high cognitive functions. The decline of its expression in aged and AD brains may relate to the degeneration of excitatory synapses.

Comparison of Protein A affinity resins for twin-column continuous capture processes: Process performance and resin characteristics.

Continuous chromatography is a promising technology for downstream processing of biopharmaceuticals. The operation of continuous processes is significantly different to batch-mode chromatography and needs comprehensive evaluation. In this work, the performances of four Protein A affinity resins were studied systematically for twin-column continuous capture processes. A model-based approach was used to evaluate the process performance (productivity and capacity utilization) under varying operation conditions, and the objective was to reveal the crucial resin properties for continuous capture. The trade-off between productivity and capacity utilization was found, and it is necessary to select appropriate resins for different feedstock and operation conditions. The capacity utilization heavily depends on mass transfer, and steep breakthrough curves are favorable for high capacity utilization. The productivity is determined by both equilibrium binding capacity and mass transfer, and the balance of feed amount and feed time is critical. Moreover, the influence of binding capacity and mass transfer on process productivity and parameter sensitivity with two important resin properties (equilibrium binding capacity qmax and effective pore diffusion coefficient De) were assessed by the model, and suitable resin parameter ranges for twin-column continuous capture were determined. The model-based approach is an effective and useful tool to evaluate the complex performance of different resins and guide the design of next-generation resins for continuous processes.

Strong Response of Stem Photosynthesis to Defoliation in Mikania micrantha Highlights the Contribution of Phenotypic Plasticity to Plant Invasiveness.

Phenotypic plasticity affords invasive plant species the ability to colonize a wide range of habitats, but physiological plasticity of their stems is seldom recognized. Investigation of the stem plasticity of invasive plant species could lead to a better understanding of their invasiveness. We performed pot experiments involving defoliation treatments and isolated culture experiments to determine whether the invasive species Mikania micrantha exhibits greater plasticity in the stems than do three non-invasive species that co-occur in southern China and then explored the mechanism underlying the modification of its stem photosynthesis. Our results showed that the stems of M. micrantha exhibited higher plasticity in terms of either net or gross photosynthetic rate in response to the defoliation treatment. These effects were positively related to an increased stem elongation rate. The enhancement of stem photosynthesis in M. micrantha resulted from the comprehensive action involving increases in the Chl a/b ratio, D1 protein and stomatal aperture, changes in chloroplast morphology and a decrease in anthocyanins. Increased plasticity of stem photosynthesis may improve the survival of M. micrantha under harsh conditions and allow it to rapidly recover from defoliation injuries. Our results highlight that phenotypic plasticity promotes the invasion success of alien plant invaders.