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Recent studies showed that patients with iron overload had increased risk of insulin resistance or diabetes. Ferroptosis is a new type of cell death mainly caused by iron-dependent oxidative damage. In the present study, we investigated potential mechanisms of iron overload induced hepatic ferroptosis and insulin resistance through in vivo and in vitro experiments. In vivo, the mice models of iron overload were established by intraperitoneal injection of iron dextran. The changes of body weight, serum ferritin and blood glucose were measured. Hematoxylin-eosin (HE) and Perl's stainings were used to observe the pathological changes and iron deposition in the liver of mice. In vitro, HepG2 cells were treated with ferric ammonium citrate (FAC, 9 mmol/L, 24 h) to establish the cell models of iron overload. The labile iron pool, cell viability, glucose consumption and glycogen contents were measured. The ultrastructure of mitochondria was observed by transmission electron microscope (TEM). The malondialdehyde (MDA) and glutathione (GSH) kits were used to detect lipid peroxidation in liver tissues of mice and HepG2 cells. RT-PCR and Western blot were used to detect the mRNA and protein expression levels of ferroptosis factors and JAK2/STAT3 signaling pathway. In this study, we used the iron chelator deferasirox in mice and HepG2 cells. Iron overload caused weight loss, elevated serum ferritin, fasting blood glucose, fasting insulin, HOMA-IR, impaired glucose tolerance, and decreased insulin sensitivity in mice. HE staining and Perls staining showed clumps of iron deposition in the liver of iron overload mice. Iron overload could reduce the glucose consumption, increase MDA contents of HepG2 cells, while reduce glycogen and GSH contents in liver tissues of mice and HepG2 cells. TEM showed deletion of mitochondrial ridge and rupture of outer membrane in HepG2 cells with iron overload. Iron chelator deferasirox could significantly improve the above indicators, which might be related to the activation of JAK2/STAT3/SLC7A11 signaling pathway and hepatic ferroptosis. Iron overload could induce hepatic ferroptosis and insulin resistance by inhibiting the JAK2/STAT3/SLC7A11 signaling pathway, and the iron chelator deferasirox might improve hepatic insulin resistance induced by iron overload. [ABSTRACT FROM AUTHOR]

Background: Central pancreatectomy is a surgical procedure for benign and low-grade malignant tumors which located in the neck and proximal body of the pancreas that facilitates the preservation of pancreatic endocrine and exocrine functions but has a high morbidity rate, especially postoperative pancreatic fistula (POPF). The aim of this systematic review and meta-analysis was to evaluate the safety and effectiveness between minimally invasive central pancreatectomy (MICP) and open central pancreatectomy (OCP) basing on perioperative outcomes. Methods: An extensive literature search to compare MICP and OCP was conducted from October 2003 to October 2023 on PubMed, Medline, Embase, Web of Science, and the Cochrane Library. Fixed-effect models or random effects were selected based on heterogeneity, and pooled odds ratios (ORs) or mean differences (MDs) with 95% confidence intervals (CIs) were calculated. Results: A total of 10 studies with a total of 510 patients were included. There was no significant difference in POPF between MICP and OCP (OR = 0.95; 95% CI [0.64, 1.43]; P = 0.82), whereas intraoperative blood loss (MD = − 125.13; 95% CI [− 194.77, -55.49]; P < 0.001) and length of hospital stay (MD = − 2.86; 95% CI [− 5.00, − 0.72]; P = 0.009) were in favor of MICP compared to OCP, and there was a strong trend toward a lower intraoperative transfusion rate in MICP than in OCP (MD = 0.34; 95% CI [0.11, 1.00]; P = 0.05). There was no significant difference in other outcomes between the two groups. Conclusion: MICP was as safe and effective as OCP and had less intraoperative blood loss and a shorter length of hospital stay. However, further studies are needed to confirm the results. [ABSTRACT FROM AUTHOR]

Postoperative pancreatic fistula (POPF) is a severe complication after distal pancreatectomy (DP); however, it is unclear how to effectively reduce the incidence. The purpose of this meta-analysis is to determine whether reinforced stapling reduces POPF after DP. From February 2007 to April 2023, a comprehensive search of electronic data and references was conducted in PubMed/Medline, Embase, Web of Science, the Cochrane Central Register of Controlled Trials, and the Cochrane Database of Systematic Reviews. In this study, the perioperative outcomes were evaluated for the reinforced stapler (RS) group and the standard stapler (SS) group in DP using Review Manager Software. Using fixed- or random-effects models, pooled odds ratios (ORs) and mean differences (MDs) with 95% confidence intervals (CIs) were calculated. In total, three randomized clinical trials (RCTs) with 425 patients and five observational clinical studies (OCS) with 318 patients were included. In pooled meta-analyses from RCTs, there was no difference between the two groups in the incidence of POPF (OR = 0.79; 95% CI [0.47,1.35]; P = 0.39), intraoperative blood loss (MD = 10.66; 95% CI [− 28.83,50.16]; P = 0.6), operative time (MD = 9.88; 95% CI [− 8.92,28.67]; P = 0.3), major morbidity (OR = 1.12; 95% CI [0.67,1.90]; P = 0.66), reoperation (OR = 0.97; 95% CI [0.41,2.32]; P = 0.95), readmission (OR = 0.99; 95% CI [0.57,1.72]; P = 0.97) or hospital stay (MD = − 0.95; 95% CI [− 5.22,3.31]; P = 0.66). However, the results of POPF and readmission were favorable for RS in the OCS group. Key findings: There is no difference in postoperative pancreatic fistula between reinforced and standard staplers in distal pancreatectomy. What is known and what is new? Reinforced staples have been shown to reduce postoperative pancreatic fistula in previous meta-analyses and observational clinical trials, but an increasing number of randomized controlled trials have not reached the same conclusion. This meta-analysis revealed that, in contrast to previous meta-analyses, reinforced stapling did nt reduce postoperative pancreatic fistula in distal pancreatectomy. What is the implication, and what should change now? Surgeons need to carefully consider the clinical role of reinforced staplers and find novel approaches to reduce postoperative pancreatic fistula in distal pancreatectomy. [ABSTRACT FROM AUTHOR]

Chronic neuroinflammation is implicated in the pathogenesis of Parkinson's disease (PD), one of the most common neurodegenerative diseases. Itaconate, an endogenous metabolite derived from the tricarboxylic acid cycle via immune‐responsive gene 1 activity, may mediate anti-inflammatory responses by activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant pathway. This study investigates the neuroprotective potential of 4-octyl itaconate (OI), a cell-permeable derivative of itaconate, in cellular models of PD. OI not only suppressed lipopolysaccharide-induced proinflammatory cascades of inducible nitric oxide synthase, cyclooxygenase-2, and cytokines release in mouse BV2 microglial cells but also activated the Nrf2 signaling pathway and its downstream targets in these cells. Conditioned medium derived from OI-treated BV2 cells protected against rotenone- and MPP+-induced neurotoxicity in Neuro 2A cells. Overall, our findings support the anti-inflammatory neuroprotective potential of OI in PD. [ABSTRACT FROM AUTHOR]

The development of preparation methods for XOFs, as a new class of organic frameworks, plays a crucial role in shaping their structures, functions, and potential applications. In this study, we presented the construction of chiral supramolecular 2D halogen-bonded organic frameworks (XOFs) through a post-synthetic modification strategy. A linear halogen-bonded organic polymer (XOP), XOP-DPBA, decorated with aldehyde groups, was initially prepared to validate the feasibility of post-synthetic modification for XOF construction and functionalization. XOP-DPBA exhibited excellent reactivity with amines, forming imine bonds. By utilizing this reactivity, a series of cross-linked 2D XOFs were efficiently prepared through post-synthetic modified cross-linking reactions. Furthermore, we successfully introduced chiral α/β-cyclodextrins (α/β-CD) into the XOF skeletons via host-guest interactions, resulting in the fabrication of chiral supramolecular 2D XOFs. These chiral XOFs displayed the induced circular dichroism (ICD) signals and assembled them into helical fibers. The post-synthetic modification strategy demonstrated its versatility and simplicity for the construction and functionalization of XOFs. [ABSTRACT FROM AUTHOR]

Lonicera macranthoides, a medicinal herb mainly distributed in southern China, is widely utilized in traditional Chinese medicine for its high yield and strong pharmacological properties rich in phenolic acids and flavonoids. However, the factors regulating plant growth and secondary metabolism in L. macranthoides remain largely unknown. In this study, the effects of seven light qualities and plant growth regulators combinations on seedlings growth and metabolite accumulation were investigated. Plants under red light produced the greatest shoot and root biomass, and promoted plant photosynthesis, as indicated by significantly greater values of net photosynthetic rate (Pn), stomatal conductance (Gs), intercellular CO2 concentration (Ci), and transpiration rate (Tr), followed by white and blue light. The plant height and stem diameter of the tissue-cultured seedlings on the medium with 1.0 mg/L 6-benzyladenine (BA) + 1.0 mg/L indole 3-butyric acid (IBA) were the highest, while the seedlings on the medium containing 2.0 mg/L BA + 0.2 mg/L IBA produced the highest amounts of phenolic acids and flavonoids. Red light dramatically enhanced the accumulation of chlorogenic acid (CGA), ferulic acid, luteoloside, and rutin compared to those under other light spectrum conditions on media with low levels of auxin (0.2 mg/L IBA). However, higher auxin levels (1.0 mg/L IBA) dramatically inhibited CGA production under red light. Moreover, blue light promoted the accumulation of chlorogenic acid and luteoloside. The results indicate that the plant growth and secondary metabolism were dramatically influenced by light quality and auxin/cytokinin combinations. Our findings provide effective strategies aiming to enhance biomass yield and bioactive compound production in L. macranthoides. [ABSTRACT FROM AUTHOR]

Alcohol consumption is a leading risk factor and increases the risk of liver diseases, cancers, tuberculosis, and injuries. The relationship between alcohol use and cardiovascular risk is complex. While it is well established that heavy alcohol use and binge drinking harm cardiovascular health, the effect of light-to-moderate alcohol consumption remains controversial. Observational studies have repeatedly confirmed the U- or J-shaped relationship between alcohol consumption and cardiovascular disease risk, with the lowest risk observed in the light-to-moderate drinking group. However, the protective effect of low-level alcohol has been challenged by recent genetic epidemiological studies with Mendelian randomization. Such studies have their own limitations, and the application of this methodology in studying alcohol has been questioned. Results from the latest Global Burden of Diseases, Injuries, and Risk Factors Study suggest that the impact of alcohol consumption on health depends on the age structure and the distribution of disease burden and underlying causes in a given population. For young adults, even small amounts of alcohol cause heath loss. For older adults facing a high burden of cardiovascular diseases, light-to-moderate alcohol consumption may improve cardiovascular health outcomes. Mechanistically, all types of alcoholic beverages, including wine, spirits, and beer, have been shown to increase the levels of high-density lipoprotein cholesterol and adiponectin, and reduce the level of fibrinogen. Nonalcoholic components of wine, especially polyphenolic compounds like resveratrol, may additionally enhance endothelial nitric oxide production, and provide antioxidant and anti-inflammatory effects. [ABSTRACT FROM AUTHOR]

Hypertensive disorders of pregnancy are complications that can lead to maternal and infant mortality and morbidity. Hypertensive disorders of pregnancy are generally defined as hypertension and may be accompanied by other end organ damages including proteinuria, maternal organ disturbances including renal insufficiency, neurological complications, thrombocytopenia, impaired liver function, or uteroplacental dysfunction such as fetal growth restriction and stillbirth. Although the causes of these hypertensive disorders of pregnancy are multifactorial and elusive, they seem to share some common vascular-related mechanisms, including diseased spiral arteries, placental ischemia, and endothelial dysfunction. Recently, preeclampsia is being considered as a vascular disorder. Unfortunately, due to the complex etiology of preeclampsia and safety concerns on drug usage during pregnancy, there is still no effective pharmacological treatments available for preeclampsia yet. An emerging area of interest in this research field is the potential beneficial effects of dietary intervention on reducing the risk of preeclampsia. Recent studies have been focused on the association between deficiencies or excesses of some nutrients and complications during pregnancy, fetal growth and development, and later risk of cardiovascular and metabolic diseases in the offspring. In this review, we discuss the involvement of placental vascular dysfunction in preeclampsia. We summarize the current understanding of the association between abnormal placentation and preeclampsia in a vascular perspective. Finally, we evaluate several studied dietary supplementations to prevent and reduce the risk of preeclampsia, targeting placental vascular development and function, leading to improved pregnancy and postnatal outcomes. [ABSTRACT FROM AUTHOR]

In this study, the structural changes of chitosan- (C) shellac (S) bio-based emulsions induced by the incorporation of pine needle essential oil (PNEO) were investigated, in addition to, the effect of enhancements in physical, functional, and antibacterial properties of coatings on egg preservation. The ability of emulsion-based coatings to combine the structural strength of the hydrophilic phase with the hydrophobicity of the lipid was interpreted. Rheological analysis indicated that an appropriate amount of PNEO endowed the coating-forming emulsions (CFE) with proper viscosity and gel properties. The C-S-PNEO3 combined with 0.6 g PNEO had the optimal application and storage stability reflected by the lowest particle size and potential. SEM observed a flatter surface morphology of the coatings with the addition of 0.4 and 0.6 g PNEO, while XRD and FTIR also revealed that PNEO improved the biocompatibility by reducing the crystallinity of the coating, while hydrogen bonds were formed between chitosan, shellac. Furthermore, the physical properties of the coating, such as water vapor permeability (WVP), gas permeability, light transmittance, and color parameters were affected by the PNEO. The results showed that the uniform texture, excellent water vapor, oxygen, ultraviolet (UV) barrier performance, and bacteriostatic effect of the C-S-PNEO3 coating contributed to egg packaging. Highlights: PNEO incorporation affected the properties of emulsions as well as the properties of coatings. Coating-forming emulsions have good structural properties and storage stability. C-S-PNEO3 was endowed with strong barrier properties and antibacterial activity for eggs preservation. PNEO had good biocompatibility with C and S determined by XRD. Prepared bio-based emulsion coatings were an emerging egg packaging material. [ABSTRACT FROM AUTHOR]

Chemical space exploration is a major task of the hit-finding process during the pursuit of novel chemical entities. Compared with other screening technologies, computational de novo design has become a popular approach to overcome the limitation of current chemical libraries. Here, we reported a de novo design platform named systemic evolutionary chemical space explorer (SECSE). The platform was conceptually inspired by fragment-based drug design, that miniaturized a "lego-building" process within the pocket of a certain target. The key to virtual hits generation was then turned into a computational search problem. To enhance search and optimization, human intelligence and deep learning were integrated. Application of SECSE against phosphoglycerate dehydrogenase (PHGDH), proved its potential in finding novel and diverse small molecules that are attractive starting points for further validation. This platform is open-sourced and the code is available at http://github.com/KeenThera/SECSE. [ABSTRACT FROM AUTHOR]

Background: Lonicera macranthoides is an important woody plant with high medicinal values widely cultivated in southern China. WRKY, one of the largest transcription factor families, participates in plant development, senescence, and stress responses. However, a comprehensive study of the WRKY family in L. macranthoides hasn't been reported previously. Objective: To establish an extensive overview of the WRKY family in L. macranthoides and identify senescence-responsive members of LmWRKYs. Methods: RNA-Seq and phylogenetic analysis were employed to identify the LmWRKYs and their evolutionary relationships. Quantitative real-time (qRT-PCR) and transgenic technology was utilized to investigate the roles of LmWRKYs in response to developmental-, cold-, and ethylene-induced senescence. Results: A total of 61 LmWRKY genes with a highly conserved motif WRKYGQK were identified. Phylogenetic analysis of LmWRKYs together with their orthologs from Arabidopsis classified them into three groups, with the number of 15, 39, and 7, respectively. 17 LmWRKYs were identified to be differentially expressed between young and aging leaves by RNA-Seq. Further qRT-PCR analysis showed 15 and 5 LmWRKY genes were significantly induced responding to tissue senescence in leaves and stems, respectively. What's more, five LmWRKYs, including LmWRKY4, LmWRKY5, LmWRKY6, LmWRKY11, and LmWRKY16 were dramatically upregulated under cold and ethylene treatment in both leaves and stems, indicating their involvements commonly in developmental- and stress-induced senescence. In addition, function analysis revealed LmWRKY16, a homolog of AtWRKY75, can accelerate plant senescence, as evidenced by leaf yellowing during reproductive growth in LmWRKY16-overexpressing tobaccos. Conclusion: The results lay the foundation for molecular characterization of LmWRKYs in plant senescence. [ABSTRACT FROM AUTHOR]

Due to catch-up growth (CUG), there are adverse effects on human health. However, there is little information about its influence on bone metabolism. This study aimed to investigate the effects of leptin on bone metabolism and formation during high-fat diet (HFD)-induced CUG. We randomly divided male Wistar rats (5 weeks old) into four groups: control (CTL), caloric restriction and normal chow (RN), caloric restriction (4 weeks), and HFD (RH), and RH + leptin antagonist (RH + LEPA). We monitored body weights, biochemical markers, and epididymal and perirenal fat in these rats. We then performed Hematoxylin and Eosin (H&E) staining to evaluate bone metabolism. We detected osteoprotegerin (OPG) and receptor activator of nuclear factor-kappa b ligand (RANKL) by qRT-PCR and immunohistochemistry (IHC). We found that HFD increased the body weights in rats. In RN, RH, and RH + LEPA groups, major biochemical markers of bone metabolism in rat serum were significantly altered. We found that epididymal and perirenal fat tissues of RH and RH + LEPA groups were higher than those in the RN group. Severe bone formation impairment in the distal diaphysis and metaphysis of the left femora and lumbar vertebra was seen in the RH group compared to RN, which was even aggravated by a leptin antagonist. OPG in the left femora and lumbar vertebra was lower in RH than the RN group. The leptin antagonist decreased OPG during CUG in the RH group, whereas RANKL expression showed an opposite alteration. During HFD-induced CUG, bone formation was mediated by OPG and RANKL and was affected by the leptin content. [ABSTRACT FROM AUTHOR]

Preeclampsia is a common medical condition during pregnancy and a major cause of maternal and prenatal mortality. The present study was conducted to investigate the effects of maternal treatment with pentaerythritol tetranitrate (PETN) in Dahl salt-sensitive rats (DSSR), a model of superimposed preeclampsia. F0 parental DSSR were treated with PETN (50 mg/kg) from the time point of mating to the end of lactation. Maternal PETN treatment improved fetal growth and had no effect on blood pressure in DSSR offspring fed with normal chow or high-salt diet. Upon high-fat diet (HFD) feeding, offspring from PETN-treated mother showed improved glucose tolerance despite similar weight gain. Unexpectedly, maternal PETN treatment significantly potentiated the HFD-induced blood pressure elevation in male DSSR offspring. Endothelium-derived hyperpolarization factor (EDHF)-mediated vasodilation was similar between NCD-fed and HFD-fed control offspring but was markedly reduced in HFD-fed PETN offspring. EDHF genes were downregulated in the vasculature of HFD-fed PETN offspring, which was associated with epigenetic changes in histone modifications. In conclusion, maternal PETN treatment in DSSR shows both beneficial and unfavorable effects. It improves fetal growth and ameliorates glucose tolerance in the offspring. Although maternal PETN treatment has no effect on blood pressure in offspring fed with normal chow or high-salt diet, the offspring is at higher risk to develop HFD-induced hypertension. PETN may potentiate the blood pressure response to HFD by epigenetic modifications of EDHF genes. Key messages: The core findings of this article suggest that maternal PETN treatment of DSSR, a rat model of a spontaneous superimposed preeclampsia, leads to • Improvement of fetal growth; • No changes of maternal blood pressure or markers of preeclampsia; • Amelioration of HFD-induced glucose intolerance in adult offspring; • No changes in blood pressure development of the offspring on normal chow or high salt-diet; • Potentiation of blood pressure elevation of the offspring on HFD. [ABSTRACT FROM AUTHOR]

Efficacy evaluation through human trials is crucial for advancing a vaccine candidate to clinics. Next-generation sequencing (NGS) can be used to quantify B cell repertoire response and trace antibody lineages during vaccination. Here, we demonstrate this application with a case study of Hecolin®, the licensed vaccine for hepatitis E virus (HEV). Four subjects are administered the vaccine following a standard three-dose schedule. Vaccine-induced antibodies exhibit a high degree of clonal diversity, recognize five conformational antigenic sites of the genotype 1 HEV p239 antigen, and cross-react with other genotypes. Unbiased repertoire sequencing is performed for seven time points over six months of vaccination, with maturation pathways characterize for a set of vaccine-induced antibodies. In addition to dynamic repertoire profiles, NGS analysis reveals differential patterns of HEV-specific antibody lineages and highlights the necessity of the long vaccine boost. Together, our study presents a quantitative strategy for vaccine evaluation in small-scale human studies. The authors provide a comprehensive characterization of the human antibody response to a licensed hepatitis E virus (HEV) vaccine, Hecolin, in four individuals over the course of six months post vaccination. They demonstrate diverse patterns of antibody response underlying the vaccine protection. [ABSTRACT FROM AUTHOR]

Impedance spectroscopy was conducted on colloidal ITO thin films that had been subjected to alternating oxygen and argon plasma treatments, followed by air annealing from 150 to 750 °C. An equivalent circuit consisting of an RC element nested within another RC element, featuring a negative resistance and a negative capacitance, fitted the data well. These results are interpreted as being due to surface plasmons that are a function of the presence of nanoporous ITO-rich regions surrounded by isolated ITO nanoparticles coated with an amorphous polymer that intertwines with the ITO-rich regions as a function of annealing treatment. [ABSTRACT FROM AUTHOR]

Biotinylated peptide-Cu2+ nanoparticles (Cu-P NPs) were synthesized via "one-pot" self-assembly. The peptide P conststs of a hydrophobic dipeptide (FF), a tripeptide (KGH), and a biotin moiety attached to the terminal amino group of the Lys residue. The Cu-P NPs contain abundant catalytically active Cu2+ ions which are liberated by acid-induced dissolution. The released Cu2+ ions catalyze the oxidization of 3,3′,5,5′-tetramethylbenzidine (TMB) by H2O2 because of their intrinsic peroxidase activity, and this results in the formation of a blue-green coloration. Based on the streptavidin–biotin interaction, the Cu-P NPs were employed to establish an enzyme-free colorimetric method for determination of prostate-specific antigen (PSA) as a model biomarker. Under optimal conditions, the linear response range is 0.001–1 ng mL−1, with a limit of detection as low as 1 pg mL−1. [ABSTRACT FROM AUTHOR]

The aim of the present study was to compare different diets used to induce obesity in a head-to-head manner with a focus on insulin resistance and vascular dysfunction. Male C57BL/6J mice were put on standard chow diet (SCD), normal-fat diet (NFD), cafeteria diet (CAF) or high-fat diet (HFD) for 12 weeks starting at the age of 6 weeks. Both CAF and HFD led to obesity (weight gain of 179% and 194%, respectively), glucose intolerance and insulin resistance to a comparable extent. In aortas containing perivascular adipose tissue (PVAT), acetylcholine-induced vasodilation was best in the NFD group and worst in the CAF group. Reduced phosphorylation of endothelial nitric oxide synthase at serine 1177 was observed in both CAF and HFD groups. Plasma coagulation activity was highest in the HFD group and lowest in the SCD group. Even the NFD group had significantly higher coagulation activity than the SCD group. In conclusions, CAF and HFD are both reliable mouse diets in inducing visceral obesity, glucose intolerance and insulin resistance. CAF is more effective than HFD in causing PVAT dysfunction and vascular dysfunction, whereas hypercoagulability was mostly evident in the HFD group. Coagulation activity was higher in NFD than NCD group. [ABSTRACT FROM AUTHOR]

Hepatitis B virus (HBV) X protein, HBx, interacts with anti-apoptotic Bcl-2 and Bcl-xL proteins through its BH3-like motif to promote HBV replication and cytotoxicity. Here we report the crystal structure of HBx BH3-like motif in complex with Bcl-xL where the BH3-like motif adopts a short α-helix to snuggle into a hydrophobic pocket in Bcl-xL via its noncanonical Trp120 residue and conserved Leu123 residue. This binding pocket is ~2 Å away from the canonical BH3-only binding pocket in structures of Bcl-xL with proapoptotic BH3-only proteins. Mutations altering Trp120 and Leu123 in HBx impair its binding to Bcl-xL in vitro and HBV replication in vivo, confirming the importance of this motif to HBV. A HBx BH3-like peptide, HBx-aa113-135, restores HBV replication from a HBx-null HBV replicon, while a shorter peptide, HBx-aa118-127, inhibits HBV replication. These results provide crucial structural and functional insights into drug designs for inhibiting HBV replication and treating HBV patients. Hepatitis B virus X protein (HBx) binds anti-apoptotic Bcl-xL through its BH3-like motif to promote viral replication. Here, the authors provide the structure of the HBx BH3-like domain and Bcl-xL, which shows an unusual mode of interaction, and identify a short peptide that inhibits HBV replication in cultured human hepatic cells. [ABSTRACT FROM AUTHOR]

Hepatitis E virus (HEV) is one of the major pathogens that cause acute viral hepatitis. The human (genotypes 1 and 2) and zoonotic (genotypes 3 and 4) groups of HEV present different epidemiology and clinical features. In this study, we developed a classification method for rapidly classifying HEV into human or zoonotic groups that combines a general antigen test with a zoonotic group-specific antigen test. Evaluation of serial samples from HEV-infected rhesus monkeys indicated that HEV antigen-positive samples can be classified using the antigen-based classification method. The antigen-based classification method was evaluated further on 55 genotyped samples from acute hepatitis E patients, including 9 human and 46 zoonotic groups. The novel method was completely consistent with the sequencing results: 9/9 for the human groups (100%, 95% confidence interval [CI] 66.4-100%) and 46/46 for the zoonotic groups (100%, 95% CI 92.3-100%). This method was also successfully used for the clustering of some samples that could not be clustered by sequencing. Compared with the sequencing-based method, this method is less time-consuming, less expensive, and less technically complex and is therefore ideal for large numbers of samples. In conclusion, this study provides a convenient and sensitive method for classifying different groups of HEV, and it has potentially important public health applications, especially in underdeveloped areas that cannot afford the high cost of nucleic acid testing. [ABSTRACT FROM AUTHOR]

The authors report on an amperometric method for the determination of glycoprotein by using an aptamer as the bioreceptor. The detection scheme is making use of aggregated citrate-capped silver nanoparticles (AgNPs) on a gold electrode. Aggregation is accomplished by addition of 4-mercaptophenylboronic acid (MPBA) which acts as a cross-linker due to the formation of Ag-S bonds and of boronate esters. A thiolated DNA aptamer was then attached to the electrode in order to capture glycoprotein. Once captured, the glycoprotein reacts with MPBA through the formation of boronate esters. The electrochemical signal is thus amplified by the formation of a network of AgNPs which act as redox reporters. To demonstrate the feasibility of the method, prostate specific antigen (PSA) was chosen as a model analyte. The detection limit for PSA is as low as 0.2 pg mL. In our preception, this method provides a powerful tool for studying the glycan function in biological and physiological processes. [ABSTRACT FROM AUTHOR]

This paper compared the potential application of gas chromatograph (GC) and differential scanning calorimetry (DSC) to verify adulteration of camellia oil (CMO) with sesame oil (SSO), sunflower oil (SFO), peanut oil (PNO), corn oil (CO) and canola oil (CNO) which are cheaper oils mixed as adulterants with CMO. DSC offers unique thermal profiling for each oil. A combination of analysis of FAs and fingerprint were applied for GC to detect the adulteration. According to a similarity calculation (with a standard below 0.9989) of included angle, the detection limit of sesame oil, sunflower oil and corn oil was 10 %, peanut oil 20 %, and rapeseed oil 30 %; for DSC, similarly CMOs had the unique fingerprint according to their DSC peak shape and thermodynamic parameters, and an adulteration contents of 5 % could be detected qualitatively. Satisfactory results were achieved from stepwise multiple linear regression analysis (SMLR) for the data of T , T , T , △ H and peak height ( H) of DSC to quantitatively predict the other five oils adulteration in CMO with R to 0.999. The average error obtained from the error analysis corresponding to SMLR was 1.2620 %. The preliminary results presented in this study suggest that DSC analysis is an attractive tool in detecting SSO, SFO, PNO, CO and CNO adulteration in CMO. [ABSTRACT FROM AUTHOR]

The authors describe an aptasensor for visual and fluorescent detection of lysozyme via an inner filter effect (IFE). The assay is based on the fact that red gold nanoparticles (AuNPs) act as powerful absorbers of the green fluorescence of CdTe because of spectral overlap. If the lysozyme-binding aptamer is adsorbed onto the surface of the AuNPs, the salt-induced aggregation of AuNPs (that leads to a color change from red to blue) does not occur and the IFE remains efficient. If lysozyme is present, it will bind the aptamer and thereby prevent its adsorption on the AuNPs. As a result, the salt-triggered aggregation of the AuNPs will occur. Consequently, color will change from red to blue, and green fluorescence will pop up because the IFE is suppressed. Under optimum conditions, fluorescence is linearly related to lysozyme concentration in the 1.0 nM to 20 nM concentration range, with a 0.55 nM limit of detection. The method is perceived to be of wider applicability in that it may be used to design other visual and fluorescent assays if appropriate aptamers are available. [ABSTRACT FROM AUTHOR]

This paper investigates the free vibration of a homogeneous Euler-Bernoulli beam with multiple transverse cracks under non-symmetric boundary conditions. The differential equation is formulated by introducing Dirac's delta function into the uniform flexural stiffness, and the close-form solution of mode shapes is then derived by applying the Laplace transform technique. The proposed method is validated against existing experimental method for damaged cantilever beams. With the validated method, a parametric study is performed to study the effect of crack numbers, damage parameters and crack locations on the natural frequencies and mode shapes for three non-symmetric boundary conditions (pinned-clamped, clamped-free shear and pinned-free shear). [ABSTRACT FROM AUTHOR]

The enzyme cleaving the beta-site amyloid precursor protein (BACE1) is a prime therapeutic target for treatment of Alzheimer's disease (AD). Hence, improved high-throughput methods for probing the activity of BACE1 are crucial with respect to the discovery of novel drugs for treatment and prevention of AD. We describe a graphene oxide (GO)-based fluorescent platform for the determination of BACE1 activity and screening of its inhibitors. It is found that covalent linkage of the fluorescein isothiocyanate (FITC)-labeled enzyme substrate (of sequence GTEEISEVNLDAEFRHDSGYKK) to GO results in quenching of the FITC fluorescence, but that on peptide cleavage by BACE1, FITC is released from the GO surface and fluorescence is restored. In contrast to other fluorescent assays, the method presented here shows much higher quenching efficiency and sensitivity, and it can be used at pH value of 4.5 where BACE1 exhibits higher activity. The inhibition of BACE1 activity by a potential inhibitor results in retarded fluorescence recovery. The half-maximum inhibition value of a well-known BACE1 inhibitor was found to be 82.7 nM, which is in agreement with the value reported earlier. In our perception, this method represents a valuable tool for screening of BACE1 inhibitors and discovery of AD drugs in a high-throughput screening format using microplate readers. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Objective: Augmenter of liver regeneration (ALR) is a growth factor that is ubiquitously expressed in multiple forms among eukaryotes. The present study focused on the role of endogenous ALR on the hypoxia/reoxygenation (H/R)-induced inflammatory response in human kidney 2 (HK-2) cells, and the underlying molecular mechanisms. Methods: To determine the relationship between exogenous and endogenous ALR, exogenous ALR was administrated to HK-2 cells, and endogenous ALR protein and mRNA expression was examined by Western blotting and quantitative real-time polymerase chain reaction (qPCR), respectively. In order to knockdown endogenous ALR expression, HK-2 cells were infected with lentiviral shRNA/ALR, after which cell viability was determined by the MTS cell viability assay. Cells were subjected to hypoxia for 6 h and reoxygenation for 12 h. Levels of monocyte chemotactic protein (MCP-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA) and qPCR. Cells were harvested, and nuclear and phosphorylated protein extracts were prepared from the HK-2 cell lysates. Nuclear factor κB (NF-κB), and phosphorylated extracellular signal-regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK) were analyzed by Western blotting. The translocation of NF-κB was detected by immunofluorescence. Results: Exogenous ALR inhibited the expression of endogenous ALR. Lentiviral shRNA/ALR markedly downregulated endogenous ALR expression, whereas there were no changes in ALR expression in lentiviral shRNA/control HK-2 cells. The results of the MTS assay showed that silencing ALR expression did not influence cell viability. H/R led to increased production of MCP-1, IL-6, and TNF-α. However, knockdown of ALR attenuated the inflammatory response via inhibition of ERK, p38, and JNK phosphorylation. The translocation of NF-κB into the nucleus was also decreased. Conclusions: These results suggest that there is a negative feedback loop involving ALR in HK-2 cells. Knockdown of ALR exerts anti-inflammatory actions via suppression of the mitogen-activated protein kinase signaling pathway. [ABSTRACT FROM AUTHOR]

Purpose: The effects of augmenter of liver regeneration (ALR) on the acute kidney injury (AKI) rats were investigated by measuring the inflammatory response associated with transcription factor nuclear factory (NF-κB) pathway. Methods: The model of AKI rats was established by occluded the renal pedicles for 60 min and then released. After that, animals were treated with ALR (100 or 200 μg/kg). All rats were killed at different time points (24, 48, 72 h). Renal function and kidney histological changes were measured. The apoptosis of tubular cells was evaluated by TdT-mediated dUTP nick end labeling assay. Cytokines and chemokines were assessed by immunohistochemistry, enzyme-linked immunosorbent assay and real-time polymerase chain reaction (RT-PCR). The NF-κB p65 protein was analyzed by immunohistochemistry and RT-PCR, respectively. Results: Ischemia reperfusion induced tubular cells necrosis and apoptosis, and ALR can significantly reduce this damages. The productions of MCP-1, IL-1β and IL-6 were lower in the group of ALR treatment, especially in the high-dose group. The inflammatory infiltrates were lower in the rats with administration of ALR. ALR mediated the level of cytokines and chemokines through inhibited the activation of NF-κB. Conclusion: ALR can improve renal function and inhibit the expression of inflammatory factors. This protects against renal ischemia reperfusion injury, which may be associated with preventing NF-κB activation in rats. [ABSTRACT FROM AUTHOR]

Oxidative stress plays an important role in cellular destruction. Augmenter of liver regeneration (ALR) is an anti-apoptotic factor that is expressed in all mammalian cells and functions as an anti-oxidant by stimulating the expression of a secretory isoform of clusterin and inhibiting reactive oxygen species (ROS) generation. Previous work from our group showed that ALR expression is upregulated in acute kidney injury (AKI) rats, and recombinant human ALR reduces tubular injury. In the present study, we used small interfering RNA (siRNA) silencing of ALR to examine its role in HO induced mitochondrial injury and apoptosis. Knockdown of ALR increased ROS levels, reduced mitochondrial membrane potential, and increased the release of mitochondrial proteins and the rate of apoptosis in response to HO. In addition, the ratio of Bax/Bcl-2 was increased in siRNA/ALR groups treated with HO. These data confirm the protective role of ALR against oxidative stress-induced mitochondrial injury and suggest a potential mechanism underlying the protective role of ALR in AKI. [ABSTRACT FROM AUTHOR]

Chloroplast markers are powerful tools for research into relationships among grapevine ( Vitis vinifera L. subsp. vinifera) cultivars. However, the high number of regions common to the chloroplast and mitochondrion genomes described in grapevine could interfere with correct phylogeny reconstruction. In this study, we established a unique chloroplast marker (UCM) set and investigated the chloroplast haplotype diversity of 17 grapevine cultivars and seven Asian Vitis species. Sequencing of four UCMs revealed four haplotype groups in grapevine based on three nucleotide substitutions, three simple sequence repeats, and one 54 nucleotide (nt) deletion in the trnC- petN region. The constructed molecular-variance parsimony and statistical parsimony networks indicated that two independent maternal lineages of grapevine cultivars originated from a core group of Asian Vitis species. The presence and absence of a newly discovered 54 nt deletion corresponded to the two maternal lineages: deletion (D) and non-deletion (ND) lineages. The D lineage consisted of three haplotypes, represented by 'Furmint', 'Gouais blanc', and 'Sauvignon blanc'. Ancient noble cultivars, such as 'Pinot noir', 'Riesling', and 'Afus ali', belong to a single haplotype group (ND) in which the deletion was absent. Our results provide compelling evidence for the dual maternal origin of grapevine, and suggest the occurrence of an ancient hybridization event. [ABSTRACT FROM AUTHOR]

Sulfonated graphene nanosheet/gold nanoparticle (SGN/Au) hybrid was synthesized by electrostatic self-assembly of anionic SGN and positively charged gold nanoparticles. Due to the well-dispersivity of SGN in aqueous solution and its adequate negative charge, Au nanoparticles were assembled uniformly on graphene surface with high distribution. With the advantages of both graphene and Au nanoparticles, SGN/Au hybrid showed enhanced electrocatalytic activity towards O reduction. Furthermore, it provided a conductive and favorable microenvironment for the glucose oxidase (GOD) immobilization and thus promoted its direct electron transfer at the glassy carbon electrode. Based on the consumption of O caused by glucose at the interface of GOD electrode modified with SGN/Au hybrid, the modified electrode displayed satisfactory analytical performance, including high sensitivity (14.55 μA mM cm), low detection limit (0.2 mM), an acceptable linear range from 2 to 16 mM, and also the prevention from the interference of some species. These results indicated that the prepared SGN/Au hybrid is a promising candidate material for high-performance glucose biosensor. [ABSTRACT FROM AUTHOR]

There is emerging evidence for an under-recognized hepatitis E virus (HEV) as a human pathogen. Among different reasons for this neglect are the unsatisfactory performance and under-utilization of commercial HEV diagnostic kits; for instance, the number of anti-HEV IgM kits marketed in China is about one-fifth of that of hepatitis A kits. Over the last two decades, substantial progress has been achieved in furthering our knowledge on the HEV-specific immune responses, antigenic features of HEV virions, and development of serological assays and more recently prophylactic vaccines. This review will focus on presenting the evidence of the importance of HEV infection for certain cohorts such as pregnant women, the key antigenic determinants of the virus, and immunogenicity and clinical efficacy conferred by a newly developed prophylactic vaccine. Robust immunogenicity, greater than 195-fold and approximately 50-fold increase of anti-HEV IgG level in seronegative and seropositive vaccinees, respectively, as well as impressive clinical efficacy of this vaccine was demonstrated. The protection rate against the hepatitis E disease and the virus infection was shown to be 100% (95% CI 75-100) and 78% (95% CI 66-86), respectively. [ABSTRACT FROM AUTHOR]

Nox4 is a member of the NADPH oxidase family, which represents a major source of reactive oxygen species (ROS) in the vascular wall. Nox4-mediated ROS production mainly depends on the expression levels of the enzyme. The present study was aimed to investigate the mechanisms of Nox4 transcription regulation by histone deacetylases (HDAC). In human umbilical vein endothelial cells (HUVEC) and HUVEC-derived EA.hy 926 cells, treatment with the pan-HDAC inhibitor scriptaid led to a marked decrease in Nox4 mRNA expression. A similar down-regulation of Nox4 mRNA expression was observed by siRNA-mediated knockdown of HDAC3. HDAC inhibition in endothelial cells was associated with enhanced histone acetylation, increased chromatin accessibility in the human Nox4 promoter region, with no significant changes in DNA methylation. In addition, we provided evidence that c-Jun played an important role in controlling Nox4 transcription. Knockdown of c-Jun with siRNA led to a down-regulation of Nox4 mRNA expression. In response to scriptaid treatment, the binding of c-Jun to the Nox4 promoter region was reduced despite the open chromatin structure. In parallel, the binding of RNA polymerase IIa to the Nox4 promoter was significantly inhibited as well, which may explain the reduction in Nox4 transcription. In conclusion, HDAC inhibition decreases Nox4 transcription in human endothelial cells by preventing the binding of transcription factor(s) and polymerase(s) to the Nox4 promoter, most likely because of a hyperacetylation-mediated steric inhibition. [ABSTRACT FROM AUTHOR]

Lysyl oxidase-like 2 (LOXL2) belongs to an amine oxidase family whose members have been implicated in crosslink formation in stromal collagens and elastin, cell motility, and tumor development and progression. Both down- and up-regulation of LOXL in tumor tissues and cancer cell lines have been described, suggesting paradoxical roles in cancer. However, LOXL2 expression and the clinical significance in non-small cell lung cancers (NSCLC) remain unresolved. Real-time PCR was performed to detect the expression of LOXL2 mRNA in lung tumor tissues (TT) and surrounding normal tissues (sNT). Moreover, the expression of the LOXL2 protein in specimens from 83 paraffin-embedded blocks was examined by immunohistochemical staining. Correlations between LOXL2 mRNA and protein expression and clinicopathological features were evaluated by statistical analysis. In the 137 patients examined, LOXL2 mRNA expression was significantly lower in lung TT than the sNT ( P < 0.05). Forty-eight specimens (48/83) showed low expression of LOXL2, as characterized by immunohistochemical staining. By statistical analysis of the correlation between LOXL2 mRNA expression and clinical features of NSCLC patients, down-regulation of Loxl-2 mRNA expression was correlated with male patients ( P = 0.008), a poorer N-stage ( P = 0.032) and a poorer pathological TNM stage ( P = 0.003). Statistical analysis of the correlation between LOXL2 protein expression and clinical features of NSCLC patients showed a statistically significant difference between low expression of the LOXL2 protein and a poorer N-stage ( P = 0.036), a higher pathological TNM stage ( P = 0.005) and poorer differentiation ( P = 0.035). When stratified by histological types, significant differences at both the mRNA and protein levels were only found for lung adenocarcinomas patients, and not for lung squamous cell carcinomas patients. The level of LOXL2 mRNA expression was found to be significantly down-regulated in NSCLC, and the lower mRNA and protein expression levels correlated with poorer differentiation, higher N-stage and advanced pathologic TNM stage in patients with lung adenocarcinomas. [ABSTRACT FROM AUTHOR]

Proteins encoded by the NAC gene family constitute one of the largest plant-specific transcription factors, which have been identified to play many important roles in both abiotic and biotic stress adaptation, as well as in plant development regulation. In the current paper, a full-length cDNA sequence of a novel wheat NAC gene, designated as TaNAC4, was isolated using in silico cloning and the reverse transcription PCR (RT-PCR) methods. TaNAC4 sharing high homology with rice OsNAC4 gene was predicted to encode a protein of 308 amino acid residues, which contained a plant-specific NAC domain in the N-terminus. Transient expression analysis indicated that the deduced TaNAC4 protein was localized in the nucleus of onion epidemical cells. Yeast one-hybrid assay revealed that the C-terminal region of the TaNAC4 protein had transcriptional activity. The expression of TaNAC4 was largely higher in the wheat seedling roots, than that in leaves and stems. TaNAC4 transcript in wheat leaves was induced by the infection of strip rust pathogen, and also by exogenous applied methyl jasmonate (MeJA), ABA and ethylene. However, salicylic acid (SA) had no obvious effect on TaNAC4 expression. Environmental stimuli, including high salinity, wounding, and low-temperature also induced TaNAC4 expression. These results indicate that this novel TaNAC4 gene functions as a transcriptional activator involved in wheat response to biotic and abiotic stresses. [ABSTRACT FROM AUTHOR]

The roles of NF-kappaB (NF-κB) expression, Bax activity and cytochrome C (Cyt C) release, apoptosis of islet cells induced by high concentration glucose were explored in vitro. Pancreatic islet cells, which were isolated from Kunming mice, were cultured with different concentrations of glucose in DMEM, and divided into the following groups: G1, G2, G3, G4, G5, and G6 groups, corresponding to the glucose concentrations of 5.6, 7.8, 11.1, 16.7, 22.5, and 27.6 mmol/L, respectively. After culture for 120 h, insulin secretion was evaluated by radioimmunoassay, and the NF-κB expression was detected by immunocytochemistry. Bax activity and Cyt C release were measured by immunofluorescence, and apoptosis was examined by Hoechst33342 assay. The results showed that in G1, G2 and G3 groups, insulin secretion was enhanced with the increase of glucose concentration, and the NF-κB expression was also increased ( P<0.05), but Bax activity, Cyt C release and apoptosis rate showed no significant difference among them. However, in G4, G5, and G6 groups, apoptosis rate of islet cells, NF-κB expression, Bax activity, and Cyt C release were all significantly increased, and insulin secretion was impaired as compared with G1, G2, and G3 groups ( P<0.05). It was concluded that the exposure of islet cells to high glucose could induce islet cells apoptosis as well as impaired insulin secretion. The NF-κB signaling pathway and mitochondria pathway in islet cells might play some roles in the progressive loss of islet cells in diabetes. The inhibition of the NF-κB expression could be an effective strategy for protecting pancreatic islet cells. [ABSTRACT FROM AUTHOR]

Marine sediments were dissolved by HNO3-HF-HClO4 in a sealed container at low pressure; HF was evaporated in an open container and salts were dissolved in HCl by heating, then transferred to 2% HNO3 solution. A total of 45 elements, including Li, Be, Sc, Ti, V, Cr, Mn, Co, Ni, Cu, Zn, Ga, Rb, Sr, Y, Zr, Nb, Mo, Cd, In, Sb, Cs, Ba, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, Hf, Ta, W, Tl, Pb, Bi, Th and U, were measured by inductively coupled plasma mass spectrometry (ICP-MS). Conditions and sample experiments showed that this procedure defines a good experimental method which has the advantages of clear interference, easy operation and reliable results. The concentrations of the 45 elements could be used for resource exploration, environmental assessment and academic research. [ABSTRACT FROM AUTHOR]

BH3-only protein Bim is a critical regulator of apoptosis and plays an essential role in mammalian development, but the characterization of Bim and its isoforms in hepatocellular carcinoma (HCC) has not been studied before. Here we investigated the expression, distribution, regulation and role of Bim isoforms in HCC cells. Fifteen Bim isoforms were identified in HCC, with six newly identified isoforms and two newly identified exons. Among of them, Bim EL, L, S, a1, a2, a3, b2, b4 and b6 are abundant isoforms according to their mRNA levels. However only Bim EL, L and S proteins could be clearly detected. Bim mRNA and protein were strongly expressed in HCC tissues compared to relevant non-tumorous regions, but the ratio of variant isoforms showed no difference between tumorous and non-tumorous tissues. Bim isoforms were differentially regulated after chemotherapeutic drug 5-Fluorouracil (5-FU) treatment. Interestingly, Bim EL, L and S, the isoforms known to induce apoptosis strongly, are the least inducible isoforms at their mRNA levels when exposed to the stress, suggesting that post-transcriptional rather than transcriptional, modulations may play a role to enhance their functions. Finally, overexpression of Bim EL, L, S and all alpha isoforms induced apoptosis in HCC cells, while overexpression of Bim beta isoforms showed no effects on cell survival after 5-FU treatment. In conclusion, Bim alpha isoforms appears to have a role in the regulation of apoptosis in HCC cells, which may contribute to not only the growth of tumor cells but also the sensitivity of HCC cells to chemotherapy. [ABSTRACT FROM AUTHOR]

As research progresses, fluorescent proteins useful for optical marking will evolve toward brighter, monomeric forms that are more diverse in color. We previously reported a new fluorescent protein from Aequorea macrodactyla, GFPxm, that exhibited many characteristics similar to wild-type green fluorescent protein (GFP). However, the application of GFPxm was limited because GFPxm expressed and produced fluorescence only at low temperatures. To improve the fluorescent properties of GFPxm, 12 variants were produced by site-directed mutagenesis and DNA shuffling. Seven of these mutants could produce strong fluorescence when expressed at 37°C. The relative fluorescence intensities of mutants GFPxm16, GFPxm18, and GFPxm19 were higher than that of EGFP (enhanced GFP) when the expression temperature was between 25 and 37°C, and mutants GFPxm16 and GFPxm163 could maintain a high fluorescence intensity even when expressed at 42°C. Meanwhile, at least 4 mutants could be successfully expressed in mammalian cell lines. The fluorescence spectra of 6 of the 12 mutants had a progressive red shift. The longest excitation-emission maximum was at 514/525 nm. In addition, 3 of the 12 mutants had two excitation peaks including an UV-excitation peak, while another mutant had only one UV-excitation peak. [ABSTRACT FROM AUTHOR]

A strategy for amplifying the signal of surface plasmon resonance (SPR) biosensors is reported. Biotinylated phenylalanine (Biotin-Phe) monomers were rapidly self-assembled into nanoparticles in a mild environment. The self-assembled nanoparticles were then used as the carriers of streptavidin-antibody complexes by the streptavidin-biotin interaction. The signal was amplified because of the high molecular weight of the nanoparticle-streptavidin-antibody conjugate. With prostate-specific antigen as a model analyte, the target concentration as low as 1 pg mL−1 was readily measured. The results of the nanoparticle-enhanced SPR biosensor for analysis of serum samples are well consistent with those achieved by the enzyme-linked immunosorbent assays. This work is valuable for designing of various optical and electronic biosensors through the streptavidin-biotin interaction. [ABSTRACT FROM AUTHOR]

Muscarinic acetylcholine receptors have been implicated as potential neuroprotective targets for glaucoma. We tested the hypothesis that the lack of a single muscarinic receptor subtype leads to age-dependent neuron reduction in the retinal ganglion cell layer. Mice with targeted disruption of single muscarinic acetylcholine receptor subtype genes (M1 to M5) and wild-type controls were examined at two age categories, 5 and 15 months, respectively. We found no differences in intraocular pressure between individual mouse groups. Remarkably, in 15-month-old mice devoid of the M1 receptor, neuron number in the retinal ganglion cell layer and axon number in the optic nerve were markedly reduced. Moreover, mRNA expression for the prooxidative enzyme, NOX2, was increased, while mRNA expression for the antioxidative enzymes, SOD1, GPx1 and HO-1, was reduced in aged M1 receptor-deficient mice compared to age-matched wild-type mice. In line with these findings, the reactive oxygen species level was also elevated in the retinal ganglion cell layer of aged M1 receptor-deficient mice. In conclusion, M1 receptor deficiency results in retinal ganglion cell loss in aged mice via involvement of oxidative stress. Based on these findings, activation of M1 receptor signaling may become therapeutically useful to promote retinal ganglion cell survival. [ABSTRACT FROM AUTHOR]

Hepatitis D virus (HDV) is a defective RNA virus that requires the presence of hepatitis B virus (HBV) for its life cycle. The in vitro HDV infection system is widely used as a surrogate model to study cellular infection with both viruses owing to its practical feasibility. However, previous methods for running this system were less efficient for high-throughput screening and large-scale studies. Here, we developed a novel method for the production of infectious HDV by adenoviral vector (AdV)-mediated transduction. We demonstrated that the AdV-based method yields 10-fold higher viral titers than the transient-transfection approach. The HDV-containing supernatant derived from AdV-infected Huh7 cells can be used as the inoculum in infectivity assays without requiring further concentration prior to use. Furthermore, we devloped a chemiluminescent immunoassay (HDV-CLEIA) to quantitatively determine intracellular HDAg with a dynamic range of 5-11,000 pg/mL. HDV-CLEIA can be used as an alternative approach to assess HDV infection. The advantages of our updated methodology were demonstrated through in vitro HDV infection of HepaRG cells and by evaluating the neutralization activity using antibodies that target various regions of the HBV/HDV envelope proteins. Together, the methods presented here comprise a novel toolbox of in vitro assays for studying HDV infection. [ABSTRACT FROM AUTHOR]