The inhibition effect of tert-butyl alcohol(TBA), identified as the·OH radical inhibitor, on the TiO_2 nano assays(TNA) photoelectrocatalytic oxidation of different organics such as glucose and phthalate was repo...The inhibition effect of tert-butyl alcohol(TBA), identified as the·OH radical inhibitor, on the TiO_2 nano assays(TNA) photoelectrocatalytic oxidation of different organics such as glucose and phthalate was reported. The adsorption performance of these organics on the TNA photoelectrode was investigated by using the instantaneous photocurrent value, and the degradation property was examined by using the exhausted reaction. The results showed that glucose exhibited the poor adsorption and easy degradation performance, phthalate showed the strong adsorption and harddegradation, but TBA showed the weak adsorption and was the most difficult to be degraded. The degradation of both glucose and phthalate could be inhibited evidently by TBA. But the effect on glucose was more obvious. The different inhibition effects of TBA on different organics could be attributed to the differences in the adsorption and the degradation property. For instance, phthalate of the strong adsorption property could avoid from the capture of·OH radicals by TBA in TNA photoelectrocatalytic process.展开更多
Objective:To evaluate the performance of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution.Methods:A cross-sectional study was conducted in HI...Objective:To evaluate the performance of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution.Methods:A cross-sectional study was conducted in HIV-infected patients aged 5-18 years receiving antiretroviral treatment with CD4 T-lymphocytes>25%or>500 cells/mm3 for at least 6 months.QuantiF ERON-TB Gold,T-SPOT.TB,and tuberculin skin test were performed in each patient.Results:A total of 50 patients were enrolled with median age of 13.7 years,CD4 counts of 753(IQR:587-989)cells/mm3.Among 27 patients with tuberculosis(16)or tuberculosis exposure(11),8(29.6%)were positive to at least one test,2(7.4%)were positive QuantiFERON-TB Gold,3(11.1%)positive T-SPOT.TB,and 7(25.9%)had tuberculin skin test≥5 mm.Among 23 patients without history of tuberculosis or exposure,all had negative interferon gamma release assays,while 2(8.7%)had positive tuberculin skin test.Conclusions:All tests had low sensitivity despite immune reconstitution.展开更多
Objective: To explore the new sources of novel bioactive compounds having pharmaceutical and agricultural interest and to search the endophytic actinobacteria from medicinal plants. Methods: NAF-1 an endophyte actinob...Objective: To explore the new sources of novel bioactive compounds having pharmaceutical and agricultural interest and to search the endophytic actinobacteria from medicinal plants. Methods: NAF-1 an endophyte actinobacteria was isolated from leaves of medicinal plant Aloe vera collected in Marrakesh, Morocco using Bennett agar as selective medium. NAF-1 was tested for its antimicrobial activity against five pathogenic bacteria such as Staphylococcus aureus PIC 53156, Micrococcus luteus ATCC381, Bacillus subtilis ATCC 14579, Pseudomonas aeruginosa DSM 50090 and Escherichia coli ATCC 8739 and four human clinic fungi belonging to the Candida, Aspergillus and Microsporum genera. Several antioxidant activities were studied such as DPPH free radical scavenging, β-carotene and linoleic acid and reducing power assays. The total of phenol and flavonoid was also calculated. Using Artemia salina shrimp assay, the cytotoxicity of NAF-1 crude extract was determined. Results: The results revealed that the actinobacteria showed a high activity(≥20 mm) against only Gram positive bacteria but it had a moderate activity(between 13 and 15 mm) against Human clinic fungi. The isolate also exhibited a LD50 of 14.20 μg/mL in the cytotoxicity assay. The result showed that the crude extract presented an interesting free radical-scavenging activity with IC50 value of(5.58 ± 0.26) μg/mL and a high value of phenolic and flavonoid compounds with(15.41 ± 0.18) μg GAE/mg extract and(11.41± 0.06) μg QE/mg extract respectively. Moreover, the taxonomic position of our endophyte actinobacteria using the morphological and physiological criteria and using 16 S r RNA gene sequence(polyphasic approach) showed that the NAF-1 isolate was similar to Streptomyces hydrogenans which was never described as an endophyte actinobacteria. Conclusions: This isolated strain appears promising resources of bioactive agents and can be exploited to produce therapeutic agents active against pathogenic disease.展开更多
HIV/AIDS is one of the most serious public health challenges globally. Despite the great efforts that are being devoted to prevent,treat and to better understand the disease,it is one of the main causes of morbidity a...HIV/AIDS is one of the most serious public health challenges globally. Despite the great efforts that are being devoted to prevent,treat and to better understand the disease,it is one of the main causes of morbidity and mortality worldwide. Currently,there are 30 drugs or combinations of drugs approved by FDA. Because of the side-effects,price and drug resistance,it is essential to discover new targets,to develop new technology and to find new anti-HIV drugs. This review summarizes the major targets and assays currently used in anti-HIV drug screening.展开更多
·Tuberculous uveitis(TBU)comprises a broad clinical spectrum of ocular manifestations,making its diagnosis challenging.Ophthalmologists usually require evidence from investigations to confirm or support a clinica...·Tuberculous uveitis(TBU)comprises a broad clinical spectrum of ocular manifestations,making its diagnosis challenging.Ophthalmologists usually require evidence from investigations to confirm or support a clinical diagnosis of TBU.Since direct isolation of the causative organism from ocular specimens has limitations owing to the small volume of the ocular specimens,resultant test positivities are low in yield.Immunodiagnostic tests,including the tuberculin skin test and interferon-gamma release assays(IGRAs),can help support a clinical diagnosis of TBU.Unlike the tuberculin skin test,IGRAs are in vitro tests that require a single visit and are not affected by prior Bacillus Calmette-Guerin vaccination.Currently,available IGRAs consist of different techniques and interpretation methods.Moreover,newer generations have been developed to improve the sensitivity and ability to detect active tuberculosis.This narrative review collates salient practice points as a reference for general ophthalmologists,such as evidence for the utilization of IGRAs in patients with suspected TBU,and summarizes basic knowledge and details of clinical applications of these tests in a clinical setting.展开更多
AIM: To investigate the presence and potency of natural enzyme inhibitors with hypoglycemic potentials amongst Eucalyptus Spp. by in vitro assays.METHODS: The leaf extracts of the three different Eucalyptus species [E...AIM: To investigate the presence and potency of natural enzyme inhibitors with hypoglycemic potentials amongst Eucalyptus Spp. by in vitro assays.METHODS: The leaf extracts of the three different Eucalyptus species [E. globulus(EG), E. citriodora(EC), E. camaldulensis(ECA)] were subjected to in vitro assay procedures to explore the prevalence of natural enzyme inhibitors(NEIs) after preliminary qualitative and quantitative phytochemical evaluations, to study their inhibitory actions against the enzymes like α-amylase, α-glucosidase, aldose reductase, angiotensin converting enzyme and dipeptidyl peptidase 4 playing pathogenic roles in type 2 diabetes. The antioxidant potential and total antioxidant capacity of the species were also evaluated.RESULTS: Major bioactive compounds like polyphenols(341.75 ± 3.63 to 496.85 ± 3.98) and flavonoids(4.89 ± 0.01 to 7.15 ± 0.02) were found in appreciable quantity in three species. Based on the IC50 values of the extracts under investigation, in all assays the effectivity was in the order of EG > ECA > EC. The results of the ferric reducing antioxidant power assay showed that the reducing ability of the species was also in the order of EG > ECA > EC. A strong correlation(R2 = 0.81-0.99) was found between the phenolic contents and the inhibitory potentials of the extracts against the targeted enzymes.CONCLUSION: These results show immense hypoglycemic potentiality of the Eucalyptus Spp. and a remarkable source of NEIs for a future phytotherapeutic approach in Type 2 diabetes.展开更多
Aim： To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. Methods： Semen samples from 87 unselected infertile patients were used t...Aim： To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. Methods： Semen samples from 87 unselected infertile patients were used to perform the following assays： （i） detection of active caspase-3 （n = 17）; （ii） integrity of the mitochondrial membrane potential （MMP） （n = 17）; （iii） externalization of phosphatidylserine （EPS） （n = 16）; and （iv） detection of intact acrosomes via CD46 （n = 37）. After the assays, 4% paraformaldehyde was added to all aliquots. The fluorescence intensity of each sample was evaluated by flow cytometry on days 0, 3, 7, 10 and 14. Results： Differences of up to ± 5% positive spermatozoa from the value measured at day 0 were estimated as acceptable deviation. The Caspase-3 FLICA^TM showed mean differences 〈 5% at day 3, 7 and 10. At day 14 the mean difference was 7.6%. In contrast, the disrupted MMP and the EPS detection showed differences 〉 5% at day 3. The CD46-FITC labeling displayed absolute differences 〈 5% CD46-positive spermatozoa at days 3, 7, 10 and 14. Conclusion： Although immediate analysis of the fluorescence signals is recommended, it is possible to evaluate caspase-3 activation up to 10 days and CD46 up to 14 days after staining of sperm. The FACS evaluation of MMP and EPS detection should be conducted on the same day.展开更多
The anti-chlorpyrifos polyclonal antibodies were obtained by using the artificial immuneantigen to immune in New Zealands white rabbits. The enzyme-tagged antibodies wereprepared by coupling horseradish peroxidase (HR...The anti-chlorpyrifos polyclonal antibodies were obtained by using the artificial immuneantigen to immune in New Zealands white rabbits. The enzyme-tagged antibodies wereprepared by coupling horseradish peroxidase (HRP) to the purified antibody with themodified sodium periodate method. The indirect competitive enzyme linked immuno-sorbentassays (ELISA) and the HRP-tagged antibody direct ELISA (E-Ab) were established, respectively.The limit of detection (LOD) for the indirect ELISA and E-Ab were 0.0033 and 0.0042 gmL-1, respectively. The linear detection ranged well from 0.005 to 2.0 g mL-1.展开更多
Arboviral encephalitis is a group of animal and human illness that is mostly caused by several distinct families of viruses including orthobunya virus, phlebovirus, flaviviruses, and the alphaviruses. Although specifi...Arboviral encephalitis is a group of animal and human illness that is mostly caused by several distinct families of viruses including orthobunya virus, phlebovirus, flaviviruses, and the alphaviruses. Although specific signs and symptoms vary by the type of central nervous system (CNS), initial signs and symptoms are very similar. Therefore rapid immunologic and molecular tools for differential diagnosis of arboviral encephalitis viruses are important for effective case management and control of the spread of encephalitis. The qRT-PCR assay, especially multiplex PCR, has the potential to produce considerable savings in time and resources in the laboratory detection. Meanwhile, the use of IC can prevent false negatives effectively by monitoring the processes of nucleic acid extraction and amplification. This report describes the development of a panel of internally controlled multiplex one-step real-time RT-PCR assays in which two virus specific-probe sets were used in the same reaction for the detection of 15 species arboviral encephalitis viruses: the comparative sensitivity of multiplex one-step qRT-PCR assays to single plex one-step qRT-PCR assays as well as one-step RT-PCR assays for detection of each viral species. And total of 150 human serum samples were detected to evaluate the multiplex one-step qRT-PCR assays. These multiplex one-step real-time RT-PCR assays with IC were evaluated in terms of sensitivity, linearity, precision, specificity, and also field samples including serum and vector. These assays can detect and differentiate arboviral encephalitis viruses by high throughput, sensitive, and specific way. It is useful for clinical management and outbreak control of arboviral encephalitis viruses and vector surveillance.展开更多
Clostridium difficile and C. perfringens are enteric pathogens affecting a variety of mammals. This study evaluated the molecular enterotoxigenicity of Clostridium swine isolates by PCRs. One hundred and ten swine fae...Clostridium difficile and C. perfringens are enteric pathogens affecting a variety of mammals. This study evaluated the molecular enterotoxigenicity of Clostridium swine isolates by PCRs. One hundred and ten swine faeces were analyzed by culture assay. The faecal samples were from sixty-seven healthy animals and 43 with gastrointestinal tract disease. C. difficile strains were PCR-screened for the presence of tcdA/tcdB and cdtA/cdtB genes. All C. perfringens isolates were tested for the characterization of the toxinotype. Overall, sixty-five swine resulted positive: 38 for C. difficile and 17 for C. perfringens. One sample tested C. perfringens and C. difficile-positive, at the same time: on the whole, 39 C. difficile strains were isolated. Thirty-eight C. difficile isolates (all from healthy animals) resulted tcdA/tcdB and cdtA/cdtB-negative by PCRs and toxins A/B-negative by immunological tests. All C. perfringens strains were type A;eight were also cpb2-positive. In the sample (diarrhoeic), with double infection, C. difficile tested tcdA/tcdB and cdtA/cdtB-positive by PCRs and toxins A/B-positive by immunoassays;C. perfringens resulted cpb2-positive. The molecular genotypeing/toxinotyping should be applied to establish a final diagnosis and to assess properly the full implications and the epidemiological impact of these findings in particular in samples of healthy animals and aid in the development of effective intervention methods for controlling clostridial disease outbreaks.展开更多
With specific designed prmers. CYP2B6 and CYP1A1 cDNA were generatecl by reverse transcrlI7tion-Polymerase chain reaction(RT-PCR )technlque Performed on total RNAs isolated frorn hum1ln liver and 3-rnethylch(,lanthren...With specific designed prmers. CYP2B6 and CYP1A1 cDNA were generatecl by reverse transcrlI7tion-Polymerase chain reaction(RT-PCR )technlque Performed on total RNAs isolated frorn hum1ln liver and 3-rnethylch(,lanthrene(3-Mtt)induc human amnion FL, cells. Cell llnes (CHL, 2B6 and CtHL-1A1 ) capableof expressing hunlan cytochome P 15O (CYP ) 2B6 and 1A1 were establishecl after transfection of corre-sponding eukaryotic reconlbinant expression plasmid with human CYP2ll6 and 1A1 cDNA lnserts respectlvely. These cell lines stably expressed the mRNAs and the enzymatic activltles cc)rresI’onding to ttYP2B6and CYP1A1, respectively’ Compared with Chinese hamster lung (CHL) cells, the n1icr()nucleus frecluencyin CHl,-2B6 cells is markedly lncreased when exPosed to nitrosamines,aflatoxln B, (AFB1) and cyclophos-Phamide (CPA). Thls is also in CHL-1A1 cells,when exposed to carcinogenic polycycllc aromatic hydrocar-bons.展开更多
Single-cell sequencing technology has become increasingly prominent in aquaculture for investigating complex biological processes,such as development,reproduction,and disease resistance in aquatic species like Nile ti...Single-cell sequencing technology has become increasingly prominent in aquaculture for investigating complex biological processes,such as development,reproduction,and disease resistance in aquatic species like Nile tilapia,an economically important fish.This technology reveals cellular heterogeneity across various biological functions.However,traditional enzymatic dissociation methods often fail to generate high-quality,intact single-cell suspensions from complex brain tissue,limiting single-cell research in this area.This study aims to optimize the isolation of single-cell nuclei from tilapia(Oreochromis niloticus)brain tissue.We systematically fine-tuned the parameters of homogenization buffer,washing buffer,and centrifugation conditions,identifying the optimal method for nuclei isolation from tilapia brain.We developed a streamlined,rapid,and efficient protocol that eliminates the need for costly equipment,such as flow cytometry and ultracentrifugation.As a proof of principle,we successfully identified the cellular heterogeneity of tilapia brain tissue using single-nucleus RNA-seq analysis.The method we developed yields intact nuclei with minimal background contamination and a broad representation of cell types,making it a reliable tool for single-cell genomic assays in aquatic fish species.展开更多
In a single step photolithography, muhi-level microfluidic device is fabricated by printing novel architectures on a film photomasks. The whole fabrication process is executed by classical PCB technology without the n...In a single step photolithography, muhi-level microfluidic device is fabricated by printing novel architectures on a film photomasks. The whole fabrication process is executed by classical PCB technology without the need to access clean room facilities. Different levels of protruding features on PCB master are produced by exposing a photomask with specifically arranged "windows and rims" architectures, followed by chemical wet etching. Poly(dimethylsiloxane)(PDMS) is then molded against the positive relief master to generate microfluidic device featured with multi-level sandbag structure and peripheral microchannels. This sandbag structure is an analog to traditional dam or weir for particle entrapment. The microstructure does not collapse when subjected to applied pressure, which is suitable for operation on elastic PDMS substrate.Typical immunocytochemcial staining assays were performed in the microdevice to demonstrate the applicability of the sandbag structure for cellular analysis. This simplified microfabrication process employs low-cost materials and minimal specialized equipment and can reproducibly produce mask lines with about 20 μm in width, which is sufficient for most microfluidic applications.展开更多
Digital enzyme-linked immunoassays(dELISA)have been successfully applied to the ultrasensitive quantification of analytes,including nucleic acids,proteins,cells,and extracellular vesicles,achieving robust detection li...Digital enzyme-linked immunoassays(dELISA)have been successfully applied to the ultrasensitive quantification of analytes,including nucleic acids,proteins,cells,and extracellular vesicles,achieving robust detection limits in complex clinical specimens such as blood,and demonstrating utility across a broad range of clinical applications.The ultrasensitivity of dELISA comes from partitioning single analytes,captured onto a microbead,into millions of compartments so that they can be counted individually.There is particular interest in using dELISA for multiplexed measurements,but generating and detecting the billions of compartments necessary to perform multiplexed ultrasensitive dELISA remains a challenge.To address this,we have developed a high-throughput,optofluidic platform that performs quantitative fluorescence measurements on five populations of microbeads,each encoded with distinct ratios of two fluorescent dyes,for digital assays.The key innovation of our work is the parallelization of droplet generation and detection,combined with time-domain encoding of the excitation sources into distinct patterns that barcode the emission signal of both dyes within each bead,achieving high throughput(6×10^(6) droplets/min)and accurate readout.Additionally,we modulate the exposure settings of the digital camera,capturing images of multiplexed beads and the droplet fluorescent substrate in consecutive frames,a method inspired by high dynamic range(HDR)photography.Our platform accurately classifies five populations of dual-encoded beads(accuracy>99%)and detects bead-bound streptavidin-horseradish peroxidase molecules in a third fluorescence channel.This work establishes the technological foundation to combine high multiplexing and high throughput for droplet digital assays.展开更多
Polyphenols are widely recognized as the effective antioxidants,which are divided into flavonoids,phenolic acids,stilbenes,lignans,tannins and so on.They could regulate internal functions and protect the body from dis...Polyphenols are widely recognized as the effective antioxidants,which are divided into flavonoids,phenolic acids,stilbenes,lignans,tannins and so on.They could regulate internal functions and protect the body from diseases related to oxidative damage.Due to the fact that their antioxidant capacity is influenced by the structure,stability and bioavailability,the detection of their bioactivity should be considered comprehensively.Currently,the methods for measuring the antioxidant capacity of phenolic compounds are divided into chemical,cell-based and in vivo assays.The chemical assays include 2,2-diphenyl-l-picrylhydrazyl(DPPHI),2,2"-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS),ferric reducing/antioxidant power(FRAP),oxygen radical absorbance capacity(ORAC),peroxyI radical scavenging capacity(PSC),which are rapid identification method,but their reaction mechanism has a great gap with the internal body response.The cell-based assays are more consistent with biological reaction,but still do not take the bioavailability into consideration.The in vivo assays,which commonly utilized Caenorhabditis elegans or rats as models,are more representative,but these methods are more complex and spend longer.This review summarizes the antioxidant evaluation methods of phenolic compounds and discusses their advantages and limitations comparatively,which could help discriminate and select the appropriate assay in the actual operation,and facilitate the development of comprehensive approaches as well.展开更多
Clustered regularly interspaced short palindromic repeats(CRISPR)is an adaptive immune system in microorganisms,utilizing CRISPR RNA(crRNA)and CRISPR-associated proteins(Cas)to eliminate invasive nucleic acids[1].Due ...Clustered regularly interspaced short palindromic repeats(CRISPR)is an adaptive immune system in microorganisms,utilizing CRISPR RNA(crRNA)and CRISPR-associated proteins(Cas)to eliminate invasive nucleic acids[1].Due to its simplicity,programmability,precise targeting,and efficient cleavage abilities,the CRISPR/Cas system has become a highly effective diagnostic tool[2].As a prominent system in CRISPR-based diagnostics,CRISPR/Cas12a enables Cas12a to cleave the target DNA(ciscleavage)and continuously trans-cleave non-specific singlestranded DNA(ssDNA)following crRNA-mediated binding to the target DNA[3,4].展开更多
Eucalyptus staigeriana essential oil(EsEO)has well-known anthelmintic activity in small ruminants.However,its volatility limits its therapeutic action.The aim of this study was to develop a water-in-oil sodium alginat...Eucalyptus staigeriana essential oil(EsEO)has well-known anthelmintic activity in small ruminants.However,its volatility limits its therapeutic action.The aim of this study was to develop a water-in-oil sodium alginatebased nanoemulsion with an effective in vitro effect on the eggs and larvae of Haemonchus contortus,a gastrointestinal parasite of sheep and goats.Four oil-in-water sodium alginate-based emulsions were prepared using a high-energy method with different proportions of Tween 80,EsEO,and sodium alginate(ALG)4%.The physical-chemical characterization included stability,particle size,zeta potential and infrared spectra.The effects of the emulsions were evaluated against H.contortus via the egg hatching test(EHT)and larval development test(LDT).The results showed that the emulsions were stable over 7 days,nanometer scale particles(218.8 to 371.5 nm)predominating and with negative zeta potentials(−28.9 to−46.9 mV).All four emulsions were more effective than EsEO in the EHT,with 50%effective concentrations(EC50)of 0.088 to 0.15 mg/mL for the emulsions and 0.308 mg/mL for EsEO.However,in the LDT,the emulsions and essential oil had similar effects,with EC50 values of 3.91 to 4.60 mg/mL for the emulsions and 4.17 mg/mL for EsEO.Emulsion 2,with low Tween 80/EsEO and ALG/EsEO ratios,was considered better overall in terms of physical,chemical and anthelmintic assessment and is a promising candidate for further in vivo assays against adult H.contortus.展开更多
Background： Tuberculosis （TB） remains a major global public health challenge. Articular T/3 is an important form of extrapulmonary tuberculosis, and its diagnosis is difficult because of the low sensitivity of trad...Background： Tuberculosis （TB） remains a major global public health challenge. Articular T/3 is an important form of extrapulmonary tuberculosis, and its diagnosis is difficult because of the low sensitivity of traditional methods. The aim of this study was to analyze the diagnostic value of T-SPOT.TB on synovial fluid for the diagnosis of articular TB. Methods： Patients with suspected articular TB were enrolled consecutively between August 2011 and December 2015. T-SPOT.TB was performed on both synovial fluid mononuclear cells （SFMCs） and peripheral blood mononuclear cells （PBMCs）. The final diagnosis of articular TB was independent of the T-SPOT.TB result. The diagnostic sensitivity, specificity, predictive value, and likelihood ratio of T-SPOT.TB on SFMCs and PBMCs were analyzed. Results： Twenty patients with suspected articular TB were enrolled. Six were diagnosed with articular TB, and 14 patients were diagnosed with other diseases. Sensitivity and specificity were 83% and 86% for T-SPOT.TB on SFMCs, and 67% and 69% for T-SPOT.TB on PBMCs, respectively. The positive predictive value （PPV） and negative predictive value （NPV） of T-SPOT.TB on SFMCs were 71% and 92%, respectively. The PPV and NPV were 50% and 82% for T-SPOT.TB on PBMCs. Conclusion： Sensitivity, specificity, and NPV of T-SPOT.TB on SFMCs appeared higher than that on PBMCs, indicating that T-SPOT. TB on SFMCs might be a rapid and accurate diagnostic test for articular TB.展开更多
基金the National High Technology Research and Development Program of China (Grant No.2009AA063003)the National Nature Science Foundation of China (No.20677039) for financial support
文摘The inhibition effect of tert-butyl alcohol(TBA), identified as the·OH radical inhibitor, on the TiO_2 nano assays(TNA) photoelectrocatalytic oxidation of different organics such as glucose and phthalate was reported. The adsorption performance of these organics on the TNA photoelectrode was investigated by using the instantaneous photocurrent value, and the degradation property was examined by using the exhausted reaction. The results showed that glucose exhibited the poor adsorption and easy degradation performance, phthalate showed the strong adsorption and harddegradation, but TBA showed the weak adsorption and was the most difficult to be degraded. The degradation of both glucose and phthalate could be inhibited evidently by TBA. But the effect on glucose was more obvious. The different inhibition effects of TBA on different organics could be attributed to the differences in the adsorption and the degradation property. For instance, phthalate of the strong adsorption property could avoid from the capture of·OH radicals by TBA in TNA photoelectrocatalytic process.
基金supported by the Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,Thailand,[Grant Number(IO)R015832028].Oxford Immunotec and Biomed diagnostics(Thailand)provided the T-SPOT.TB test kit
文摘Objective:To evaluate the performance of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution.Methods:A cross-sectional study was conducted in HIV-infected patients aged 5-18 years receiving antiretroviral treatment with CD4 T-lymphocytes>25%or>500 cells/mm3 for at least 6 months.QuantiF ERON-TB Gold,T-SPOT.TB,and tuberculin skin test were performed in each patient.Results:A total of 50 patients were enrolled with median age of 13.7 years,CD4 counts of 753(IQR:587-989)cells/mm3.Among 27 patients with tuberculosis(16)or tuberculosis exposure(11),8(29.6%)were positive to at least one test,2(7.4%)were positive QuantiFERON-TB Gold,3(11.1%)positive T-SPOT.TB,and 7(25.9%)had tuberculin skin test≥5 mm.Among 23 patients without history of tuberculosis or exposure,all had negative interferon gamma release assays,while 2(8.7%)had positive tuberculin skin test.Conclusions:All tests had low sensitivity despite immune reconstitution.
文摘Objective: To explore the new sources of novel bioactive compounds having pharmaceutical and agricultural interest and to search the endophytic actinobacteria from medicinal plants. Methods: NAF-1 an endophyte actinobacteria was isolated from leaves of medicinal plant Aloe vera collected in Marrakesh, Morocco using Bennett agar as selective medium. NAF-1 was tested for its antimicrobial activity against five pathogenic bacteria such as Staphylococcus aureus PIC 53156, Micrococcus luteus ATCC381, Bacillus subtilis ATCC 14579, Pseudomonas aeruginosa DSM 50090 and Escherichia coli ATCC 8739 and four human clinic fungi belonging to the Candida, Aspergillus and Microsporum genera. Several antioxidant activities were studied such as DPPH free radical scavenging, β-carotene and linoleic acid and reducing power assays. The total of phenol and flavonoid was also calculated. Using Artemia salina shrimp assay, the cytotoxicity of NAF-1 crude extract was determined. Results: The results revealed that the actinobacteria showed a high activity(≥20 mm) against only Gram positive bacteria but it had a moderate activity(between 13 and 15 mm) against Human clinic fungi. The isolate also exhibited a LD50 of 14.20 μg/mL in the cytotoxicity assay. The result showed that the crude extract presented an interesting free radical-scavenging activity with IC50 value of(5.58 ± 0.26) μg/mL and a high value of phenolic and flavonoid compounds with(15.41 ± 0.18) μg GAE/mg extract and(11.41± 0.06) μg QE/mg extract respectively. Moreover, the taxonomic position of our endophyte actinobacteria using the morphological and physiological criteria and using 16 S r RNA gene sequence(polyphasic approach) showed that the NAF-1 isolate was similar to Streptomyces hydrogenans which was never described as an endophyte actinobacteria. Conclusions: This isolated strain appears promising resources of bioactive agents and can be exploited to produce therapeutic agents active against pathogenic disease.
文摘HIV/AIDS is one of the most serious public health challenges globally. Despite the great efforts that are being devoted to prevent,treat and to better understand the disease,it is one of the main causes of morbidity and mortality worldwide. Currently,there are 30 drugs or combinations of drugs approved by FDA. Because of the side-effects,price and drug resistance,it is essential to discover new targets,to develop new technology and to find new anti-HIV drugs. This review summarizes the major targets and assays currently used in anti-HIV drug screening.
文摘·Tuberculous uveitis(TBU)comprises a broad clinical spectrum of ocular manifestations,making its diagnosis challenging.Ophthalmologists usually require evidence from investigations to confirm or support a clinical diagnosis of TBU.Since direct isolation of the causative organism from ocular specimens has limitations owing to the small volume of the ocular specimens,resultant test positivities are low in yield.Immunodiagnostic tests,including the tuberculin skin test and interferon-gamma release assays(IGRAs),can help support a clinical diagnosis of TBU.Unlike the tuberculin skin test,IGRAs are in vitro tests that require a single visit and are not affected by prior Bacillus Calmette-Guerin vaccination.Currently,available IGRAs consist of different techniques and interpretation methods.Moreover,newer generations have been developed to improve the sensitivity and ability to detect active tuberculosis.This narrative review collates salient practice points as a reference for general ophthalmologists,such as evidence for the utilization of IGRAs in patients with suspected TBU,and summarizes basic knowledge and details of clinical applications of these tests in a clinical setting.
文摘AIM: To investigate the presence and potency of natural enzyme inhibitors with hypoglycemic potentials amongst Eucalyptus Spp. by in vitro assays.METHODS: The leaf extracts of the three different Eucalyptus species [E. globulus(EG), E. citriodora(EC), E. camaldulensis(ECA)] were subjected to in vitro assay procedures to explore the prevalence of natural enzyme inhibitors(NEIs) after preliminary qualitative and quantitative phytochemical evaluations, to study their inhibitory actions against the enzymes like α-amylase, α-glucosidase, aldose reductase, angiotensin converting enzyme and dipeptidyl peptidase 4 playing pathogenic roles in type 2 diabetes. The antioxidant potential and total antioxidant capacity of the species were also evaluated.RESULTS: Major bioactive compounds like polyphenols(341.75 ± 3.63 to 496.85 ± 3.98) and flavonoids(4.89 ± 0.01 to 7.15 ± 0.02) were found in appreciable quantity in three species. Based on the IC50 values of the extracts under investigation, in all assays the effectivity was in the order of EG > ECA > EC. The results of the ferric reducing antioxidant power assay showed that the reducing ability of the species was also in the order of EG > ECA > EC. A strong correlation(R2 = 0.81-0.99) was found between the phenolic contents and the inhibitory potentials of the extracts against the targeted enzymes.CONCLUSION: These results show immense hypoglycemic potentiality of the Eucalyptus Spp. and a remarkable source of NEIs for a future phytotherapeutic approach in Type 2 diabetes.
文摘Aim： To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. Methods： Semen samples from 87 unselected infertile patients were used to perform the following assays： （i） detection of active caspase-3 （n = 17）; （ii） integrity of the mitochondrial membrane potential （MMP） （n = 17）; （iii） externalization of phosphatidylserine （EPS） （n = 16）; and （iv） detection of intact acrosomes via CD46 （n = 37）. After the assays, 4% paraformaldehyde was added to all aliquots. The fluorescence intensity of each sample was evaluated by flow cytometry on days 0, 3, 7, 10 and 14. Results： Differences of up to ± 5% positive spermatozoa from the value measured at day 0 were estimated as acceptable deviation. The Caspase-3 FLICA^TM showed mean differences 〈 5% at day 3, 7 and 10. At day 14 the mean difference was 7.6%. In contrast, the disrupted MMP and the EPS detection showed differences 〉 5% at day 3. The CD46-FITC labeling displayed absolute differences 〈 5% CD46-positive spermatozoa at days 3, 7, 10 and 14. Conclusion： Although immediate analysis of the fluorescence signals is recommended, it is possible to evaluate caspase-3 activation up to 10 days and CD46 up to 14 days after staining of sperm. The FACS evaluation of MMP and EPS detection should be conducted on the same day.
文摘The anti-chlorpyrifos polyclonal antibodies were obtained by using the artificial immuneantigen to immune in New Zealands white rabbits. The enzyme-tagged antibodies wereprepared by coupling horseradish peroxidase (HRP) to the purified antibody with themodified sodium periodate method. The indirect competitive enzyme linked immuno-sorbentassays (ELISA) and the HRP-tagged antibody direct ELISA (E-Ab) were established, respectively.The limit of detection (LOD) for the indirect ELISA and E-Ab were 0.0033 and 0.0042 gmL-1, respectively. The linear detection ranged well from 0.005 to 2.0 g mL-1.
文摘Arboviral encephalitis is a group of animal and human illness that is mostly caused by several distinct families of viruses including orthobunya virus, phlebovirus, flaviviruses, and the alphaviruses. Although specific signs and symptoms vary by the type of central nervous system (CNS), initial signs and symptoms are very similar. Therefore rapid immunologic and molecular tools for differential diagnosis of arboviral encephalitis viruses are important for effective case management and control of the spread of encephalitis. The qRT-PCR assay, especially multiplex PCR, has the potential to produce considerable savings in time and resources in the laboratory detection. Meanwhile, the use of IC can prevent false negatives effectively by monitoring the processes of nucleic acid extraction and amplification. This report describes the development of a panel of internally controlled multiplex one-step real-time RT-PCR assays in which two virus specific-probe sets were used in the same reaction for the detection of 15 species arboviral encephalitis viruses: the comparative sensitivity of multiplex one-step qRT-PCR assays to single plex one-step qRT-PCR assays as well as one-step RT-PCR assays for detection of each viral species. And total of 150 human serum samples were detected to evaluate the multiplex one-step qRT-PCR assays. These multiplex one-step real-time RT-PCR assays with IC were evaluated in terms of sensitivity, linearity, precision, specificity, and also field samples including serum and vector. These assays can detect and differentiate arboviral encephalitis viruses by high throughput, sensitive, and specific way. It is useful for clinical management and outbreak control of arboviral encephalitis viruses and vector surveillance.
文摘Clostridium difficile and C. perfringens are enteric pathogens affecting a variety of mammals. This study evaluated the molecular enterotoxigenicity of Clostridium swine isolates by PCRs. One hundred and ten swine faeces were analyzed by culture assay. The faecal samples were from sixty-seven healthy animals and 43 with gastrointestinal tract disease. C. difficile strains were PCR-screened for the presence of tcdA/tcdB and cdtA/cdtB genes. All C. perfringens isolates were tested for the characterization of the toxinotype. Overall, sixty-five swine resulted positive: 38 for C. difficile and 17 for C. perfringens. One sample tested C. perfringens and C. difficile-positive, at the same time: on the whole, 39 C. difficile strains were isolated. Thirty-eight C. difficile isolates (all from healthy animals) resulted tcdA/tcdB and cdtA/cdtB-negative by PCRs and toxins A/B-negative by immunological tests. All C. perfringens strains were type A;eight were also cpb2-positive. In the sample (diarrhoeic), with double infection, C. difficile tested tcdA/tcdB and cdtA/cdtB-positive by PCRs and toxins A/B-positive by immunoassays;C. perfringens resulted cpb2-positive. The molecular genotypeing/toxinotyping should be applied to establish a final diagnosis and to assess properly the full implications and the epidemiological impact of these findings in particular in samples of healthy animals and aid in the development of effective intervention methods for controlling clostridial disease outbreaks.
文摘With specific designed prmers. CYP2B6 and CYP1A1 cDNA were generatecl by reverse transcrlI7tion-Polymerase chain reaction(RT-PCR )technlque Performed on total RNAs isolated frorn hum1ln liver and 3-rnethylch(,lanthrene(3-Mtt)induc human amnion FL, cells. Cell llnes (CHL, 2B6 and CtHL-1A1 ) capableof expressing hunlan cytochome P 15O (CYP ) 2B6 and 1A1 were establishecl after transfection of corre-sponding eukaryotic reconlbinant expression plasmid with human CYP2ll6 and 1A1 cDNA lnserts respectlvely. These cell lines stably expressed the mRNAs and the enzymatic activltles cc)rresI’onding to ttYP2B6and CYP1A1, respectively’ Compared with Chinese hamster lung (CHL) cells, the n1icr()nucleus frecluencyin CHl,-2B6 cells is markedly lncreased when exPosed to nitrosamines,aflatoxln B, (AFB1) and cyclophos-Phamide (CPA). Thls is also in CHL-1A1 cells,when exposed to carcinogenic polycycllc aromatic hydrocar-bons.
基金supported by the National Natural Science Foundation of China(32200414,32373113)Natural Science Foundation of Shanghai(23ZR1426800).
文摘Single-cell sequencing technology has become increasingly prominent in aquaculture for investigating complex biological processes,such as development,reproduction,and disease resistance in aquatic species like Nile tilapia,an economically important fish.This technology reveals cellular heterogeneity across various biological functions.However,traditional enzymatic dissociation methods often fail to generate high-quality,intact single-cell suspensions from complex brain tissue,limiting single-cell research in this area.This study aims to optimize the isolation of single-cell nuclei from tilapia(Oreochromis niloticus)brain tissue.We systematically fine-tuned the parameters of homogenization buffer,washing buffer,and centrifugation conditions,identifying the optimal method for nuclei isolation from tilapia brain.We developed a streamlined,rapid,and efficient protocol that eliminates the need for costly equipment,such as flow cytometry and ultracentrifugation.As a proof of principle,we successfully identified the cellular heterogeneity of tilapia brain tissue using single-nucleus RNA-seq analysis.The method we developed yields intact nuclei with minimal background contamination and a broad representation of cell types,making it a reliable tool for single-cell genomic assays in aquatic fish species.
文摘In a single step photolithography, muhi-level microfluidic device is fabricated by printing novel architectures on a film photomasks. The whole fabrication process is executed by classical PCB technology without the need to access clean room facilities. Different levels of protruding features on PCB master are produced by exposing a photomask with specifically arranged "windows and rims" architectures, followed by chemical wet etching. Poly(dimethylsiloxane)(PDMS) is then molded against the positive relief master to generate microfluidic device featured with multi-level sandbag structure and peripheral microchannels. This sandbag structure is an analog to traditional dam or weir for particle entrapment. The microstructure does not collapse when subjected to applied pressure, which is suitable for operation on elastic PDMS substrate.Typical immunocytochemcial staining assays were performed in the microdevice to demonstrate the applicability of the sandbag structure for cellular analysis. This simplified microfabrication process employs low-cost materials and minimal specialized equipment and can reproducibly produce mask lines with about 20 μm in width, which is sufficient for most microfluidic applications.
基金funding from the following sources:National Human Genome Research Institute(RM1-HG-010023)National Cancer Institute(R21CA236653,R33CA278551)+2 种基金National Institute of Mental Health(R33-NIMH-118170)National Institute of Allergy and Infectious Diseases(R33-AI-147406)National Defense Science and Engineering Graduate Fellowship.
文摘Digital enzyme-linked immunoassays(dELISA)have been successfully applied to the ultrasensitive quantification of analytes,including nucleic acids,proteins,cells,and extracellular vesicles,achieving robust detection limits in complex clinical specimens such as blood,and demonstrating utility across a broad range of clinical applications.The ultrasensitivity of dELISA comes from partitioning single analytes,captured onto a microbead,into millions of compartments so that they can be counted individually.There is particular interest in using dELISA for multiplexed measurements,but generating and detecting the billions of compartments necessary to perform multiplexed ultrasensitive dELISA remains a challenge.To address this,we have developed a high-throughput,optofluidic platform that performs quantitative fluorescence measurements on five populations of microbeads,each encoded with distinct ratios of two fluorescent dyes,for digital assays.The key innovation of our work is the parallelization of droplet generation and detection,combined with time-domain encoding of the excitation sources into distinct patterns that barcode the emission signal of both dyes within each bead,achieving high throughput(6×10^(6) droplets/min)and accurate readout.Additionally,we modulate the exposure settings of the digital camera,capturing images of multiplexed beads and the droplet fluorescent substrate in consecutive frames,a method inspired by high dynamic range(HDR)photography.Our platform accurately classifies five populations of dual-encoded beads(accuracy>99%)and detects bead-bound streptavidin-horseradish peroxidase molecules in a third fluorescence channel.This work establishes the technological foundation to combine high multiplexing and high throughput for droplet digital assays.
基金supported by the National Key Research and Development Plan for the Fourteenth Five Year(2022YFD2100803)the National Natural Science Foundation of China(32202040)+1 种基金the First Batch of Liaoning“Unveiling Leader”Scientific and Technological Projects(2021JH1/10400036)the Key Laboratory of Marine Fishery Resources Exploitment&Utilization of Zhejiang Province(SL2022015).
文摘Polyphenols are widely recognized as the effective antioxidants,which are divided into flavonoids,phenolic acids,stilbenes,lignans,tannins and so on.They could regulate internal functions and protect the body from diseases related to oxidative damage.Due to the fact that their antioxidant capacity is influenced by the structure,stability and bioavailability,the detection of their bioactivity should be considered comprehensively.Currently,the methods for measuring the antioxidant capacity of phenolic compounds are divided into chemical,cell-based and in vivo assays.The chemical assays include 2,2-diphenyl-l-picrylhydrazyl(DPPHI),2,2"-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS),ferric reducing/antioxidant power(FRAP),oxygen radical absorbance capacity(ORAC),peroxyI radical scavenging capacity(PSC),which are rapid identification method,but their reaction mechanism has a great gap with the internal body response.The cell-based assays are more consistent with biological reaction,but still do not take the bioavailability into consideration.The in vivo assays,which commonly utilized Caenorhabditis elegans or rats as models,are more representative,but these methods are more complex and spend longer.This review summarizes the antioxidant evaluation methods of phenolic compounds and discusses their advantages and limitations comparatively,which could help discriminate and select the appropriate assay in the actual operation,and facilitate the development of comprehensive approaches as well.
基金supported by the Natural Science Foundation of Xiamen,China(3502Z202373019)National Key R&D Program of China(2023YFD1800502)+5 种基金National Natural Science Foundation of China(31971369)Natural Science Foundation of Fujian Province(2019J02004)CAMS Innovation Fund for Medical Sciences(2019RU022)the Fundamental Research Funds for the Central Universities(20720220006)the Fundamental Research Funds for the Central Universities(20720220005)the Science and Technology Planning Project of Guangdong Province of China(B1212030009)。
文摘Clustered regularly interspaced short palindromic repeats(CRISPR)is an adaptive immune system in microorganisms,utilizing CRISPR RNA(crRNA)and CRISPR-associated proteins(Cas)to eliminate invasive nucleic acids[1].Due to its simplicity,programmability,precise targeting,and efficient cleavage abilities,the CRISPR/Cas system has become a highly effective diagnostic tool[2].As a prominent system in CRISPR-based diagnostics,CRISPR/Cas12a enables Cas12a to cleave the target DNA(ciscleavage)and continuously trans-cleave non-specific singlestranded DNA(ssDNA)following crRNA-mediated binding to the target DNA[3,4].
基金supported by Conselho Nacional de Desenvolvimento Cientifico eTecnologico(CNPq)(grant No.142165/2018-2)Mr.Araujo-Filho had a doctoral scholarship and Dr.Bevilaqua has a research fellowship(grant No.305911/2019-8)from CNPq.
文摘Eucalyptus staigeriana essential oil(EsEO)has well-known anthelmintic activity in small ruminants.However,its volatility limits its therapeutic action.The aim of this study was to develop a water-in-oil sodium alginatebased nanoemulsion with an effective in vitro effect on the eggs and larvae of Haemonchus contortus,a gastrointestinal parasite of sheep and goats.Four oil-in-water sodium alginate-based emulsions were prepared using a high-energy method with different proportions of Tween 80,EsEO,and sodium alginate(ALG)4%.The physical-chemical characterization included stability,particle size,zeta potential and infrared spectra.The effects of the emulsions were evaluated against H.contortus via the egg hatching test(EHT)and larval development test(LDT).The results showed that the emulsions were stable over 7 days,nanometer scale particles(218.8 to 371.5 nm)predominating and with negative zeta potentials(−28.9 to−46.9 mV).All four emulsions were more effective than EsEO in the EHT,with 50%effective concentrations(EC50)of 0.088 to 0.15 mg/mL for the emulsions and 0.308 mg/mL for EsEO.However,in the LDT,the emulsions and essential oil had similar effects,with EC50 values of 3.91 to 4.60 mg/mL for the emulsions and 4.17 mg/mL for EsEO.Emulsion 2,with low Tween 80/EsEO and ALG/EsEO ratios,was considered better overall in terms of physical,chemical and anthelmintic assessment and is a promising candidate for further in vivo assays against adult H.contortus.
文摘Background： Tuberculosis （TB） remains a major global public health challenge. Articular T/3 is an important form of extrapulmonary tuberculosis, and its diagnosis is difficult because of the low sensitivity of traditional methods. The aim of this study was to analyze the diagnostic value of T-SPOT.TB on synovial fluid for the diagnosis of articular TB. Methods： Patients with suspected articular TB were enrolled consecutively between August 2011 and December 2015. T-SPOT.TB was performed on both synovial fluid mononuclear cells （SFMCs） and peripheral blood mononuclear cells （PBMCs）. The final diagnosis of articular TB was independent of the T-SPOT.TB result. The diagnostic sensitivity, specificity, predictive value, and likelihood ratio of T-SPOT.TB on SFMCs and PBMCs were analyzed. Results： Twenty patients with suspected articular TB were enrolled. Six were diagnosed with articular TB, and 14 patients were diagnosed with other diseases. Sensitivity and specificity were 83% and 86% for T-SPOT.TB on SFMCs, and 67% and 69% for T-SPOT.TB on PBMCs, respectively. The positive predictive value （PPV） and negative predictive value （NPV） of T-SPOT.TB on SFMCs were 71% and 92%, respectively. The PPV and NPV were 50% and 82% for T-SPOT.TB on PBMCs. Conclusion： Sensitivity, specificity, and NPV of T-SPOT.TB on SFMCs appeared higher than that on PBMCs, indicating that T-SPOT. TB on SFMCs might be a rapid and accurate diagnostic test for articular TB.
