Bioassay Studies Research Articles

Preparation and characterization of silica nanoparticles (sol–gel & river sand): A comparative toxicity studies against seed weevil (Sitophilus oryzae L). and effect on agro-morphological characteristics of maize seeds

The present study aimed to compare the entomotoxic efficacy of silica nanoparticles obtained through sol–gel and river sand against Sitophilus oryzae L. by keeping insecticidal dust (malathion 5D) as a control. Physicochemical characterization results revealed that silica nanoparticles (SiO2 NPs) were spherical in shape. Bioassay studies exhibited the LD50 values as 0.44 mg/100 g for sol–gel synthesized SiO2 NPs and 0.92 mg/100 g for river sand based SiO2 NPs. Cent percent mortality was brought about by sol–gel synthesized SiO2 NPs at the dose of 3 mg after 4 days of exposure whereas only 73.33 ± 1.57 per cent mortality was observed for river sand based SiO2 NPs at the dose of 9 mg after 4 days. Further, sol–gel synthesized SiO2 NPs [LD90 (6.64 mg/100 g maize seed)] enhanced the seedling characteristics i.e., (%) germination (98 ± 1.72), seedling length (49.71 ± 0.13 cm), vigour index (4871.58) and dry matter production (4.00 ± 0.01 g) compared to river sand based SiO2 NPs.

Relative Efficacy of Some Chitin Synthesis Inhibitors on the Mortality of Okra Jassid, <i>Amrasca biguttula biguttula</i> (Ishida)

In this laboratory study, three chitin synthesis inhibitors (CSIs) <i>viz.</i> Tacoma 40SC (Buprofezin), Heron 5EC (Lufenuron) and Pyrifen 10.8EC (Pyriproxifen) were evaluated with three concentrations of each through different application methods <i>viz</i>. direct or topical, indirect or leaf-dip and combined (Direct + Indirect). Data on jassids mortality was collected at 3 and 7 days after treatment (DAT) application. Our present study showed that all of the CSIs had significant effect on the mortality of jassid. In all cases, mortality was clearly dose, time and application method dependent. It has been seen that higher concentrations were found to be more effective than lower concentrations. Moreover, significant level of mortality was found at 3DAT but reached to the peak level at 7DAT in case of all selected CSIs. Bioassay study has showed that all the selected CSIs became able to enter the insect body through contact as well as stomach action to disrupt moulting process by inhibiting chitin synthesis that confirmed the contact and systemic actions of the selected CSIs. Moreover, protocols developed in this study for jassids collection and their safe transferring inside the petridishes would be a useful and convenient approach for the researchers.

Honey bee larval culture in vitro: gut emptying determines the transition from larva to prepupa and recombinant AccApidaecin improves antibacterial activity.

In vitro rearing of honey bee larvae is ideal for bioassay studies; no honey bee stable cell lines are available. Inconsistency of internal development staging of reared larvae and a susceptibility to contamination are common problems encountered. Standardized protocols on rearing larvae in vitro to make the larvae growth and development more similar to that of natural colonies are necessary to ensure the accuracy of experimental results and promote honey bee research as a model organism. Here, we concluded that when larval fasting weight was >160 mg, the time point of gut emptying can be defined as the critical point separating the larval and prepupal stages. In this way, we can conduct precise studies on the prepupal stage, such as organ remodeling during metamorphosis. Simultaneously, we further verified that recombinant AccApidaecin in genetic engineered bacteria added to the larval diet upregulated antibacterial peptide gene expression, and did not stimulate the stress response in larvae, nor did it affect the pupation rate or eclosion rate. This demonstrated that feeding recombinant AccApidaecin can enhance the individual antibacterial ability at the molecular level.

Ethnobotanical Study of Traditional Medicinal Plants Used for the Treatment of Infectious Diseases by Local Communities in Traditional Authority (T/A) Mbelwa, Mzimba District, Northern Region, Malawi

Local communities in Mzimba District, Malawi, have limited access to healthcare services and often rely on traditional medical practice and medicinal plants (MPs) for most of their medical care. However, phytomedicines’ use has not been well documented. This study aimed to identify and document medicinal plants and the associated ethnobotanical knowledge. Ethnobotanical data were collected in seven localities (19 villages) in the T/A Mbelwa, Mzimba, from May to June 2021. Forty traditional healers, herbalists, and farmers selected purposively and by snowball sampling were interviewed through semi-structured interviews, field observations, group discussions, and guided field walks. Quantitative indices, viz. relative frequency of citation (RFC), use value (UV), relative importance (RI) values, informant consensus factors (ICFs), and fidelity levels (FLs), were used to analyze the data. Eighty MPs belonging to 43 families and 77 genera were recorded. The Leguminosae family showed the highest number of species (16), followed by Solanaceae, Rubiaceae, and Phyllanthaceae. Trees (35 species) and roots (62%) accounted for the most significant habit and part, respectively. Washing (29%) was the most common preparation method. The most cited plant was Zanthoxylum chalybeum (RFC = 0.80, UV = 0.28, RI = 1.66), followed by Cassia abbreviata (RFC = 0.68, UV = 0.35, RI = 1.50). Respiratory disorders showed the highest ICF (0.53), followed by general and unspecified disorders (0.31). Z. chalybeum, C. abbreviata, and Oldfieldia dactylophylla showed maximum FLs (100%) for treating malaria and dysentery. Phytochemical, bioassay, toxicity, and conservation studies are needed to assess medicinal plants’ safety, efficacy, and quality as steps toward discovering new promising therapeutic leads without neglecting conservation programs for their sustainable utilization.

Characterization and field evaluation of Spilosoma obliqua nucleopolyhedrosis virus (SpobNPV) CRIJAF1 strain against jute hairy caterpillar, Spilosoma obliqua (Walker) infesting jute, Corchorus olitorius Linn.

BackgroundSpilosoma (= Spilarctia) obliqua (Walker) (Lepidoptera: Arctiidae), once considered as a sporadic pest, is now considered as a major and polyphagous pest widely distributed throughout India. Recent outbreaks of this pest were manifested in both jute and sunnhemp. During a survey in and around adjoining villages of I.C.A.R-C.R.I.J.A.F, Barrackpore, an epizootic was observed in the field population of S. obliqua caused by a nucleopolyhedrosis virus. Hence, an attempt was made to isolate, characterize using electron microscopy and assess the efficacy of S. obliqua nucleopolyhedrosis virus under laboratory and field conditions.ResultsElectron microscopic studies revealed typical baculovirus occlusion bodies of type nucleopolyhedrosis virus with tetrahedral and triangular in shape with the size ranging from 2.55 to 2.90 µm, with an average size of about 2.72 µm. Bioassay studies showed the larval mortality at low concentration of SpobNPV, i.e., 2.42 × 104 POBs/ml. The virus-infected S. obliqua was identified through the amplification of polyhedrin, polh gene sequences with the available sequences in public database, GenBank, NCBI. The sequence data generated for the polyhedrin gene were deposited in GenBank, and accession number was obtained, MN648213. Field experiment revealed 50.52, 63.25 and 82.91% larval reductions, respectively, at 2, 4 and 7 days after spray of S. obliqua nucleopolyhedrosis virus.ConclusionThe present study revealed that foliar application of S. obliqua nucleopolyhedrosis virus was very effective in causing mortality against S. obliqua larvae and thus can be used as an effective microbial bioagent in the integrated pest management of the caterpillar, S. obliqua.

A New ent-kaurene Diterpenoid Isolated from Leaves of Espeletia semiglobulata Cuatrec. and its Potential Antimicrobial Activity

The fraction of the neutral extraction of the leaves of Espeletia semiglobulata Cuatrec. was subjected to chromatographic separation, it yielded four ent-kaurene type diterpenoids: three known [ent-kaur-16-en-19-al (I), ent-kaur-18-nor-16-en-4-ol (III), ent-kaur-16-en-19-ol (IV)] and a new one elucidated as ent-kaur-3-acetoxy-15-ene (II), based on the physicochemical and spectroscopic data of FTIR, GC-MS, and 1D and 2D NMR. These compounds were subjected to antimicrobial bioassay studies. This new ent-kaurene showed a significant inhibition potential against the growth of gram negative bacterial strains [Escherichia coli (ATCC 25922): 8 mm, Klebsiella pneumoniae (ATCC 23357): 10 mm, Pseudomonas aeruginosa (ATCC 27853): 8 mm], it also showed inhibition against the growth of fungal strain (Candida krusei: 8 mm), at a 2 mg/mL concentration. The compounds (I), (III) and (IV) failed to show any significant results in the antimicrobial screening against five bacterial strains [Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 29212), Klebsiella pneumoniae (ATCC 23357), Escherichia coli (ATCC 25922) y Pseudomonas aeruginosa (ATCC 27853)] and one fungal strain [Candida krusei (ATCC 6558)]. These results reveal a remarkable natural structure-activity relationship of the ent-kaurene core regarding the C-3 position (A ring of perhydrophenanthrene unit), whose oxygenation or addition of a hydrogen bond acceptor or donor group, improves the antimicrobial activity.

Ethnobotanical study of plants used for treating intestinal worms in Ibadan, Nigeria

Poverty and poor sanitation in sub-Saharan Africa are the main reasons for parasitic infection prevalence. Medicinal herbs have been sources of natural and cheaper remedies for the treatment of many parasitic infections, including intestinal parasites. The study aimed to document medicinal herbs used in treating intestinal worm infections in the Ibadan metropolis through an ethnobotanical survey. The three main traditional herb markets, Oje, Oja Oba and Iwo Road, in Ibadan city, were purposively selected for the survey. The respondents, who were mainly herb sellers, were interviewed using structured questionnaires. The information obtained included demographic information, names of plants used in the treatment of worms, plant parts, dosage and method of preparation. The data collected was analysed using descriptive statistics and the Relative Frequency Citation (RFC), Use Value (UV) and Informant Consensus Factor (ICF) of the mentioned plants were calculated. The survey revealed 45 plants that are used in the treatment of intestinal worms in Ibadan. These plants belong to 31 families. The most frequently mentioned plants with their RFCs were Cryptolepis sanguinoleta (Lindl.) Schltr (0.39) and Aristolochia albida Duch (0.39) while most of the plants belong to the Families Apocynaceae and Euphorbiaceae. The roots (25%) and the leaves (18.75%) were the most commonly used plant parts. Most of the plants were indicated to be used singly while some were used in combinations of two or more. The study has revealed the medicinal plants that are used against intestinal parasites. Thus, bioassay studies to confirm the claims should be carried out to establish the activities of these plants.

Antibacterial and Antifungal Activities of Acacia modesta, Achyranthe saspara and Solanum surattense used in Folk Medicine

Background: The antibiotic resistance is a grave concern across the globe due to inappropriate and over usage of antibiotics without prescription which could be fatal for the humans in the long run. Therefore, the need of the hour is to look for more effective and economical options for treatment worldwide to reduce antibiotic resistance.&#x0D; Objective: This study was targeted to assess the antibacterial and antifungal properties of leaf extracts of three plants (Achyranthes aspara, Acacia modesta and Solanum surattense) against Escherichia coli, Salmonella and Malassezia species.&#x0D; Methods: The bioactive components were extracted using methanol and ethanol (70%) as solvents. The agar-well diffusion method was employed to assess the antibacterial and antifungal activities of leaf extract. The inhibitory zones were recorded. The bioassay studies of the crude extracts were undertaken at three different concentrations i.e., 30.0, 40.0, and 50.0 mg/ml.&#x0D; Results and Conclusion: The ethanol extracts of Achyranthes aspara had shown the highest growth inhibitory effect at a concentration of 50.0 mg/ml with an inhibition zone of 13.2mm against E. coli. Ethanolic leaf extracts of solanum surattense had shown maximum inhibition effects at a concentration of 50.0 mg/ml with 12.5mm zones of inhibition against Salmonella. Acacia modesta did not show good results against any bacterial and fungal strains as compared to the other two plants.

Proximate assessment and bioassay of breakfast cereals produced from blends of acha (Digitaria exilis), mungbean (Vigna eadiata) and cashew nut (Anarcadium occidentale Linn) flours

Breakfast cereals were formulated from blends of acha, fermented mungbean and cashew nut (undefatted and defatted) flours. The undefatted and defatted cashew nut flours were used at different levels of substitution (10, 20, 30, 40 and 50 %) with the best blend of acha and mungbean (80:20) flour which was determined by sensory evaluation. Breakfast cereals were produced by toasting (170 oC) a dry heat treatment process to gelatinize and semi-dextrinize the starch in order to generate dry ready to eat products. The samples were subjected to proximate and bioassay analyses. The breakfast cereal was used for a four week bioassay study using eighty four (84) healthy male albino rats weighing 60 - 90 g. The proximate composition showed: crude protein 10.24-20.10 %, moisture content 6.01-8.07 %, crude fat 3.60-24.51 %, crude fibre 4.02-7.07 %, ash 3.39-4.80 % and carbohydrate 46.4- 68.08 %. Feed intake was measured daily while weight gain was measured weekly. Results after the study showed; feed intake 234.22 – 344.55 g, weight gain 34.14 – 80.25 g, feed efficiency 0.14-0.25 and protein efficiency 0.99-1.89. Feeding experiment showed positive increase in weight when the rats were fed the formulated breakfast products. This implies that the products could support growth. No adverse growth rate was observed. Feed intake was also acceptable and compared well with the stable diet for rats. Feed efficiency of the samples compared well with the control diet.

Cardio-Protective Effect of the Leaf Extract of Andrographis paniculata in Isoproterenol-Induced Myocardial Infarction

Background: Nowadays, myocardial infarction has been regarded as one of the chronic diseases with increasing mortality rate worldwide. The use of medicinal plants in the treatment of this chronic disease is gaining wide acceptance globally.&#x0D; Materials and Methods: This study was carried out to evaluate the cardio-protective effect of the leaf extract of A. paniculata in isoproterenol-induced myocardial infarction. Fresh green leaves of A paniculata were collected from the Faculty of Agriculture farmland, Nnamdi Azikiwe University, Awka, Nigeria. The plant was identified and authenticated at the Department of Botany, Nnamdi Azikiwe University and a voucher specimen was deposited at the herbarium accordingly. The shredded, air-dried sample was then pulverized and weighed. Solvent-solvent (ethanol and water) (7:3) was used for extraction via maceration for 72 hr. The filtrate was evaporated to dryness to obtain the ethanol extract which was used for further bioassay study. The bioactive constituents of the plant extract were quantitatively analyzed by Gas chromatography mass spectrometry (GC-MS). The animals were administered with the extract of A. paniculata orally for seven days at a divided dose of 100 mg/kg, 200 mg/kg and 400 mg/kg body weights. On the eight day, myocardial infarction was induced through subcutaneous administration of isoproterenol at a dose of 150 mg/kg/day diluted in 2 ml of saline on two consecutive days. Subsequently, the blood pressures were monitored and blood collected for bioassay studies.&#x0D; Results: The results of the study showed that the leaf extract of A. paniculata was rich in 2,5-Octadecadiynoic acid, methyl ester (28.21%); 1,2,3,5-Cyclohexanetetrol,(1à,2á,3à,5á)- (15.10 %) and 10-12-Pentacosadiynoic acid (13.05%). The findings also showed a significant decrease (p&gt;0.05) in the Mean arterial blood pressure, heart rate, aspartate transaminase, alanine transaminase, creatinine kinase and lactate dehydrogenase activities of the treatment group compared with the untreated control group while the antioxidant (superoxide dismutase, catalase and glutathione) activities were significantly increased in the treatment group, compared with the untreated control group.&#x0D; Conclusion: The findings of this work have shown that leaf of A. paniculata was rich in bioactive compounds which could be synthesized to produce plant based products to combat cardiovascular diseases especially myocardial infarction.

Natural products-based: Synthesis and antifungal activity evaluation of novel L-pyroglutamic acid analogues.

Botanical pesticides are one of the sources of third-generation pesticides, which have received much attention at home and abroad in recent years due to their degradable and pollution-free advantages in nature. This article explored a concise approach toward synthesizing a series of novel L-pyroglutamic acid analogues from L-hydroxyproline. Furthermore, bioassay studies of these sulfonyl ester derivatives against Pyricularia oryzae, Fusarium graminearum, Alternaria brassicae, Valsa mali, and Alternaria alternariae showed moderate antifungal activity. For instance, C08a and C08l provide potential lead agents for controlling Fusarium graminearum because of their inhibitory activity.

Chemical Management of Spilosoma obliqua Walker in Greengram

Background: Greengram the most important pulse crops in India suffers a lot due to infestation of many lepidopteran caterpillars mainly at its reproductive stage. Spilosoma obliqua Walk. Arctiidae, Lepidoptera commonly called Bihar hairy caterpillar (BHC) is categorized as the most devastating. Methods: Field efficacy of nine new generation popular insecticides were tested in coastal saline zone of West Bengal against BHC in greengram (IPM 2-3) during 2021-2022. Leaf dip bioassay study of those selected insecticides were also conducted to compute the LC50 value against the field collected as well as laboratory reared F2 generation 3rd instar larvae of BHC. Result: All the test insecticides showed lethal effect towards BHC but the magnitude of protection over control was significantly higher in spinosad (70.92%) followed by flubendiamide (70.35%). Pyriproxifen+fenpropathrin (29.92%) was proved to be the least effective treatment followed by pyriproxyfen+bifenthrin (31.91%). As per LC50 value (ppm) flubendiamide showed the highest toxicity against BHC in respect of laboratory reared caterpillar and field collected caterpillar (4.49 and 6.71 respectively), while pyriproxifen+fenpropathrin the lowest (624 and 637 ppm). BHC was recorded as potent defoliator which was reflected in yield, while pod damage is not so pronounced.

Traditional Use, Phytochemistry and Pharmacological Activities of Four Dalbergia Species (Dalbergia Sissoo, Dalbergia Odorifera, Dalbergia Melanoxylon and Dalbergia Lactea Vatke): A Review

This review study concerned to survey traditional use and scientific reports for four Dalbergia species namely: Dalbergia sisso, Dalbergia melanoxylon, Dalbergia odorifera and Dalbergia lactea vatke. Genus Dalbergia (Fabaceae or Leguminosae) is an important plant in traditional use. For example, in different part of the world these plants have been used traditionally for the treatment of blood diseases, syphilis, stomach problems, dysentery, nose disorders, ulcers, skin diseases, abdominal pains and anthelmintic.&#x0D; Phytochemical studies on different parts of these plants have indicated the presences of varieties secondary metabolites except Dalbergia lactea vatke. The widely reported are flavonoids, Cinnamyl flavan, terpenoids, and benzofuran among other things. On the other hand the bioassay studies on crude extracts and pure isolates have shown significant anti-inflammatory, anti-cancer, anti-oxidant, anti-microbial, anti-diabetic and anti-analgesic activities. However there is no literature survey performed on Dalbergia lactea vatke.

Amendment of mycorrhizae and its residual effect on growth and yield of maize (Zea mays) hybrids in coastal land

Abstract. Rustikawati, Suprijono E, Murcitro BG, Herison C, Sitohang SJ. 2022. Amendment of mycorrhizae and its residual effect on growth and yield of maize (Zea mays) hybrids in coastal land. Biodiversitas 23: 5600-5605. Maize (Zea mays L.) is known sensitive to less optimal environmental conditions. Maize cultivation in coastal land has low productivity due to the availability of low nutrients and the ability to hold groundwater. The addition of mycorrhizal fungi (MF) to the coastal land is expected to improve the root environment suitable for plants. This study aimed to evaluate growth and yield of maize hybrids grown in coastal areas amended with MF and examining its residual effect. The newly developed hybrids, CT34 and CT17, and a commercial check cultivar, BISI-18, were grown on a coastal land with the application of mycorrhizae at the rate of 0, 5, and 10 g.plant-1 in a factorial experiment with three replications. Observations were made on plant height, stem diameter, number of leaves, leaf length, leaf width, leaf greenness, root fresh weight, ear length, ear diameter, number of grain rows, number of grain.ear-1, and grain weight.plant-1. Data were analyzed for ANOVA, and the mean comparisons were conducted with DMRT at a=5%. The results showed that there was no interaction effect between the maize hybrids and the rate of mycorrhizae on any observed variables. The new hybrid of CT34 showed almost similar growth to that of the check cultivar, BISI 18, but the yield was significantly lower. CT34 showed better performance than CT17 on all variables. The application of MF significantly increased stem diameter and root fresh weight, which in turn increased maize yields in the coastal land. Increasing the mycorrhizal rate up to 10 g.plant-1 in coastal lands was ineffective in increasing maize productivity compared to 5 g.plant-1. In a bioassay study on the mycorrhizal residues in the second season, the treatment of 10 g.plant-1 significantly increased maize growth by more than 100% compared to without mycorrhizae.

Laboratory evaluation of chemicals against Giant African Snail, Achatina fulica bowdich and bio-assay studies on silkworm, Bombyx mori (L.)

Laboratory evaluation of chemicals against Giant African Snail, Achatina fulica bowdich and bio-assay studies on silkworm, Bombyx mori (L.)

Olfactory enrichment using a maternal pheromone improved post-weaning pig performance and behavior.

The post-weaning environment in commercial pig nursery buildings is typically devoid of both the sow and her semiochemicals. Among other factors, the loss of maternal odors may contribute to post-weaning stress. In this work, we report four studies (studies 1-4) using 192 weaned pigs to evaluate the effects of a maternal pheromone (MP) containing Myristic Acid and Skatole on weaned pig behavior and performance. Study 1 examined behavior among weaned pigs with a focus on lying, aggression and feeding behaviors. All studies used body weight gain and the percentage of pigs that lost weight the first 7 days after weaning as key response variables. The MP stimulated early feeding behavior and reduced aggressive behaviors among weaned pigs (p < 0.01). In study one, pigs were over 4 kg heavier 16 weeks post-weaning when the MP was present for 48 h post-weaning compared to control pigs (p = 0.05). The three other studies used a 7-days bioassay to assess the best dose and form of delivery of the MP among weaned pigs. The single measure that responded uniformly was the percentage of pigs that lost body weight in the first 7 days after weaning. Among control pigs in four separate studies, 25% of the weaned pigs in each study lost body weight by 7 days after weaning, while in 3 of the 4 bioassay studies, 0% of MP-exposed pigs lost weight after weaning (one study had 6% of pigs losing body weight with the MP). This MP effect is both highly statistically significant and, if found on commercial farms, would have a large positive economic benefit. Simultaneously, having fewer pigs losing weight and exhibiting less aggressive behavior will improve the health and welfare of weaned pigs. Providing biologically-relevant olfactory enrichment can improve pig health and welfare.

Systematic evidence map (SEM) template: Report format and methods used for the US EPA Integrated Risk Information System (IRIS) program, Provisional Peer Reviewed Toxicity Value (PPRTV) program, and other "fit for purpose" literature-based human health analyses.

Systematic evidence maps (SEMs) are gaining visibility in environmental health for their utility to serve as problem formulation tools and assist in decision-making, especially for priority setting. SEMs are now routinely prepared as part of the assessment development process for the US Environmental Protection Agency (EPA) Integrated Risk Information System (IRIS) and Provisional Peer Reviewed Toxicity Value (PPRTV) assessments. SEMs can also be prepared to explore the available literature for an individual chemical or groups of chemicals of emerging interest. This document describes the typical methods used to produce SEMs for the IRIS and PPRTV Programs, as well as "fit for purpose" applications using a variety of examples drawn from existing analyses. It is intended to serve as an example base template that can be adapted as needed for the specific SEM. The presented methods include workflows intended to facilitate rapid production. The Populations, Exposures, Comparators and Outcomes (PECO) criteria are typically kept broad to identify mammalian animal bioassay and epidemiological studies that could be informative for human hazard identification. In addition, a variety of supplemental content is tracked, e.g., studies presenting information on in vitro model systems, non-mammalian model systems, exposure-level-only studies in humans, pharmacokinetic models, and absorption, distribution, metabolism, and excretion (ADME). The availability of New Approach Methods (NAMs) evidence is also tracked (e.g., high throughput, transcriptomic, in silico, etc.). Genotoxicity studies may be considered as PECO relevant or supplemental material, depending on the topic and context of the review. Standard systematic review practices (e.g., two independent reviewers per record) and specialized software applications are used to search and screen the literature and may include the use of machine learning software. Mammalian bioassay and epidemiological studies that meet the PECO criteria after full-text review are briefly summarized using structured web-based extraction forms with respect to study design and health system(s) assessed. Extracted data is available in interactive visual formats and can be downloaded in open access formats. Methods for conducting study evaluation are also presented which is conducted on a case-by-case basis, depending on the usage of the SEM.

C-Series Coelenterazine-Driven Bioluminescence Signature Imaging.

The present study introduces a unique BL signature imaging system with novel CTZ analogues named "C-series." Nine kinds of C-series CTZ analogues were first synthesized, and BL intensity patterns and spectra were then examined according to the marine luciferases. The results show that the four CTZ analogues named C3, C4, C6, and C7, individually or collectively luminesce with completely distinctive BL spectral signatures and intensity patterns according to the luciferases: Renilla luciferase (RLuc), NanoLuc, and artificial luciferase (ALuc). The signatural reporters were multiplexed into a multi-reporter system comprising RLuc8.6-535SG and ALuc16. The usefulness of the signatural reporters was further determined with a multi-probe system that consists of two single-chain probes embedding RLuc8 and ALuc23. This study is a great addition to the study of conventional bioassays with a unique methodology, and for the specification of each signal in a single- or multi-reporter system using unique BL signatures and patterns of reporter luciferases.

Label-Free SERS Detection of Protein Damage in Organelles under Electrostimulation with 2D AuNPs-based Nanomembranes as Substrates.

Proteins as the material basis of life are the main undertakers of life activities. However, it is difficult to identify the related proteins in organelles during stimuli-induced stress responses in cells and remains a great challenge in early diagnosis and treatment of disease. Here, proteins in the cell nucleus and mitochondria of cells under the electrical stimulation (ES) process were collected and sensitively detected based on label-free surface-enhanced Raman spectroscopy (SERS) by using AuNP-based nanomembranes as high-performance SERS substrates. Due to the existence of rich "hot spots" on the 2D plasmonic sensing platform, high-quality SERS spectra of proteins were obtained with superior sensitivity and repeatability. From the SERS analyses in vitro, it was found that the conformation of some proteins in the two kinds of organelles from cancerous HCT-116 cells (compared with normal NCM-460 cells) changed significantly and the expression levels of tyrosine, phenylalanine, and tryptophan were significantly promoted during the stimulation process. Although currently the exact proteins are still unknown, the damage of proteins in the organelles of cells at the amino acid level under ES can be revealed by the method. The developed plasmonic SERS sensing platform would be promising for bioassay and cell studies.

Antibacterial and Anticancer Properties of Microbispora sp., AL22: An Endophyte of Alpinia galanga (L.) Willd.

&lt;b&gt;Background and Objective:&lt;/b&gt; The AL22 strain was isolated from the rhizosphere soil of &lt;i&gt;Alpinia galanga&lt;/i&gt; (L.) Willd (Zingiberaceae) and identified as &lt;i&gt;Microbispora&lt;/i&gt; sp., by analysing its morphology, chemotaxonomy and 16S rDNA sequence. Previous studies demonstrated the bactericidal effects of its crude extract against &lt;i&gt;Bacillus cereus&lt;/i&gt;, &lt;i&gt;Bacillus subtilis&lt;/i&gt;, &lt;i&gt;Staphylococcus aureus&lt;/i&gt; and methicillin-resistant &lt;i&gt;Staphylococcus aureus&lt;/i&gt;. The present study aimed to isolate the major compounds and evaluate their biological properties. &lt;b&gt;Materials and Methods:&lt;/b&gt; Silica gel column chromatography and thin-layer chromatography were used for the purification and identification of 3,4-dihydro-lactucin (compound &lt;b&gt;1&lt;/b&gt;) and umbelliferone (compound &lt;b&gt;2&lt;/b&gt;) by NMR and mass spectrometry, respectively. Antibacterial and anticancer activities were carried out. &lt;b&gt;Results:&lt;/b&gt; The bioassay studies illustrated that compound &lt;b&gt;1&lt;/b&gt; had antibacterial activity against gram-positive bacteria, with its minimum inhibitory concentration and minimum bactericidal concentration of 16-32 and 64-128 μg mL&lt;sup&gt;&lt;/sup&gt;&lt;sup&gt;1&lt;/sup&gt;, respectively. The crude extract and purified compounds showed weak cytotoxic activity on the L929 and Vero cells with IC&lt;sub&gt;50&lt;/sub&gt; values >512.00 μg mL&lt;sup&gt;&lt;/sup&gt;&lt;sup&gt;1&lt;/sup&gt;. The cytotoxicity of compound &lt;b&gt;1&lt;/b&gt; was observed in the MDA-MB-231 and HeLa cells with IC&lt;sub&gt;50&lt;/sub&gt; values of 37.62 and 75.34 μg mL&lt;sup&gt;&lt;/sup&gt;&lt;sup&gt;1&lt;/sup&gt;, respectively, while its IC&lt;sub&gt;50&lt;/sub&gt; value against the HepG2 cells was 456.67 μg mL&lt;sup&gt;&lt;/sup&gt;&lt;sup&gt;1&lt;/sup&gt;. &lt;b&gt;Conclusion:&lt;/b&gt; These findings showed that compound &lt;b&gt;1&lt;/b&gt; of &lt;i&gt;Microbispora&lt;/i&gt; sp., AL22 exhibited antibacterial and anticancer activities. Extensive studies on 3,4-dihydro-lactucin could lead to the development of beneficial approaches for managing bacterial infections and cancer.