Disposable Sensor Chips with Molecularly Imprinted Carbon Paste Electrodes for Monitoring Anti-Epileptic Drugs.

Epilepsy is a neurological disorder that affects millions of people worldwide. Anti-epileptic drugs (AEDs) are critical for their management. However, the therapeutic window is narrow, and traditional laboratory-based therapeutic drug monitoring (TDM) methods can be time consuming and unsuitable for point-of-care testing. To address this issue, we developed a disposable sensor chip based on molecularly imprinted polymer-modified carbon paste electrodes (MIP-CPs) for the TDM of AEDs such as phenobarbital (PB), carbamazepine (CBZ), and levetiracetam (LEV). In this work, functional monomers (methacrylic acid) and crosslinking monomers (methylene bisacrylamide and ethylene glycol dimethacrylate) were copolymerized in the presence of the AED template and grafted on the graphite particles by simple radical photopolymerization. The grafted particles were mixed with silicon oil, dissolving ferrocene as a redox marker to make the MIP-carbon paste (CP). Disposable sensor chips were fabricated by packing the MIP-CP into the base made of poly (ethylene glycol terephthalate) (PET) film. The sensor's sensitivity was determined using differential pulse voltammetry (DPV), carried out on a single sensor chip for each operation. Linearity was obtained from 0-60 µg/mL in PB and LEV and 0-12 µg/mL in CBZ, covering their respective therapeutic range. The time taken for each measurement was around 2 min. The experiment using whole bovine blood and bovine plasma indicated that the existence of species that interfered had a negligible effect on the test's sensitivity. This disposable MIP sensor provides a promising approach for point-of-care testing and facilitating the management of epilepsy. Compared with existing tests, this sensor offers a faster and more accurate way to monitor AEDs, which is crucial for optimizing therapy and improving patient outcomes. Overall, the proposed disposable sensor chip based on MIP-CPs represents a significant advancement in AED monitoring, with the potential for rapid, accurate, and convenient point-of-care testing.

Two unusual conjugated fatty acids, parinaric acid and α-eleostearic acid, are present in several Impatiens species, but not in congener Hydrocera triflora.

Parinaric and α-eleostearic acids are unusual conjugated fatty acids. Unusual fatty acids, in general, are known to have roles in defense response; however, the role of parinaric acid in I. balsamina is not known, nor is it known whether it occurs in different species of Impatiens or its closest monotypic relative, Hydrocera triflora (L.) Wight & Arn. The aim of the study was to (a) characterize the fatty acid composition of 21 species of Impatiens and H. triflora and (b) determine whether parinaric and α-eleostearic acids are present in these taxa and, if so, (c) whether there is interspecific and intraspecific variation in parinaric acid content. Fatty acid profiling was done using gas chromatography and mass spectrometry (GC-MS). To uncover taxonomic patterns of variation in fatty acids, principal component analysis and hierarchical cluster analysis were performed. The major fatty acids in Impatiens were found to be palmitic (5.57-20.85%), stearic (2.86-21.61%), oleic (2.79-28.99%), linoleic (C18:2Δ9,12, 2.04-26.64%), α-linolenic (C18:3∆9,12,15; 11.07-53.99%), and four forms of parinaric acid (5.93-70.21%). Genus Impatiens contains two unusual conjugated fatty acids- parinaric and α-eleostearic, however these are absent in closely related H. triflora. This study reports the presence of four different forms of parinaric acid in Impatiens for the first time. Some species (I. mengtszeana, I. racemulosa and I. oppositifolia) were found to contain very high levels (> 50%) of parinaric acid and they might be useful for various biomedical and industrial applications. Apparently, the presence of parinaric acid is a characteristic of Impatiens. Significant variations were found in the amount and forms of parinaric acid. We propose the potential application of parinaric acid and α-eleostearic acid as chemotaxonomic markers for Impatiens. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01194-4.

Interaction between Cymbidium aloifolium and Apis cerana: Incidence of an outlier in modular pollination network of oil flowers.

So far, oil-rewarding flowers are known to be pollinated only by oil-collecting bees, which gather and use lipids for larval feed and nest building. As honeybees do not have oil-collecting appendages on their legs, they have not been associated with pollination of such flowers. In a predominantly Apis pollinated and food deceptive clade of wild Cymbidiums, we investigated the reproductive strategy of Cymbidium aloifolium, hitherto unknown for its floral oil reward. Our study demonstrates the requisites for establishment of mutualistic interaction between the oil flower and Apis cerana indica, a corbiculate bee. Success in pollination requires learning by honeybees to access the food reward, thereby displaying cognitive ability of the pollinator to access the customized reward. Morphometric matching between orchid flowers and the pollinator, and that between pollinia and stigmatic cavity also appear to be essential in the pollination success. Absence of pollinator competition and prolonged flower-handling time are suggested to promote floral constancy. The present study highlights the need to explore the spectrum of pollination rewards pursued by honeybees, which may include unconventional composition of floral resources.

Functional characterization and expression profile of microsomal FAD2 and FAD3 genes involved in linoleic and α-linolenic acid production in Leucas cephalotes.

The genus Leucas belongs to Lamiaceae, and has attained more attention due to the presence of unusual allenic fatty acids called laballenic and phlomic acid in majority of its species. This genus has been known since traditional medicinal times and has numerous economical, nutritional, and industrial properties. So far genetic, molecular and biochemical analyses of lipid metabolism and fatty acid biosynthetic pathway in Leucas has not been reported. The objective of this study is to identify, isolate, analyze expression profiles, and functionally characterize the membrane-associated desaturases responsible for unsaturated fatty acid accumulation in Leucas cephalotes. Full-length LcFAD2 and LcFAD3 cDNAs were isolated and expressed in Saccharomyces cerevisiae BY4741 for functional characterization. Substrate feeding assay using S. cerevisiae confirmed that the LcFAD2 enzyme catalyzes desaturation of both palmitoleic (16:1∆9) and oleic (18:1∆9) acids to form palmitolinoleic (16:2∆9,12) and linoleic (18:2∆9,12) acids respectively. As a contrast, the heterologous activity of LcFAD2 enzyme in S. cerevisiae led to the synthesis of palmitolinoleic (16:2∆9,12) acid, an unusual fatty acid that is not found naturally in Leucas cephalotes. While the LcFAD3 enzyme catalyzed linoleic acid (18:2∆9,12) into α-linolenic acid (18:3∆9,12,15). Furthermore, transcript abundance of LcFAD2 and LcFAD3 cDNAs were estimated from various plant parts such as roots, shoots, leaves, petals and developing seeds. Our results have shown that the differential transcriptional activity of LcFAD2 and LcFAD3 desaturase genes differs significantly in developing seeds, petals, leaves, stems, and roots of L. cephalotes. Furthermore, for the industrial production of these essential fatty acids, namely, linoleic and α-linolenic acid, FAD2 and FAD3 enzyme activity could be exploited from this upcoming significant oil plant, Leucas cephalotes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01016-z.

Justicia adhatoda reveals two morphotypes with possible functional significance.

Justicia adhatoda L. (Acanthaceae), an Old-World species of Justicia, is found in almost all geographical regions of India. Indian botanists have persistently used two accepted synonyms of J. adhatoda, namely, Adhatoda vasica and Adhatoda zeylanica, treating them as names of separate species, but without considering or making any reference to variation of forms in the species. Here, different aspects of variation-phenotypic, genotypic, and distributional-in Indian populations of J. adhatoda were studied to determine whether the two names might have been used to designate distinguishable forms of the species. We conducted field studies in different regions of India, laboratory studies of diverse phenotypic traits in experimental plots (anatomical, biochemical, reproductive, and morphometric), and a preliminary study of genetic variation using homologous cytochrome P450 gene fragments. We assessed herbarium samples from across India and the taxonomic literature for pointers indicating the presence of distinguishable forms. Population-level phenotypic and genetic variation pointed to the presence of two distinct morphotypes of the species, which separately tend to occur in dry and wet regions. Each form retains its original phenotype, either when the two forms are transplanted and cultivated together, or when found growing in regions (presumed introduced) outside its normal distributional range. Morphological studies and metabolic profiling (leaf and seed fatty acids, wax load and wax composition in leaf) suggest functional adaptation of the two forms, one to drier and the other to wetter regions. We could distinguish these forms in herbarium specimens dating back to 1821, but neither herbarium specimens nor the taxonomic literature reveal any reference to two forms. We propose that the forms be recognized as two distinct morphotypes of Justicia adhatoda.

COVID-19: Emergence of Infectious Diseases, Nanotechnology Aspects, Challenges, and Future Perspectives.

Wuhan, a city of China, is the epicenter for the pandemic outbreak of coronavirus disease-2019 (COVID-19). It has become a severe public health challenge to the world and established a public health emergency of international worry. This infectious disease has pulled down the economy of almost all top developed nations. The coronaviruses (CoVs) known for various epidemics caused time to time. Infectious diseases such as severe acute respiratory syndrome (SARS) and middle east respiratory syndrome (MERS), followed by COVID-19, are all coronaviruses led outbreaks that scourged the history of mankind. CoVs evolved themselves to more infectious, transmissible, and more pandemic with time. To prevent the spread of the SARS-CoV-2, many countries have ordered the complete lockdown to combat the outbreak. This paper briefly discussed the historical background of CoVs and the evolution of human coronaviruses (HCoVs), the case studies and the development of their antiviral medications. The viral infection encountered with present-day challenges and futuristic approaches with the help of nanotechnology to minimize the spread of infectious viruses. The antiviral drugs and their clinical advances, along with herbal medicines for viral inhibition and immunity boosters, are described. Elaboration of tables related to CoVs for the compilation of the literature has been adopted for the better understanding.

Fatty acid profiling and multivariate analysis in the genus Leucas reveals its nutritional, pharmaceutical and chemotaxonomic significance.

Genus Leucas with about 41 species found in India, is an Asian genus with separation from its close relatives in Africa based on phylogenetic evidence. Present study represents the only comprehensive phytochemical investigation on this genus. We have analyzed the seed fatty acid compositions of 26 species and five varieties of Leucas for nutritional, pharmaceutical and chemotaxonomic perspectives. The fatty acids and their composition in seeds of Leucas species, collected from different geographical regions in India, were analyzed by gas chromatography and mass spectrometry. Significant variations have been observed in fatty acid profiles among species and their varieties. We observed major fatty acids as palmitic, stearic, oleic, linoleic and laballenic acid; whereas myristic, palmitoleic, cis-vaccenic, linolenic, eicosanoic, eicosenoic, phlomic and docosanoic acid were detected in minor quantities. Laballenic and phlomic acids are unusual allenic fatty acids found in few Lamiaceae members from order Lamiales. Laballenic acid, a proven molecule of pharmaceutical importance, was observed in all the Leucas species studied. Three species of Leucas; L. helianthimifolia, L. ciliata var. vestita and L. hirta were found to contain ≥40% laballenic acid and can act as potential source for isolation of pharmaceutical compounds. This study also reports the presence of another allenic fatty acid, phlomic acid, in several Leucas species. Principal component analysis and hierarchical cluster analysis showed a distinct separation among the species based on abundance of similar fatty acids. The fatty acid profile appears to be overlapping at higher level and does not support separation of Asian Leucas from its African relatives and the inclusion of Asian taxa in morphologic sections. However, hierarchical clustering of L. helianthimifolia, L. ciliata var. vestita and L. hirta supported treatment under the morphologic section Astrodon. Multivariate analysis on the chemometric data also supported this cluster as the most prominent source of medicinally useful laballenic acid. Based on the FAs profile, a reconsideration of species boundaries in L. ciliata and L. marrubioides species complex is proposed.

A modified protocol yields high-quality RNA from highly mucilaginous Dioscorea tubers.

Extraction of RNA from Dioscorea is difficult because of rich mucilage and secondary metabolite content. High-quality RNA is required for RT-PCR and transcriptome analysis. Different protocols and common extraction kits were used for RNA extraction in Dioscorea, but the results were not satisfactory. A CTAB based protocol with lithium chloride precipitation yielded good quality RNA. The RNA isolated using this protocol was successfully used for RT-PCR and transcriptome sequencing experiments. Mucilage content varies at different developmental stages of Dioscorea and the present protocol was effective in isolating RNA from such samples. Although the protocol was originally modified for tuber tissues, it can be used also for extraction of RNA from rhizome, root, shoot and leaf.