RESUMO
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.
Assuntos
Grafite , Impressão Molecular , Animais , Bovinos , Humanos , Carbono , Polímeros , Eletrodos , Impressão Molecular/métodos , Técnicas Eletroquímicas/métodos , Limite de DetecçãoRESUMO
The implant survival largely depends on the mechanical setting in which they work as an independent entity. The implant design, number and position markedly affect the treatment plan. Deficient bone quantity and quality and presence of a vital anatomical landmark have often led the practitioners to conduct researches to find newer ways of implant insertion. One such technique is the 'All-On-4' concept, an alternative to the conventional implant therapy; lessens the amount of strain to the alveolar bone by increasing the antero-posterior spread with distal tilting the posterior implant. It is a cost-effective procedure that decreases the treatment time and the morbidity rate allowing a higher patient quality of life. The article demonstrates clinical cases describing rehabilitation of completely edentulous arches using the All-On-4 concept.
Assuntos
Perda do Osso Alveolar , Implantes Dentários , Carga Imediata em Implante Dentário , Arcada Edêntula , Humanos , Arcada Edêntula/cirurgia , Qualidade de Vida , Perda do Osso Alveolar/cirurgia , Carga Imediata em Implante Dentário/métodos , Maxila/cirurgia , Implantação Dentária Endóssea , Resultado do TratamentoRESUMO
Indian lotus (Nelumbo nucifera) is one of the dominant aquatic plants cultivated in Dal Lake, situated at 1586 m above mean sea level (MSL) in the northeast of Srinagar, Kashmir. Despite their economic and ecological role, the microbial communities associated with the lotus plant are still unexplored. In this study, we investigated the prokaryotic communities on surfaces of different lotus microhabitats (roots, rhizome, leaves, flowers, and fruits), lake water, and sediments using 16S rRNA gene amplicon sequencing. Overall, prokaryotic diversity decreased significantly on the surface of lotus microhabitats in comparison to the lake water and sediments. Among the microhabitats of lotus, roots and leaves harbored more diverse communities in comparison to rhizomes, fruits, and flowers. A total of 98 genera were shared by lotus and the Dal Lake sediments and water. However, significant differences were found in their relative abundance; for example, Pseudomonas was the most dominant genus on the majority of lotus microhabitats. On the other hand, Flavobacterium was highly abundant in the lake water, while a higher abundance of Acinetobacter was recorded in sediments. Additionally, we also noted the presence of potential human pathogenic genera including Escherichia-Shigella, Enterobacter, Pantoea, Raoultella, Serratia, and Sphingomonas on the lotus microhabitats. Predicted functions of prokaryotic communities revealed a higher abundance of genes associated with nutrient uptake in the microhabitats of the lotus. This study offered first-hand information on the prokaryotic communities harbored by lotus plants and water and sediments of the Dal Lake and demonstrated the adaptation of diverse communities to microhabitats of lotus.
Assuntos
Nelumbo , Humanos , Nelumbo/genética , Lagos , RNA Ribossômico 16S/genética , Altitude , ÁguaRESUMO
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.
RESUMO
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.
RESUMO
PURPOSE: We describe here a multicentric community-dwelling cohort of older adults (>60 years of age) established to estimate incidence, study risk factors, healthcare utilisation and economic burden associated with influenza and respiratory syncytial virus (RSV) in India. PARTICIPANTS: The four sites of this cohort are in northern (Ballabgarh), southern (Chennai), eastern (Kolkata) and western (Pune) parts of India. We enrolled 5336 participants across 4220 households and began surveillance in July 2018 for viral respiratory infections with additional participants enrolled annually. Trained field workers collected data about individual-level and household-level risk factors at enrolment and quarterly assessed frailty and grip strength. Trained nurses surveilled weekly to identify acute respiratory infections (ARI) and clinically assessed individuals to diagnose acute lower respiratory infection (ALRI) as per protocol. Nasal and oropharyngeal swabs are collected from all ALRI cases and one-fifth of the other ARI cases for laboratory testing. Cost data of the episode are collected using the WHO approach for estimating the economic burden of seasonal influenza. Handheld tablets with Open Data Kit platform were used for data collection. FINDINGS TO DATE: The attrition of 352 participants due to migration and deaths was offset by enrolling 680 new entrants in the second year. All four sites reported negligible influenza vaccination uptake (0.1%-0.4%), low health insurance coverage (0.4%-22%) and high tobacco use (19%-52%). Ballabgarh had the highest proportion (54.4%) of households in the richest wealth quintile, but reported high solid fuel use (92%). Frailty levels were highest in Kolkata (11.3%) and lowest in Pune (6.8%). The Chennai cohort had highest self-reported morbidity (90.1%). FUTURE PLANS: The findings of this cohort will be used to inform prioritisation of strategies for influenza and RSV control for older adults in India. We also plan to conduct epidemiological studies of SARS-CoV-2 using this platform.
Assuntos
COVID-19 , Influenza Humana , Infecções por Vírus Respiratório Sincicial , Vírus Sincicial Respiratório Humano , Infecções Respiratórias , Vírus , Idoso , Humanos , Índia/epidemiologia , Lactente , Influenza Humana/epidemiologia , Infecções Respiratórias/epidemiologia , SARS-CoV-2RESUMO
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.
RESUMO
AIM AND OBJECTIVE: To compare between mineral trioxide aggregate (MTA) mixed with water and water-based gel regarding shear bond strength with resin-modified glass ionomer cement (RMGIC) and composite. METHODS AND MATERIALS: In this study, 40 blocks of cylindrical shape were prepared with acrylic. These blocks were divided into four groups with each group consisting of 10 blocks: group-1A: MTA + distilled water + composite, group-1B: MTA + distilled water + RMGIC, group-2A: MTA + polymer + composite, and group-2B: RMGIC + MTA + polymer. After that, a universal testing machine was used for the measurement of shear bond strength. The acrylic blocks were placed under this machine. A blade with a knife-edge was used to provide a crosshead speed of 1 mm/minute. This was continued till bond of MTA in both forms (distilled water/gel) and restorative material failed. RESULTS: It was observed that a statistically significant difference was found between MTAw + composite and MTAg + composite resin but no statistically significant difference between MTAw + RMGIC and MTAg + RMGIC with p ≥ 0.05. It was found that a statistically significant difference was present between the RMGIC and composite groups within the same MTA type with p ≤ 0.05. CONCLUSION: It was concluded from the present study that MTA with a water-based gel has a better shear bond strength than composite resin and RMGIC materials. CLINICAL SIGNIFICANCE: It has been found that MTA has different properties when it is mixed with polymer and water. Very few studies have been conducted in the past to compare MTA mixed with water and water-based gel regarding the shear bond strength with RMGIC and composite.
Assuntos
Colagem Dentária , Cimentos de Ionômeros de Vidro , Compostos de Alumínio , Compostos de Cálcio , Resinas Compostas , Combinação de Medicamentos , Teste de Materiais , Óxidos , Resistência ao Cisalhamento , Silicatos , ÁguaRESUMO
AIM: To compare the shear bond strength of composites to the sodium ascorbate hydrogel-treated bleached enamel surfaces. MATERIALS AND METHODS: Sixty-six extracted human maxillary first premolars were sectioned mesiodistally to obtain two nonocclusal surfaces (n = 132). The specimen in the experimental group (n = 80) were divided into four treatment groups depending upon bleaching protocols used: group I (10% FGM home bleach), group II (22% Pola Night home bleach), group III (37% light-activated in-office bleach Pola Office+), and group IV (35% Pola office in-office bleach without light activation). Specimens were subjected to the 1-minute application twice with sodium ascorbate hydrogel and were further divided into two subgroups "a" (acetone-based) and "b"(ethanol-based) depending upon the bonding agents used. The specimens of the control group (n = 52) were further divided into two treatment groups, that is, positive control and negative control. The specimens were tested using universal testing machine. RESULTS: The result of the present study revealed that the specimens bleached with 10% FGM home bleach showed the greatest shear bond strength and specimens bleached with 35% in-office bleach Pola Office showed the lowest shear bond strength. CONCLUSION: A twice 1-minute application of sodium ascorbate hydrogel was effective to reverse the deleterious results of bleaching on enamel shear bond strength. Bonding agent containing acetone as a solvent (Prime and Bond NT) in this present study showed greater shear bond strength values than the bonding agent containing ethanol as a solvent (Adper Single Bond). STATEMENT OF CLINICAL RELEVANCE: The shear bond strength of composite restorative materials could be improved by applying sodium ascorbate gel to the bleached enamel surfaces. HOW TO CITE THIS ARTICLE: Pathak K, Kumar P, Choudhary A, et al. Comparative Analysis of Shear Bond Strength of Composites to the Sodium Ascorbate Hydrogel-treated Bleached Enamel Surfaces: An In vitro Analysis. Int J Clin Pediatr Dent 2021;14(6):741-747.
RESUMO
OBJECTIVE: This study aimed to evaluate the surface roughness of three flowable esthetic restorative materials after exposure to sports/energy drinks and alcoholic beverages. MATERIALS AND METHODS: A total of 210 specimens of dimension (2cm diameter and 2 mm thickness) with giomer, compomer, and composite (70 samples with each esthetic material) were made with the help of plastic rings. The prepared samples were tested in six experimental sports/energy drinks (beer, whiskey, vodka, Gatorade, Red Bull, and Sting) and distilled water was considered as the control group. Profilometric analyses of all samples were recorded before immersing into the experimental and control solutions. Then, the samples were stored in the experimental and control group solutions for 5min for 30 days. The profilometric analysis was repeated after 30 days and records were statistically analyzed. RESULTS: Flowable composite showed the minimum surface roughness, whereas the flowable compomer showed the maximum surface roughness in the present test conditions. When the erosive potential of the test solutions was evaluated, surface roughness values were more for sports/energy drinks when compared to that of alcoholic beverages. CONCLUSION: All the sports/energy drinks and alcoholic beverages evaluated in this study altered the surface roughness of the tested restorative materials. The effects ranged from slight to a markedly negative impact on the surface roughness of the test restorative materials.
RESUMO
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.
Assuntos
Justicia/anatomia & histologia , Fenótipo , Ecossistema , Variação Genética , Índia , Justicia/química , Folhas de Planta/químicaRESUMO
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.
RESUMO
AIM: The present study aimed to evaluate and compare the applicability of Demirjian, Willems, and Haavikko methods of dental age estimation in southeastern (Dravidian ethnicity) and northwestern regions (Aryan ethnicity) of the Indian population. MATERIALS AND METHODS: The study includes the orthopantomographs (OPGs) of 303 individuals (173 males and 130 females) of age ranging from 5 to 14 years. The participants in the present research were evaluated under two study groups: group I: participants of the southeastern region and group II: participants of the northwestern region. Dental age was calculated using Demirjian, Willems, and Haavikko methods and compared with the chronologic age of each participant. The accuracy of dental age estimation methods was evaluated by mean absolute error. One-way analysis of variance (ANOVA) and Student's t tests were used to test the significant difference between the chronologic age and estimated dental ages. RESULTS: The Demirjian method showed overestimation, while Willems and Haavikko methods showed underestimation for boys and girls in both the ethnic groups of Indian population. The Willems method of dental age estimation showed comparatively more accurate and reliable results in both the ethnic groups of the Indian population. CONCLUSION: The dental age estimation by Demirjian, Willems, and Haavikko methods showed no significant variation between the different ethnicities of the Indian population. CLINICAL SIGNIFICANCE: The present research will be helpful in pedodontic, orthodontic, and forensic investigations for accurate and reliable dental age estimation in different parts of Indian population. HOW TO CITE THIS ARTICLE: Chaudhry K, Talwar M, Vanga NRV, et al. A Comparative Evaluation of Three Different Dental Age Estimation Methods in India: A Test of Ethnic Variability. Int J Clin Pediatr Dent 2020;13(1):16-20.
RESUMO
AIM: This study evaluates epoxy resin-based sealers after their final set, immersed in Endosolve-R or xylene for 1-2 minutes, for its easy removal mechanically after softening. MATERIALS AND METHODS: Sixty Teflon molds were grouped with 20 samples in each of the three commercially available sealers, i.e., AH 26, AH Plus, and Adseal. The sealers were put in the specific molds after their manipulation as per the instructions given in the literature by the manufacturer. They were allowed to harden for 2 weeks at 37°C in 100% humidity. Two subgroups, A-Xylene and B-Endosolv-R, of 10 samples each, were formed from 20 set specimens based on solvents to which they were immersed for 1 and 2 minutes, respectively. The data obtained was subjected to the Mauchly's test one-way ANOVA and two-way ANOVA for analysis. RESULTS: It was proved that for all the sealers immersed in solvents, there was a significant reduction in the mean Vickers hardness as the time increases. There was a significant difference in the initial hardness between the mentioned sealers with AH plus showing the highest followed by AH 26 and Adseal showing the lowest. AH Plus and Adseal sealers were softened by xylene after 2 minutes of their initial microhardness (p < 0.001); least effect was seen on AH 26. After 2 minutes, Endosolv-R softened initial microhardness of all the three sealers (p < 0.001). CONCLUSION: It was concluded that Endosolv-R was more effective in softening the epoxy-based resin sealer than xylene, after 2 minutes of exposure. HOW TO CITE THIS ARTICLE: Tyagi S, Choudhary E, Choudhary A, et al. A Comparative Evaluation of Two Commonly Used GP Solvents on Different Epoxy Resin-based Sealers: An In Vitro Study. Int J Clin Pediatr Dent 2020;13(1):35-37.
RESUMO
[This corrects the article DOI: 10.1371/journal.pone.0093102.].
RESUMO
INTRODUCTION: Increasing age, long-term deleterious habits, and increased bone resorption leading to the gingival recession have become a significant reason for poor geriatric oral health among which root caries are the most prevalent ones among the Indian population. METHODOLOGY: A cross-sectional study was conducted among the geriatric population above 60 years of age and those who gave voluntary consent for participating in the study were included in the study and were briefed about the questionnaire. The self-administered questionnaire was given and complete oral examination was done. Results were analyzed using the SPSS 22 version using descriptive analysis and Chi-square test. RESULT: Percentage of deleterious habits amongst males was higher than that of females i.e. 56%. Thus, males were reported with root caries more than females. The percentage of married people suffering from root caries was 85%. The most prevalent occupation among the geriatric population was found to be the farmers with 37%. Religion with maximum percentage and frequency was found to be Hinduism with 84%. Educational statuses with 6th to 12th standards were reported with maximum root caries, i.e. 55%. CONCLUSION: The study showed that criteria including income, gender, marital status, occupation, religion, and educational status were significant in assessing the prevalence of root caries. Whereas the risk factors such as habits, methods of cleaning, and brushing techniques influenced the root caries among the geriatric population.
RESUMO
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.
Assuntos
Ácidos Graxos/análise , Lamiaceae/química , Cromatografia Gasosa-Espectrometria de Massas , Índia , Lamiaceae/genética , FilogeniaRESUMO
Auxin levels are tightly regulated within the plant cell, and its storage in the isolated cavity of proteins is a measure adopted by cells to maintain the availability of auxin. We report the first crystal structure of Wrightia tinctoria 11S globulin (WTG) in complex with Indole-3-acetic acid (IAA), an auxin, at 1.7 Å resolution. WTG hexamers assemble as a result of the stacking interaction between the hydrophobic surfaces of two trimers, leaving space for the binding of charged ligands. The bound auxin is stabilized by non-covalent interactions, contributed by four chains in each cavity. The presence of bound ligand was confirmed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and high-resolution mass spectrometry (HRMS). Here, we hypothesize that the cleavage of globulins by endopeptidases leads to the movement of the hydrophilic loop region from the surface to the periphery, leaving space for the binding of auxin, and promotes hexamer formation. As the process of germination proceeds, there is a change in the pH, which induces the dissociation of the hexamer and the release of auxin. The compact hexameric assembly ensures the long-term, stable storage of the hormone. This suggests a role for globulin as a novel player in auxin homeostasis.
Assuntos
Apocynaceae/metabolismo , Globulinas/química , Globulinas/metabolismo , Ácidos Indolacéticos/química , Ácidos Indolacéticos/metabolismo , Apocynaceae/química , Apocynaceae/fisiologia , Sítios de Ligação , Cristalografia por Raios X , Endopeptidases/metabolismo , Germinação , Modelos Moleculares , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Ligação Proteica , Conformação Proteica , Estrutura Quaternária de Proteína , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
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.
RESUMO
BACKGROUND: Improvements in both performance and cost for next-generation sequencing (NGS) have spurred its rapid adoption for clinical applications. We designed and optimized a pan-cancer target-enrichment panel for 51 well-established oncogenes and tumor suppressors, in conjunction with a bioinformatic pipeline informed by in-process controls and pre- and post-analytical quality control measures. METHODS: The evaluation of this workflow consisted of sequencing mixtures of intact DNA to establish analytical sensitivity and precision, utilization of heuristics to identify systematic artifacts, titration studies of intact and FFPE samples for input optimization, and incorporation of orthogonal sequencing strategies to increase both positive predictive value and variant detection. We also used 128 FFPE samples to assess clinical accuracy and incorporated the previously described quantitative functional index (QFI) for sample qualification as part of detailing complete system performance. RESULTS: We observed a concordance correlation coefficient of 0.99 between the observed versus expected percent variant at 250 ng input across 4 independent sequencing runs. A subset of the systematic variants were confirmed to be barely detectable on an independent sequencing platform (Wilcox signed-rank test p-value <10(-16)), and the incorporation of orthogonal sequencing strategies increased the harmonic mean of sensitivity and positive predictive value of mutation detection by 41%. In one cohort of FFPE tumor samples, coverage and inter-platform concordance were positively correlated with the QFI, emphasizing the need for pre-analytical sample quality control to reduce the risk of false positives and negatives. In a separate cohort of FFPE samples, the 51-gene panel achieved 78% sensitivity (95% CI = 56.3, 92.5) with 100% PPV (95% CI = 81.5, 100.0) based on known mutations at 7.9% median abundance. By sequencing specimens using an orthogonal NGS technology, sensitivity was improved to 87.0% (95% CI = 66.4,97.2) while maintaining PPV. CONCLUSIONS: The results highlight the value of process integration in a comprehensive targeted NGS system, enabling both discovery and diagnostic applications, particularly when sequencing low-quality cancer specimens.