Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature

Abstract The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.

Resumo A esporulação de Didymella bryoniae in vitro é de grande importância para estudos que requerem inóculo puro e em grandes quantidades. Assim, os objetivos deste estudo foram identificar a melhor condição para esporulação de D. bryoniae combinando diferentes espectros de luz (luz UV-A ou UV-B, luz branca e escuro contínuo) com distintos meios de cultura (PDA, V8, ML e PDAB) e, avaliar a sobrevivência do fungo armazenado a -20°C ao longo do tempo. As amostras de fungo só esporularam quando submetidas ao tratamento com luz UV-B, independentemente do meio de cultura. Maior aparecimento de esporos do tipo conídio foi observado no meio PDAB, e a menor produção ocorreu no meio ML. Estruturas reprodutivas, como peritécios e picnídeos, foram observadas em todos os meios de cultura. No entanto, houve uma variação considerável na quantidade de cada estrutura entre os diferentes meios de cultura. Os meios ML e V8 apresentaram maior número de peritécios e os meios PDA e PDAB apresentaram maior proporção de picnídeos em relação aos peritécios. A duração do armazenamento a -20°C não afetou o crescimento micelial ou a taxa de crescimento micelial. Em conclusão, a luz UV-B é essencial para a esporulação de D. bryoniae in vitro. Além disso, a composição do meio de cultura influencia o tipo de estrutura fúngica produzida, bem como o tamanho e a quantidade dos esporos. O congelamento a -20°C é uma técnica eficiente que pode ser usada para armazenar D. bryoniae por pelo menos cinco meses sem perda de viabilidade

Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature / Fornecendo inóculo para estudos de Didymella bryoniae: o efeito do espectro de luz e armazenamento em baixa temperatura

Abstract The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus' survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores' size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.

Resumo A esporulação de Didymella bryoniae in vitro é de grande importância para estudos que requerem inóculo puro e em grandes quantidades. Assim, os objetivos deste estudo foram identificar a melhor condição para esporulação de D. bryoniae combinando diferentes espectros de luz (luz UV-A ou UV-B, luz branca e escuro contínuo) com distintos meios de cultura (PDA, V8, ML e PDAB) e, avaliar a sobrevivência do fungo armazenado a -20°C ao longo do tempo. As amostras de fungo só esporularam quando submetidas ao tratamento com luz UV-B, independentemente do meio de cultura. Maior aparecimento de esporos do tipo conídio foi observado no meio PDAB, e a menor produção ocorreu no meio ML. Estruturas reprodutivas, como peritécios e picnídeos, foram observadas em todos os meios de cultura. No entanto, houve uma variação considerável na quantidade de cada estrutura entre os diferentes meios de cultura. Os meios ML e V8 apresentaram maior número de peritécios e os meios PDA e PDAB apresentaram maior proporção de picnídeos em relação aos peritécios. A duração do armazenamento a -20°C não afetou o crescimento micelial ou a taxa de crescimento micelial. Em conclusão, a luz UV-B é essencial para a esporulação de D. bryoniae in vitro. Além disso, a composição do meio de cultura influencia o tipo de estrutura fúngica produzida, bem como o tamanho e a quantidade dos esporos. O congelamento a -20°C é uma técnica eficiente que pode ser usada para armazenar D. bryoniae por pelo menos cinco meses sem perda de viabilidade

Poly-ε-caprolactone based nanoparticles for delivery of genistein in melanoma treatment

Abstract We developed poly-ε-caprolactone (PCL)-based nanoparticles containing D-α-tocopherol polyethylene glycol-1000 succinate (TPGS) or Poloxamer 407 as stabilizers to efficiently encapsulate genistein (GN). Two formulations, referred to as PNTPGS and PNPol, were prepared using nanoprecipitation. They were characterized by size and PDI distribution, zeta potential, nanoparticle tracking analysis (NTA), GN association (AE%), infrared spectroscopy (FT-IR), and differential scanning calorimetry (DSC). PNTPGS-GN exhibited a particle size of 141.2 nm, a PDI of 0.189, a zeta potential of -32.9 mV, and an AE% of 77.95%. PNPol-GN had a size of 146.3 nm, a better PDI than PNTPGS-GN (0.150), a less negative zeta potential (-21.0 mV), and an AE% of 68.73%. Thermal and spectrometric analyses indicated that no new compounds were formed, and there was no incompatibility detected in the formulations. Cellular studies revealed that Poloxamer 407 conferred less toxicity to PCL nanoparticles. However, the percentage of uptake decreased compared to the use of TPGS, which exhibited almost 80% cellular uptake. This study contributes to the investigation of stabilizers capable of conferring stability to PCL nanoparticles efficiently encapsulating GN. Thus, the PCL nanoparticle proposed here is an innovative nanomedicine for melanoma therapy and represents a strong candidate for specific pre-clinical and in vivo studies.

Carotenoids from marine bacteria: A natural antioxidant and UV protectant

For the cosmetics and pharmaceutical sectors, marine resources are a promising supply of organic substances. Marine organisms have evolved particular metabolites to combat threats to their survival, reproduction, and the easier storage, movement, and turnover of essential biological materials in habitats with extreme conditions like high salinity, low temperature, or intense pressure.Several newly conducted research studies on pigments, such as violacein, astaxanthin, canthaxanthin, zeaxanthin, rubrolone, and carotenoid derivatives of bacteria from marine sources, confirm their effective radical scavenging activity. Carotenoids are tetraterpene, lipophilic bioactive pigments synthesized by archaea, algae, photosynthetic bacteria, and plants. Carotenoids are divided into two categories, oxygenated carotenoids (xanthophylls) having oxygen as a functional group, and hydrocarbon carotenoids (carotene) made of carbon and hydrogen atoms Carotenoids are important because of their functional properties, which include their usage in the food industry as color additives and natural antioxidants, as well as chemotaxonomic markers and pharmaceuticals. The present review aims to describe 1) carotenoids from marine bacteria 2) the antioxidant properties of carotenoids 3) the Exposure consequences of ultraviolet radiation 4) the protective role of carotenoids. Photoaging, sunburn, and skin cancer are all possible side effects of excessive UV exposure. Carotenoids have potential use in the pharmaceuticals and cosmetic industry as anti-aging, photoprotective, skin-whitening, anticancer, and immunity boosters because of their antioxidant and UV protection properties.

Feasibility in the use of the UV-Visible region for the characterization of glucose in deionized water using Arduino 1 / Viabilidad en el uso de la región UV-Visible para la caracterización de glucosa en agua desionizada mediante Arduino 1

ABSTRACT With an estimated approximately 2 million deaths per year, diabetes is one of the top 5 deadliest noncommunicable diseases globally. Although this disease is not fatal, the degradation of the patient's health due to a bad plan to control their glucose levels can have a fatal outcome. In order to lay the foundations for the development of a device that allows estimating glucose levels in some body fluid, we present the results obtained not only for the estimation of glucose in deionized water, but also describe the development and configuration of the created device. After analyzing 50 signals obtained from 5 different glucose concentrations, the feasibility of using the developed device for the analysis is evident, since, considering the K-Nearest Neighbors (KNN) algorithm, all the signals were associated correctly to the glucose group to which they belong.

RESUMEN Con un estimado de aproximadamente 2 millones de muertes por año, la diabetes es una de las 5 enfermedades no transmisibles más mortales a nivel mundial. Aunque esta enfermedad no es mortal, el deterioro de la salud del paciente por un mal plan para controlar sus niveles de glucosa puede tener un desenlace fatal. Con el fin de sentar las bases para el desarrollo de un dispositivo que permita estimar los niveles de glucosa en algún fluido corporal, presentamos los resultados obtenidos no solo para la estimación de glucosa en agua desionizada, sino que también describimos el desarrollo y configuración del dispositivo creado. Luego de analizar 50 señales obtenidos a partir de 5 concentraciones de glucosa diferentes, se evidencia la factibilidad de utilizar el dispositivo desarrollado para el análisis, ya que, considerando el algoritmo K-Nearest Neighbors (KNN), todas las señales se asociaron correctamente al grupo de glucosa al que pertenecen.

Knowledge regarding reverse osmosis (R.O) waste water utilization among general public in urban areas

Introduction: Reverse osmosis (RO) is a water purification process that uses a partial permeable membrane to remove ions, unwanted molecules and larger particles from drinking water. In reverse osmosis, an applied pressure is used to overcome osmotic pressure, a colligative property that is driven by chemical potential differences of the solvent, a thermodynamic parameter. In the process of reverse osmosis the amount of water that is drained is a concern area for the people using the R.O. filtration device in their household because it wasted about 70% of the water to purify just one liter of water. This R.O. waste water can be utilized for various purposes such as washing vehicle like car bike etc, cleaning toilet this study is aimed to assess the knowledge reverse osmosis waste water utilization among general public at Indore. Objectives: 1. To assess the pretest knowledge regarding reverse osmosis (R.O) waste water utilization among general public. 2. To assess the posttest knowledge regarding reverse osmosis waste water utilization among general public. 3. To evaluate the effectiveness of structured teaching program on reverse osmosis (R.O) waste water utilization among general public. Hypothesis: H1- there will be significant difference between pretest and posttest knowledge who received structured teaching program regarding the utilization of waste R.O water. Materials and Methods: Quantitative, pre-experimental, one group pretest posttest design was adopted for the study. Total of 60 general public selected by using simple randomized sampling technique was used. Description of Tools: Structured knowledge questionnaire. Result: Data was analyzes using descriptive and inferential statistics. Major Finding: In the pre-test majority of the sample (44 out of 60, 73.3%) had inadequate knowledge and in the post-test, majority (54 out of 60, 90%) had adequate knowledge regarding reverse osmosis. A paired‘t’ test was done and it showed a‘t’ value of 22.34 at 0.05 level of significance, this indicates the effectiveness of structured teaching programme in enhancing the knowledge of the general public. There was no association found between the mean pre-test knowledge of the general public. There was no association found between the mean pre-test knowledge scorer with the selected socio-demographic variable such as age (?2 = 8.643), gender (?2 = 4.455), education qualification (?2 = 4.706), Occupation (?2 = 2.531), number of family member (?2 = 5.653) and previous knowledge about reverse osmosis filter water (?2 =0.393). Conclusion: There is a significant difference between the mean pre-test and post-test knowledge score among general public regarding reverse osmosis waste water utilization.

Evaluation of effects of riboflavin and/or ultraviolet-A on survival of rat limbal epithelial stem cells in ex-vivo

Purpose: To investigate the effects of riboflavin and/or ultraviolet?A (UV?A) irradiation on the cell viability of ex?vivo?cultured rat limbal stem cells (LSCs). Methods: LSCs of male Wistar rats (N = 12 eyes) were cultured, and immunofluorescence staining was performed to evaluate them. After characterization, these cells were assigned to four groups of control (C), a group that was exposed to UV?A radiation (UV), a group that was treated with riboflavin (R), and a group that cotreated with both UV?A and riboflavin (UV+R). To determine the cell viability of LSCs, these cells were subjected to MTT assay on days 1, 3, and 7 after exposure to UV?A and/or riboflavin. The duration of exposure to UV?A and riboflavin was similar to levels used during the conventional corneal collagen cross?linking procedure. Results: Compared with the viable cells in the control group, there was a significant decrease (P < 0.0001) in the number of LSCs in the UV group during all study days. In the R group, the level of viable LSCs was as same as the level of viable LSCs in the C group. Combined treatment with UV?A plus riboflavin significantly decreased the survival of LSCs on days 1 and 3 (P < 0.0001, P < 0.001, respectively) compared with the control group. Interestingly, in the UV+R group, the photosensitizing effect of riboflavin significantly decreased the cytotoxic effect of UV irradiation 7 days after exposure. Conclusion: These results suggest that the administered UV energy in the presence or absence of riboflavin can damage LSCs. Likewise, riboflavin could decrease the toxic effect of UVA on LSCs

Preservation of phenolic compounds on dried leaf infusion of Bauhinia forficata Link

Abstract Bauhinia forficata Link aqueous extract is usually recommended as a phytomedicine to reduce blood glucose levels and its biological activity has been linked to the presence of phenolic compounds from B. forficata preparations. Several drying processes are used in the production of dry herbal extracts, which may influence the chemical composition and efficacy of final herbal medicines. Due to significant chemical changes, defining appropriate drying processes is essential for phytopharmaceutical drug development. In view of this, we analyzed dried B. forficata leaf infusion (BFLI) extracts by HPLC-UV-MSn, followed by molecular networking analysis to evaluate the chemical profiles from dried extracts yielded by freeze-and spray-drying processes. The main metabolites detected included 11 ferulic/isoferulic acid derivatives and 13 glycosylated flavonoids. The qualitative chemical profiles were alike for both drying processes, whereas the relative abundance of some flavonoids was higher using spray-drying. Taken together, our results showed that freeze-and spray-drying preserved the phenolic profile of BFLI and suggested that spray-drying may be the most suitable to obtain its dried products. Along with studying the chemical profiles of dried herbal extracts, evaluating the influence of drying processes on the quality and chemical profiles of final products is pivotal and may benefit future research.

Qualitative and quantitative study of constituents in Lysionoti Herba based on UHPLC-Q-Exactive Orbitrap HRMS and HPLC-UV / 中国中药杂志

Ultra-high performance liquid chromatography-quadrupole-Exactive Orbitrap high resolution mass spectrometry(UHPLC-Q-Exactive Orbitrap HRMS) was employed to systematically analyze the chemical constituents in Lysionoti Herba, and high perfor-mance liquid chromatography-ultraviolet(HPLC-UV) to determine the content of main compounds. A Synergi~(TM) Hydro-RP 100 Å colu-mn(2 mm×100 mm, 2.5 μm) was used for gradient elution with acetonitrile-0.1% aqueous formic acid as the mobile phase at a flow rate of 0.2 mL·min~(-1) and a column temperature of 40 ℃. MS and MS/MS were conducted with electrospray ionization(ESI) in both positive and negative modes. The chemical components in Lysionoti Herba were identified by comparison with the retention time and mass spectra of reference compounds and the relevant mass spectral data reported in MS databases and relevant literature. Furthermore, the content of five constituents(neochlorogenic acid, chlorogenic acid, forsythoside B, acteoside, and nevadensin) in different Lysiono-ti Herba samples was simultaneously determined by HPLC-UV at the wavelength of 330 nm. A total of 84 compounds were identified in Lysionoti Herba, including 27 flavonoids, 20 phenylethanoid glycosides, 5 amino acids, 18 organic acids, 1 alkaloid, 6 nucleosides, and 7 others. The content of neochlorogenic acid, chlorogenic acid, forsythoside B, acteoside, and nevadensin showed good linear relationship(r>0.999) with the peak area within certain concentration ranges, which were 3.22-102.90, 12.84-410.82, 31.63-1 012.01, 25.00-800.11, and 4.08-130.51 μg·mL~(-1), respectively. The instrument precision, method repeatability, and solution stability all met requirement, and the average recovery rate was 97.31%-100.2%, with RSD ranging from 0.95% to 2.4%. The content of the five components varied among different Lysionoti Herba samples collected from different regions of Guizhou, and the average content of forsythoside B was the highest. The established qualitative method can rapidly and efficiently identify the chemical components of Lysionoti Herba, and the developed HPLC-UV method can simultaneously determine the content of five components in a simple, ra-pid, and accurate manner, providing a scientific basis for the quality evaluation of Lysionoti Herba.

Plants with antioxidant activity and their potential use as sun protector agents in Costa Rica

SUMMARY Aims: To carry out a bibliographic review related to plants available in Costa Rica that have demonstrated antioxidant power and a sun protection factor (SPF) suitable to be used in sunscreen products. Methods: The bibliographic review was carried out using different descriptors and by consulting different databases. Results: Information about antioxidant power about acerola, avocado, carrots, passion fruit, moringa, banana, pumpkin and amaranth, guava, matico, ginkgo, blackberry, mango, coffee was obtained. The information obtained suggests that the mentioned plants could be used for the formulation of sunscreens. Conclusion: Sun protection products should be used every day to prevent skin damage; some sunscreens produce allergic reactions, so it is necessary to investigate natural options to be used in sun protection products. Some natural products exhibit SPF values that allow their incorporation as sun protection agents, adjuvants, or enhancers in sunscreens.

Objetivo: llevar a cabo una investigación bibliográfica sobre plantas disponibles en Costa Rica que han demostrado su poder antioxidante y un factor de protector solar (SPF) apto para ser utilizados en protectores solares. Métodos: la revisión bibliográfica se realizó utilizando diferentes descriptores y mediante la consulta en diferentes bases de datos. Resultados: se obtuvo información sobre el poder antioxidante de las plantas: acerola, aguacate, zanahoria, maracuyá, moringa, banano, calabaza y amaranto, guayaba, matico, ginkgo, mora, mango, café. La información obtenida sugiere que las plantas mencionadas podrían ser utilizadas para la formulación de protectores solares. Conclusiones: los protectores solares deben ser utilizados diariamente para prevenir el daño sobre la piel. Algunos de estos productos generan reacciones alérgicas y por esta razón es necesario investigar posibles opciones naturales para ser incorporadas en bloqueadores solares. Algunos productos naturales exhiben valores de SPF que permiten su uso como agentes protectores solares, adyuvantes o potenciadores en los bloqueadores solares.

Objetivo: realizar uma pesquisa bibliográfica sobre plantas disponíveis na Costa Rica que demonstraram poder antioxidante e um fator de proteção solar (FPS) adequado para uso em protetores solares. Métodos: a revisão bibliográfica foi realizada usando diferentes descritores e consultando diferentes bases de dados. Resultados: foram obtidas informações sobre o poder antioxidante das plantas: acerola, abacate, cenoura, maracujá, moringa, banana, abóbora e amaranto, goiaba, matico, ginkgo, amora, manga, café. As informações sugerem que as referidas plantas poderiam ser utilizadas para a formulação de protetores solares. Conclusões: protetores solares devem ser usados diariamente para prevenir danos à pele. Alguns desses produtos geram reações alérgicas e por isso é necessário investigar possíveis opções naturais para serem incorporadas aos protetores solares. Alguns produtos naturais apresentam valores de FPS que permitem seu uso como protetores solares, adjuvantes ou potencializadores em protetores solares.

Etnofarmacología de plantas utilizadas por pacientes diabéticos tz'utujiles en Santiago Atitlán, Sololá, Guatemala / Ethnopharmacology of plants used by Tz'utujil diabetic patients in Santiago Atitlán, Sololá, Guatemala

La diabetes mellitus tipo 2 es una afección crónica con una elevada prevalencia en las comunidades rurales e indígenas de Guatemala, en las cuales se ha reportado el uso de plantas medicinales para su tratamiento. Con el propósito de conocer qué plantas son utilizadas en la población para esta afección, se realizó un estudio etnofarmacológico con 36 sujetos de la población tz'utujil de Santiago Atitlán, Sololá, por medio de un análisis antropológico y farmacológico-toxicológico de entrevistas semiestructuradas y cuestionarios abiertos. Se identificaron 19 plantas, y se calculó el índice de consenso de enfermedad (ICE) y el valor de uso (VU) para evaluar el conocimiento y la importancia relativa de cada especie; ambos valores ayudan a comprender la relevancia cultural, es decir, la importancia de las especies medicinales para la población. Las cinco plantas con mayor relevancia cultural fueron Neurolaena lobata R.Br. ex Cass. (ICE=0.4074, VU=0.0084), Artemisia absinthium L. (ICE=0.1481, VU=0.0027), Psidium guajava L. (ICE=0.1111, VU=0.0023), Tanacetum parthenium (L.) Sch.Bip. (ICE=0.0741, VU=0.0015) y Coffea arabica L. (ICE=0.0741, VU=0.0015). Se determinó que N. lobata y P. guajava tienen un alto potencial para tratar la diabetes, C. arabica y T. parthenium pueden provocar efectos adversos e interacciones con ciertos medicamentos y A. absinthium puede ser peligrosa para su consumo. No obstante, se requieren estudios de los metabolitos secundarios aislados responsables de la actividad antidiabética para establecer dosis seguras, efectos adversos, interacciones y toxicología.

Type 2 diabetes mellitus is a chronic disease with a high prevalence in rural and indigenous communities of Guatemala, in which the use of medicinal plants for its treatment has been reported. In order to know which plants are used by the population for this condition, an ethnopharmacological study was carried out with 36 individuals from the Tz'utujil population of Santiago Atitlán, Sololá, through an anthropological and pharmacological-toxicological analysis of semistructured interviews and open questionnaires. Nineteen plants were identified, and the disease consensus index (DCI) and the value in use (UV) were calculated to assess the knowledge and relative importance of each species; both values help to understand the cultural relevance, that is, the importance of medicinal species for the population. The five plants with the higher cultural relevance were Neurolaena lobata R.Br. ex Cass. (DCI = 0.4074, UV = 0.0084), Artemisia absinthium L. (DCI = 0.1481, UV = 0.0027), Psidium guajava L. (DCI = 0.1111, UV = 0.0023), Tanacetum parthenium (L.) Sch.Bip. (DCI = 0.0741, UV = 0.0015) and Coffea arabica L. (DCI = 0.0741, UV = 0.0015). It was determined that N. lobata and P. guajava have a high potential to treat diabetes, C. arabica and T. parthenium can cause adverse effects and interactions with certain medications, and A. absinthium can be dangerous if consumed. However, further studies of the isolated secondary metabolites responsible for antidiabetic activity are required to establish safe doses, adverse effects, interactions and toxicology

Photocatalytic destruction of Escherichia coli in water over rGO/TiO2 nanocomposites

In the present study, Graphene-TiO2 catalysts are prepared by solvothermal method with varied graphene concentrations (1%, 2.5% and 5%). The prepared nanocomposites were characterized by FTIR, Raman and TEM. The photocatalytic activity towards the destruction of Escherichia coli in water under UV and UV-visible irradiations were studied. Graphene-TiO2 nano composite destructs the bacteria significantly at higher rates than unmodified TiO2 and graphene. The results indicates that, at the beginning, the inactivation of E. coli cells is more due to the generation of reactive oxygen species (ROS) like, OH, H2O2, and O2– . Among all samples, the nano composite containing 2.5 wt.% of graphene exhibits a complete E. coli destruction in a minimum irradiation time of 15 and 20 min under UV–Visible and UV light irradiation respectively. The high photocatalytic activity is achieved with the optimum loading concentration of 2.5 wt. % graphene on titania

Mosquito larvicidal potential of Solanum xanthocarpum leaf extract derived silver nanoparticles and its bio-toxicity on non-target aquatic organism

Background & objectives: Mosquitoes are insects of public health importance that act as a vector to transmit various vector-borne diseases in humans including dengue, malaria, filariasis and yellow fever. The continually employed synthetic insecticides have developed resistance in mosquitoes. Nano-based botanical insecticides can be considered as the best alternative due to several advantages like being simple, non-pathogenic, biodegradable and safe to the environment. The present work reported the maximum larvicidal potential of green synthesized silver nanoparticles (AgNPs) derived from the leaf extract of Solanum xanthoearpum against the third instar larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus over its crude leaf extract. Methods: The synthesis of AgNPs was done by adding leaf extract into silver nitrate solution in a conical flask. The characterization of AgNPs was done using different techniques such as UV-Vis, SEM, TEM, XRD, DLS and SAED. FT-IR analysis was done to find out the compound responsible for bio-reduction of silver nitrate. Larvicidal activity of AgNPs was checked against An. stephensi, Ae. aegypti, and Cx. quinquefasciatus according to WHO standard protocol and toxicity was evaluated against Poecilia reticulate. Results: A change in colour was observed indicating the synthesis of AgNPs which was further confirmed by a strong surface plasmon resonance peak at 421nm under the UV-Vis spectrum. SEM and TEM micrographs exhibited that the most common shape of AgNPs was spherical. XRD spectrum showed crystalline nature of silver nanoparticles. FT-IR spectrum showed the presence of various functional groups such as carboxyl and hydroxyl which might be responsible for bio-reduction and capping of silver nanoparticles. Further, silver nanoparticles were very effective against An. stephensi, Ae. aegypti, and Cx. quinquefasciatus with LC50 and LC90 values of 1.90, 2.36, 2.93, 3.82, 4.31 and 7.63 ppm, respectively, as compared to aqueous leaf extract after 72 h of exposure and were non-toxic against non-target organism P. retieulata. Interpretation & eonelusion: From the above finding, it can be concluded that fabricated AgNPs can be promising eco-friendly tools for controlling mosquito vectors.

Investigation of the influence of clathrate hydrate crystals on the structuring of homeopathic high dilutions

High dilutions (HD) of drugs used in homeopathy are mostly too dilute to contain original drug molecules. But evidences support their specific biological and therapeutic effects. The reason behind this is thought to be water structure characteristic of the original drug. Spectroscopic studies indicate that the specific water structure in HDs can be resolved into free water molecules, hydrogen bonding strength of water hydroxyl, number of hydrogen bonds and clathrate hydrate crystals (CHC). HDs are prepared in EtOH water solution by serial dilution and mechanical agitation, and are called potencies. The objective of the present study is to further confirm the presence of CHCs in the two potencies of three drugs. Electronic spectra of the HDs of the potencies indicate two broad peaks and marked difference in intensities of absorption. Furior Transform Infrared (FT-IR) spectra of the test potencies and their control show difference in intensity shift and contour shape of OH stretching and bending bands. All the experimental data indicate the presence of CHCs in varying amounts in the test potencies.

Physicochemical and UV/VIS measurements of UHD solutionsof water, potentized water,and different potencies of organic substance

Conventional science regards the study of UHD (highly homeopathically potentized) solutions as pseudo-science. However, an increasing number of rigorous scientific investigations demonstrate differences in physicochemical and physical characteristics of such solutions. Strictly chemically regarded, they correspond to highly distilled water. Our research team developed a system of physicochemical and UV spectrographic measurements, whereby the differences may be consistently confirmed with high statistical significance.Methods:For measurement of the physicochemical parameters,we usedpH,electrical conductivity,and oxidation-reduction potential (ORP)sensors.For UV/VIS spectroscopymeasurements,we useda Macherey-Nagelspectrophotometer.For UHD research, we used two batches of substances: Russian (R) and Brazilian (B). In R, distilled water (W) was used for dilutions and potencies as follows: potentized water (W cH9), potentized dilution of antibodies to interferon-gamma (Abs IFNγ cH9), the same original substance in the form of a mixture of potencies (Abs IFNγ cH12, cH30, cH50, shortly Abs IFNγ mix). Furthermore, we prepared higher potencies of the substances (supplementary potentiation) in a specially prepared solution and measured their characteristics. In B, the solution was used for further dilutions and potencies as follows: potentized water (W cH1) and Glyphosate potencies (Gly cH6 => cH8, cH30 => cH 32, and cH200 => cH 202).For direct or post-hoc analysis, we used Wilcoxon signed-rank test, two-tailed.Results:UV-VIS spectroscopy (R): measurements of received liquids potentized for further cH1 show statistically significant differences between all substances, except between water W and W cH9 at 260 nm. Significant differences (p-values) were as follows: Abs IFNγ mixvs. W= 0.007; Abs IFNγ mixvs.W cH9=0.008; Abs IFNγ cH9vs. W=0.044; Abs IFNγ cH9vs.W cH9= 0.026; Abs IFNγ mixvs.W cH9= 0.007; W vs.W cH9= 0.506.Physicochemical measurements:R: measurementsdemonstrated statistical difference only in pH (Abs IFNγ mix towards all others). Significant differences (p-values) were as follows: Abs IFNγ mixvs. W=0.022; Abs IFNγ mixvs.W cH9=0.005; Abs IFNγ mixvs.Abs IFNγ cH9=0.025. After supplementarypotentiation, we obtained a more conspicuous picture with many statistical differences in conductivity and ORP, ranging from p= 0.001 to 0.046.A difference between water and potentized water has also been demonstrated.B: the measurements demonstrated statistical differences mainly in pH between Gly cH8 and the rest and between Gly cH202 and W cH1.Significant differences (p-values) were as follows:Gly cH8vs. Gly cH32 =0.027; Gly cH8vs. Gly cH202 = 0.011; Gly cH8vs. W cH1= 0.014; Gly cH202vs. W cH1= 0.034.Conclusion:UV/VIS at wavelength 260 nm Abs IFNγ mix discloses a pattern similar to exclusion zone (EZ)water at 270 nm.By additional potentiation and with physicochemical measurements, we obtained higher statistical differences than in the original dilutions.In contrast, UV/VIS spectroscopy showed more conspicuous results without additional potentiation. However, the very act of succussion becomes very distinct

Immunohistochemical and mutational status of telomerase reverse transcriptase in conjunctival squamous cell carcinoma

Purpose: Mutations in human telomerase reverse transcriptase (TERT) are associated with increased telomerase activity in cutaneous melanomas. Conjunctival squamous cell carcinoma, also referred to as ocular surface squamous cell carcinoma, is cancer on the surface of the eye. Recent studies have identified UV signature mutations in TERT promoters in ocular melanoma and ocular surface squamous neoplasia. However, its immunohistochemical status has not been reported in ocular surface squamous cell carcinoma. This study aimed to explore the immunohistochemical and mutational status of TERT in ocular surface SCC. Methods: The immunohistochemical expression of TERT and mutational status of TERT promoter was evaluated in 19 ocular surface squamous cell carcinoma cases. Conjunctival melanoma tissue was used as a positive control. Results: The cytoplasmic overexpression of TERT was detected in 11/19 (57%), and TERT promoter mutations were identified in 6/19 (31%) of ocular surface squamous cell carcinoma. Out of these, 66% had a C228T mutation, and 33% had a C250T mutation. The TERT expression was found to be associated with a high (?T3) AJCC category (P = 0.023), and TERT immunoexpression was significantly correlated with reduced disease?free survival (P = 0.024, log?rank analysis) in ocular surface squamous cell carcinoma patients. Conclusion: The present study demonstrates that TERT promoter mutations with UV signatures are frequent in ocular surface squamous cell carcinoma. The increased expression of TERT could be of biological significance in aggressive ocular surface squamous cell carcinoma.

Chemical denaturants induced folding unfolding pathway of the recombinant zebrafish dihydrofolate reductase

Denaturation of proteins plays a crucial part in cellular activities. In this study, we have investigated the folding unfolding pathways of zebrafish dihydrofolate reductase (zDHFR) in presence of different chemical denaturants which were found to be an influential factor for the refolding yield by UV-visible spectrophotometric analysis. The activity change of zDHFR has been observed in presence of three different denaturants like Acetic Acid (AcOH), Sodium Dodecyl Sulphate (SDS), and Ethanol (C2H5OH). Spectrophotometric analysis reveals that protein unfolded completely at different concentrations and times by these denaturants. The spontaneous refolding experiments of chemically denatured zDHFR were also conducted to verify the spontaneous refolding yield. These investigations have helped us to decipher a picture about the denaturants contributing to achieving the refolding yield. We observed that acetic acid is a stronger denaturant among all, and the spontaneous refolding yields were higher from SDS denaturation. In the light of the above findings, higher spontaneous refolding yields were obtained from the low concentration of denaturants.

In vitro skin retention and drug permeation study of Tongluo-Qutong rubber plaster by UPLC/UV/MS/MS

Abstract Tongluo-Qutong rubber plaster (TQRP), a typical Chinese patent medicine that contains 13 different herbal remedies, is widely used in clinical practice for the treatment of cervical spondylosis and osteoarthritis. However, due to a lack of in vitro transdermal studies, the active ingredients of TQRP have not been fully elucidated. This presents a huge obstacle for quality evaluation, pharmacokinetic studies and clinical safety assessment of TQRP. In this work, a UPLC/UV/MS/MS method was established and validated to evaluate five analytes in TQRP. The validation demonstrated linearity (r > 0.99), specificity (no co-eluting peaks at the retention times of the analytes), and precision (RSD < 15%) within acceptable parameters. A skin permeation study was performed to determine the concentrations of drugs delivered to the dermis. The 24-hour cumulative permeation of ferulic acid, aleo-emodin, emodin and piperine were 303.68, 709.31, 671.06 and 25561.01 ng/cm2, respectively. According to the fitting data of the TQRP active components, skin permeation was mainly due to a combination of passive diffusion and drug release after matrix erosion

A study on 2-(2'-hydroxyphenyl) benzoxazoles derivatives as potential organic UV filters, Part I

Abstract Damage resulting from the incidence of ultraviolet (UV) radiation on the skin is common nowadays, with UVB (290-320 nm) and UVA (320-400 nm) radiation responsible for photoaging, sunburn and carcinogenesis. For this reason, sunscreens represent products of growing interest to prevent such damage. However, there are few organic filters marketed worldwide with photostability and effectiveness at wavelengths greater than 340 nm (long UVA), which justifies the exploration for new compounds. In this work, we determined the photostability and sun protection factor (SPF) of three 2-(2'-hydroxyphenyl)benzoxazole derivative dyes in order to develop new organic UV filters. UV-vis spectrophotometry has high level of reproducibility when compared with in vivo human clinical methods. Solubility determinations were performed in different solvents. The compounds absorbed UVA and UVB radiation, with maximum absorption wavelengths ranging from 336 to 374 nm. Photostability was evaluated using a solar simulator (3 J.m2.s-1 UVA radiation) for a maximum of 3 h. The 2-(amino-2'-hydroxyphenyl) benzoxazoles showed higher photostability than the acetylated derivative under the evaluated conditions. The three benzoxazoles presented SPF values of around 40 and preliminary results indicate that they show suitable properties to act as good chemical filters in photoprotective formulations.

Determination of sunset yellow, allura red, and fast green using a novel magnetic nanoadsorbent modified with Elaeagnus angustifolia based on magnetic solid-phase extraction by HPLC

Abstract Sunset yellow (SY), allura red (AR) and fast green (FG) are frequently used in commercial food products, although they are considered to be hazardous to public health due to their toxic efficacy and high exposure risk potency. In this study, a new, rapid, and reliable method based on a magnetic solid-phase extraction (MSPE) was developed for the simultaneous determination of SY, AR, and FG. Fe3O4 modified with Elaeagnus angustifolia was used for the first time as an adsorbent (Fe3O4-EA) in MSPE. It was characterized with scanning electron microscopy, Brunauer Emmet Teller surface area analysis and X-ray diffraction. MSPE parameters were optimized in terms of pH, adsorption, and elution time and elution volume. High-performance liquid chromatography was used for dye quantitation. Analytical separation was performed by applying ammonium acetate buffer, acetonitrile, and methanol as the mobile phase to a C18 reverse-phase analytical column. Intraday and inter-day repeatability of the method performed at the concentration of 0.2, 1.0 and 2.0 µg/mL exhibited <8.1% RSD (n=3). The limit of detection values was between 0.05-0.1 µg/mL. The adsorption data of SY, AR and FG on Fe3O4-EA were fitted with the Langmuir model with qmax values of 45.0, 70.4 and 73.0 mg/g, respectively.