RNA sequencing-based exploration of the effects of blue laser irradiation on mRNAs involved in functional metabolites of D. officinales

Dendrobium officinale Kimura et Migo (D. officinale) has promising lung moisturizing, detoxifying, and immune boosting properties. Light is an important factor influencing functional metabolite synthesis in D. officinale. The mechanisms by which lasers affect plants are different from those of ordinary light sources; lasers can effectively address the shortcomings of ordinary light sources and have significant interactions with plants. Different light treatments (white, blue, blue laser) were applied, and the number of red leaves under blue laser was greater than that under blue and white light. RNA-seq technology was used to analyze differences in D. officinale under different light treatments. The results showed 465, 2,107 and 1,453 differentially expressed genes (DEGs) in LB-B, LB-W and W-B, respectively. GO, KEGG and other analyses of DEGs indicated that D. officinale has multiple blue laser response modes. Among them, the plasma membrane, cutin, suberine and wax biosynthesis, flavone and flavonol biosynthesis, heat shock proteins, etc. play central roles. Physiological and biochemical results verified that blue laser irradiation significantly increases POD, SOD, and PAL activities in D. officinale. The functional metabolite results showed that blue laser had the greatest promoting effect on total flavonoids, polysaccharides, and alkaloids. qPCR verification combined with other results suggested that CRY DASH, SPA1, HY5, and PIF4 in the blue laser signal transduction pathway affect functional metabolite accumulation in D. officinale through positively regulated expression patterns, while CO16 and MYC2 exhibit negatively regulated expression patterns. These findings provide new ideas for the efficient production of metabolites in D. officinale.

Social Responsibility of Economic Units and the Well-Being of Society in the Tourism Sector: Example of Accommodation Facility

Corporate social responsibility (CSR) assumes that companies should justify their existence with services for various stakeholders and not with profit alone. There is a strong emphasis on the implications of CSR for managing human resources in hotels and other accommodation services, supporting local communities, and promoting environmental sustainability. This article aims to present the adjustment of business entities providing accommodation services in Poland to the CSR principles. Two aspects were examined: the environmental aspect, i.e., waste segregation and the use of economic energy receivers, while the social aspect included engaging employees, as well as investing in improving their qualifications towards pro-ecological awareness. The study was conducted on a sample of 207 owners and managers of economic entities in the accommodation sector in Poland. The Statistica software was used to analyze the obtained results, including Chi2 statistics and correspondence analysis. Based on the obtained results, it can be concluded that the employees of accommodation centers usually deal with the final segregation of waste because they believe that tourists do not follow the rules of proper segregation. Most hotels are equipped with bins for all fractions of waste. The affiliation of a center to a specific macroregion in Poland was not statistically significant in terms of waste segregation practices. Only in the southern macroregion of Poland, ordinary light bulbs are used in accommodation establishments, which account for 0.97% of the surveyed population. In other regions, ¾ establishments use energy-saving receivers in all rooms. Only 1/3 of the facilities invested in developing their staff by organizing training courses toward sustainable development. The most popular subjects were topics related to waste segregation, energy and water saving, and environmental protection.

MO182CRYSTALLINE LIGHT CHAIN CAST NEPHROPATHY IS ASSOCIATED WITH EARLY MORTALITY IN PATIENTS WITH MULTIPLE MYELOMA

Background and Aims Light chain cast nephropathy is the most common paraprotein-associated kidney lesion in patients with multiple myeloma (MM). Rarely, light chain cast nephropathy could show crystalline appearance in patients with multiple myeloma, also known as crystalline light chain cast nephropathy. We here report the first retrospective study of crystalline light chain cast from a single centre. Method All native kidney biopsies were retrospectively studied in the Peking University First Hospital from 2000 to 2020. Newly diagnosed MM patients with were enrolled. Patients with light chain cast nephropathy at least one cast with crystalline appearance were identified as crystalline light chain cast nephropathy (Figure 1, n = 8), others were identified as ordinary light chain cast nephropathy (n = 18). Results The cohort of crystalline light chain cast nephropathy consisted of 6 men and 2 women with a median age of 59.5 (range, 41-73) years. All patients suffered from advanced multiple myeloma (1 with ISS staging II, 7 with ISS staging III) and acute kidney injury with a median eGFR of 5.59 (range, 2.27-26.04) mL/min/1.73m2. All patients except 1 required emergency dialysis at admission. Microhematuria was presented in 3 patients. Median proteinuria was 2.13 (rang, 0.83-3.59) g/24h and median serum albumin was 38.2 (30.7-46.7) g/L. No one presented with nephrotic syndrome. Monoclonal immunoglobulin, detected in all patients on serum protein immunofixation electrophoresis, was λ alone in 5 patients, κ alone in 1 patient, IgG λ in 1 patient, IgA λ in 1 patient. The 8 patients were followed up with a median time of 8 (range, 2-24) months. Three patients received VAD chemotherapy and 5 patients received bortezomib based regimens. At the time of last follow-up, 2 of 7 patients who needed emergency dialysis got rid of dialysis and the rest remained dialysis-dependent. Five patients died with a median time of 5 (range, 2-19) months, 2 patients achieved partial remission and 1 patient achieved complete remission. There was no significant difference in clinical features, treatments and main outcomes between crystalline light chain cast nephropathy patients and ordinary light chain cast nephropathy patients (Table 1). However, crystalline light chain cast nephropathy patients had higher early mortality than ordinary light chain cast nephropathy patients (50.0% vs 11.1%, p = 0.03). Conclusion Crystalline light chain cast nephropathy patients usually presented with acute kidney injury requiring emergency dialysis. Although various types of monoclonal immunoglobulin were detected, there was a dominance of the λ isotype. Compared to ordinary light chain cast nephropathy patients, crystalline light chain cast nephropathy patients had higher early mortality.

Pharmacognostic, phytochemical, biological and spectroscopic analyses of Capparis decidua (Forsk.) Edgew root and stem bark

Purpose: To investigate the pharmacognostic, phytochemical, biological and spectroscopic analyses of Capparis decidua (Forsk.) Edgew root and stem bark.Methods: Plant material (root and stem bark) was collected, authenticated, shade-dried and extracted by maceration using methanol as a solvent separately. Powder microscopy was performed using a binocular microscope. Fluorescence, physico-chemical analysis and phytochemical screening for the presence of secondary metabolites were performed using standard methods. Brine shrimp lethalitybioassay was carried out using Artemia salina bioassay, while enzymatic modulatory study was performed by α-amylase inhibition assay. Microscopic analysis was carried out with scanning electron microscopy. Spectroscopic analysis was performed by Fourier transform infrared spectroscopy (FTIR).Results: Powder microscopy showed the presence of different cellular structures. Various colors were observed under ultraviolet (UV) and ordinary light when treated with different reagents. Phytochemical screening revealed the presence of alkaloids, tannins, saponins and flavonoids but phenol and cardiac glycosides were absent from both extracts. The root bark of the plant showed significant brine shrimp lethality activity.Conclusion: Capparis decidua (Forsk.) Edgew root and stem bark contain a variety of bioactive compounds that have medicinal and therapeutic potentials. Therefore, further investigations are required to elucidate their pharmacological properties. Keywords: Capparis decidua (Forsk.) Edgew, Phytochemical, Spectroscopy, α-Amylase

Recent advances in understanding the molecular basis of melanogenesis in melanocytes

Melanin pigments are responsible for human skin and hair color, and they protect the body from harmful ultraviolet light. The black and brown melanin pigments are synthesized in specialized lysosome-related organelles called melanosomes in melanocytes. Mature melanosomes are transported within melanocytes and transferred to adjacent keratinocytes, which constitute the principal part of human skin. The melanosomes are then deposited inside the keratinocytes and darken the skin (a process called tanning). Owing to their dark color, melanosomes can be seen easily with an ordinary light microscope, and melanosome research dates back approximately 150 years; since then, biochemical studies aimed at isolating and purifying melanosomes have been conducted. Moreover, in the last two decades, hundreds of molecules involved in regulating melanosomal functions have been identified by analyses of the genes of coat-color mutant animals and patients with genetic diseases characterized by pigment abnormalities, such as hypopigmentation. In recent years, dynamic analyses by more precise microscopic observations have revealed specific functions of a variety of molecules involved in melanogenesis. This review article focuses on the latest findings with regard to the steps (or mechanisms) involved in melanosome formation and transport of mature melanosomes within epidermal melanocytes. Finally, we will touch on current topics in melanosome research, particularly on the "melanosome transfer" and "post-transfer" steps, and discuss future directions in pigment research.

THU0406 IDENTIFICATION OF INTRACELLULAR VACUOLES IN SYNOVIAL FLUID WITH CALCIUM PYROPHOSPHATE AND MONOSODIUM URATE CRYSTALS

Background:Synovial fluid analysis using polarized microscopy is the gold standard for the diagnosis of crystal-related arthritis. In our experience, we have noted that, when calcium pyrophosphate (CPP) crystals are observed, they sometimes appear within intracellular vacuoles. However, this phenomenon is not seen in those samples containing monosodium urate (MSU) crystals. This finding has been scantly reported in the literature, but may be useful in clinical practice to ensure accurate crystal identification.Objectives:Our study aims to assess whether the presence of vacuoles contributes to identifying the type of crystal, and also to gauge the frequency of their presentation.Methods:We conducted an observational study in a rheumatology unit between February and June of 2019. Synovial fluids containing CPP or MSU crystals, obtained in daily clinical practice, were consecutively included for analysis. Two observers simultaneously analyzed the presence of vacuoles by ordinary light and phase contrast microscopy in less than 24 hours after their extraction, using a microscope equipped with two viewing stations. The primary study variable was to determine whether CPP and MSU crystals are seen inside intracellular vacuoles, and to calculate the frequency of this finding for each type of crystal, estimating their 95% confidence interval (95% CI) and comparing rates using Fisher’s exact test.Results:Twenty-one samples were obtained. Data is given in the Table. MSU crystals were present in 7 (33.3%) and CPP crystals in 14 (66.6%). Interestingly, none of the MSU samples showed crystal-containing vacuoles (95% CI 0-35.4%). On the contrary, cytoplasmic vacuoles containing crystals were present in all of the CPP samples (95% CI 78.5-100%). The findings were confirmed by phase-contrast microscopy. Differences were statistically significant (p<0.001).Table.SAMPLES ACCORDING TO TYPE OF MICROCRYSTAL(n=21)SAMPLES WITH VACUOLS(UNDER ORDINARY LIGHT)SAMPLES WITH VACUOLS(UNDER PHASE CONTRAST)CPP (14; 66.6%)14 (100%)(95%CI 78.5-100%)14 (100%)(95%CI 78.5-100%)MSU (7; 33.3%)0 (0%)(95%CI 0-35.4%)0 (0%)(95%CI 0-35.4%)Conclusion:The presence of vacuoles may be a useful and easy way to differentiate MSU and CPP crystals when performing synovial fluid microscopy in clinical practice, since it appears to be a distinctive feature in CPP crystal fluids.References:[1]Kohn NN, Hughes RE, McCarty DJ Jr, Faires JS. The significance of calcium phosphate crystals in the synovial fluid of arthritic patients: the «pseudogout syndrome». II. Identification of crystals. Ann InternMed. 1962 May;56:738-45.[2]Pascual E, Sivera F, Andrés M. Synovial Fluid Analysis for Crystals. CurrOpRheumatol 2011;23:161-169.[3]McCarty DJ, Koopman WJ. Arthritis and allied conditions: A textbook of rheumatology, volumen 1. Lea &amp;Febiger. 1993.[4]Pascual E, Sivera F. Synovial fluid crystal Analysis. En Gout and other crystal arthropathies. Terkeltaub R ed. Elsevier; 2012: p.20-34.[5]Hwang HS, Yang CM, Park SJ, Kim HA. Monosodium Urate Crystal-Induced Chondrocyte Death via Autophagic Process. Int J Mol Sci. 2015 Dec 8;16(12):29265-77.Image 1. Microscopy with ordinary light. Cells with cytoplasmic vacuoles are observed, as well as abundant intra and extracellular CPP crystals.Image 2. Microscopy with phase contrast technique. Cells with intracellular vacuoles are observed inside which have microcrystals with parallelepiped morphology, compatible with CPP.Disclosure of Interests: :None declared

THU0414 ORDINARY LIGHT MICROSCOPY IS ABLE TO IDENTIFY MOST CRYSTAL-CONTAINING SYNOVIAL FLUIDS

Background:Optical microscopy remains the gold standard for the diagnosis of crystal arthropathies. The complete protocol consists of three phases. In the first stage, microscopy with simple light provides information on the morphology of the crystal. The second stage, polarized light, allows detecting the intensity of the birefringence. Finally, with the first-order red compensator, the type of elongation is detected, positive for calcium pyrophosphate (CPP) crystals and negative for monosodium urate (MSU) crystals. Finally, with the obtained data, the presence and type of crystals is concluded.Objectives:Analyze the validity and agreement of each stage of microscopy regarding the conclusion, emphasizing ordinary light microscopy.Methods:Fifty consecutive samples of synovial fluid obtained in routine clinical practice were independently analyzed under the compensated polarized microscope by 5 observers blinded to clinical data (250 observations in total). Each observer recorded the presence and type of crystals at each stage and reached a conclusion after gathering all the information. To estimate the diagnostic yield of each microscope stage, sensitivity, specificity and positive and negative predictive values, as well as the accuracy (number of correct observations/number of total observations), were calculated; also, the total weighted kappa was used to assess the degree of agreement with the complete protocol.Results:Main results of the study are shown in Table 1. Regarding diagnostic yield, ordinary light microscopy showed excellent sensitivity, specificity and predictive values, similar to the results noted with simple and compensated polarized microscopy.Table 1.In parentheses, 95% confidence intervals.AccuracySensitivitySpecificityPositive predictive valueNegative predictive valueKappaOrdinary light96.8%(93.8-98.4)97.2%(93.1-98.9)96.2%(90.7-98.5)97.2%(93.1-98.9)96.2%(90.7-98.5)0.954(0.919-0.989)Simple polarized light92.0%(88.0-94.8)84.1%(76.8-89.5)100%(97.0-100)100%(96.5-100)86.1%(79.5-90.8)0.874(0.821-0.927)Compensated polarized light97.6 %(94.9-98.9)95.5%(89.8-98.0)99.3%(96.1-99.9)99.1%(94.8-99.8)96.5%(92.1-98.5)0.962(0.933-0.992)Diagnoses established by ordinary light microscopy matched conclusions (accuracy) in 242/250 (96.8%) observations. Discrepant cases were crystals missed under ordinary light in 4 cases (3 MSU, 1 CPP), and 4 samples with CPP crystals initially seen but later concluded their absence. Interestingly, lowest accuracy was seen with simple polarization; CPP crystals were not detected in 20 out of 93 observations with CPP (21.5%). The accuracy of compensated polarized light was similar to ordinary light. On 5 occasions no crystals were seen but finally they were present (1 MSU, 4 CPP); on the contrary, CPP was registered in one observation but the conclusion indicated no crystals.Regarding agreement with the complete protocol, the kappa with simple light is 0.954, similar to compensated polarized light (0.962), while simple polarized light showed the lowest agreement (0.874).Conclusion:Ordinary light microscopy is enough to correctly reach the majority of diagnoses, with a very high degree of agreement with the complete protocol. Results were comparable to using a compensated polarized microscopy. Thus, if a microscope with polarizer and first-order compensator was not available, using ordinary light would be enough on most occasions. Polarized light microscopy better identifies MSU crystals, but over 20% of CPP crystals were missed at this stage, reinforcing the value of the ordinary light microscopy.Acknowledgments:Thanks to Loreto Carmona for the help with the statistical aspects.Disclosure of Interests: :None declared

Flight to Light Response of Red Pumpkin Beetle (Aulacophora africana Weise) to Differently Coloured Light-emitting Diode and Incandescent Bulb Lights

Red pumpkin beetle (Aulacophora africana Weise) is an important defoliator and vector of pathogens to its numerous crop hosts. Control had largely been by synthetic insecticides with their attendant consequences on man and the environment thus necessitating scientific studies on environmental-friendly management strategies. The experiment was conducted in the Research Farm of Federal University Wukari in the month of May 2019 with the aim of evaluating the attractiveness of A. africana to Light-emitting diode (LED) and Incandescent Light bulb colours. Five colours (red, yellow, green, blue and white) were used for the study. Each colour light was properly projected on 2 metre vertical screen (made of white polyethene) placed one meter above the ground. A setup without bulb served as the control. The light traps were arranged in a completely randomized design (CRD) in 6 replicates and ran simultaneously for six hours (1800 to 2400hrs). The pumpkin beetles attracted were collected in tubs containing soapy water. A. africana collected were counted and recorded according to bulb type and colour. Samples were identified at the Insect Museum of Ahmadu Bello University, Zaria. Among the Incandescent bulbs, White colour was most attractive to A. africana (4.30±0.38) while red attracted the least (0.71±0.01). Among LED bulbs, Blue was most attractive (3.99±1.01) while Red also attracted the least (0.78±0.03). Overall, LED attracted more pumpkin beetles than Incandescent bulb even though Student Newman Keul’s test indicates that the difference between them was due to random variation (p = 0.16). Correlation and regression analyses indicated increase in insect attraction with increased light intensity. The results, therefore, suggest that white Incandescent or blue LED bulb colours can be incorporated into insecticidal light traps to suppress their population/attract them away from host plants or fixed into ordinary light traps to harvest the insect for scientific studies.

A Study on Establishment of Phytochemical Analysis of Quality Parameters and Fluorescence Analysis of Costus spicatus- rhizome extract Medicinal Plants a Well Known Tropical Folklore Medicine

Objective: To evaluate the diagnostic pharmacognostical characters of Costus spicatus in rhizomes extract along with their physico-chemical parameters and fluorescence analysis. Methods: The pharmacognostical characters were determined in terms of macroscopy, microscopy, Powder microscopy, rhizome constant, fluorescence analysis and Preliminary phytochemical investigation. Result: The fluorescence characteristics of rhizome powdered extracts were determined under UV radiation of long and short wavelengths and ordinary visible light. When the powdered drug and extracts were treated with different reagents and observed under UV and ordinary light, they emitted various colour radiations. The colour change for the rhizome powder and individual extract were distinctive and reproducible revealing the solvent properties to the phytoconstituents and data is present. The moister content was calculated through the loss on drying method and was found to be 1.43%. The swelling and foaming index values were recorded. The phytochemical investigation indicates the presence of the organic phytoconstituents such as alkaloids, carbohydrate, flavonoids, protein, amino acids, phenols, tannin, glycosides, steroids and inorganic constituents like iron, phosphate, sulphate and chlorides. Conclusion: Various pharmacognostical and physicochemical parameters have pivotal roles in identification, authentication and establishment of quality parameters of the species. Keywords: Costus spicatus, Physico-Chemical parameters and fluorescence analysis.