Anthurium Fragrance, Genetic and Biochemical Studies

The total of 147 Anthurium species and hybrids at the University of Hawaii and the Missouri Botanical Garden germplasm collections was evaluated for the presence or absence of scent, type of fragrance, time of emission, daily occurrence, developmental stage of scent emission, color of spathe and spadix. A majority of inflorescences (76%) emitted scent. Scent was categorized as citrus, fishy, floral, foul, fruity, menthol, minty, pine, spicy and sweet. There was no relationship between scent production or scent quality with flower color or botanical section. A plurality of inflorescences emitted scent during the morning (45%) and at the pistillate stage (77%). Fragrance life of unharvested inflorescences varied from 2 to 3 days up to 4 weeks, whereas that of harvested inflorescences was only 1 or 2 days. F1 progenies of crosses between fragrant x fragrant and non-fragrant x fragrant parents were studied to determine whether a single gene or more than one gene governed presence of scent. Progenies from 24 crosses were tested by Chi-square analysis for a single dominant or recessive trait. None produced expected segregation ratios for a single gene trait, indicating that multiple genes likely govern presence of fragrance in Anthurium. Progeny also segregated for fragrance quality and included non-parental scents. Fragrance of seven Anthurium species and ten hybrids was analyzed by gas chromatography and mass spectrometry. Nineteen monoterpenes (lipids) and some alcohols, aldehydes and esters were identified. Limonene and 1,8-cineole were common to most samples along with a-pinene, p-pinene, myrcene and linalool. Hybrid UH1299, emitting a sweet and floral scent all day, showed fluctuation in amounts and types of compound during the daily cycle. Tepals were associated with fragrance production in Anthurium. Histological comparison between tepals of fragrant and non-fragrant spadices showed lipids and starches present in both fragrant and non-fragrant samples. However, in fragrant samples, the amount of lipids was significantly greater than that of non-fragrant ones, whereas the amount of starch was significantly greater in non-fragrant samples compared with fragrant ones. These data support the hypothesis that high levels of lipids were associated with fragrance production

GENERATION OF MOUSE INDUCED PLURIPOTENT STEM CELLS BY PROTEIN TRANSDUCTION.

Somatic cell reprogramming has generated enormous interest after the first report by Yamanaka and his coworkers in 2006 on the generation of induced pluripotent stem cells (iPSCs) from mouse fibroblasts. Here we report the generation of stable iPSCs from mouse fibroblasts by recombinant protein transduction (Klf4, Oct4, Sox2 and c-Myc), a procedure designed to circumvent the risks caused by integration of exogenous sequences in the target cell genome associated with gene delivery systems. The recombinant proteins were fused in frame to the GST tag for affinity purification and to the TAT-NLS polypeptide to facilitate membrane penetration and nuclear localization. We performed the reprogramming procedure on embryonic fibroblasts from inbred (C57BL6) and outbred (ICR) mouse strains. The cells were treated with purified proteins four times, at 48-hour intervals, and cultured on mitomycin C treated MEF (mouse embryonic fibroblast) cells in complete embryonic stem cell medium until colonies formed. The iPSCs generated from the outbred fibroblasts exhibited similar morphology and growth properties to embryonic stem (ESC) cells and were sustained in an undifferentiated state for more than 20 passages. The cells were checked for pluripotency-related markers (Oct4, Sox2, Klf4, cMyc, Nanog) by immunocytochemistry and by RT-PCR. The protein iPSCs (piPSCs) formed EBs and subsequently differentiated towards all three germ layer lineages. Importantly the piPSCs could incorporate into the blastocyst and led to variable degrees of chimerism in newborn mice. These data show that recombinant purified cell-penetrating proteins are capable of reprogramming mouse embryonic fibroblasts to iPSCs. We also demonstrated that the cells of the generated cell line satisfied all the requirements of bona fide mouse ESC cells: form round colonies with defined boundaries; have a tendency to attach together with high nuclear/cytoplasmic ratio; express key pluripotency markers; and are capable of in vitro differentiation into ecto-, endo-, and mesoderm, and in vivo chimera formation

Toxicity response of Chlorella microalgae to glyphosate herbicide exposure based on biomass, pigment contents and photosynthetic efficiency

The extensive use of glyphosate (N-(phosphonomethyl) glycine) herbicide in agriculture is accompanied by the risk of environmental contamination of aquatic ecosystems. In this study, the effects of glyphosate at different concentrations (50–500 µg ml-1) on three Chlorella species including Chlorella ellipsoidea, Chlorella sorokiniana and Chlorella vulgaris especially in relation to the biomass, pigment contents and photosynthetic efficiency were assessed. After treatment for 24 hr, the acute toxicity results showed that C. vulgaris (IC50 = 449.34 ± 6.20 µg ml-1) was more tolerant to glyphosate than C. ellipsoidea (IC50 = 288.23 ± 23.53 µg ml-1) and C. sorokiniana (IC50 = 174.28 ± 0.50 µg ml-1). After a 72-hr chronic toxicity treatment with glyphosate, glyphosate concentrations decreased to 400–500 µg ml-1 in C. ellipsoidea, 200–300 µg ml-1 in C. sorokiniana and 200–500 µg ml-1 in C. vulgaris respectively. During 24-hr acute toxicity exposure to glyphosate, the pigment contents and maximum quantum efficiency of photosystem II (Fv/Fm) decreased as the concentration of glyphosate increased. Overall, the biomass, pigment contents and photosynthetic efficiency presented a high positive correlation. It is worthwhile to mention that our study provides detailed information on the toxicity and sensitivity of these Chlorella species to glyphosate

GC-MS profiling, anti-oxidant and anti-diabetic assessments of extracts from microalgae Scenedesmus falcatus (KU.B1) and Chlorella sorokiniana (KU.B2)

Microalgae are a potentially valuable source in the food, pharmaceutical and nutraceutical sectors. While biological activities surveys have investigated the pharmaceutical properties of a few microalgae species, there are not many reports covering biological activity studies. This study was carried out to identify the metabolites by gas chromatography-mass spectrometry and evaluate the anti-oxidant, anti-diabetic properties of green algae extracts, Chlorella sorokiniana (KU.B2) and Scenedesmus falcatus (KU.B1). A total of 51 different chemical constituents were detected and tentatively identified. The primary compounds in both microalgae extracts included (R)-2-hexanol (38.67% in C. sorokiniana and 23.53% in S. falcatus), n-hexadecanoic acid (13.58% in C. sorokiniana and 18.94% in S. falcatus) and octadecanoic acid (22.30% in C. sorokiniana and 32.67% in S. falcatus). According to the profiling results, the C. sorokiniana extract exhibited greater anti-oxidant activity, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging (IC50 = 480.30 ±?14.85 µg ml-1), nitric oxide (NO) radical scavenging (562.73 ±?3.52 µg mL-1) and ferric reducing anti-oxidant power (FRAP) of 58.51 ± 2.42 mgTE g-1. Comparatively, the C. sorokiniana extract had higher contents of alpha-glucosidase and alpha-amylase (IC50 = 491.22 ± 78.41 and 2,817.00 ±143.04 µg mL-1, respectively) than the S. falcatus extract. This first report demonstrated anti-diabetic effect of both extracts on diabetic enzymes. The results confirm microalgae's anti-oxidant and anti-diabetic properties and suggest their potential benefits in cosmeceutical, nutraceutical and pharmaceutical applications

Effect of Different Mechanical Properties of Core Build-up Materials on the Root Furcation of A Severely damaged Primary Molar: A Finite Element Analysis

Objectives: To assess the von Mises stress and stress distribution pattern on the root furcation of a severely damaged primary molar restored with different core build-up materials and stainlesssteel crown (SSC). Materials and Methods: The finite element analysis was used to investigate stresses induced in the tooth structures included a sound primary molar and severely damaged primary molars restored with four different core-build up materials, including flowable composite core build-up, bulkfill resin composite, RMGIC and nano-RMGIC. The maximum von Mises stress was used to representthe internal load induced in the model. Results: Overall maximum von Mises stresses was the highest in the sound tooth. However, when focusing on apico-cervical aspect, all restored primary molars showed higher maximum von Mises stress than the sound tooth. The stress distribution pattern of each group was similar, except for the nano-RMGIC group that showed high stress concentrated at the tooth furcation and the buccal aspect of the root furcation. From the ratio of its tensile strength and the maximum von Mises stress, the nano-RMGIC possessed the highest fracture resistance, followed by bulk-fill composite, RMGIC and flowable composite core group, respectively. Discussion: Although nano-RMGIC possessed the highest fracture resistance, it showed an unfavorable stress distribution pattern, which caused high stress at the root furcation. The bulk-fill composite possessed not only high fracture resistance but also favorable stress distribution. Conclusion: The present study introduces crucial information that could lead to an alternative treatment for severely damaged primary molar. Our findings recommend bulk-fill composite as a potential core build-up material. > ว.ทันต.มศว ปีที่ 16 ฉบับที่ 1 พ.ศ. 2566 หน้า 62-77. > SWU Dent J. 2023;16(1): 62-77

Analysis of Intersectional Hybrids of Dendrobium by RAPD Technique

ABSTRACT Thirty interspecific hybridizations of inter-or intra-section of Dendrobium were made. Five crosses showed good compatibility and could yield hybrids as follows: 2 intersectional hybridizations, 1) section Phalaenanthe: Den. phalaenopsis (D017) × section Formosae: Den. draconis (D022) and 2) section Dendrobium: Den. finlayanum (D030) × section Formosae: Den. cariniferum (D018); and 3 intrasectional hybridizations, 1) section Dendrobium: D030 × Den. nobile (D031), 2) section Formosae: Den. trigonopus (D037) × D022, and 3) D037 × Den. infundibulum (D034). The greatest number of plantlets (1,250 plantlets) was obtained from cross D037 × D022 whereas the least number (3 plantlets) was found in cross D037 × D034. After transplanting for 6 months, survival rate of D037 × D034 was 100% whereas no survival plant was found in cross D030 × D018. Parents and hybrids of 5 crosses were analyzed by RAPD using 21 decamer primers and the number of primers generating polymorphic patterns of each cross varied. RAPD markers were able to differentiate parents in inter-and intra-sectional crosses and these markers could confirm the interspecific hybrids

Free Radical Scavenging Properties and Induction of Apoptotic Effects of Fa Fraction Obtained after Proteolysis of Bioactive Peptides from Microalgae Synechococcus sp. VDW

Svrha ovog istraživanja bila je odrediti antioksidacijsku aktivnost bioaktivnih peptida izoliranih iz stanica mikroalge Synechococcus sp. VDW nakon 21 dana uzgoja. Hidrolizati proteina iz mikroalge Synechococcus sp. VDW dobiveni su pomoću tripsina, te su zatim pročišćeni ultrafiltracijom pomoću membrana veličine od 10, 5 i 3 kDa. Frakcija proteina FA molekularne mase od 3 kDa imala je najveću sposobnost uklanjanja slobodnih radikala 2,2\u27-azino-bis(3-etilbenzotiazolin-6-sulfonske kiseline) (ABTS), IC50 vrijednosti od (11,5±0,3) μg/mL, i 2,2\u27-difenil-1-pikrilhidrazila (DPPH), IC50 vrijednosti od (13,6±0,2) μg/mL. Frakcija proteina FA razdvojena je pomoću HPLC kromatografije obrnutih (reverznih) faza u četiri podfrakcije (F1–4). Subfrakcija F4 imala je maksimalni učinak uklanjanja ABTS radikala od (3,55±0,61) %, pa je odabrana za daljnju analizu pomoću metode ESI-Q-TOF-MS/MS, temeljene na sekvenciranju peptida de novo. Identificirano je pet peptida s antioksidacijskim učinkom, od kojih je najveću sposobnost uklanjanja ABTS radikala imao peptid AILESYSAGKTK. Nadalje, frakcija FA imala je snažan citotoksični učinak na stanične linije humanih karcinoma, pogotovo staničnu liniju raka debelog crijeva (SW620), pri čemu je IC50 vrijednost bila (106,6±21,5) μg/mL, no nije imala citotoksični učinak na netransformiranu staničnu liniju Wi38. Uzrokovala je početak apoptoze u staničnoj liniji SW620 nakon 24, 48 i 72 sata, s najvećom aktivnosti kaspaza 3, 8 i 9 nakon 72 sata. Dobiveni rezultati pokazuju da se mikroalga Synechococcus sp. VDW može upotrijebiti za proizvodnju prirodnih antitumorskih lijekova.This study aims to determine the antioxidant activity of bioactive peptides derived from Synechococcus sp. VDW cells cultured for 21 days. Synechococcus sp. VDW protein hydrolysates were prepared with trypsin and purified by ultrafiltration with molecular mass cut-off membranes of 10, 5 and 3 kDa. The M<3 kDa (FA) fraction had the highest 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2’-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities, with IC50 values of (11.5±0.3) and (13.6±0.2) µg/mL, respectively. The FA fraction was separated by reversed phase HPLC to yield four subfractions (F1–4). The F4 subfraction showed the highest maximum ABTS radical scavenging activity (3.55±0.61) % and it was selected for further analysis by electrospray ionisation quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS/MS) based on de novo peptide sequencing. Five antioxidant peptides were identified, of which AILESYSAGKTK had the highest ABTS radical scavenging activity. Furthermore, the FA fraction showed high cytotoxic activities against human cancer-derived cell lines, especially the colon cancer cell line (SW620) with an IC50 value of (106.6±21.5) µg/mL, but not the untransformed Wi38 cell line. The FA fraction activated the apoptotic pathway in SW620 cells after treatment for 24, 48 and 72 h, with the highest activities of caspases-3, -8 and -9 being observed after treatment for 72 h. These findings suggested that microalgae Synechococcus sp. VDW may be used to develop natural anticancer drugs

Effects of regional citrate anticoagulation on thrombin generation, fibrinolysis and platelet function in critically ill patients receiving continuous renal replacement therapy for acute kidney injury: a prospective study

Background: Regional citrate anticoagulation (RCA) is recommended for continuous renal replacement therapy (CRRT). However, filter life varies and premature filter clotting can occur. The aims of this explorative prospective study were to investigate the effects of RCA on thrombin generation, fibrinolysis and platelet function in critically ill patients receiving CRRT, to compare clotting parameters between systemic and intra-circuit blood samples, and to screen participants for coagulation disorders. We recruited critically ill adult patients admitted to a 30-bedded Intensive care unit in a tertiary care hospital who required CRRT with RCA for acute kidney injury (AKI). Patients with pre-existing thrombotic, bleeding tendencies or a CRRT duration less than 48 h were excluded. We measured coagulation and thrombophilia parameters at baseline. Thrombin generation, D-dimer and platelet function were measured pre-CRRT and at 12, 24, 36, 48 and 72 h after commencing CRRT using blood samples taken from the arterial line and the circuit. Results: At baseline, all eleven patients (mean age 62.4 years, 82% male) had Factor VIII and von Willebrand Factor concentrations above reference range and significantly increased peak thrombin generation. During CRRT, there were no significant changes in systemic maximum peak thrombin generation, time to peak thrombin generation, fibrinogen, D-dimer and platelet function analysis. We observed no significant difference between paired samples taken from the patient's arterial line and the circuit. Conclusions: Critically ill patients with AKI requiring CRRT are hypercoagulable. Citrate used for anticoagulation during CRRT does not affect thrombin generation, D-dimer or platelet function. Systemic clotting parameters reflect intra-circuit results. Trial registration: ClinicalTrials.gov Identifier: NCT02486614. Registered 01 July 2015—Registered after recruitment of first patient. https://clinicaltrials.gov/ct2/show/NCT0248661

Acute kidney injury prevalence, progression and long-term outcomes in critically ill patients with COVID-19: a cohort study

BACKGROUND: There are limited data on acute kidney injury (AKI) progression and long-term outcomes in critically ill patients with coronavirus disease-19 (COVID-19). We aimed to describe the prevalence and risk factors for development of AKI, its subsequent clinical course and AKI progression, as well as renal recovery or dialysis dependence and survival in this group of patients. METHODS: This was a retrospective observational study in an expanded tertiary care intensive care unit in London, United Kingdom. Critically ill patients admitted to ICU between 1st March 2020 and 31st July 2020 with confirmed SARS-COV2 infection were included. Analysis of baseline characteristics, organ support, COVID-19 associated therapies and their association with mortality and outcomes at 90 days was performed. RESULTS: Of 313 patients (70% male, mean age 54.5 ± 13.9 years), 240 (76.7%) developed AKI within 14 days after ICU admission: 63 (20.1%) stage 1, 41 (13.1%) stage 2, 136 (43.5%) stage 3. 113 (36.1%) patients presented with AKI on ICU admission. Progression to AKI stage 2/3 occurred in 36%. Risk factors for AKI progression were mechanical ventilation [HR (hazard ratio) 4.11; 95% confidence interval (CI) 1.61-10.49] and positive fluid balance [HR 1.21 (95% CI 1.11-1.31)], while steroid therapy was associated with a reduction in AKI progression (HR 0.73 [95% CI 0.55-0.97]). Kidney replacement therapy (KRT) was initiated in 31.9%. AKI patients had a higher 90-day mortality than non-AKI patients (34% vs. 14%; p < 0.001). Dialysis dependence was 5% at hospital discharge and 4% at 90 days. Renal recovery was identified in 81.6% of survivors at discharge and in 90.9% at 90 days. At 3 months, 16% of all AKI survivors had chronic kidney disease (CKD); among those without renal recovery, the CKD incidence was 44%. CONCLUSIONS: During the first COVID-19 wave, AKI was highly prevalent among severely ill COVID-19 patients with a third progressing to severe AKI requiring KRT. The risk of developing CKD was high. This study identifies factors modifying AKI progression, including a potentially protective effect of steroid therapy. Recognition of risk factors and monitoring of renal function post-discharge might help guide future practice and follow-up management strategies. Trial registration NCT04445259