Immunomodulatory Factors in Primary Endometrial Cell Cultures Isolated from Cancer and Noncancerous Human Tissue–Focus on RAGE and IDO1

Background: Immune modulatory factors like indoleamine 2,3-dioxygenase 1 (IDO1) generating kynurenine (Kyn) and receptor for advanced glycation end-products (RAGE) contribute to endometrial and cancer microenvironment. Using adequate experimental models is needed to learn about the significance of these molecular factors in endometrial biology. In this paper we study IDO1 activity and RAGE expression in the in vitro cultured primary human endometrial cells derived from cancerous and noncancerous tissue. Methods: The generated primary cell cultures from cancer and noncancerous endometrial tissues were characterized using immunofluorescence and Western Blot for expression of endometrial and cancer markers. IDO1 activity was studied by Kyn quantification with High Performance Liquid Chromatography with Diode Array Detector. Results: The primary cultures of endometrial cells were obtained with 80% success rate and no major genetic aberrations. The cells retained in vitro expression of markers (mucin MUC1 and HER2) or immunomodulatory factors (RAGE and IDO1). Increased Kyn secretion was associated with cancer endometrial cell culture in contrast to the control one. Conclusions: Primary endometrial cells express immune modulatory factors RAGE and IDO1 in vitro associated with cancer phenotype of endometrium.

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Phytochemical variability during vegetation of Chamerion angustifolium (L.) Holub genotypes derived from in vitro cultures

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-021-02154-8 ◽

2021 ◽

Author(s):

Mariola Dreger ◽

Katarzyna Seidler-Łożykowska ◽

Milena Szalata ◽

Artur Adamczak ◽

Karolina Wielgus

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

High Performance ◽

Array Detector ◽

Reproduction Rate ◽

Full Spectrum ◽

Breeding Programs ◽

Chamerion Angustifolium ◽

Oenothein B

AbstractThe purpose of the study was to evaluate Chamerion angustifolium (L.) Holub genotypes for preliminary selection and further breeding programs aimed at obtaining a suitable industrial form for the pharmaceutical applications. Clonally propagated plants representing 10 genotypes of Ch. angustifolium were regenerated under in vitro conditions, hardened and planted in the field. Studies included an evaluation of shoot proliferation, phytochemical assessment of in vitro and ex vitro plants as well as investigations of intraspecies variability regarding four phenological stages: vegetative, beginning of blooming, full blooming, and green fruit phases. Quantitative and qualitative analyses of bioactive compounds were performed using high-performance liquid chromatography coupled with diode array detector and tandem mass spectrometer (HPLC–DAD–MS/MS) and high-performance liquid chromatography (HPLC) methods. The efficiency of shoot multiplication varied between genotypes from 8.12 to 21.48 shoots per explant. A high reproduction rate (> 20 shoots per explant) was recorded for four lines (PL_45, PL_44, PL_58, DE_2). Plants grown in vitro synthesized oenothein B (11.2–22.3 mg g−1 DW) and caffeic acid derivatives. Plants harvested from field contained the full spectrum of polyphenols characteristic for this species, and oenothein B and quercetin 3-O-glucuronide were the most abundant. The maximal content of oenothein B was determined in the vegetative phase of fireweed, while some flavonoids were found in the highest amount in full blooming phase. The results of analysis of variance indicated significant differences among genotypes in oenothein B, 3-O-caffeoylquinic acid and flavonoids accumulation in four phenological phases. PL_44 plants were characterized by high content of oenothein B and quercetin 3-O-glucuronide as well as a relatively high level of other flavonoids. Based on our phytochemical and micropropagation studies, PL_44 genotype was the best candidate for early selection and further breeding programs.

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In Vitro Culture of Rosmarinus officinalis L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production

Pharmaceuticals ◽

10.3390/ph14080747 ◽

2021 ◽

Vol 14 (8) ◽

pp. 747

Author(s):

Eder Villegas Sánchez ◽

Mariana Macías-Alonso ◽

Soraya Osegueda Robles ◽

Lisset Herrera-Isidrón ◽

Hector Nuñez-Palenius ◽

...

Keyword(s):

High Performance ◽

Emerging Infectious Diseases ◽

Rosmarinus Officinalis ◽

Wild Plant ◽

Array Detector ◽

Phase System ◽

Naphthaleneacetic Acid ◽

Temporary Immersion System ◽

Temporary Immersion

Emerging infectious diseases have become a major global problem with public health and economic consequences. It is an urgent need to develop new anti-infective therapies. The natural diterpene carnosol exhibit a wide variety of interesting antibacterial and antiviral properties, and it is considered a theoretical inhibitor of COVID-19 Mpro. However, this compound is present in the family Lamiaceae in low quantities. To obtain carnosol in concentrations high enough to develop pharmacological studies, we evaluated the efficiency of a micropropagation protocol of Rosmarinus officinalis using a solid medium and a temporary immersion system (TIS), as well as the effect of 6-benzylaminopurine (6-BAP) and α-naphthaleneacetic acid (NAA) on the growth of shoots. Moreover, we developed and validated an analytical method to quantify carnosol using the H-point standard additions method in the high-performance liquid chromatography diode array detector (HPLC-DAD). After 30 days of culture, TIS produced the maximum number of shoots per explant (24.33 ± 1.15) on a liquid medium supplemented with 6-BAP at 5.0 mg L−1. Next, we also evaluated the effect of immersion time and frequency for TIS. After 72 days of culture, the best results were obtained with an immersion cycle of 1 min every 12 h, yielding 170.33 ± 29.40 shoots. The quantification of carnosol on the samples was performed at a flow rate of 1.2 mL min−1 using binary isocratic mobile phase system 60:40 (v/v) 10 mM formic acid (pH 3.0) (A) and acetonitrile (B) on a reverse-phase column. The content of carnosol in the in vitro cultures was around 8-fold higher than in the wild plant. The present study represents an efficient alternative method to obtain carnosol for its pre-clinical and clinical development.

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Antiproliferative Activity and Antioxidant Potential of Extracts of Garcinia gardneriana

Molecules ◽

10.3390/molecules25143201 ◽

2020 ◽

Vol 25 (14) ◽

pp. 3201

Author(s):

Simone da Cunha Demenciano ◽

Magalli Costa Barbosa Lima e Silva ◽

Caroline Almeida Farias Alexandrino ◽

Wilson Hino Kato Junior ◽

Patrícia de Oliveira Figueiredo ◽

...

Keyword(s):

Vitamin C ◽

Antiproliferative Activity ◽

High Performance ◽

Ethanolic Extract ◽

Antioxidant Potential ◽

Array Detector ◽

Chloroform Fraction ◽

Chemical Profile ◽

Polycarboxylic Acids

The aim of this study was to evaluate the antiproliferative activity, the antioxidant potential, and the chemical profile obtained from the whole fruit and from leaves of Garcinia gardneriana, a fruit tree from Brazilian Cerrado. To determine in vitro antiproliferative activity, the following neoplastic cell lines were considered, along with an immortalized nontumor cell line. The antioxidant potential was determined, and the evaluation of antiradical air activity was performed. The levels of vitamin C and carotenoids were determined. The chemical profile was analyzed by high-performance liquid chromatography coupled to a diode array detector and a mass spectrometer using electrospray ionization interface. The chloroform fraction of the leaf showed antioxidant activity. The vitamin C content had lower values in fruits and higher in leaves. The content of carotenoids for fruits and leaves was expressive. The ethanolic extract and the hexane and chloroform fractions of fruits were active in all neoplastic lines tested. The leaves showed cytotoxic activity in the hexane fraction in the breast carcinoma line. The analysis of data obtained verified the presence of dimers, monomers, and tetramers of hexoses, polycarboxylic acids, xanthones, flavonoids, biflavonoids, and benzophenones.

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Schisandra rubriflora Plant Material and In Vitro Microshoot Cultures as Rich Sources of Natural Phenolic Antioxidants

Antioxidants ◽

10.3390/antiox9060488 ◽

2020 ◽

Vol 9 (6) ◽

pp. 488

Author(s):

Agnieszka Szopa ◽

Michał Dziurka ◽

Sebastian Granica ◽

Marta Klimek-Szczykutowicz ◽

Paweł Kubica ◽

...

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Phenolic Compounds ◽

Antioxidant Capacity ◽

Phenolic Acids ◽

High Performance ◽

Total Phenolic ◽

Array Detector ◽

Plant Materials

Schisandra rubriflora is a dioecious, underestimated medicinal plant species known from traditional Chinese medicine. The present study was aimed at characterising the polyphenolic profile composition and the related antioxidant capacity of S. rubriflora fruit, stem and leaf and in vitro microshoot culture extracts. Separate analyses of material from female and male specimens were carried out. This study was specifically aimed at detailed characterisation of the contribution of phenolic compounds to overall antioxidant activity using ultra-high-performance liquid chromatography with a photodiode array detector coupled to electrospray ionization ion trap mass spectrometry (UHPLC-DAD-ESI-MS3) and a high-performance liquid chromatography-diode array detector (HPLC-DAD). Using UHPLC-DAD-ESI-MS3, twenty-seven phenolic compounds from among phenolic acids and flavonoids were identified. Concentrations of three phenolic acids (neochlorogenic, chlorogenic and cryptochlorogenic acids) and eight flavonoids (hyperoside, rutoside, isoquercitrin, guaijaverin, trifolin, quercetin, kaempferol, and isorhamnetin) were determined using HPLC-DAD using reference standards. The highest total phenolic content was confirmed for the stem and leaf extracts collected in spring. The contents of phenolic compounds of in vitro biomasses were comparable to that in the fruit extracts. The methanolic extracts from the studied plant materials were evaluated for their antioxidant properties using various in vitro assays, namely free radicals scavenging estimation using 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), ferric-reducing antioxidant power (FRAP) and cupric-reducing antioxidant capacity (CUPRAC) as well as QUick, Easy, New, CHEap, and Reproducible CUPRAC (QUENCHER-CUPRAC) assays. A close relationship between the content of polyphenolic compounds in S. rubriflora and their antioxidant potential has been documented.

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A Pilot Study of a Natural Food Supplement as New Possible Therapeutic Approach in Chronic Kidney Disease Patients

Pharmaceuticals ◽

10.3390/ph13070148 ◽

2020 ◽

Vol 13 (7) ◽

pp. 148 ◽

Cited By ~ 2

Author(s):

Annalisa Noce ◽

Alessio Bocedi ◽

Margherita Campo ◽

Giulia Marrone ◽

Manuela Di Lauro ◽

...

Keyword(s):

Chronic Kidney Disease ◽

Kidney Disease ◽

High Performance ◽

Glutathione Transferase ◽

Food Supplement ◽

Array Detector ◽

Natural Food ◽

Inflammatory Status

The identification of natural bioactive compounds, able to counteract the abnormal increase of oxidative stress and inflammatory status in chronic degenerative non-communicable diseases is useful for the clinical management of these conditions. We tested an oral food supplement (OFS), chemically characterized and evaluated for in vitro and in vivo activity. Vitamin C, analyzed by High Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD), was 0.19 mg/g in rosehip dry extract and 15.74 mg/capsule in the OFS. The identification of polyphenols was performed by HPLC-DAD; the total antioxidant capacity was assessed by Folin–Ciocalteu test. Total polyphenols were 14.73 mg/g gallic acid equivalents (GAE) for rosehip extract and 1.93 mg/g GAE for OFS. A total of 21 chronic kidney disease (CKD) patients and 10 healthy volunteers were recruited. The evaluation of routine laboratory and inflammatory parameters, erythrocyte glutathione transferase (e-GST), human oxidized serum albumin (HSAox), and assessment of body composition were performed at two different times, at baseline and after 5 weeks of OFS assumption. In the study, we highlighted a significant decrease of traditional inflammatory biomarkers (such as C-reactive protein, erythrocyte sedimentation rate, platelet to lymphocyte ratio) and other laboratory parameters like e-GST, azotaemia, and albuminuria after OFS treatment in CKD patients. Moreover, we demonstrated a lipid profile improvement in CKD patients after OFS supplementation.

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Acantholippia salsoloides: Phytochemical Composition and Biological Potential of a Thujonic Population

Natural Product Communications ◽

10.1177/1934578x19858542 ◽

2019 ◽

Vol 14 (6) ◽

pp. 1934578X1985854

Author(s):

Liliana Celaya ◽

Carmen Viturro ◽

Luís R. Silva

Keyword(s):

Essential Oil ◽

High Performance ◽

Micrococcus Luteus ◽

Pharmaceutical Formulations ◽

In Vitro Assays ◽

Array Detector ◽

Biological Potential ◽

Polar Extracts ◽

Hydroethanolic Extract

Acantholippia salsoloides (Verbenaceae) is an aromatic plant widespread in the Andean region. The infusion (leaves and flowers) is widely used as a digestive stimulant as well as for the treatment of various diseases in traditional medicine. A. salsoloides attributes its common name “rica-rica” to the fresh and sweet fragrance of the plant. In this work, 2 different polar extracts and the essential oil of a selected rica-rica population were studied. The phenolic composition was analyzed by high-performance liquid chromatography diode array detector; the essential oil profile was determined by gas-chromatography ion-trap mass spectrometry/flame ionization detection. For all extracts, the antibacterial potential was performed by in vitro assays; the antioxidant and α-glucosidase inhibition were determined in decoction and hydroethanolic extracts. The volatile profile allowed the identification of 26 volatile compounds, β-thujone (84%) being the major one in this rica-rica population. Eighteen phenolic compounds were identified; isoferulic acid (16%-18%) and cynaroside (45%-47%) were the larger ones. In a general way, the hydroethanolic extract was more active against Staphylococcus aureus and Micrococcus luteus (minimum inhibitory concentrations= 0.3- 1.3 mg/mL). Both polar extracts have strong antiradical activities although decoction extract proved to be more active against DPPH· (half-maximal inhibitory concentration [IC50] =36 µg/mL) and O2•− (IC50 =28 µg/mL) while hydroethanolic extract shows higher action over α-glucosidase (IC50 =217 µg/mL). The results suggest that A. salsoloides leaves and flowers may be an interesting source of natural antioxidants, antidiabetics, or antimicrobials, and could be used in dietary supplements, medicinal products and pharmaceutical formulations.

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Heteropterys tomentosa A. Juss: Toxicological and adaptogenic effects in experimental models

Nutrition and Health ◽

10.1177/0260106017729908 ◽

2017 ◽

Vol 23 (4) ◽

pp. 289-298 ◽

Cited By ~ 1

Author(s):

Géssica Alves Fraga ◽

Sikiru Olaitan Balogun ◽

Emilly Della Pascqua ◽

Ruberlei Godinho de Oliveira ◽

Guilherme Botelho ◽

...

Keyword(s):

Oral Administration ◽

High Performance ◽

Scientific Evidence ◽

Endurance Performance ◽

Experimental Models ◽

Time To Exhaustion ◽

Oral Toxicity ◽

Exercise Load

Background: The constant pursuit of improved athletic performance characterizes high-performance sport and the use of medicinal plants as dietary supplements is becoming widespread among athletes to enhance long-term endurance performance. Aim: The present study evaluated the toxicity of Heteropterys tomentosa (HEHt) and its acute adaptogenic effects. Methods: The in vitro safety profile was evaluated on CHO-k1 cells using the alamar Blue assay, at concentrations ranging from 3.125 to 200 µg/mL. In vivo acute oral toxicity was conducted in male and female mice with oral administration of graded doses of HEHt from 400 to 2000 mg/kg. A subchronic oral toxicity study was completed by oral administration of HEHt (50, 200 or 1000 mg/kg) and vehicle for 30 days in male Wistar rats. Clinical observations and toxicological related parameters were determined. Blood was collected for biochemical and hematological analyses, while histological examinations were performed on selected organs. Thereafter, an adaptogenic test consisting of progressive loads until exhaustion was conducted in rats ( n = 5/group) orally pre-treated with the vehicle and HEHt (25, 100 or 400 mg/kg). Results: HEHt exhibited no cytotoxic effects on the CHO-k1 cells and, apparently, no acute toxicity in mice and no subchronic toxicity in rats. An ergogenic effect was observed only at the dose of 25 mg/kg compared with the vehicle in relation to time to exhaustion and exercise load ( p = .011 and .019, respectively). HEHt is safe at up to 400 mg/kg, contains astilbin and taxifolin as the major phytochemical compounds, and exhibited a potential adaptogenic effect. Conclusions: These results justify its anecdotal usage as a tonic, show that the hydroethanolic maceration of the root does not cause toxicity, and provide scientific evidence of its potential as a source of new adaptogenic substance(s).

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Characterizing the relationship between the chemical structures of drugs and their activities on primary cultures of pediatric solid tumors

Current Medicinal Chemistry ◽

10.2174/0929867328666210419134708 ◽

2021 ◽

Vol 28 ◽

Author(s):

Saw Simeon ◽

Ghita Ghislat ◽

Pedro Ballester

Keyword(s):

Solid Tumors ◽

Cell Cultures ◽

Primary Cultures ◽

Cancer Recurrence ◽

Secondary Resistance ◽

Chemical Structures ◽

Pediatric Solid Tumors ◽

Orthotopic Xenografts ◽

The Relationship

Background: Despite continued efforts to develop new treatments, there is an urgent need to discover new drug leads to treat tumours exhibiting primary or secondary resistance to exiting drugs. Cell cultures derived from patient-derived orthotopic xenografts are promising pre-clinical models to better predict drug response in cancer recurrence. Objective: The aim of the study was to investigate the relationship between the physiochemical properties of drugs and their in vitro potency as well as identifying chemical scaffolds biased towards selectivity or promiscuity of such drugs. Methods: The bioactivities of 158 drugs screened against cell cultures derived from 30 cancer orthotopic patient-derived xenograft (O-PDX) models were considered. Drugs were represented by physicochemical descriptors and chemical structure fingerprints. Supervised learning was employed to model the relationship between features and in vitro potency. Results: Drugs with in vitro potency for alveolar rhabdomyosarcoma and osteosarcoma tend to have a higher number of rings, two carbon-hetero bonds and halogens. Selective and promiscuous scaffolds for these phenotypic targets were identified. Highly-predictive models of in vitro potency were obtained across these 30 targets, which can be applied to unseen molecules via a webserver (https://rnewbie.shinyapps.io/Shobek-master). Conclusion: It is possible to identify privileged chemical scaffolds and predict the in vitro potency of unseen molecules across these 30 targets This information and models should be helpful to select which molecules to screen against these primary cultures of pediatric solid tumors.

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Validation and Application of a New Reversed Phase HPLC Method forIn VitroDissolution Studies of Rabeprazole Sodium in Delayed-Release Tablets

Journal of Analytical Methods in Chemistry ◽

10.1155/2013/976034 ◽

2013 ◽

Vol 2013 ◽

pp. 1-8 ◽

Cited By ~ 4

Author(s):

Md. Saddam Nawaz

Keyword(s):

High Performance ◽

Method Development ◽

High Performance Liquid Chromatographic ◽

Reversed Phase ◽

Hplc Method ◽

Array Detector ◽

Relative Standard ◽

Quality Control Tool ◽

Rabeprazole Sodium

The purpose of this study was to develop and validate a new reversed phase high performance liquid chromatographic (RP-HPLC) method to quantifyin vitrodissolution assay of rabeprazole sodium in pharmaceutical tablet dosage form. Method development was performed on C 18,100×4.6 mm ID, and 10 μm particle size column, and injection volume was 20 μL using a diode array detector (DAD) to monitor the detection at 280 nm. The mobile phase consisted of buffer: acetonitrile at a ratio of 60 : 40 (v/v), and the flow rate was maintained at 1.0 mL/min. The method was validated in terms of suitability, linearity, specificity, accuracy, precision, stability, and sensitivity. Linearity was observed over the range of concentration 0.05–12.0 μg/mL, and the correlation coefficient was found excellent >0.999. The method was specific with respect to rabeprazole sodium, and the peak purity was found 99.99%. The method was precise and had relative standard deviations (RSD) less than 2%. Accuracy was found in the range of 99.9 to 101.9%. The method was robust in different variable conditions and reproducible. This proposed fast, reliable, cost-effective method can be used as quality control tool for the estimation of rabeprazole sodium in routine dissolution test analysis.

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