Cytomegalovirus detected by qPCR in iris and ciliary body of immunocompetent corneal donors.

BACKGROUND: Cytomegalovirus (CMV) can cause a wide panel of ocular infections. The involvement of CMV as a cause of anterior uveitis in the immunocompetent patient is recent and remains poorly understood. OBJECTIVE: To investigate the presence of CMV in anterior uveal tissues of immunocompetent corneal donors. STUDY DESIGN: We collected aqueous humor, iris, and ciliary body from both eyes of 25 donors died at the Limoges University Hospital between January 2020 and July 2021. CMV serology was determined for all patients from post-mortem blood sample. Ocular tissues were split in 2 fragments for qPCR and 2 for histological analysis. CMV genomes copies were quantified by Multiplex qPCR after DNA extraction. RESULTS: 16 of 25 patients (64%) displayed positive CMV serology, with a median age of 67 years. Viremia was positive in 3 of 16 (19%) CMV-positive patients. No CMV DNA copies were found from the aqueous humor samples. CMV DNA was detected in iris and ciliary body of 28 of 32 eyes of seropositive donors, and 5 of 18 eyes of seronegative donors. The median viral copy number [IQR] was 2.41 × 102 [8.91 × 101 - 1.01 × 103] copies/1 × 106 cells in the CMV-positive group and 0.00 [0.00 - 3.54 × 102] copies/1 × 106 cells in the CMV-negative group (p<0.001). Histology and immunohistochemistry did not reveal any CMV lesions from any sample. CONCLUSION: CMV DNA was found in iris and ciliary body of immunocompetent seropositive patients, but also, although less frequently, from seronegative donors. These results highlight mechanisms of infection, latency and reactivation of CMV in ocular tissues.

Biogenic Silver Nanoparticles from Iris tuberosa as Potential Preservative in Cosmetic Products.

Biogenic-silver nanoparticles emerge as new nanosilver platforms that allow us to obtain silver nanoparticles via "green chemistry". In our study, biogenic-silver nanoparticles were obtained from Iris tuberosa leaf extract. Nanoparticles were characterized by a UV-vis spectroscopy, dynamical light scattering technique. The transmission electron microscope revealed spheric and irregular nanoparticles with 5 to 50 nm in diameter. Antimicrobial properties were evaluated against typical microbial contaminants found in cosmetic products, showing high antimicrobial properties. Furthermore, natural moisturizing cream was formulated with biogenic-silver nanoparticles to evaluate the preservative efficiency through a challenge test, indicating its promising use as preservative in cosmetics.

Oligostilbenes extracts from Iris lactea Pall. var. chinensis (Fisch.) Koidz improve lipid metabolism in HFD/STZ-induced diabetic mice and inhibit adipogenesis in 3T3-L1 cells.

The present study investigated the anti-diabetic effects of Oligostilbenes extracts (Olie) from Iris lactea Pall. var. chinensis (Fisch.) Koidz (I. lactea) and the potential mechanisms, in high-fat-diet (HFD)-induced diabetic mice and 3T3-L1 adipocytes. Olie are a group of major active extracts from I. lactea that have been used as nutraceutical because of their antioxidant activity. Six-week Olie treatment improved fasting blood glucose levels, as well as blood lipid profiles in HFD/streptozocin (STZ)-induced diabetic mice, compared with non-treated mice. Olie treatment upregulated the levels of phosphorylated of AMPK and lipolysis-related proteins, while the hepatic expression of ACC and FAS in diabetic mice was inhibited. In cultured 3T3-L1 cells, Olie (2-15 µg/mL) treatment dose-dependently suppressed the differentiation into mature adipocytes and lowered cellular lipid accumulation. Consistently, Olie reduced expression of adipogenic transcription factors including CCAAT/enhancer-binding protein ß (C/EBPß) and peroxisome proliferator-activated receptor γ (PPARγ). In addition, mitochondrial function in 3T3-L1 adipocytes was improved after Olie treatment. Taken together, our findings indicate that a lipid-lowering effect of Olie in HFD/STZ-induced diabetic mice and adipogenesis/ lipogenesis suppressing effect in 3T3-L1 cells, via regulating the expression of lipid metabolism-related proteins.

Molecular and biochemical mechanisms of human iris color: A comprehensive review.

Eye color is determined as a polymorphism and polygenic trait. Brown is the most common eye color in the world, accounting for about 79%, blue eye color for about 8-10%, hazel for 5%, and green for 2%. Rare-colored eyes include gray and red/violet. Different factors are involved in determining eye color. The two most important factors are the iris pigment and the way light is scattered from the iris. Gene expression determines the iris pigmentation and how much melanin is present in the eye, which is the number of melanin subunits that identify eye color. The genes involved in the pigmentation of single-nucleotide polymorphism (SNP) have a significant role; and even some genes are included only in the eye color through SNP. MicroRNAs also affect melanocyte synthesis, which is usually affected by the downregulation of essential genes involved in pigmentation. In this study, we assess the biochemical pathways of melanin synthesis, and the role of each gene in this pathway also has been examined in the signaling pathway that stimulates melanin synthesis.

Protective effects of Belamcandae Rhizoma against skin damage by ameliorating ultraviolet-B-induced apoptosis and collagen degradation in keratinocytes.

Ultraviolet-B light (UV-B) is a major cause of skin photoaging, inducing cell death and extracellular matrix collapse by generating reactive oxygen species (ROS). Belamcandae Rhizoma (BR), the rhizome of Belamcanda chinensis Leman, exhibits antioxidant properties, but it remains unknown whether BR extract ameliorates UV-B-induced skin damage. In this study, we evaluated the effects of a standardized BR extract on UV-B-induced apoptosis and collagen degradation in HaCaT cells. BR was extracted using four different methods. We used radical-scavenging assays to compare the antioxidative activities of the four extracts. Cells were irradiated with UV-B and treated with BR boiled in 70% (vol/vol) ethanol (BBE). We measured cell viability, intracellular ROS levels, the expression levels of antioxidative enzymes, and apoptosis-related and collagen degradation-related proteins. The irisflorentin and tectorigenin levels were measured via high-performance liquid chromatography. BBE exhibited the best radical-scavenging and cell protective effects of the four BR extracts. BBE inhibited intracellular ROS generation and induced the synthesis of antioxidative enzymes such as catalase and glutathione. BBE attenuated apoptosis by reducing the level of caspase-3 and increasing the Bcl-2/Bax ratio. BBE reduced the level of matrix metalloproteinase-1 and increased that of type I collagen. The irisflorentin and tectorigenin contents were 0.23% and 0.015%, respectively. From these results, BBE ameliorated UV-B-induced apoptosis and collagen degradation by enhancing the expression of antioxidative enzymes. It may be a useful treatment for UV-B-induced skin damage.

Belamchinanes A-D from Belamcanda chinensis: Triterpenoids with an Unprecedented Carbon Skeleton and Their Activity against Age-Related Renal Fibrosis.

Belamchinanes A-D (1-4), four triterpenoids with an unprecedented skeleton, were isolated from the seeds of Belamcanda chinensis and fully characterized. The structures of belamchinanes feature a 4/6/6/6/5 polycyclic system in which a four-membered carbocyclic ring bridges the C-1 and C-11 positions of a classical triterpenoid framework. A plausible pathway for the biosynthesis of 1-4 is proposed. Biological studies reveal that 1-4 dose-dependently protect age-related renal fibrosis in vitro .

Three-step monitoring of glycan and galectin profiles in the anterior segment of the adult chicken eye.

A histochemical three-step approach is applied for processing a panel of sections that covers the different regions of fixed anterior segment of the adult chicken eye. This analysis gains insight into the presence of binding partners for functional pairing by galectin/lectin recognition in situ. Glycophenotyping with 11 fungal and plant lectins (step 1) revealed a complex pattern of reactivity with regional as well as glycan- and cell-type-dependent differences. When characterizing expression of the complete set of the seven adhesion/growth-regulatory chicken galectins immunohistochemically (step 2), the same holds true, clearly demonstrating profiles with individual properties, even for the CG-1A/B paralogue pair. Testing this set of labeled tissue lectins as probes (step 3) detected binding sites in a galectin-type-dependent manner. The results of steps 2 and 3 reflect the divergence of sequences and argue against functional redundancy among the galectins. These data shape the concept of an in situ network of galectins. As consequence, experimental in vitro studies will need to be performed from the level of testing a single protein to work with mixtures that mimic the (patho)physiological situation, a key message of this report.

Secondary Glaucoma due to Iridescent Crystalline Particles Masquerading as Refractory Hypertensive Uveitis in an Eye With Irradiated Iris Melanoma.

PURPOSE: We report a previously unrecognized mechanism of secondary glaucoma due to iridescent crystalline particles released from an irradiated iris melanoma. It masqueraded as refractory hypertensive uveitis following uncomplicated phacoemulsification. MATERIALS AND METHODS: A 58-year-old gentleman had an iris melanoma that underwent successful regression following irradiation with proton beam radiotherapy. Three years later an uncomplicated phacoemulsification with intraocular lens implant was performed and subsequently the patient presented with apparently "refractory hypertensive uveitis." Closer examination identified unique iridescent crystalline particles originating from a disintegrating tumor and dispersing within the anterior chamber and drainage angle. The patient developed a unilateral secondary open-angle glaucoma attributable to these particles. Ultrasound biomicroscopy of the anterior segment confirmed absence of tumor recurrence or intrascleral spread and systemic investigations ruled out distant metastases. RESULTS: The intraocular pressure was refractory to maximal medical treatment, but was eventually controlled with trans-scleral diode laser cyclo-photocoagulation. CONCLUSIONS: This is the first report of a secondary glaucoma attributable to trabecular blockage with iridescent crystalline particulate material released from a disintegrating, previously irradiated, iris melanoma. Proton beam radiotherapy and possibly phacoemulsification may have played a role in triggering the release of these previously undescribed particles from the atrophied tumor surface. This unique mechanism of secondary glaucoma needs to be kept in mind in such rare cases. Trans-scleral cyclodiode laser may be used as a good initial option in such cases to minimize potential risk of tumor seeding with incisional glaucoma surgery.

A novel culture method reveals unique neural stem/progenitors in mature porcine iris tissues that differentiate into neuronal and rod photoreceptor-like cells.

Iris neural stem/progenitor cells from mature porcine eyes were investigated using a new protocol for tissue culture, which consists of dispase treatment and Matrigel embedding. We used a number of culture conditions and found an intense differentiation of neuronal cells from both the iris pigmented epithelial (IPE) cells and the stroma tissue cells. Rod photoreceptor-like cells were also observed but mostly in a later stage of culture. Neuronal differentiation does not require any additives such as fetal bovine serum or FGF2, although FGF2 and IGF2 appeared to promote neural differentiation in the IPE cultures. Furthermore, the stroma-derived cells were able to be maintained in vitro indefinitely. The evolutionary similarity between humans and domestic pigs highlight the potential for this methodology in the modeling of human diseases and characterizing human ocular stem cells.

PTP1B inhibitors from the seeds of Iris sanguinea and their insulin mimetic activities via AMPK and ACC phosphorylation.

To find PTP1B inhibitors from natural products, two new compounds (1 and 2), along with nine known compounds (3-11), were isolated from a methanol-soluble extract of Iris sanguinea seeds. The structures of compounds 1 and 2 were determined based on extensive spectroscopic data analysis including UV, IR, NMR, and MS. The IC50 value of compound 5 on protein tyrosine phosphatase 1B (PTP1B) inhibitory activity is 7.30±0.88µM with a little activity compared to the IC50 values of the tested positive compound. Compound 5 significantly enhanced glucose uptake and activation of pACC, pAMPK and partially Erk1/2 signaling. These results suggest that compound 5 from Iris sanguinea seeds are utilized as both PTP1B inhibitors and regulators of glucose uptake. These beneficial effects could be applied to treat metabolic diseases such as diabetes and obesity.

PROTECTIVE EFFECT OF IRIS GERMANICA L. IN Β-AMYLOID-INDUCED ANIMAL MODEL OF ALZHEIMER'S DISEASE.

BACKGROUND: Alzheimer's disease (AD) is the most common cause of dementia that is an irretrievable chronic neurodegenerative disease. In the current study, we have examined the therapeutic effects of Iris germanica extract on Amyloid ß (Aß) induced memory impairment. MATERIALS AND METHODS: Wistar rats were divided into five groups of 8 per each. Groups were as followed: control group which were normal rats without induction of AD, Aß group which received Aß (50 ng/side), iris 100 group which received Aß + Iris (100 mg/kg), iris 200 group which received Aß + Iris (200 mg/kg), and iris 400 group which received Aß + Iris (400 mg/kg). AD was established by intrahippocampal injection of 50 ng/µl/side Aß1-42. The day after surgery, animals in treatment groups received different doses of the aqueous extract of Iris by gavage for 30 days. Morris water maze test (MWM) was performed to assess the effects of I. germanica on learning and memory of rats with Aß induced AD. RESULTS: Data from MWM tests, including escape latency and traveled distance, demonstrated that I. germanica extract could markedly improve spatial memory in comparison to control. Moreover, the plant had a significantly better effect on the performance of AD rats in the probe test. CONCLUSION: I. germanica extract can successfully reverse spatial learning dysfunction in an experimental model of AD. Further neuro psyco-pharmacological studies are mandatory to reveal the mechanism of action of this natural remedy in the management of AD symptoms.

Polycycloiridals A-D, Four Iridal-Type Triterpenoids with an α-Terpineol Moiety from Iris tectorum.

Polycycloiridals A-D, four novel iridals with an unprecedented α-terpineol moiety resulting from cyclization of the homofarnesylside chain, were isolated from the ethanol extract of rhizomes of Iris tectorum. Their structures were elucidated on the basis of comprehensive spectroscopic analysis. The absolute configuration of 1 was determined by the modified Mosher's method and comparison of experimental and calculated electronic circular dichroism (ECD) spectrum. A possible biosynthetic pathway was postulated.

Comparative pharmacokinetic profiles of tectorigenin in rat plasma by UPLC-MS/MS after oral administration of Iris tectorum Maxim extract and pure tectoridin.

Iris tectorum Maxim, a well-known herb medicine, is commonly used for treatment of inflammation, cough, and pharyngitis for a long time in China. Tectoridin, main active ingredient of Iris tectorum Maxim, is often used for its quality control. This study was aimed to analyze the pharmacokinetic profile of tectorigenin (the metabolite of tectoridin) after oral administration of I. tectorum Maxim extract, and to compare the pharmacokinetic characterization of tectorigenin after oral administration of I. tectorum Maxim extract (ITME) and pure tectoridin (PT) in rats. In addition, a simple, reliable and sensitive UPLC-MS/MS method was developed for determination of tectorigenin in rat plasma, using kaempferol as internal standard. The processed samples were separated on a Poroshell 120 SB-C18 column and detected by positive electrospray ionization in multiple reaction monitoring (MRM) mode. The method validation results indicated that the established method was simple, specific and reliable. The pharmacokinetic results showed that the plasma concentration of tectorigenin in ITME group was much higher than that of the PT group (p<0.01). Moreover, compared to PT group, t1/2 value and AUC(0-∞) value were also notably increased in ITME group (p<0.01). In conclusion, potential interaction exists between those chemical components in ITME, and the co-existing components in ITME could notably promote the absorption of tectoridin in rats, however, the exact compound(s) which enhance the absorption of tectoridin should be investigated in future study.

A metabolomic approach to quality determination and authentication of raw plant material in the fragrance field. Iris rhizomes: a case study.

This study aimed to discriminate 22 samples of commercial Iris rhizomes (orris root) by species and origin (Iris germanica (Morocco), I. albicans (Morocco), I. pallida (Morocco), I. pallida (China), I. pallida (Italy)) by applying a strategy derived from those adopted in metabolomics. The specimens' fingerprints from conventional analysis methods (LC-UV and/or LC-MS) were unable to provide clear discrimination. A strategy combining UHPLC/TOF-HRMS, in positive and negative modes, with multivariate statistical methods was therefore applied. Exact mass/retention time (EMRT) pairs obtained by UHPLC-TOF/HRMS were successfully submitted to statistical processing by principal component analysis (PCA), partial least square discriminant analysis (PLS-DA), and then orthogonal partial least square-discriminant analysis (OPLS-DA), to extract the discriminating EMRT pairs through their trend views. 146 EMRT pairs were selected on the basis of their trend views, because they significantly varied, and 104 of them were included to discriminate between species and origins. 32 of them were tentatively identified as discriminating markers (flavonoids, isoflavonoids, triterpenoids, benzophenone derivatives and related glycosides …) from the reference database created on the basis of Iris genus components reported in the literature: eight of them specific for I. albicans, four for I. germanica, five for I. pallida (Italy), five for I. pallida (China), and ten for I. pallida (Morocco). The reliability of this strategy was confirmed by identifying species and origin of two unknown samples submitted to the same analytical procedure.

New isoflavones with cytotoxic activity from the rhizomes of Iris germanica L.

Two new compounds, 5-methoxy-3',4'-dihydroxy-6,7-methylenedioxy-4H-1-benzo-pyran-4-one(iriskashmirianin A) (1) and 5,3'-dihydroxy-3-(4'-ß-D-glucopyranosyl)-6,7-methylenedioxy-4H-1-benzo-pyran-4-one (germanaism H) (2), along with eight known compounds (3-10), were isolated from the rhizomes of Iris germanica L. The cytotoxicities of these compounds were tested using Ehrlich's ascites carcinoma (EAC) cancer cell line by 3-(4, 5-dimethylthiazole-2-yl)-2, 5-diphenyltetrazoli-umbromide (MTT) and ATP assays. The results showed that these compounds possessed antiproliferative effects on EAC cell line. Among them, compound 1 possessed the best cytotoxic activity with IC50 ± SD of 20.9 ± 2.7 and 4.3 ± 0.9 µM for MTT and ATP assay methods, respectively.

Osteogenic activity of yellow flag iris (Iris pseudacorus) extract modulating differentiation of osteoblasts and osteoclasts.

Bone integrity is maintained through a balance between bone formation by osteoblasts and bone resorption by osteoclasts. Imbalance of the process results in metabolic bone diseases such as osteoporosis. This study investigated the yellow flag iris extract (YFIE) and revealed its anti-osteoporotic effects in osteoblastic MC3T3-E1 mouse cells and RAW 264.7 murine macrophages. When osteoblasts were treated with 1-20 µg/ml YFIE in an osteogenic medium, the bone nodule formation by calcium deposits was markedly enhanced during differentiation. Consistently, YFIE stimulated alkaline phosphatase activity and collagen type I secretion with a substantial effect on osteoblast proliferation. On the other hand, RAW 264.7 macrophages were pre-incubated with 1-20 µg/ml YFIE for 5 days in the presence of receptor activator of nuclear factor-κB ligand (RANKL). Non-toxic YFIE markedly attenuated the differentiation of macrophages to multi-nucleated osteoclasts. YFIE diminished RANKL-elevated tartrate-resistant acid phosphatase activity and bone resorption. In addition, the YFIE treatment retarded RANKL-induced cathepsin K production and carbonic anhydrase II expression, both of which are involved in bone resorption. Therefore, YFIE potentially posesses therapeutic agents that may prevent osteoporosis through promoting bone formation and reducing bone resorption.

Characteristics of neurological Wilson's disease without Kayser-Fleischer ring.

BACKGROUND: Although Kayser-Fleischer ring has been regarded as a key diagnostic feature of neurologic Wilson's disease, there have been previous reports of neurologic Wilson's disease patients without Kayser-Fleischer ring. We assessed the characteristics of neurologic Wilson's disease patients without Kayser-Fleischer ring. METHODS: We enrolled neurologic Wilson's disease patients from 4 university hospitals by review of medical records in this study. Patients with neurologic Wilson's disease were diagnosed based on the neurologic symptoms and international scoring system for the diagnosis of Wilson's disease. All subjects were divided into two groups according to the presence of a Kayser-Fleischer ring. We compared demographic data, laboratory findings and imaging findings of the liver and brain between the two groups. RESULTS: There were 12 (26.7%) patients without Kayser-Fleischer ring out of a total of 45 neurologic Wilson's disease patients. The Wilson's disease patients without Kayser-Fleischer ring demonstrated a higher ceruloplasmin concentration and serum copper level than those with Kayser-Fleischer ring. In addition, liver cirrhosis and typical signal changes in brain magnetic resonance imaging were less common in neurologic Wilson's disease patients without Kayser-Fleischer ring. CONCLUSION: Based on our results, the absence of Kayser-Fleischer ring can be regarded as a form of neurologic Wilson's disease with less copper involvement. In addition, it is important to understand these features and to perform further investigations if patients without Kayser-Fleischer ring are suspected of having neurologic Wilson's disease.

Pro-apoptotic effects of tectorigenin on human hepatocellular carcinoma HepG2 cells.

AIM: To investigate the effects of tectorigenin on human hepatocellular carcinoma (HCC) HepG2 cells. METHODS: Tectorigenin, one of the main components of rhizome of Iris tectorum, was prepared by simple methods, such as extraction, filtration, concentration, precipitation and recrystallization. HepG2 cells were incubated with tectorigenin at different concentrations, and their viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptosis was detected by morphological observation of nuclear change, agarose gel electrophoresis of DNA ladder, and flow cytometry with Hoechst 33342, Annexin V-EGFP and propidium iodide staining. Generation of reactive oxygen species was quantified using DCFH-DA. Intracellular Ca(2+) was monitored by Fura 2-AM. Mitochondrial membrane potential was monitored using Rhodamine 123. Release of cytochrome c from mitochondria to cytosol was detected by Western blotting. Activities of caspase-3, -8 and -9 were investigated by Caspase Activity Assay Kit. RESULTS: The viability of HepG2 cells treated by tectorigenin decreased in a concentration- and time-dependent manner. The concentration that reduced the number of viable HepG2 cells by 50% (IC(50)) after 12, 24 and 48 h of incubation was 35.72 mg/L, 21.19 mg/L and 11.06 mg/L, respectively. However, treatment with tectorigenin at 20 mg/L resulted in a very slight cytotoxicity to L02 cells after incubation for 12, 24 or 48 h. Tectorigenin at a concentration of 20 mg/L greatly inhibited the viability of HepG2 cells and induced the condensation of chromatin and fragmentation of nuclei. Tectorigenin induced apoptosis of HepG2 cells in a time- and dose-dependent manner. Compared with the viability rate, induction of apoptosis was the main mechanism of the anti-proliferation effect of tectorigenin in HepG2 cells. Furthermore, tectorigenin-induced apoptosis of HepG2 cells was associated with the generation of reactive oxygen species, increased intracellular [Ca(2+)](i), loss of mitochondrial membrane potential, translocation of cytochrome c, and activation of caspase-9 and -3. CONCLUSION: Tectorigenin induces apoptosis of HepG2 cells mainly via mitochondrial-mediated pathway, and produces a slight cytotoxicity to L02 cells.