Cold and Heat in Acute Coronary Syndrome： A Review / 中国实验方剂学杂志

Cold and heat belong to the eight-principal syndrome differentiation of traditional Chinese medicine， which can reflect the rise and fall of Yin and Yang in the body and the Yin and Yang nature of the disease. At present， traditional Chinese medicine has an inconsistent understanding of cold and heat in acute coronary syndrome. The emphasis on pathogenic factors of cold and heat is biased， and the elements of cold and heat syndrome are not fully reflected in the scale. Therefore， the literature has been reviewed from the perspectives of etiology， pathogenesis， symptom elements， and test signs with drugs. From the perspective of etiology， both cold evil and heat evil can increase the risk of acute coronary syndrome. It was previously believed that acute coronary syndrome occurs frequently in cold climates such as winter and spring. Based on this understanding， hot weather can also induce acute coronary syndrome， and different temperatures have different effects on patients of different ages and with different underlying diseases. In addition， artificial pathogenic factors such as excessive consumption of cold food and refrigeration air conditioners were added. From the perspective of pathogenesis， on the basis of the traditional ''asthenia in origin and asthenia in superficiality'' and ''phlegm stagnation''， it is found that Yin-cold and fire-heat can both cause paralysis of the heart chakra and pain induced by the blockage. The pathogenesis of acute coronary syndrome characterized by heat stagnation and coldness featuring heartburn should be distinguished from gastroesophageal reflux disease. Moreover， the pathogenesis of Yin cold coagulation and pulse stagnation and wind obstruction are different. The acute coronary syndrome is in line with the wind characteristics of frequent changes and can be treated with wind medicine. From the perspective of syndrome elements， the syndrome elements such as cold condensation， heat accumulation， and toxicity are analyzed， and the use of basic syndrome elements and their combination forms facilitates clinical and scientific research. In addition， according to the test sign with the drug， it can be seen that the attributes of cold and heat of traditional Chinese medicine prescriptions for acute coronary syndrome can be explained according to the temperature-sensitive transient receptor potential （TRP） ion channel， thus proving the pathogenesis of cold and heat of acute coronary syndrome.

Research progress of TRPV4 in ocular pathophysiological function / 国际眼科杂志(Guoji Yanke Zazhi)

Transient receptor potential vanilloid receptor 4(TRPV4)is a non-selective cation channel responsible for sensing changes in cell swelling, temperature, mechanical stretch, shear stress and osmotic pressure by regulating transmembrane calcium signaling and thereby influencing gene expression, cell morphology, and cytoskeletal construction. TRPV4 is widely expressed throughout the body. Intraocularly, TRPV4 is functionally expressed in the cornea, lens, ciliary body, trabecular meshwork and retina. In this article, the expression and physiopathological functions of TRPV4 in various tissues of the eye were described. With the in-depth study of TRPV4 in ocular pathophysiological functions, TRPV4 may become a potential drug target in corneal injury repair, glaucoma and retinal angiogenesis, but further in-depth study is still needed.

Cold bandage contact lens use reduces post-photorefractive keratectomy or corneal collagen-crosslinking pain perception in patients

Purpose: To compare post?operative pain perception using bandage contact lens (BCL) stored at 2–8?C (Cold BCL, CL?BCL) or room temperature (23 – 25?C, RT?BCL) after photorefractive keratectomy (PRK) or corneal collagen?crosslinking (CXL) and determine status of nociception associated factors. Methods: In this prospective interventional study, 56 patients undergoing PRK for refractive correction and 100 keratoconus (KC) undergoing CXL were recruited following approval from the institutional ethics committee with informed consent. Patients undergoing bilateral PRK received RT?BCL on one eye and CL?BCL on the other. Pain was graded by Wong–Baker scoring on the first post?operative day (PoD1). Expression of transient receptor potential channels (TRPV1, TRPA1, TRPM8), calcitonin gene?related peptide (CGRP) and IL?6 was measured in cellular content from used BCLs collected on PoD1. Equal number of KC patients received RT?BCL or CL?BCL post?CXL. Pain was graded by Wong–Baker scoring on PoD1. Results: Pain scores on PoD1 were significantly (P < 0.0001) reduced in subjects receiving CL?BCL (Mean ± SD: 2.6 ± 2.1) compared to RT?BCL (6.0 ± 2.4) post?PRK. 80.4% of subjects reported reduced pain scores with CL?BCL. 19.6% reported no change or increased pain scores with CL?BCL. TRPM8 expression was significantly (P < 0.05) increased in BCL of subjects reporting reduced pain with CL?BCL compared to those who did not. Pain scores on PoD1 were significantly (P < 0.0001) reduced in subjects receiving CL?BCL (3.2 ± 2.1) compared to RT?BCL (7.2 ± 1.8) post?CXL. Conclusion: The simple approach of using a cold BCL post?operatively substantially reduced pain perception and could overcome post?operative pain?related limited acceptance of PRK/CXL.

Expression and functions of transient receptor potential channels in liver diseases

Liver diseases constitute a major healthcare burden globally, including acute hepatic injury resulted from acetaminophen overdose, ischemia-reperfusion or hepatotropic viral infection and chronic hepatitis, alcoholic liver disease (ALD), non-alcoholic fatty liver disease (NAFLD) and hepatocellular carcinoma (HCC). Attainable treatment strategies for most liver diseases remain inadequate, highlighting the importance of substantial pathogenesis. The transient receptor potential (TRP) channels represent a versatile signalling mechanism regulating fundamental physiological processes in the liver. It is not surprising that liver diseases become a newly explored field to enrich our knowledge of TRP channels. Here, we discuss recent findings revealing TRP functions across the fundamental pathological course from early hepatocellular injury caused by various insults, to inflammation, subsequent fibrosis and hepatoma. We also explore expression levels of TRPs in liver tissues of ALD, NAFLD and HCC patients from Gene Expression Omnibus (GEO) or The Cancer Genome Atlas (TCGA) database and survival analysis estimated by Kaplan-Meier Plotter. At last, we address the therapeutical potential and challenges by pharmacologically targeting TRPs to treat liver diseases. The aim is to provide a better understanding of the implications of TRP channels in liver diseases, contributing to the discovery of novel therapeutic targets and efficient drugs.

Inhibition of intracellular proton-sensitive Ca2+-permeable TRPV3 channels protects against ischemic brain injury

Ischemic brain stroke is pathologically characterized by tissue acidosis, sustained calcium entry and progressive cell death. Previous studies focusing on antagonizing N-methyl-d-aspartate (NMDA) receptors have failed to translate any clinical benefits, suggesting a non-NMDA mechanism involved in the sustained injury after stroke. Here, we report that inhibition of intracellular proton-sensitive Ca2+-permeable transient receptor potential vanilloid 3 (TRPV3) channel protects against cerebral ischemia/reperfusion (I/R) injury. TRPV3 expression is upregulated in mice subjected to cerebral I/R injury. Silencing of TRPV3 reduces intrinsic neuronal excitability, excitatory synaptic transmissions, and also attenuates cerebral I/R injury in mouse model of transient middle cerebral artery occlusion (tMCAO). Conversely, overexpressing or re-expressing TRPV3 increases neuronal excitability, excitatory synaptic transmissions and aggravates cerebral I/R injury. Furthermore, specific inhibition of TRPV3 by natural forsythoside B decreases neural excitability and attenuates cerebral I/R injury. Taken together, our findings for the first time reveal a causative role of neuronal TRPV3 channel in progressive cell death after stroke, and blocking overactive TRPV3 channel may provide therapeutic potential for ischemic brain injury.

Mechanism of Combination of Pungent Dispersing Bitter Evacuant in Treatment of Asthma Based on Theory of Five Flavor Compatibility / 中国实验方剂学杂志

Bronchial asthma， a chronic inflammatory airway disease， belongs to the category of wheezing disease in the system of traditional Chinese medicine （TCM）. The wheezing symptom of this disease is mainly caused by the imbalance of lung Qi. According to the theory of five flavor compatibility， the Chinese medicinal materials with five different flavors （pungent， bitter， sour， sweet， and salty） can be combined to produce new functions. The pungent medicinal materials have dispersing effect and the bitter medicinal materials have discharging effect， which are important components in the theory of five flavor compatibility. Pungent herbs and bitter herbs can relieve the adverse lung Qi， occupying an important position in the current medication for the treatment of asthma. However， there is still a lack of in-depth analysis of the TCM theory and mechanism of the compatibility of pungent herbs and bitter herbs in the treatment of asthma. The molecular mechanisms of action of pungent herbs and bitter herbs are closely related to transient receptor potential （TRP） channels and bitter taste receptors （TAS2Rs）， respectively. Ca2+ signaling has been recognized in the process of asthma and is involved in the development of multiple symptoms of asthma. The TRP channels and TAS2Rs located on the cell membrane have been proved to directly regulate the intracellular Ca2+ signal and play a role in the treatment of asthma. Therefore， the dispersing effect of pungent herbs and the discharging effect of bitter herbs may be realized through the Ca2+ signaling pathway mediated by TRPs/TAS2Rs. We summarized the theoretical understanding and modern studies of pungent herbs dispersing lung Qi and bitter herbs discharging lung Qi， aiming to explain the internal relationship and mechanism of the compatibility of pungent herbs and bitter herbs in the treatment of asthma from the perspective of TCM theory and modern medicine. The compatibility of pungent herbs and bitter herbs based on the theory of five flavor compatibility for the treatment of asthma has a solid theoretical basis of TCM， and its mechanism can be verified by modern research. Therefore， it may be a main research direction in the treatment of asthma by Chinese medicinal herbs in the future.

Genetically-engineered "all-in-one" vaccine platform for cancer immunotherapy

An essential step for cancer vaccination is to break the immunosuppression and elicit a tumor-specific immunity. A major hurdle against cancer therapeutic vaccination is the insufficient immune stimulation of the cancer vaccines and lack of a safe and efficient adjuvant for human use. We discovered a novel cancer immunostimulant, trichosanthin (TCS), that is a clinically used protein drug in China, and developed a well-adaptable protein-engineering method for making recombinant protein vaccines by fusion of an antigenic peptide, TCS, and a cell-penetrating peptide (CPP), termed an "all-in-one" vaccine, for transcutaneous cancer immunization. The TCS adjuvant effect on antigen presentation was investigated and the antitumor immunity of the vaccines was investigated using the different tumor models. The vaccines were prepared

Effect of Houpu Mahuangtang on Airway Inflammation and Expression of TRPA1, TRPV1 mRNA and Protein in Asthmatic Mice / 中国实验方剂学杂志

<b>Objective::To investigate the effect of Houpu Mahuangtang on airway inflammation and expressions of gene and proteins of Transient receptor potential ankyrin 1, vanilloid 1 (TRPA1, TRPV1) in asthmatic mice. <b>Method::Sixty female Balb/c mice were randomly divided into six groups: control group, model group, low, medium and high-dose Houpu Mahuangtang groups (7, 14, 28 g·kg^-1) and dexamethasone group (0.002 4 g·kg^-1), with 10 mice in each group. A mouse model of asthma was replicated by sensitizing and challenging with ovalbumin. The changes of enhanced pause (Penh) following acetylcholine chloride (ACh) inhalation were detected. The pathological changes of the lung tissues were observed. The number of eosinophils (EOS) in peripheral blood and the percentage of EOS in bronchoalveolar lavage fluid (BALF) were detected. Interleukin (IL)-4, IL-13, prostaglandin D₂ (PGD₂) and substance P (SP) were detected by enzyme-linked immuno sorbent assay (ELISA). The expressions of TRPA1 and TRPV1 gene and protein in lung tissues were detected by Quantitative Real-time polymerase chain reaction (Real-time PCR) and Western blot. <b>Result::Compared with control group, mice of model group showed significantly increased in Penh following ACh inhalation (<italic>P</italic><0.05, <italic>P</italic><0.01), EOS in blood and the percentage of EOS in BALF (<italic>P</italic><0.01). Histopathological changes in lungs showed inflammatory cell infiltration and bronchial mucosa damage. The levels of IL-4, IL-13, PGD₂ and SP in BALF and the expressions of TRPA1, TRPV1 mRNA and protein in lung tissues significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01). Compared with model group, treatment groups had significant effects in decreasing Penh, relieving lung injury, reducing EOS count in blood and the percentage of EOS in BALF (<italic>P</italic><0.05, <italic>P</italic><0.01), reducing IL-4, IL-13, PGD₂ and SP levels in BALF (<italic>P</italic><0.05, <italic>P</italic><0.01), as well as down-regulating TRPA1 and TRPV1 mRNA and protein expressions in lung tissues (<italic>P</italic><0.05, <italic>P</italic><0.01). <b>Conclusion::Houpu Mahuangtang could reduce airway inflammation, airway responsiveness. In addition to the reduction of levels of Th2 related cytokines, the mechanism of Houpu Mahuangtang might be related to the regulation of TRPA1, TRPV1 mRNA and protein expressions, and the decrease of associated neurofactor levels.

Analysis of TRPA1 antagonist, A-967079, in plasma using high-performance liquid chromatography tandem mass-spectrometry / 药物分析学报

The noxious effects from exposure to toxic inhalation hazards (TIHs, such as isocyanates, chlorine, etc.) are known to be triggered by the activation of transient receptor potential ankyrin 1 (TRPA1) ion channel. Antagonists of TRPA1 have shown near complete attenuation of the noxious effects from TIH exposure. One of the TRPA1 antagonists, (1E,3E)-1-(4-fluorophenyl)-2-methyl-1-pentene-3-one oxime (A-967079), has shown impressive efficacy, high selectivity, high potency, and oral bioavailability. Although a vali-dated method to quantify A-967079 in biological matrices is vital for the further development of A-967079 as a therapeutic agent, no method for its analysis from any matrix is currently available. Hence, a rapid and simple HPLC-MS/MS method was developed and validated to quantify A-967079 in rabbit plasma. The method presented here features an excellent LOD of 25 nM and a wide linear range (0.05-200 μM), with good accuracy and precision (100 ± 10.5％ and <14.2％ relative standard deviation, respectively). The stability of A-967079 in plasma was excellent for most of the storage conditions evaluated. The method was successfully applied to determine A-967079 from treated animals and it may facilitate the development of this TRPA1 antagonist as a therapeutic agent against the noxious effects of TIH exposure.

Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests / Expressão e análise antigênica da proteína RTP36 recombinante da amostra São Paulo de Ehrlichia canis para testes sorológicos

Abstract Ehrlichia canis is the main etiological agent of canine monocytic ehrlichiosis (CME), a globally canine infectious disease. In Brazil, CME is considered to be endemic, and its prevalence can reach 65% in some states. The diagnosis of ehrlichiosis is important for treatment and epidemiological purposes. The E. canis TRP36 (Tandem Repeat Protein) protein elicits the earliest acute-phase antibody response observed during the course of the disease. This study aimed to generate the recombinant TRP36 protein from E. canis São Paulo strain and to evaluate its potential as a tool for the serologic diagnosis of CME. The E. canis São Paulo isolate was cultivated in DH82 lineage cells, and its genomic DNA was obtained. The bacterial DNA fragment encoding the entire ORF of TRP36 was cloned into the pBAD/Thio-TOPO vector and transformed into Escherichia coli DH10B competent cells with the trp36-bearing plasmid for protein expression. To evaluate the protein antigenicity, 16 canine serum samples were previously tested (by PCR and the commercial SNAP®4Dx® serological test). The results were in accordance with the SNAP®4Dx® test. Experiments using this recombinant protein as an antigen, targeting the development of a serologic test based on ELISA methodology, are the next step to produce a reliable, affordable and useful diagnostic tool for CME in Brazil.

Resumo Ehrlichia canis é o principal agente etiológico da erliquiose monocítica canina (EMC), uma doença infecciosa canina globalmente dispersa. No Brasil, a EMC é considerada endêmica, e a infecção pode atingir 65% em cães em alguns estados. O diagnóstico de erliquiose é importante para fins de tratamento e epidemiológicos. A proteína TRP36 de E. canis leva a uma resposta humoral com produção de anticorpos em fase aguda, encontrada durante o curso da doença. O objetivo deste estudo foi obter a proteína TRP36 recombinante da amostra São Paulo de E. canis e avaliar seu potencial como ferramenta para o diagnóstico sorológico da CME. O isolado de E. canis São Paulo foi cultivado em células da linhagem DH82 e o DNA genômico foi obtido. O fragmento de DNA bacteriano que codifica toda a ORF de TRP36 foi clonado no vetor pBAD / Thio-TOPO e transformado em células competentes Escherichia coli DH10B, com o plasmídeo portador de trp36 para expressão de proteínas. Para avaliar a antigenicidade da proteína, 16 amostras de soro canino foram previamente analisadas (por PCR e teste sorológico comercial SNAP®4Dx®). Os resultados estavam de acordo com o teste SNAP®4Dx®. Os experimentos que utilizam essa proteína recombinante como antígeno, visando ao desenvolvimento de um teste sorológico baseado no ELISA, são o próximo passo para produzir um teste de diagnóstico confiável, acessível e útil para o diagnóstico da EMC no Brasil.

Comparison of ranibizumab alone versus ranibizumab with targeted retinal laser for branch retinal vein occlusion with macular edema

Purpose: To determine the effect of ultra-widefield fluorescein angiography (UWFFA)-guided targeted retinal photocoagulation (TRP) in branch retinal vein occlusion (BRVO) with macular edema after intravitreal Ranibizumab (RBZ). Methods: 33 eyes of 32 treatment naïve patients diagnosed as BRVO with macular edema were prospectively randomized to 0.5 mg Ranibizumab only (RBZ group) (n = 17) or Ranibizumab with UWFFA-guided laser (RBZ + TRP group) (n = 16). Both groups received three injections at monthly intervals and PRN henceforth. RBZ + TRP group additionally underwent UWFFA-guided TRP of peripheral capillary nonperfusion areas 1 week post injection. Outcome measures included improvement in visual acuity, central subfoveal thickness (CST), and the number of injections required with a minimum follow-up of 9 months. Results: Both groups showed significant improvement in mean BCVA (25.7 ± 8.19 letters, P < 0.001 vs. 23.38 ± 7.56 letters, P < 0.001; in RBZ and RBZ + TRP group, respectively) and reduction in mean central subfoveal thickness (379.12 ± 242.7 ?m, P < 0.001 vs. 253.75 ± 137.9 ?m, P < 0.001 in RBZ and RBZ + TRP group, respectively) at 9 months. The number of injections in the RBZ group (5.76 ± 1.3) was significantly greater than RBZ + TRP (4.06 ± 0.99) (P < 0.001). Both groups had significant improvement in contrast sensitivity and mean deviation on visual fields; however, the difference between the groups was not significant (P = 0.62 and P = 0.79, respectively). Conclusion: UWFFA-guided TRP reduced the number of injections of Ranibizumab in patients having BRVO with macular edema, while maintaining similar benefits in the improvement of BCVA, central subfoveal thickness without deleterious effect on the visual field, and contrast sensitivity.

Clinical and genetic findings of two cases with Apert syndrome / Hallazgos clínicos y genéticos de dos casos con síndrome de Apert

Abstract Background: Craniosynostosis is described as the premature fusion of cranial sutures that belongs to a group of alterations which produce an abnormal phenotype. Case report: Two unrelated female patients with clinical findings of Apert syndrome-characterized by acrocephaly, prominent frontal region, flat occiput, ocular proptosis, hypertelorism, down-slanted palpebral fissures, midfacial hypoplasia, high-arched or cleft palate, short neck, cardiac anomalies and symmetrical syndactyly of the hands and feet-are present. In both patients, a heterozygous missense mutation (c.755C>G, p.Ser252Trp) in the FGFR2 gene was identified. Conclusions: Two cases of Apert syndrome are described. It is important to recognize this uncommon entity through clinical findings, highlight interdisciplinary medical evaluation, and provide timely genetic counseling for the family.

Resumen Introducción: Las craneosinostosis se describen como la fusión prematura de las suturas craneales y resultan un grupo de alteraciones que producen un fenotipo anormal. Caso clínico: En este informe de casos se presentan dos pacientes de sexo femenino no emparentadas con hallazgos clínicos del síndrome de Apert, caracterizado por acrocefalia, región frontal prominente, occipucio plano, proptosis ocular, hipertelorismo, fisuras palpebrales hacia abajo, hipoplasia mediofacial, paladar alto o hendido, cuello corto, cardiopatía congénita y sindactilia simétrica en manos y pies. En ambas pacientes se identificó una mutación cambio de sentido en heterocigosis (c.755C>G, p.Ser252Trp) en el gen FGFR2. Conclusiones: Se presentan dos casos de síndrome de Apert. Es importante reconocer a través de los hallazgos clínicos esta entidad infrecuente, resaltar la evaluación médica interdisciplinaria y proporcionar un oportuno asesoramiento genético a la familia.

Anti-inflammatory effects of volatile oils of Desmodium styracifolium via regulation of TRP channels / 中草药

Objective To screen out the key chemical constituents and target protein of essential oil of Desmodium styracifolium for its anti-inflammatory effect. Methods Steam distillation method was used to extract the volatile oils from D. styraci folium, and its chemical constituents were identified by GC-MS, and the relative content of chemical constituents was determined by peak area normalization. The small molecule ligand library was established based on Traditional Chinese Medicine Systems Pharmacology (TCMSP). Reverse target prediction was conducted online using Swiss Target Prediction, the anti-inflammatory pathways were screened by KOBAX 3.0, conducting energy match between the key small molecular and the target protein in the TRP channels by molecular docking (SYBYL2.1). Construction of chemical constituents-targets network model was based on Cytoscape 3.5.1. Results A total of 48 chromatographic peaks were detected from D. styracifolium volatile oils, and 33 kinds of compound structure were determined by searching in mass spectral database and document retrieval, which account for 90.1% of total volatile oils. There were 17 key chemical constituents, and 88 target proteins were selected. TRP channels included 11 potential targets. Through molecular docking, we found that the phytol, hentriacontane, farnesyl acetone, and squalene were the key anti-inflammatory chemical constituents of D. styraci folium volatile oils. TPRV1 (transient receptor potential cation channel, subfamily V, member 1), PRKCB (protein kinase C, beta), and PRKCD (protein kinase C, delta) (degree > 10) are the key anti-inflammatory target protein. Conclusion We preliminarily select the key anti-inflammatory target and active constituents of D. styraci folium volatile oils from this study, and this research provides the theoretical basis for the development and application of its products.

Development of a novel vector for cloning and expressing extremely toxic genes in Escherichia coli

Background: Escherichia coli has been widely used as a host to clone and express heterologous genes. However, there are few vectors available for cloning and expressing extremely toxic genes, which limits further basic and applied research on extremely toxic proteins. Results: In this study, a novel vector pAU10 was constructed in E. coli. pAU10 utilizes the combination of the efficient but highly repressible T7-lacO promoter/operator and the strong rrnBT2 transcriptional terminator upstream of the T7 promoter to strictly control unwanted transcription of the extremely toxic gene; in addition, the trp promoter/operator is oriented opposite to the T7 promoter to control the production of the antisense RNA that may block the translation of leaky mRNA. Without the supplementation of IPTG and L-tryptophan in the culture medium, transcription of the extremely toxic gene by the T7 promoter is highly repressed, and the trp promoter produces the antisense RNA, which strictly prevents unwanted expression of the extremely toxic protein in E. coli. With the supplementation of IPTG and L-tryptophan, the T7 promoter efficiently transcribes the extremely toxic gene, and the trp promoter does not produce the antisense RNA, ensuring efficient expression of the extremely toxic protein in E. coli. Tight regulation and efficiency of expression of an extremely toxic gene cloned in the vector pAU10 were confirmed by cloning and expressing the restriction endonuclease-encoding gene bamHI without its corresponding methylase gene in E. coli JM109(DE3). Conclusion: pAU10 is a good vector used for cloning and expressing extremely toxic genes in E. coli.

Inhibition of two stages of melanin synthesis by sesamol,sesamin and sesamolin

Objective: To investigate the antimelanogenesis properties of three sesame compounds-sesamol,sesamin and sesamolin via two stages of melanin synthesis vis-`a-vis sunscreen function and enzyme inhibition in melanoma cell line in order to search for alternative depigmenting agents. Methods: Antimelanogenic effects of sesame lignans were assessed in SK-MEL2 compared with the reference depigmenting agents, kojic acid and β-arbutin, in order to evaluate:(a)the sunscreen function of sesamol,sesamin and sesamolin by measurement of UV absorbtion property; (b) the inhibition of tyrosinase activity through mushroom and cellular tyrosinase; and (c) the effect on melanin content and melanogenic protein expression(tyrosinase,TRP-1 and TRP-2)by Western blot analysis;and(d)the toxicity of sesamol,sesamin and sesamolin to cells using cell cytotoxicity assay. Results: The results showed that sesamin, sesamolin and sesamol exerted satisfiable sunscreen function by absorbed UVB at 290 nm.Sesamol exhibited the highest inhibition of mushroom tyrosinase activity, but lipophilic sesamolin exhibited the highest cellular tyrosinase inhibition (IC50of 1.6 μM) followed by sesamin, sesamol, and kojic acid, respectively.The order from high to low inhibition of melanin pigment was detected in the SK-MEL2 treated with sesamolin, sesamin, sesamol, kojic acid, and β-arbutin, respectively.Sesamolin and sesamin successfully inhibited cellular tyrosinase activity and respectively decreased TRP-1/TRP-2 (36%/15%) and TRP-1 levels (16%), thereby inhibiting melanogenesis via antityrosinase activity. No cytotoxicity to SK-MEL2 or Vero (normal) cell lines was observed at the lignan concentrations that exerted an anti-melanogenic effect. Conclusions: Three sesame lignans prevent melanin synthesis through 2 stages: (a) by blocking melanin-induction and(b)by interrupting melanogenic enzyme production.This study provides evidence that sesamol, sesamin and sesamolin are potential for anti-melanogenesis agents.

Inhibition of two stages of melanin synthesis by sesamol, sesamin and sesamolin

Objective To investigate the antimelanogenesis properties of three sesame compounds-sesamol, sesamin and sesamolin via two stages of melanin synthesis vis-à-vis sunscreen function and enzyme inhibition in melanoma cell line in order to search for alternative depigmenting agents. Methods Antimelanogenic effects of sesame lignans were assessed in SK-MEL2 compared with the reference depigmenting agents, kojic acid and β-arbutin, in order to evaluate: (a) the sunscreen function of sesamol, sesamin and sesamolin by measurement of UV absorbtion property; (b) the inhibition of tyrosinase activity through mushroom and cellular tyrosinase; and (c) the effect on melanin content and melanogenic protein expression (tyrosinase, TRP-1 and TRP-2) by Western blot analysis; and (d) the toxicity of sesamol, sesamin and sesamolin to cells using cell cytotoxicity assay. Results The results showed that sesamin, sesamolin and sesamol exerted satisfiable sunscreen function by absorbed UVB at 290 nm. Sesamol exhibited the highest inhibition of mushroom tyrosinase activity, but lipophilic sesamolin exhibited the highest cellular tyrosinase inhibition (IC

Food Intake and Thermogenesis in Adipose Tissue

Brown adipose tissue (BAT) is regarded as a key target for developing interventions to prevent and treat obesity and age-related diseases. In addition, uncoupling pro tein 1 (UCP1)

Structural insights into the regulatory mechanism of the Pseudomonas aeruginosa YfiBNR system

YfiBNR is a recently identified bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) signaling system in opportunistic pathogens. It is a key regulator of biofilm formation, which is correlated with prolonged persistence of infection and antibiotic drug resistance. In response to cell stress, YfiB in the outer membrane can sequester the periplasmic protein YfiR, releasing its inhibition of YfiN on the inner membrane and thus provoking the diguanylate cyclase activity of YfiN to induce c-di-GMP production. However, the detailed regulatory mechanism remains elusive. Here, we report the crystal structures of YfiB alone and of an active mutant YfiB(L43P) complexed with YfiR with 2:2 stoichiometry. Structural analyses revealed that in contrast to the compact conformation of the dimeric YfiB alone, YfiB(L43P) adopts a stretched conformation allowing activated YfiB to penetrate the peptidoglycan (PG) layer and access YfiR. YfiB(L43P) shows a more compact PG-binding pocket and much higher PG binding affinity than wild-type YfiB, suggesting a tight correlation between PG binding and YfiB activation. In addition, our crystallographic analyses revealed that YfiR binds Vitamin B6 (VB6) or L-Trp at a YfiB-binding site and that both VB6 and L-Trp are able to reduce YfiB(L43P)-induced biofilm formation. Based on the structural and biochemical data, we propose an updated regulatory model of the YfiBNR system.

Canales iónicos Receptores de Potencial Transitorio y su papel protagónico en la terapia analgésica / Transient receptor potential ion channels and their leading role in analgesic therapy

En la búsqueda de nuevas opciones terapéuticas para el tratamiento del dolor, se ha llegado al descubrimiento de canales iónicos que actúan como receptores y están presentes en neuronas nociceptoras aferentes primarias. Entre estos receptores, se encuentran los canales iónicos receptores de potencial transitorio que regulan las vías involucradas en el dolor y la nocicepción. Se realizó una revisión actualizada de los principales canales iónicos receptores de potencial transitorio implicados en la fisiopatología del dolor. Se hace una reseña histórica del descubrimiento de estas moléculas y sus estudios avanzados. A continuación se revisan las diferentes familias de estos canales con su clasificación, nomenclatura, estructura y funciones celulares. También se hace un recuento de la relación de estos canales con la analgesia, así como el mecanismo de acción de algunos analgésicos que actúan sobre ellos. Finalmente, se detallan importantes consideraciones a tomar en cuenta, que pudieran influir sobre la utilización de estos medicamentos en la clínica. Por tal motivo, el trabajo procura ser una revisión que abarque el rol de los canales TRP como nuevas dianas farmacológicas en el tratamiento del dolor.

In the search for new therapeutic options for pain treatment, new ion channels have been discovered, which act as receptors and are present in primary afferent nociceptor neurons. Some of them are the transient receptor potential ion channels that regulate the pathways involved in pain and nociception. An updated review of the main transient potential receptors ion channels involved in pain physiopathology and a historical review of the discovery of these molecules and advanced studies on them were also made. Then the different families of these channels with their classification, nomenclature, structure and cell functions were also examined. An account of the relationships of these channels with analgesia as well as the mechanism of action of some analgesics acting upon them was also presented. Finally, important considerations were given, which should be taken into account since they might influence on the clinical use of these drugs. For these reasons, the paper intends to be a review covering the role of the transient receptor potential channels as new pharmacological targets in the pain treatment.

Role of the Trp-disulfide Triads in the UV Light Induced Degradation of a Monoclonal Antibody scFv.

Proteins are targets for photodegradation due to absorption of incident light by endogenous chromophores, e.g aromatic side chains. In this work we study the role of Trp-disulfide triads in the light induced loss of immunoglobulin activity. Study Design: We investigated a single chain variable fragment (scFv) of the Trp-disulfide triad containing monoclonal antibody 82D6A3. The scFv binds to von Willebrand factor (VWF) and upon illumination with near UV-B-light the scFv partially loses its binding capacity to VWF. In order to relate this observed degeneration to the specific Trp-disulfide triads, we mutated W35(VL) and W36(VH) which are in direct contact with the disulfide