Antifungal and antiviral products of marine organisms.

Marine organisms including bacteria, fungi, algae, sponges, echinoderms, mollusks, and cephalochordates produce a variety of products with antifungal activity including bacterial chitinases, lipopeptides, and lactones; fungal (-)-sclerotiorin and peptaibols, purpurides B and C, berkedrimane B and purpuride; algal gambieric acids A and B, phlorotannins; 3,5-dibromo-2-(3,5-dibromo-2-methoxyphenoxy)phenol, spongistatin 1, eurysterols A and B, nortetillapyrone, bromotyrosine alkaloids, bis-indole alkaloid, ageloxime B and (-)-ageloxime D, haliscosamine, hamigeran G, hippolachnin A from sponges; echinoderm triterpene glycosides and alkene sulfates; molluscan kahalalide F and a 1485-Da peptide with a sequence SRSELIVHQR; and cepalochordate chitotriosidase and a 5026.9-Da antifungal peptide. The antiviral compounds from marine organisms include bacterial polysaccharide and furan-2-yl acetate; fungal macrolide, purpurester A, purpurquinone B, isoindolone derivatives, alterporriol Q, tetrahydroaltersolanol C and asperterrestide A, algal diterpenes, xylogalactofucan, alginic acid, glycolipid sulfoquinovosyldiacylglycerol, sulfated polysaccharide p-KG03, meroditerpenoids, methyl ester derivative of vatomaric acid, lectins, polysaccharides, tannins, cnidarian zoanthoxanthin alkaloids, norditerpenoid and capilloquinol; crustacean antilipopolysaccharide factors, molluscan hemocyanin; echinoderm triterpenoid glycosides; tunicate didemnin B, tamandarins A and B and; tilapia hepcidin 1-5 (TH 1-5), seabream SauMx1, SauMx2, and SauMx3, and orange-spotted grouper ß-defensin. Although the mechanisms of antifungal and antiviral activities of only some of the aforementioned compounds have been elucidated, the possibility to use those known to have distinctly different mechanisms, good bioavailability, and minimal toxicity in combination therapy remains to be investigated. It is also worthwhile to test the marine antimicrobials for possible synergism with existing drugs. The prospects of employing them in clinical practice are promising in view of the wealth of these compounds from marine organisms. The compounds may also be used in agriculture and the food industry.

In vitro refolding process of bovine allergen ß-lactoglobulin by Multispectroscopic method.

OBJECTIVE: To characterize the relationship between the refolding process of recombinant bovine ß-lactoglobulin and its immunoreactivity for clinical purposes. To establish a spectral method which examine the extent of recombinant allergen renaturation. METHODS: The refolding process of recombinant bovine ß-lactoglobulin was investigated by using circular dichroism, fluorescence and synchronous fluorescence spectra. IgE-binding capacity of recombinant protein was analyzed by ELISA. In addition, bioinformatic methods were used to explain the spectral characteristics and analyze the relationship between the conformational changes and the immunoreactivity of the protein during renaturation in vitro. RESULTS: Renaturation of recombinant bovine ß-lactoglobulin resulted in a more compact structure resembling the natural counterpart with stronger IgE-binding capacity. CONCLUSION: The degree of protein renaturation correlated with the IgE-binding capacity of the protein. Results from this study may be of help for food allergy therapy and development of vaccination in the future.

[Cloning, expression and purification of arginine kinase from Blattella germanica and its immune activity].

OBJECTIVE: To clone and express the arginine kinase (AK) gene of Blattella germanica and analyze its immune activity. METHODS: The cDNA of AK was cloned using specific primers from the total RNA of Blattella germanica The open reading frame (ORF) of AK was cloned into pET-28A vector, and expressed in Escherichia coli BL21(DE3) with IPTG induction. The recombinant protein was purified by Ni2+ chelating affinity chromatography. The recombinant protein was detected by SDS-PAGE, and its immune activity was analyzed by Western blotting. RESULTS: The cloned cDNA ORF sequence (GenBank accession No. FJ514482) contained 1071bp and encoded 356 amino acids. Its sequence homology with the published one (GenBank accession No. EU429466) was 97.2% at nucleotide level. The recombinant containing recombinant plasmid pET-28a-AK expressed a soluble protein of AK (Mr 45 000) after being induced with IPTG. The recombinant AK protein was recognized by sera of allergic patients, indicating that the recombinant AK protein has an adequate response activity. CONCLUSION: The AK gene of Blattella germanica has been cloned and the recombinant AK protein has been confirmed with immune activity.

Strain and Electric Field Controllable Schottky Barriers and Contact Types in Graphene-MoTe2 van der Waals Heterostructure.

Two-dimensional (2D) transition metal dichalcogenides with intrinsically passivated surfaces are promising candidates for ultrathin optoelectronic devices that their performance is strongly affected by the contact with the metallic electrodes. Herein, first-principle calculations are used to construct and investigate the electronic and interfacial properties of 2D MoTe2 in contact with a graphene electrode by taking full advantage of them. The obtained results reveal that the electronic properties of graphene and MoTe2 layers are well preserved in heterostructures due to the weak van der Waals interlayer interaction, and the Fermi level moves toward the conduction band minimum of MoTe2 layer thus forming an n type Schottky contact at the interface. More interestingly, the Schottky barrier height and contact types in the graphene-MoTe2 heterostructure can be effectively tuned by biaxial strain and external electric field, which can transform the heterostructure from an n type Schottky contact to a p type one or to Ohmic contact. This work provides a deeper insight look for tuning the contact types and effective strategies to design high performance MoTe2-based Schottky electronic nanodevices.

Narrow-band ultraviolet B phototherapy for early stage mycosis fungoides.

Recently there have been some reports concerned the treatment of early stage mycosis fungoides (MF) with narrow-band ultraviolet B (NB-UVB) phototherapy. In most of the previous reports, NB-UVB phototherapy was given three times a week on non-consecutive days. Our aim was to evaluate the effect of a twice weekly regimen of NB-UVB phototherapy in the treatment of early-stage MF. Eight patients with early stage MF received NB-UVB phototherapy twice weekly. Six patients (75%) had a complete response in a mean of 23.4 treatments, two (25%) had a partial response. Upon discontinuation of treatment, four patients with complete response relapsed in a mean time to relapse of 5 months. The twice weekly regimen of NB-UVB phototherapy is effective and well-tolerated in the treatment of early stage MF.

Regulation of haptoglobin expression in a human keratinocyte cell line HaCaT by inflammatory cytokines and dexamethasone.

BACKGROUND: Haptoglobin (Hp) is one of the acute-phase proteins. Recent studies have demonstrated that Hp exerts immunoregulatory and anti-inflammatory actions and may be one of the inhibitory factors of immune reactions in the skin. In this study we investigated the regulation of Hp expression in a human keratinocyte cell line HaCaT by various cytokines and glucocorticoid. METHODS: HaCaT cells were cultured with IL-6 (50 ng/ml), TNF-alpha (20 ng/ml), IFN-gamma (20 ng/ml) or IL-4 (20 ng/ml) with or without 1 micromol/L dexamethasone in 6-well plates for 12, 24 and 48 hours. Both the cells and the supernatants were collected to detect the changes of Hp expression by reverse-transcription PCR, ELISA and immunohistochemistry. RESULTS: The results showed that Hp expression were elevated at both the mRNA and protein level by the combination of IL-6, TNF-alpha or IL-4 with dexamethasone, whereas the three cytokines alone did not upregulate the Hp expression. IFN-gamma showed no effect on the Hp expression in HaCaT cells. CONCLUSIONS: These findings suggest that different inflammatory cytokines as well as glucocorticoid may be involved in the regulation of Hp expression in keratinocytes, and this may be one of the negative feedback mechanisms in inflammatory skin diseases.

[Cloning, expression and purification of allergen arginine kinase from Periplaneta americana and its allergic activity].

OBJECTIVE: To clone the gene of arginine kinase (AK) from Periplaneta americana, produce its recombinant protein and investigate its allergenicity. METHOD: The cDNA of AK was cloned using specific primers from the total RNA of P. americana. The cloned gene was inserted into pMD18-T vector and digested by BamHI and HindIII. The cDNA was sequenced and subcloned into pET-28a expression vector. The cloned AK cDNA gene was expressed in Escherichia coli BL21 (DE3) by IPTG induction. The recombinant AK (rAK) was purified by metal (Ni2+) chelating affinity chromatography. Its allergenicity was examined by both Western blotting and enzyme-linked immunosorbent assay (ELISA). RESULT: The cloned cDNA ORF sequence (Accession no. EU429466) contained 1068 bp and encoded 365 amino acids. Its sequence homology with the published one (Accession no. AY563004) was 99.9% at nucleotide level. The allergen rAK was highly expressed in E. coli BL21 (DE3) as a soluble protein mainly with the molecular weight of about Mr 45000 under induction of IPTG and purified by 6-His-tag purification system. Both in the non-denaturalization and denaturalization conditions, the recombinant allergen was identified as its affinity to IgE antibodies from the cockroach-allergic patient sera by Western blotting and ELISA. CONCLUSION: The recombinant cockroach arginine kinase has been obtained with proper allergenicity.

TMPyP4 promotes cancer cell migration at low doses, but induces cell death at high doses.

TMPyP4 is widely considered as a potential photosensitizer in photodynamic therapy and a G-quadruplex stabilizer for telomerase-based cancer therapeutics. However, its biological effects including a possible adverse-effect are poorly understood. In this study, whole genome RNA-seq analysis was used to explore the alteration in gene expression induced by TMPyP4. Unexpectedly, we find that 27.67% of changed genes were functionally related to cell adhesion. Experimental evidences from cell adhesion assay, scratch-wound and transwell assay indicate that TMPyP4 at conventional doses (≤0.5 µM) increases cell-matrix adhesion and promotes the migration of tumor cells. In contrast, a high dose of TMPyP4 (≥2 µM) inhibits cell proliferation and induces cell death. The unintended "side-effect" of TMPyP4 on promoting cell migration suggests that a relative high dose of TMPyP4 is preferred for therapeutic purpose. These findings contribute to better understanding of biological effects induced by TMPyP4 and provide a new insight into the complexity and implication for TMPyP4 based cancer therapy.

[Optimization of microemulsion containing vinpocetine and its physicochemical properties].

AIM: To prepare the microemulsion of vinpocetine in order to increase solubility and its in vitro transdermal delivery by using appropriate proportion of oil, surfactant (S), cosurfactant (CoS) and water. The formulation of proportion was optimized. The physicochemical properties and the skin irritation of the microemulsion was studied. METHODS: Pseudo-tertiary phase diagrams were prepared to obtain the concentration ratio of components of the microemulsion. Using simplex lattice method, the formulation of microemulsion was optimized and the physicochemical properties including pH, viscosity, refractive index, conductivity and particle size distribution were examined. The MTT method was applied to test the skin irritation of the microemulsion on the Hacat cell. RESULTS: The diagrams showed that the areas of O/W microemulsion increased with the increasing ratio of surfactant to cosurfactant (S/CoS). The predicted values from simplex lattice system were close to that of the experiment. The property of optimized microemulsion showed to be stable behavior and not irritant to Hacat cell. CONCLUSION: The drug solubility and in vitro perscutaneous permeation flux of the optimized microemulsion was improved significantly and the irritation study showed that optimized microemulsion was safe as an ideal transdermal delivery system.

Determination of the enthalpy of vaporization and prediction of surface tension for ionic liquid 1-alkyl-3-methylimidazolium propionate [C(n)mim][Pro](n = 4, 5, 6).

With the use of isothermogravimetrical analysis, the enthalpies of vaporization, Δ(g)lH(o)m(T(av)), at the average temperature, T(av) = 445.65 K, for the ionic liquids (ILs) 1-alkyl-3-methylimidazolium propionate [C(n)mim][Pro](n = 4, 5, 6) were determined. Using Verevkin's method, the difference of heat capacities between the vapor phase and the liquid phase, Δ(g)lC(p)(o)m, for [C(n)mim][Pro](n = 2, 3, 4, 5, 6), were calculated based on the statistical thermodynamics. Therefore, with the use of Δ(g)lC(p)(o)m, the values of Δ(g)lH(o)m(T(av)) were transformed into Δ(g)lH(o)m(298), 126.8, 130.3, and 136.5 for [C(n)mim][Pro](n = 4, 5, 6), respectively. In terms of the new scale of polarity for ILs, the order of the polarity of [C(n)mim][Pro](n = 2, 3, 4, 5, 6) was predicted, that is, the polarity decreases with increasing methylene. A new model of the relationship between the surface tension and the enthalpy of vaporization for aprotic ILs was put forward and used to predict the surface tension for [C(n)mim][Pro](n = 2, 3, 4, 5, 6) and others. The predicted surface tension for the ILs is in good agreement with the experimental one.

Genetic sequence characteristics of varicella-zoster virus epidemic strains in Changchun City from 2021 to 2022 / 中国生物制品学杂志

@#Objective To identify the genotypes and analyze the molecular characteristics of varicella-zoster virus(VZV)endemic strains in Changchun from 2021 to 2022.Methods A total of 59 patients with varicella or herpes zoster treated in China Japan Union Hospital of Jilin University in Changchun from 2021 to 2022 were selected as the research objects,which were identified for the serum VZV-specific antibodies by fluorescent antibody to membrane antigen(FAMA)method.The viral DNA was extracted from herpetic fluid of the confirmed patients,and multiple open reading frames(ORFs)including ORF1,ORF12,ORF16,ORF17,ORF21,ORF22,ORF37,and ORF54 of VZV were amplified by PCR,which was genotyped according to the single nucleotide polymorphisms(SNPs) of the ORFs.Furthermore,multiple ORFs,ORF38,ORF54 and ORF62 were identified by using restriction fragment length polymorphism(RFLP)to distinguish clinical strains and vaccine strains.Results 59 serum samples were positive,indicating that all the patients were infected with VZV.Among the 59 clinical samples,4 samples were completely matched with Clade 2 genotype,4 samples showed 2 SNPs,including ORF1(SNP790)mutation of C→A and ORF12(SNP18 082)mutation of T→C,and 51 samples showed ORF12(SNP18 082)mutation of T→C.All 59 VZV clinical isolates were PstⅠ+BglⅠ+BglⅠ+SmaⅠ+SmaⅠ-,which was different from the vaccine strain PstⅠ-,which was different from the vaccine strain PstⅠ-BglⅠ-BglⅠ+SmaⅠ+SmaⅠ+.Conclusion All the VZV endemic strains in Changchun from 2021 to 2022 were Clade 2 genotype,and no vaccine strain was found to be pathogenic.

Pharmacotherapy approaches to antifungal prophylaxis.

INTRODUCTION: Invasive fungal infection (IFI) is a serious problem due to difficulties in early diagnosis and high mortality. Different approaches are adopted for the treatment and management of IFI, including prophylactic, empiric, preemptive and directed strategies. AREAS COVERED: This paper reviews the type of pharmacotherapy used for antifungal prophylaxis in infants with extremely low birth weights, pediatric patients with cardiac disease, preterm neonates, pediatric oncology patients, adult cancer patients with neutropenia, adult patients with hematologic malignancy, hematopoietic stem-cell transplantation recipients, organ transplant recipients, HIV-infected patients, immunosuppressed patients treated with moderate or high doses of corticosteroids, and patients with invasive fusariosis, candidemia, invasive candidiasis, systemic mycoses and immunocompromised patients. EXPERT OPINION: Azole drugs are the drugs most often used in cost-effective antifungal prophylaxis of patients with conditions such as immunodeficiency and cancer, which render them highly susceptible to IFI. Fluconazole is the most outstanding example. However, there are many azoles with different pharmacological characteristics that the physician can choose from. Echinocandins have favorable characteristics that make them useful for treating Candida infections. Antibodies, or their engineered derivatives directed against cell-wall polysaccharides and glycopeptides, and some protein epitopes of Candida albicans, appear to be a promising novel approach for prophylaxis against Candida infection and deserve further in-depth investigations.

A two-site monoclonal antibody immunochromatography assay for rapid detection of peanut allergen Ara h1 in Chinese imported and exported foods.

Food allergen labeling has not yet been implemented in China. Therefore, a gold immunochromatography assay (GICA) was developed using two monoclonal antibodies (mAb) against the peanut allergen Ara h1. The GICA was specific for standard peanut samples with a sensitivity of 10ng/ml. Peanut protein traces extracted from 124 food products imported and exported by China Customs were easily and rapidly detected by GICA. 68 food samples originally labeled as containing peanuts were positive for Ara h1 and 54 food samples labeled as not containing peanuts were negative for Ara h1, indicating that the labels from the manufacturers were accurate. However, 2 food samples labeled as not containing peanuts tested positive for Ara h1. The present GICA provides a fast, simple, semi-quantitative method for the determination of peanut allergens in foods. This detection system can be used to ensure the safety of food imported and exported by China Customs.

An unusual case of autoimmune hepatitis in a patient with adult-onset Still's disease.

Adult-onset Still's disease (AOSD) is an autoimmune disease characterized by spiking fever, evanescent rash, arthritis, serositis, and liver involvement. Although autoimmune hepatitis (AIH) has been reported in association with various autoimmune diseases, AOSD-associated AIH is rare. We herein report an unusual case of AIH in a patient with AOSD.