Therapy of Pseudoxanthoma Elasticum: Current Knowledge and Future Perspectives

Pseudoxanthoma elasticum (PXE) is a rare, genetic, metabolic disease with an estimated prevalence of between 1 per 25,000 and 56,000. Its main hallmarks are characteristic skin lesions, development of choroidal neovascularization, and early-onset arterial calcification accompanied by a severe reduction in quality-of-life. Underlying the pathology are recessively transmitted pathogenic variants of the ABCC6 gene, which results in a deficiency of ABCC6 protein. This results in reduced levels of peripheral pyrophosphate, a strong inhibitor of peripheral calcification, but also dysregulation of blood lipids. Although various treatment options have emerged during the last 20 years, many are either already outdated or not yet ready to be applied generally. Clinical physicians often are left stranded while patients suffer from the consequences of outdated therapies, or feel unrecognized by their attending doctors who may feel uncertain about using new therapeutic approaches or not even know about them. In this review, we summarize the broad spectrum of treatment options for PXE, focusing on currently available clinical options, the latest research and development, and future perspectives.

Defining matrix Gla protein expression in the Dunkin-Hartley guinea pig model of spontaneous osteoarthritis

Background Matrix Gla (γ-carboxyglutamate) protein (MGP) is considered a strong inhibitor of ectopic calcification, and it has been associated with OA severity, although not conclusively. We utilized male Dunkin-Hartley (DH) guinea pigs to investigate the expression of MGP throughout aging and disease pathogenesis in a spontaneous model. Method Twenty-five male DH guinea pigs were obtained and nurtured to several timepoints, and then randomly and equally divided by age into five subgroups (1-, 3-, 6-, 9-, and 12-months, with the 1-month group as the reference group). DH guinea pigs in each group were euthanized at the designated month-age and the left or right medial tibial plateaus cartilages were randomly excised. OA severity was described by modified Mankin Score (MMS) at microscopy (Safranin O/Fast Green stain). Proteomic evaluation using isobaric tags for relative and absolute quantification (iTRAQ) was performed to validate the age-related changes in the MGP profiles, and immunohistochemistry (IHC) methods were applied for semi-quantitative determination of MGP expression in articular cartilage. Results The histopathologic findings validated the increasing severity of cartilage degeneration with age in the DH guinea pigs. The MMS showed significant, stepwise (every adjacent comparison P < 0.05) disease progression with month-age. The iTRAQ indicated that MGP levels increased significantly with advancing age (P < 0.05), as supported by the IHC result (P < 0.05). Conclusion Increased expression of MGP in male DH guinea pigs was present throughout aging and disease progression and may be link to increased OA severity. Further studies are needed to investigate and confirm the association between MGP levels and OA severity.

Molecular Docking of Anti Helicobacter pylori Antibiotics and Proton Pump Inhibitor: A Single Center Survey

Helicobacter pylorus (H. pylori) is a deadly bacterium responsible for significant worldwide Gastric Cancer (GC) related mortality. The present study aimed to screen all the anti-microbial drugs used to eradicate H .pylori infection and to identify the most efficient drug by using computational methods through molecular docking analysis. The 3-D structure of protein chorismate synthase of H. pylori was downloaded from the Protein data bank (PDB) online browser. The x-ray crystallography structures of 13 common drugs used against H.pylori infection were also downloaded from the drug bank. We screened all 13 common drugs through molecular docking to know the most efficient binding interaction between the diverse ligand-protein complexes. The results were further compared with clinical survey data from the patients with diverse gastrointestinal H. pylori infected cases. Among the screened compounds, by in-silico approach we found that fluoroquinolone (FLRQ) and tetracycline (TET) manifested more significant interactions with chorismate synthase (CS) protein along with binding energies of -9.2 and -8.1 kcal/mole respectively. Further, the drugs were also corroborated with the survey data from patients with varied gastrointestinal disorders in our study. With this computational study, we could find FLRQ and TET may be the most efficient drug for H. pylori treatment, which can be tried in case of anti H. Pylori treatment failure due to resistance. Hence, effective inter-analysis between the experimental and computational approaches is crucial to build up a strong inhibitor.

Assessment of fitness and vector competence of a New Caledonia wMel Aedes aegypti strain before field-release

Background Biological control programs involving Wolbachia-infected Aedes aegypti are currently deployed in different epidemiological settings. New Caledonia (NC) is an ideal location for the implementation and evaluation of such a strategy as the only proven vector for dengue virus (DENV) is Ae. aegypti and dengue outbreaks frequency and severity are increasing. We report the generation of a NC Wolbachia-infected Ae. aegypti strain and the results of experiments to assess the vector competence and fitness of this strain for future implementation as a disease control strategy in Noumea, NC. Methods/principal findings The NC Wolbachia strain (NC-wMel) was obtained by backcrossing Australian AUS-wMel females with New Caledonian Wild-Type (NC-WT) males. Blocking of DENV, chikungunya (CHIKV), and Zika (ZIKV) viruses were evaluated via mosquito oral feeding experiments and intrathoracic DENV challenge. Significant reduction in infection rates were observed for NC-wMel Ae. aegypti compared to WT Ae. aegypti. No transmission was observed for NC-wMel Ae. aegypti. Maternal transmission, cytoplasmic incompatibility, fertility, fecundity, wing length, and insecticide resistance were also assessed in laboratory experiments. Ae. aegypti NC-wMel showed complete cytoplasmic incompatibility and a strong maternal transmission. Ae. aegypti NC-wMel fitness seemed to be reduced compared to NC-WT Ae. aegypti and AUS-wMel Ae. aegypti regarding fertility and fecundity. However further experiments are required to assess it accurately. Conclusions/significance Our results demonstrated that the NC-wMel Ae. aegypti strain is a strong inhibitor of DENV, CHIKV, and ZIKV infection and prevents transmission of infectious viral particles in mosquito saliva. Furthermore, our NC-wMel Ae. aegypti strain induces reproductive cytoplasmic incompatibility with minimal apparent fitness costs and high maternal transmission, supporting field-releases in Noumea, NC.

Mechanical Control of Cell Migration by the Metastasis Suppressor Tetraspanin CD82/KAI1

The plasma membrane is a key actor of cell migration. For instance, its tension controls persistent cell migration and cell surface caveolae integrity. Then, caveolae constituents such as caveolin-1 can initiate a mechanotransduction loop that involves actin- and focal adhesion-dependent control of the mechanosensor YAP to finely tune cell migration. Tetraspanin CD82 (also named KAI-1) is an integral membrane protein and a metastasis suppressor. Its expression is lost in many cancers including breast cancer. It is a strong inhibitor of cell migration by a little-known mechanism. We demonstrated here that CD82 controls persistent 2D migration of EGF-induced single cells, stress fibers and focal adhesion sizes and dynamics. Mechanistically, we found that CD82 regulates membrane tension, cell surface caveolae abundance and YAP nuclear translocation in a caveolin-1-dependent manner. Altogether, our data show that CD82 controls 2D cell migration using membrane-driven mechanics involving caveolin and the YAP pathway.

Comparative Pathogenesis, Genomics and Phylogeography of Mousepox

Ectromelia virus (ECTV), the causative agent of mousepox, has threatened laboratory mouse colonies worldwide for almost a century. Mousepox has been valuable for the understanding of poxvirus pathogenesis and immune evasion. Here, we have monitored in parallel the pathogenesis of nine ECTVs in BALB/cJ mice and report the full-length genome sequence of eight novel ECTV isolates or strains, including the first ECTV isolated from a field mouse, ECTV-MouKre. This approach allowed us to identify several genes, absent in strains attenuated through serial passages in culture, that may play a role in virulence and a set of putative genes that may be involved in enhancing viral growth in vitro. We identified a putative strong inhibitor of the host inflammatory response in ECTV-MouKre, an isolate that did not cause local foot swelling and developed a moderate virulence. Most of the ECTVs, except ECTV-Hampstead, encode a truncated version of the P4c protein that impairs the recruitment of virions into the A-type inclusion bodies, and our data suggest that P4c may play a role in viral dissemination and transmission. This is the first comprehensive report that sheds light into the phylogenetic and geographic relationship of the worldwide outbreak dynamics for the ECTV species.

Understanding and Efficiently Manipulating Environmental Stress Caused by Metal Ions to Improve Ethanol Fermentation

The inconsistent quality of molasses directly influences ethanol production, particularly due to contamination by metal ions that causes severe problems and reduces production efficiency. This research focused on calcium (Ca2+), potassium (K+), and magnesium (Mg2+) ions that are common in molasses. The key objective was to understand clearly the effect of ions on ethanol fermentation and Saccharomyces cerevisiae performance. Individual ions and ion mixtures were studied in sucrose solution and in molasses. The results showed that severe stress could be ordered as Ca2+>K+>Mg2+ and the adverse effect was greater when the ion concentration increased. Ca2+ was a strong inhibitor while trace amounts of Mg2+ produced a positive effect. To achieve the greatest efficiency in ethanol production using molasses in the substrate preparation, Ca2+ should not exceed 0.18% (w/w) prior to fermentation and the final sugar concentration should be 20–25% (w/v), as adjusting the addition of sucrose will result in a suitable yeast medium. Pretreatment and dilution were the best practices for ion removal, with Ca2+ being clearly decreased. Furthermore, determination of the composition and ion concentration in molasses are essential initial steps that must be routinely applied to ensure that the knowledge gained and the efficient techniques investigated can both be used to improve ethanol production.

Drug interactions in hospital prescriptions in Denmark: Prevalence and associations with adverse outcomes

Importance: While the beneficial effects of medications are numerous, drug-drug interactions may lead to adverse drug reactions that are preventable causes of morbidity and mortality. Objective: To quantify the prevalence of potential drug-drug interactions in drug prescriptions at Danish hospitals, estimate the risk of adverse outcomes associated with discouraged drug combinations, and highlight the patient types (defined by the primary diagnosis of the admission) that appear to be more affected. Design: Cross-sectional (descriptive part) and cohort study (adverse outcomes part). Setting: Hospital electronic health records from two Danish regions (approx. 2.5 million people) from January 2008 through June 2016. Participants: Inpatients receiving two or more medications during their admission. Exposure: Concomitant prescriptions of potentially interacting drugs as per the Danish Drug Interaction Database. Main outcome and measure: Descriptive part: prevalence of potential drug-drug interactions in general and discouraged drug pairs in particular during admissions. Adverse outcomes part: post-discharge all-cause mortality rate, readmission rate and length-of-stay. Results: Among 2,886,227 hospital admissions (945,475 patients; median age 62 years [IQR: 41-74]; 54% female; median number of drugs 7 [IQR: 4-11]), patients in 1,836,170 admissions were exposed to at least one potential drug-drug interaction (659,525 patients; median age 65 years [IQR: 49-77]; 54% female; median number of drugs 9 [IQR: 6-13]), and in 27,605 admissions to a discouraged drug pair (18,192 patients; median age 68 years [IQR: 58-77]; female 46%; median number of drugs 16 [IQR: 11-22]). Meropenem-valproic acid (HR: 1.5, 95% CI: 1.1-1.9), domperidone-fluconazole (HR: 2.5, 95% CI: 2.1-3.1), imipramine-terbinafine (HR: 3.8, 95% CI: 1.2-12), agomelatine-ciprofloxacin (HR: 2.6, 95% CI: 1.3-5.5), clarithromycin-quetiapine (HR: 1.7, 95% CI: 1.1-2.7), and piroxicam-warfarin (HR: 3.4, 95% CI: 1-11.4) were associated with elevated mortality. Confidence interval bounds of pairs associated with readmission were close to 1; length-of-stay results were inconclusive. Conclusions and Relevance: Well-described potential drug-drug interactions are still missed and alerts at point of prescription may reduce the risk of harming patients; prescribing clinicians should be alert when using strong inhibitor/inducer drugs (i.e. clarithromycin, valproic acid, terbinafine) and prevalent anticoagulants (i.e. warfarin and NSAIDs) due to their great potential for dangerous interactions. The most prominent CYP isoenzyme involved in mortality and readmission rates was 3A4.

Assessment of Chronic Supraphysiological Glucocorticoid Dosing Needs of Patients With Adrenal Insufficiency

Background: Patients who require higher than replacement glucocorticoid doses to avoid symptoms of adrenal insufficiency (AI) may have inadequate adherence or abnormal drug absorption or metabolism. The goal of this study was to identify why excessive glucocorticoid doses were needed in patients with ongoing AI symptoms. Methods: We performed pharmacokinetic (PK) analysis of glucocorticoid plasma concentrations (prednisone, prednisolone and dexamethasone measured by tandem mass spectrometry, Mayo Laboratories, and cortisol by immunoassay, Clinical Center) in 8 AI patients after weight-based oral hydrocortisone (HC) dose (1), IV HC 20 mg and /or prednisone 5 mg PO. The time (Tmax) to maximal plasma concentrations (Cmax), time to a 50% decrease in concentration (T1/2), elimination rate (ER) and area under the concentration curve (AUC) were determined using MATLAB and SimBiology, and compared to literature reference ranges (RR) (1,2). Results: Patients included one man; six had secondary AI due to previous supraphysiologic hydrocortisone or prednisone treatment and two had primary autoimmune AI. One of the latter was appropriately replaced with thyroid hormone. No patient was taking any medication known to be a strong inhibitor or inducer of CYP3A4 and none were taking oral estrogens. To study the potential contribution of intestinal absorption to abnormal pharmacokinetics, serum cortisol values were compared to expected values at 3.5 or 4 hours after a weight-based oral dose of HC in eight patients (1); 7 patients had values at the 50 - 80th centile of expected values. The eighth patient’s cortisol level at 4 hours after 5 mg HC was 30.3 nmol/L, below the 10th centile. She then underwent the same sampling with a 15 mg dose, with values also at the 10th centile. To uncover any discrepancy between PK oral and IV HC administration, four patients, including the patient with a low 4 hour value (LowHC4h) underwent sampling after 20 mg hydrocortisone, IV (2). Tmax and Cmax were within the RR in all four patients, while one patient had a faster T1/2 but an AUC similar to others. The LowHC4h patient had a dexamethasone level 8 hours after a 1 mg dose that was also within the RR and was maintained on dexamethasone. All others were eventually able to be weaned to a conventional glucocorticoid replacement dose. Conclusions: Evaluation of oral and IV HC PK may be useful in patients suspected of having abnormal absorption of oral glucocorticoids. Ref: 1. Mah PM et al. Clin Endo 61:367,20042. Thomson AH et al. Clin Endo 66:789,2007

Applying Nanotechnology in the Synthesis of Benzimidazole Derivatives: A Pharmacological Approach

Benzimidazoles are classified as a category of heterocyclic compounds. They possess an essential structural feature of 6-membered benzene fused to 5-membered imidazole moiety. Molecules having benzimidazole motifs confirmed promising utility in organic and scientific studies. Various pharmacological residences were explored with a strong inhibitor of numerous enzymes. They are concerned with being efficient antidiabetic, anticancer, antimicrobial, antiparasitic, analgesics, antiviral and antihistaminic agents. Moreover, they can be utilized in cardiovascular disease, neurology, endocrinology, and ophthalmology. The multiple activities for benzimidazole compounds have emerged due to their stability, bioavailability, and good-sized organic activity. Modifications of some organic polymers were carried out by utilizing different azole moieties. This review is devoted to mention some of the various current techniques for the synthesis of benzimidazole derivatives and their pharmacological residences with representing numerous derivatives.