In vitro and in silico assessment of the developability of a designed monoclonal antibody library

Despite major advances in antibody discovery technologies, the successful development of monoclonal antibodies (mAbs) into effective therapeutic and diagnostic agents can often be impeded by developability liabilities, such as poor expression, low solubility, high viscosity and aggregation. Therefore, strategies to predict at the early phases of antibody development the risk of late-stage failure of antibody candidates are highly valuable. In this work, we employ the in silico solubility predictor CamSol to design a library of 17 variants of a humanized mAb predicted to span a broad range of solubility values, and we examine their developability potential with a battery of commonly used in vitro and in silico assays. Our results demonstrate the ability of CamSol to rationally enhance mAb developability, and provide a quantitative comparison of in vitro developability measurements with each other and with more resource-intensive solubility measurements, as well as with in silico predictors that offer a potentially faster and cheaper alternative. We observed a strong correlation between predicted and experimentally determined solubility values, as well as with measurements obtained using a panel of in vitro developability assays that probe non-specific interactions. These results indicate that computational methods have the potential to reduce or eliminate the need of carrying out laborious in vitro quality controls for large numbers of lead candidates. Overall, our study provides support to the emerging view that the implementation of in silico tools in antibody discovery campaigns can ensure rapid and early selection of antibodies with optimal developability potential.     

Genome-Scale Model of Rhizopus microsporus: Metabolic integration of a fungal holobiont with its bacterial and viral endosymbionts

Rhizopus microsporus often lives in association with bacterial and viral symbionts that alter its biology. This fungal model represents an example of the complex interactions established among diverse organisms in functional holobionts. We constructed a Genome-Scale Model (GSM) of the fungal-bacterial-viral holobiont (iHol). We employed a constraint-based method to calculate the metabolic fluxes to decipher the metabolic interactions of the symbionts with their host. Our computational analyses of iHol simulate the holobiont's growth and the production of the toxin rhizoxin. Analyses of the calculated fluxes between R. microsporus in symbiotic (iHol) versus asymbiotic conditions suggest that changes in the lipid and nucleotide metabolism of the host are necessary for the functionality of the holobiont. Glycerol plays a pivotal role in the fungal-bacterial metabolic interaction, as its production does not compromise fungal growth, and Mycetohabitans bacteria can efficiently consume it. Narnavirus RmNV-20S and RmNV-23S affected the nucleotide metabolism without impacting the fungal-bacterial symbiosis. Our analyses highlighted the metabolic stability of Mycetohabitans throughout its co-evolution with the fungal host. We also predicted changes in reactions of the bacterial metabolism required for the active production of rhizoxin. This iHol is the first GSM of a fungal holobiont.     

Praziquantel-encapsulated niosomes against Schistosoma mansoni with reduced sensitivity to praziquantel

Introduction: Praziquantel (PZQ) is the only commercially available drug for schistosomiasis. The current shortage of alternative effective drugs and the lack of successful preventive measures enhance its value. The increase in the prevalence of PZQ resistance under sustained drug pressure is, therefore, an upcoming issue.Objective: To overcome the tolerance to PZQ using nanotechnology after laboratory induction of a Schistosoma mansoni isolate with reduced sensitivity to the drug during the intramolluscan phase.Materials and methods: Shedding snails were treated with PZQ doses of 200 mg/kg twice/ week followed by an interval of one week and then repeated twice in the same manner. The success of inducing reduced sensitivity was confirmed in vitro via the reduction of cercarial response to PZQ regarding their swimming activity and death percentage at different examination times.Results: Oral treatment with a single PZQ dose of 500 mg/kg in mice infected with cercariae with reduced sensitivity to PZQ revealed a non-significant reduction (35.1%) of total worm burden compared to non-treated control mice. Orally inoculated PZQ- encapsulated niosomes against S. mansoni with reduced sensitivity to PZQ successfully regained the pathogen’s sensitivity to PZQ as evidenced by measuring different parameters in comparison to the non-treated infected animals with parasites with reduced sensitivity to PZQ. The mean total worm load was 1.33 ± 0.52 with a statistically significant reduction of 94.09% and complete eradication of male worms. We obtained a remarkable increase in the percentage reduction of tissue egg counts in the liver and intestine (97.68% and 98.56%, respectively) associated with a massive increase in dead eggs and the complete absence of immature stages.Conclusion: PZQ-encapsulated niosomes restored the drug sensitivity against laboratory- induced S. mansoni adult worms with reduced sensitivity to PZQ.     

Comparative conventional preventive strategies for insect pest of okra

The use of natural substances for pest control in agriculture is economically viable. It benefits both the human being and the environment due to its low persistence and toxicity. Therefore, the biopesticidal potential of three- plants-derived extracts (clove [Syzygium aromaticum], Hing [Hing (Asafetida)], and Wood Ash [Eucalyptus globulas]) was evaluated against different ' 'insect's pests on five okra varieties. All the treatments were sprayed at two stages, 1st before flowering and 2nd at the fruit-bearing stage. The results of the 24 h pre-spray revealed that the mean density of Aphis gossypii, Erias insulana, and Bemisia tabaci were significantly lower on a Shehzadi variety. However, among the treatments mean density of the A. gossypii and E. insulana after 1st and 2nd treatments were substantially more bass with E. globulas. Moreover, the Mean density of aphids was significantly lower after 72 h and 1-week time intervals. Furthermore, after 1st and 2nd treatments, the B. tabaci was considerably lower with hing on Shehzadi variety. It was found in the present study that the yield of five okra varieties was affected significantly by the application of the three treatments-pesticides. Among the various treatments, the application with E. globulas recorded a considerably higher crop yield. Therefore, clove, hing and E. globulas could be effective as alternative pest management methods. Furthermore, biopesticides generally are encouraged since they can proffer the solution of controlling insect pests without any environmental concern.     

Potential Role of circRNA-HIPK3/microRNA-124a Crosstalk in the Pathogenesis of Rheumatoid Arthritis

Background:Circular RNA-HIPK3 (CircHIPK3) has been shown to be aberrantly expressed in a variety of diseases, contributing to disease initiation and progression. The aim of the present study is to investigate the role of the circHIPK3 RNA/microRNA-124a interaction in the pathogenesis of rheumatoid arthritis (RA).Methods:This study included 79 RA patients and 30 control individuals. The patients involved were classified according to the disease activity score (DAS28) into mild (24 patients), moderate (24 patients), and severe (31 patients). Serum samples were collected to estimate the relative gene expression of circHIPK3 RNA and its target gene microRNA-124a by quantitative real time-PCR. Moreover, ELISA was used to detect the serum levels of monocyte chemoattractant protein-1 (MCP-1). Routine laboratory estimation of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and rheumatoid factor (RF) was also done.Results:In all grades of RA groups, there was a significantly substantial elevation of circHIPK3 RNA gene expression, with subsequent downregulation of miRNA-124a when compared to the control group. CircHIPK3 and microRNA-124a expression have been established to be inversely linked. Also, estimation of serum levels of MCP-1, ESR, CRP, and RF exhibited a significant increase in all grades of RA as compared to the control group.Conclusion:CircHIPK3 and microRNA-124a might be regarded as key players in the pathogenesis of RA. The cross-talk between them appears to be responsible for inducing joint inflammation by increasing MCP-1 production. Targeting circHIPK3 and microRNA-124a, and their downstream adaptor molecules, poses a new challenge for RA therapy.Key Words: Circhipk3, Circular RNA, Microrna-124a, Rheumatoid arthritis     

Health Service Utilization among Syrian Refugees with Chronic Health Conditions in Jordan

IntroductionThe influx of Syrian refugees into Jordan presents an immense burden to the Jordanian health system, particularly in treating chronic health conditions. This study was undertaken to assess utilization of health services for chronic health conditions among Syrian refugees in non-camp settings.MethodsA survey of Syrian refugees in Jordan was undertaken in June 2014 to characterize health seeking behaviors and issues related to accessing care for hypertension, diabetes, cardiovascular diseases, chronic respiratory diseases, and arthritis. A cluster design with probability proportional to size sampling was used to attain a nationally representative sample of 1550 non-camp Syrian refugee households.ResultsOf 1363 cases with a chronic health condition diagnosis, 84.7% had received care in Jordan. Public facilities faced a heavy burden serving over half (53.9%) of care-seekers; the remainder received care in the private (29.6%) and NGO/charity (16.6%) sectors. Individuals with non-communicable diseases (NCDs) in the central region of Jordan and with arthritis had the lowest rates of care-seeking when compared to other regions and conditions. Overall, 31.6% of care-seekers had an out-of-pocket payment for the most recent care-seeking event which averaged 18.8 USD (median = 0 USD), excluding cost of medications.DiscussionForced displacement presents major challenges to those with NCDs, which have the potential to seriously impact both the quality of life and life expectancy amongst refugees. NCD patterns among Syrian refugees indicate the importance of continuing support to public sector services in Jordan to adequately meet expanding needs and ensure appropriate prevention and control of priority NCDs.     

Electron microscope studies on the development of the external zone of the mouse median eminence

Summary The development of the external zone of the median eminence of the mouse was studied in the electron microscope. The examination follows the development of the embryo from the 15th day of the gestation period and the juvenile growth until 24 days of age.Single terminals of the tubero-infundibular neurons of the external zone were found to extend to the outer basement membrane of the perivascular space of the portal primary capillary plexus in the 16 day-old embryo. In the 18 day-old embryo a narrow external zone has developed. Organization of the external zone into the adult pattern is accomplished at the age of three to four weeks. Small agranular as well as large granular vesicles are present in the tubero-infundibular nerve terminals even in the 16 day-old embryo.Changes in the organization of the nerve endings along the outer perivascular basement membrane in relation to the ependymal vascular feet were considered.