Leishmanicidal Metabolites from Cochliobolus sp., an Endophytic Fungus Isolated from Piptadenia adiantoides (Fabaceae)

Protozoan parasites belonging to genera Leishmania and Trypanosoma are the etiological agents of severe neglected tropical diseases (NTDs) that cause enormous social and economic impact in many countries of tropical and sub-tropical areas of the world. In our screening program for new drug leads from natural sources, we found that the crude extract of the endophytic fungus Cochliobolus sp. (UFMGCB-555) could kill 90% of the amastigote-like forms of Leishmania amazonensis and inhibit by 100% Ellman's reagent reduction in the trypanothione reductase (TryR) assay, when tested at 20 µg mL−1. UFMGCB-555 was isolated from the plant Piptadenia adiantoides J.F. Macbr (Fabaceae) and identified based on the sequence of the internally transcribed spacer (ITS) regions of its ribosomal DNA. The chromatographic fractionation of the extract was guided by the TryR assay and resulted in the isolation of cochlioquinone A and isocochlioquinone A. Both compounds were active in the assay with L. amazonensis, disclosing EC50 values (effective concentrations required to kill 50% of the parasite) of 1.7 µM (95% confidence interval = 1.6 to 1.9 µM) and 4.1 µM (95% confidence interval = 3.6 to 4.7 µM), respectively. These compounds were not active against three human cancer cell lines (MCF-7, TK-10, and UACC-62), indicating some degree of selectivity towards the parasites. These results suggest that cochlioquinones are attractive lead compounds that deserve further investigation aiming at developing new drugs to treat leishmaniasis. The findings also reinforce the role of endophytic fungi as an important source of compounds with potential to enter the pipeline for drug development against NTDs

Meibomian Gland Probing Stimulates a Proliferative Epithelial Response Resulting in Duct Regeneration

Steven L Maskin,&ast; Claire Toland&ast; Dry Eye and Cornea Treatment Center, Tampa, FL, USA&ast;These authors contributed equally to this workCorrespondence: Steven L Maskin, Dry Eye and Cornea Treatment Center, 3001 West Swann Ave, Tampa, FL, 33609, USA, Tel +1 813 875 0000, Email [email protected]: To demonstrate that the meibomian gland ductal basement membrane and basal epithelial cell layer are in continuity with and may derive from lid margin orifice-associated rete ridge epithelial/basement membrane structures (OARREBS) and to characterize changes in the distal duct microanatomy after meibomian gland probing (MGP) using in vivo confocal microscopy (IVCM).Patients and Methods: Pre/post-MGP IVCM examinations were performed on upper lids. Thirty-six identical glands from 20 lids of 16 patients (49.24 ± 17.11 y/o with 13:3 F:M) were identified, analyzed, and compared to control cases. Statistical analyses were performed using ImageJ software and IBM SPSS version 27. All MGPs were performed within 12 weeks of the initial examination. Post-MGP follow-up exams occurred at 5.03 ± 4.48 months.Results: Post-MGP images showed more superficially organized OARREBS with accelerated and more superficial basement membrane formation, and an average increase of 32.2%, 25.4%, 32.04%, 77.7%, and 81.3% in duct wall epithelial cell layers (DWECL) (p < 0.001, compared to control (CTC) p < 0.001), distal duct wall thickness (DWT) (p < 0.001, CTC p < 0.001), proximal DWT (p < 0.001, CTC p < 0.001), distal lumen area (p < 0.001, CTC p = 0.037), and proximal lumen area (p < 0.001, CTC p = 0.007), respectively. The increase in the distal DWT and lumen area correlated with the months of follow-up (p = 0.004 and p = 0.010, respectively). Immediate post-MGP imaging revealed the probe track confined to the ductal epithelial compartment.Conclusion: MGP appears to stimulate a proliferative epithelial response characterized by an accelerated more superficial formation of ductal basement membrane with increased DWECL as well as DWT and lumen area at two separate duct foci. These findings suggest activation of lid margin meibomian gland precursor cells and confirm that MGP stimulates an epithelial regenerative phenomenon, not a fibrotic one.Keywords: meibomian gland probing, rete ridges, dry eye, meibomian gland dysfunction, precursor cells, meibomian gland regeneratio

Harmonizing international trials of early goal-directed resuscitation for severe sepsis and septic shock: methodology of ProCESS, ARISE, and ProMISe

ProCESS/ARISE/ProMISe methodology writing committee members: Sandra L. Peake and Patricia Williams for the University of Adelaide, South Australia.PURPOSE To describe and compare the design of three independent but collaborating multicenter trials of early goal-directed resuscitation for severe sepsis and septic shock. METHODS We reviewed the three current trials, one each in the USA (ProCESS: protocolized care for early septic shock), Australasia (ARISE: Australasian resuscitation in sepsis evaluation), and the UK (ProMISe: protocolised management in sepsis). We used the 2010 CONSORT (consolidated standards of reporting trials) statement and the 2008 CONSORT extension for trials assessing non-pharmacologic treatments to describe and compare the underlying rationale, commonalities, and differences. RESULTS All three trials conform to CONSORT guidelines, address the same fundamental questions, and share key design elements. Each trial is a patient-level, equal-randomized, parallel-group superiority trial that seeks to enroll emergency department patients with inclusion criteria that are consistent with the original early goal-directed therapy (EGDT) trial (suspected or confirmed infection, two or more systemic inflammatory response syndrome criteria, and refractory hypotension or elevated lactate), is powered to detect a 6–8 % absolute mortality reduction (hospital or 90-day), and uses trained teams to deliver EGDT. Design differences appear to primarily be driven by between-country variation in health care context. The main difference between the trials is the inclusion of a third, alternative resuscitation strategy arm in ProCESS. CONCLUSIONS Harmonization of study design and methods between severe sepsis trials is feasible and may facilitate pooling of data on completion of the trials.The ProCESS/ARISE/ProMISe Methodology Writing Committe