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Cysteine proteases, also known as thiol proteases, are a class of nucleophilic proteolytic enzymes containing cysteine residues in the enzymatic domain. These proteases generally play a pivotal role in many biological reactions, such as catabolic functions and protein processing, in all living organisms. They specifically take part in many important biological processes, especially in the absorption of nutrients, invasion, virulence, and immune evasion of parasitic organisms from unicellular protozoa to multicellular helminths. They can also be used as parasite diagnostic antigens and targets for gene modification and chemotherapy, as well as vaccine candidates, due to their species and even life-cycle stage specificity. This article highlights current knowledge on parasitic cysteine protease types, biological functions, and their applications in immunodiagnosis and chemotherapy.

The human enteric bacterial pathogen Salmonella enterica causes approximately 1.35 million cases of food borne illnesses annually in the United States. Of these salmonellosis cases, almost half are derived from the consumption of fresh, raw produce. Although epiphytic S . enterica populations naturally decline in the phyllosphere, a subset of phytophagous insects have recently been identified as biological multipliers, consequently facilitating the growth of bacterial populations. We investigated whether tomato leaves with macroscopic feeding damage, caused by infestation of adult Western flower thrips ( Frankliniella occidentalis ), support higher S . enterica populations. To explore this hypothesis, we assessed S . enterica populations in response to thrips feeding by varying insect density, plant age, and the gender of the insect. As a reference control, direct leaf damage analogous to thrips feeding was also evaluated using directed, hydraulic pressure. In a supplementary set series of experiments, groups of F . occidentalis infested tomato plants were later inoculated with S . enterica to determine how prior insect infestation might influence bacterial survival and persistence. Following an infestation period, leaves visibly damaged by adult F . occidentalis supported significantly higher S . enterica populations and resulted in greater amounts of electrolyte leakage (measured as electrical conductivity) than leaves lacking visible feeding damage. Plant age did not significantly influence S . enterica populations or estimates of electrolyte leakage, independent of initial infestation. Additionally, the gender of the insect did not uniquely influence S . enterica population dynamics. Finally, applications of aggressive water bombardment resulted in more electrolyte leakage than leaves damaged by F . occidentalis , yet supported comparable S . enterica populations. Together, this study indicates that F . occidentalis feeding is one of the many potential biological mechanisms creating a more habitable environment for S . enterica .

Trichomonas tenax is a flagellated protozoan that inhabits the human and canine oral cavity in patients with poor oral hygiene and periodontal disease. The loop-mediated isothermal amplification (LAMP) assay could provide clinicians with a quick, cheap and reliable diagnostic test used for the detection of T. tenax in various settings. In this study, we aimed to develop a LAMP assay that can detect T. tenax with high sensitivity and specificity. A set of LAMP primers were specifically designed to detect the ITS and 5.8S rRNA gene of T. tenax. The newly developed LAMP assay was 1000 times more sensitive than conventional PCR. The limit of detection of the LAMP assay was 10 fg of genomic DNA, or 0.2–1 cell. Moreover, the LAMP assay was specific, resulting in no cross-reaction even with a closely related protozoan T. vaginalis or other microorganisms (Streptococcus pyogenes, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, and Candida albicans) used. The present LAMP assay can be performed directly without prior DNA extraction, making the assay an easy, fast, cheap, specific and sensitive diagnostic tool for the detection of T. tenax at the point-of-care of both medical and veterinary clinics in developed and developing countries.

Trichomonas tenax is a flagellated protozoan parasite found in the oral cavities of humans and animals and has been associated with periodontal disease, the most prevalent inflammatory disease affecting them all. Studies have shown that T. tenax can cause damage to mammalian cells and secretes virulent proteins, such as cysteine. It is presently considered zoonotic. Despite the few studies that have been done, the pathogenicity of this oral protozoan is still not fully understood. A database search was performed in July 2022 using PubMed and Google Scholar to retrieve data eligible for this study. PRISMA-ScR guidelines were followed to conduct this scoping review. A total of 321 articles were found with 87 included in this review after applying the exclusion criteria. Due to its increasing prevalence worldwide in both humans and dogs, detecting and elucidating the pathogenicity of this parasite is paramount for effective global control and prevention of periodontal disease. However, there is a paucity in the literature on this neglected zoonotic trichomonad, which is in large contrast to the closely related human pathogen T. vaginalis. Here, we comprehensively review the history, morphology and reproduction, host, prevalence, diagnosis, pathogenicity, control, and prevention of T. tenax. Hopefully, this article will call attention to both medical and veterinary professionals as well as epidemiologists on this most neglected and zoonotic protozoan. More epidemiological and clinical studies need to be conducted on T. tenax to gain a better understanding of its pathogenicity, to increase the chances of developing effective drugs to aid in the control of this oral parasite, and reduce the spread of periodontal disease worldwide.

In the original publication [...]

Trichomonas tenax is a flagellated protozoan that inhabits the human and canine oral cavity in patients with poor oral hygiene and periodontal disease. The loop-mediated isothermal amplification (LAMP) assay could provide clinicians with a quick, cheap and reliable diagnostic test used for the detection of T. tenax in various settings. In this study, we aimed to develop a LAMP assay that can detect T. tenax with high sensitivity and specificity. A set of LAMP primers were specifically designed to detect the ITS and 5.8S rRNA gene of T. tenax. The newly developed LAMP assay was 1000 times more sensitive than conventional PCR. The limit of detection of the LAMP assay was 10 fg of genomic DNA, or 0.2–1 cell. Moreover, the LAMP assay was specific, resulting in no cross-reaction even with a closely related protozoan T. vaginalis or other microorganisms (Streptococcus pyogenes, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, and Candida albicans) used. The present LAMP assay can be performed directly without prior DNA extraction, making the assay an easy, fast, cheap, specific and sensitive diagnostic tool for the detection of T. tenax at the point-of-care of both medical and veterinary clinics in developed and developing countries.

Time and again, Murphy has turned his back on small business owners in favor of big out-of-town businesses. The Fifth/Forbes corridor project is the latest example (\"A Deal for Fifth/Forbes,\" Oct. 3). Ignoring how much of this was done in private, there is no evidence that I've seen that the people of Pittsburgh are clamoring for the businesses Murphy wants to bring in. Instead of rewarding the businesses that have been the lifeblood of Downtown for so long, Murphy wants to relegate them to the outer edges and replace them with the flavor of the week. The question shouldn't be whether Pittsburgh can put together an attractive financial package to lure the retailer Nordstrom. The question should be, do the people of Pittsburgh want a Nordstrom store?

Since the 1970s, the ocean has absorbed almost all of the additional energy in the Earth system due to greenhouse warming. However, sparse observations limit our knowledge of where ocean heat uptake (OHU) has occurred and where this heat is stored today. Here, we equilibrate a reanalysis-forced ocean-sea ice model, using a spin-up that improves on earlier approaches, to investigate recent OHU trends basin-by-basin and associated separately with surface wind trends, thermodynamic properties (temperature, humidity and radiation) or both. Wind and thermodynamic changes each explain ~ 50% of global OHU, while Southern Ocean forcing trends can account for almost all of the global OHU. This OHU is enabled by cool sea surface temperatures and sensible heat gain when atmospheric thermodynamic properties are held fixed, while downward longwave radiation dominates when winds are fixed. These results address long-standing limitations in multidecadal ocean-sea ice model simulations to reconcile estimates of OHU, transport and storage. This study improves on limitations of the most commonly used spin-up approach for ocean-sea ice models. The authors find that, over the last 50 years, atmospheric changes over the Southern Ocean have driven almost all of the global ocean heat uptake.

Detailed computational and structural analysis of a large data set of non-peptidic macrocycles revealed particular functional groups, substituents and molecular properties that are critical for dictating cellular permeability. Macrocycles are of increasing interest as chemical probes and drugs for intractable targets like protein–protein interactions, but the determinants of their cell permeability and oral absorption are poorly understood. To enable rational design of cell-permeable macrocycles, we generated an extensive data set under consistent experimental conditions for more than 200 non-peptidic, de novo –designed macrocycles from the Broad Institute's diversity-oriented screening collection. This revealed how specific functional groups, substituents and molecular properties impact cell permeability. Analysis of energy-minimized structures for stereo- and regioisomeric sets provided fundamental insight into how dynamic, intramolecular interactions in the 3D conformations of macrocycles may be linked to physicochemical properties and permeability. Combined use of quantitative structure–permeability modeling and the procedure for conformational analysis now, for the first time, provides chemists with a rational approach to design cell-permeable non-peptidic macrocycles with potential for oral absorption.