Non-Systemic Drugs: A Critical Review

Enteric viral infection in human and animal.

Protein crystallography has the potential to accelerate drug discovery greatly. High-resolution structures of membrane proteins of pharmaceutical interest open new perspectives in drug design. Recent structural data obtained for cyclooxygenases, monoamine oxidase, squalene cyclase, rhodopsine, porins, aquaporins, and ABC transporters are presented and briefly discussed.

This observational study was conducted to evaluate the effect of transition cow health on pregnancy per artificial insemination (P/AI) and pregnancy loss (PL) in cows submitted to a Double-Ovsynch protocol (DO) for first service. Lactating Holstein cows (n = 15,041) from one commercial dairy farm in northern Germany between January 2015 to December 2021 were enrolled into a modified Double-Ovsynch protocol (GnRH, 7 d later PGF2α, 3 d later GnRH, 7 d later GnRH, 7 d later PGF2α, 24 h later PGF2α, 32 h later GnRH, and 16 to 18 h later timed artificial insemination) for first service at 72 ± 3 d in milk. Pregnancy was diagnosed at 32 and 60 d post-AI via transrectal ultrasonography. Pregnancy loss was defined as the proportion of cows diagnosed pregnant 32 d post-artificial insemination that were diagnosed nonpregnant 60 d post-artificial insemination. Health-related events (i.e., milk fever [MF], hyperketonemia [KET], retained fetal membranes [RFM], metritis, mastitis, left displaced abomasum [LDA]) were assessed by farm personnel using standard operating procedures. Multivariable logistic regression was used for testing potential associations between transition cow health event occurrence and outcome variables, including P/AI and PL. Three separate models were built for cows in first lactation, second lactation, and ≥third lactation. Overall, 20.0% (885/4,430), 34.9% (1,391/3,989), and 53.9% (3,570/6,622) of cows had at least one transition cow health event for first, second, and ≥third lactations, respectively. The most prevalent transition cow health event for first-lactation cows was metritis (10.7%; [473/4,430]), whereas second-lactation cows suffered mostly from mastitis (16.6%; [664/3,989] and KET (16.6%; [661/3,989]), and cows with ≥third lactations were mostly affected by KET (33.2%; [2,198/6,622]). We observed a negative association between inflammatory disorders (i.e., RFM, metritis, mastitis) and P/AI in all cows irrespective of parity. Metabolic disorders (i.e., MF, KET, LDA) were negatively associated with P/AI only in multiparous cows. Irrespective of parity, only uterine diseases (i.e., RFM, metritis) were significantly associated with PL. These results show that enrolling cows into a fertility protocol, such as DO, cannot overcome the carryover effects of inflammatory and metabolic disorders on P/AI and PL and highlight the importance of optimizing transition cow health as a prerequisite for achieving high fertility in a DO protocol.

Harnessing the antibacterial and immunological properties of mucosal-associated invariant T cells in the development of novel oral vaccines against enteric infections

Uric acid can be generated in the gastrointestinal (GI) tract from the breakdown of nucleotides ingested in the diet or from purines released from host cells as a result of pathogen-induced cell damage. Xanthine oxidase (XO) is the enzyme that converts hypoxanthine or xanthine into uric acid, a reaction that also generates hydrogen peroxide. It has been assumed that the product of XO responsible for the pro-inflammatory effects of this enzyme is hydrogen peroxide. Recent literature on uric acid, however, has indicated that uric acid itself may have biological effects. We tested whether uric acid itself has detectable pro-inflammatory effects using an in vivo model using ligated rabbit intestinal segments (“loops”) as well as in vitro assays using cultured cells. Addition of exogenous uric acid increased the influx of heterophils into rabbit intestinal loops, as measured by myeloperoxidase activity. In addition, white blood cells adhered avidly to uric acid crystals, forming large aggregates of cells. Uric acid acts as a leukocyte chemoattractant in the GI tract. The role of uric acid in enteric infections and in non-infectious disorders of the GI tract deserves more attention.

The identification of modules in protein structures has major relevance in structural biology, with consequences in protein stability and functional classification, adding new perspectives in drug design. In this work, we present the comparison between a topological (spectral clustering) and a geometrical (k-means) approach to module identification, in the frame of a multiscale analysis of the protein architecture principles. The global consistency of an adjacency matrix based technique (spectral clustering) and a method based on full rank geometrical information (k-means) give a proof-of-concept of the relevance of protein contact networks in structure determination. The peculiar "small-world" character of protein contact graphs is established as well, pointing to average shortest path as a mesoscopic crucial variable to maximize the efficiency of within-molecule signal transmission. The specific nature of protein architecture indicates topological approach as the most proper one to highlight protein functional domains, and two new representations linking sequence and topological role of aminoacids are demonstrated to be of use for protein structural analysis. Here we present a case study regarding azurin, a small copper protein implied in the Pseudomonas aeruginosa respiratory chain. Its pocket molecular shape and its electron transfer function have challenged the method, highlighting its potentiality to catch jointly the structure and function features of protein structures through their decomposition into modules.

Microbes in Gastrointestinal Health and Disease

Sunflower trypsin inhibitor-1 (SFTI-1) and Momordica cochinchinensis trypsin inhibitor-II (MCoTI-II) are potent protease inhibitors comprising a cyclic backbone. Elucidation of structure-activity relationships for SFTI-1 and MCoTI-II was used to design inhibitors with enhanced inhibitory activity. An analog of MCoTI-II is one of the most potent inhibitors of matriptase. These results provide a solid basis for the design of selective peptide inhibitors of matriptase with therapeutic potential. The type II transmembrane serine protease matriptase is a key activator of multiple signaling pathways associated with cell proliferation and modification of the extracellular matrix. Deregulated matriptase activity correlates with a number of diseases, including cancer and hence highly selective matriptase inhibitors may have therapeutic potential. The plant-derived cyclic peptide, sunflower trypsin inhibitor-1 (SFTI-1), is a promising drug scaffold with potent matriptase inhibitory activity. In the current study we have analyzed the structure-activity relationships of SFTI-1 and Momordica cochinchinensis trypsin inhibitor-II (MCoTI-II), a structurally divergent trypsin inhibitor from Momordica cochinchinensis that also contains a cyclic backbone. We show that MCoTI-II is a significantly more potent matriptase inhibitor than SFTI-1 and that all alanine mutants of both peptides, generated using positional scanning mutagenesis, have decreased trypsin affinity, whereas several mutations either maintain or result in enhanced matriptase inhibitory activity. These intriguing results were used to design one of the most potent matriptase inhibitors known to date with a 290 pm equilibrium dissociation constant, and provide the first indication on how to modulate affinity for matriptase over trypsin in cyclic peptides. This information might be useful for the design of more selective and therapeutically relevant inhibitors of matriptase.

Gentamicin sulphate is an antibiotic belonging to the aminoglycosides and to class III of the Biopharmaceutical Classification System (BCS). Gentamicin sulphate is highly water soluble, but has very low intestinal permeability. The wide use is because of its broad spectrum of activity. In the current study, the suitability of administering gentamicin sulphate orally for local action against susceptible gastrointestinal tract (GIT) bacteria was investigated. The possibility of the drug escaping into the systemic circulation even in the presence of some permeation enhancers was ascertained. Representatives of potential GIT bacteria pathogens were evaluated for their susceptibility to the drug at concentrations obtainable in the GIT using standard microbiological methods. Dose levels that will inhibit these potential bacteria pathogens were also established, as well as the frequency of their administration. Different batches of oral capsules of 250 mg gentamicin sulphate were formulated and their release profiles ascertained using standard methods. The results showed that the selected representatives of the GIT potential pathogenic bacteria were all susceptible to gentamicin sulphate. The drug at its plausible dosage levels of 14.28 mg/kg (1,000 mg/70 kg), 10.71 mg/kg (750 mg/70 kg) and 7.14 mg/kg (500 mg/70 kg) did not cross the mucosal barrier into the systemic circulation even in the presence of some permeation enhancers. The drug's frequency of administration were found to be on 8-hourly bases. Gentamicin sulphate (250 mg) granules formulated with polyethylene glycol (PEG 4000) as granulating aid were quick drying because the granules were not hygroscopic. The formulated gentamicin sulphate capsule batch released enough concentration of the drug that inhibits the test organism within 2 min of dissolution. The above stated doses are acceptable in the dosage form design; it is possible to formulate non-absorbable oral gentamicin sulphate dosage form for local activity in the GIT using existing conventional solid dosage formulating equipment.

Manipulation of the Microbiota for Treatment of IBS and IBD—Challenges and Controversies

Effects of activated charcoal and atropine for the removal of organophosphorus compounds, which remain in the gastrointestinal tract or have already been absorbed into the systemic circulation, were investigated in rats. Activated charcoal extensively adsorbed the organophosphates fenitrothion, tolelofos methyl, piperophos and salithion, and its immediate administration after oral ingestion of fenitrothion remarkably reduced serum fenitrothion levels, but had no effect on the serum levels of the compound which had been absorbed from the gastrointestinal tract. Thus, all of the organophosphorus compounds were poorly exsorbed (0.002-0.39% of the dose in 120 min) from the blood into the intestinal lumen probably due to their extensive protein binding and large distribution volumes. Atropine inhibited absorption of fenitrothion in the perfusion in-situ and also delayed the absorption of the compound in-vivo, but had no significant effect on exsorption of fenitrothion. The serum fenitrothion levels on treatment with both atropine and charcoal significantly decreased compared with those of the control. We conclude that, oral activated charcoal will not be able to enhance the elimination of organophosphorus compounds which have already been absorbed into the systemic circulation, but constituted useful method for the removal of the compounds remaining in the gastrointestinal tract because of its excellent adsorptive capacity.

A review for the anti-inflammatory effects of paeoniflorin in inflammatory disorders

Human mast cells (MCs) and eosinophils were first described and named by Paul Ehrlich. These cells have distinct myeloid progenitors and differ morphologically, ultrastructurally, immunologically, biochemically, and pharmacologically. However, MCs and eosinophils play a pivotal role in several allergic disorders. In addition, these cells are involved in autoimmune disorders, cardiovascular diseases, and cancer. MCs are distributed throughout all normal human tissues, whereas eosinophils are present only in gastrointestinal tract, secondary lymphoid tissues, and adipose tissue, thymus, mammary gland, and uterus. However, in allergic disorders, MCs and eosinophils can form the “allergic effector unit.” Moreover, in several tumors, MCs and eosinophils can be found in close proximity. Therefore, it is likely that MCs have the capacity to modulate eosinophil functions and vice versa. For example, interleukin 5, stem cell factor, histamine, platelet-activating factor (PAF), prostaglandin D2 (PGD2), cysteinyl leukotrienes, and vascular endothelial growth factors (VEGFs), produced by activated MCs, can modulate eosinophil functions through the engagement of specific receptors. In contrast, eosinophil cationic proteins such as eosinophil cationic protein and major basic protein (MBP), nerve growth factor, and VEGFs released by activated eosinophils can modulate MC functions. These bidirectional interactions between MCs and eosinophils might be relevant not only in allergic diseases but also in several inflammatory and neoplastic disorders.

The field of deep learning has witnessed dramatic and rapid progress in the past several years, largely driven by the availability of massive datasets and increased computational power. Although promising advances have been made, the implementation of deep learning in drug design is still in the vigorously developing stage. Here we summarize the frontiers and emerging applications of deep learning in drug design. We provide the background of deep learning and the architecture of several important deep neural networks for drug design and discovery, including convolutional neural networks, recurrent neural networks, transformer, deep auto‐encoder, generative adversarial networks, graph neural networks, and flow‐based neural networks. We give some examples of tasks for which these different deep neural networks are suitable. The successful employment of deep learning in drug design and discussing its further opportunities are reviewed, such as molecular representation, drug‐likeness prediction, de novo molecular generation, ligand‐protein interaction prediction, reaction, and retrosynthetic prediction. Current challenges and further possibilities are proposed to help accelerate the progress of the field, such as model interpretation, the requirement for large quantities of training datasets, hyperparameters tuning, and open source.

The emerging role of endothelin-1 in the pathogenesis of subchondral bone disturbance and osteoarthritis