Proteinase Inhibitor Research Articles

The biological function of Serpinb9 and Serpinb9-based therapy.

Recent breakthroughs in discovering novel immune signaling pathways have revolutionized different disease treatments. SERPINB9 (Sb9), also known as Proteinase Inhibitor 9 (PI-9), is a well-known endogenous inhibitor of Granzyme B (GzmB). GzmB is a potent cytotoxic molecule secreted by cytotoxic T lymphocytes and natural killer cells, which plays a crucial role in inducing apoptosis in target cells during immune responses. Sb9 acts as a protective mechanism against the potentially harmful effects of GzmB within the cells of the immune system itself. On the other hand, overexpression of Sb9 is an important mechanism of immune evasion in diseases like cancers and viral infections. The intricate functions of Sb9 in different cell types represent a fine-tuned regulatory mechanism for preventing immunopathology, protection against autoimmune diseases, and the regulation of cell death, all of which are essential for maintaining health and responding effectively to disease challenges. Dysregulation of the Sb9 will disrupt human normal physiological condition, potentially leading to a range of diseases, including cancers, inflammatory conditions, viral infections or other pathological disorders. Deepening our understanding of the role of Sb9 will aid in the discovery of innovative and effective treatments for various medical conditions. Therefore, the objective of this review is to consolidate current knowledge regarding the biological role of Sb9. It aims to offer insights into its discovery, structure, functions, distribution, its association with various diseases, and the potential of nanoparticle-based therapies targeting Sb9.

Epidemic trend, genetic characteristics, and transmission networks of HIV-1 among treatment-naive men who have sex with men in Hebei province, China.

Homosexual transmission has contributed greatly to the current HIV-1 epidemic in Hebei province, China. Dolutegravir (DTG) will be conditionally used as a component of free antiretroviral therapy (ART) according to manual for national free anti-AIDS treatment drugs (2023 edition) issued by China in June 2023. However, current genetic characteristics and pretreatment drug resistance (PDR) to proteinase inhibitors (PIs), reverse transcriptase inhibitors (RTs) and integrase strand transfer inhibitors (INSTIs) of HIV-1 in this population have remained unclear. Serial consecutive cross-sectional analyses for HIV- 1 infection trend, genetic characteristics, PDR and molecular transmission networks were conducted from 2018 to 2022. All of participants were HIV-1- infected MSM newly diagnosed at the HIV surveillance points (HSPs) in Hebei, China. Evidence of PDR was confirmed using the world health organization (WHO) list for surveillance of drug resistance mutations. In this study, a total of 14 HIV-1 subtypes were circulating in the HSPs of Hebei province, China. CRF01_ AE (51.9%, 350/675), CRF07_BC (30.4%, 205/675), B (6.2%, 42/675) and URFs (5.8%, 39/675) were the four most predominant subtypes among MSM. And, CRF07_BC (r > 0) and URFs (r > 0) indicated an increasing trend, respectively; however, CRF01_AE (r < 0) showed a decline trend. The overall prevalence of HIV-1 PDR showed a substantial increase from 6.3% in 2018 to 7.9% in 2022. The prevalence of NNRTI-PDR was the highest (5.8%, 39/675), followed by INSTIs (2.4%, 16/675), NRTIs (0.6%, 4/675) and PIs (0.3%, 2/675). Furthermore, extensive HIV-1 strains bearing PDR were circulating in the MSM population via molecular transmission networks for major HIV-1 subtypes, especially CRF01_AE and CRF07_BC. Our findings reflect that HIV-1 epidemic in the MSM population is complex and severe in Hebei, China. Therefore, it is urgent for us to implement more effective intervention measures to limit the further dissemination of HIV-1, especially the spread of HIV-1 INSTI-PDR strains.

Determination of antioxidant and anti-inflammatory properties of Salvia aethiopis/sclarea and synthesized silver nanoparticles

Aim: In the study, the phenolic compounds of Salvia aethiopis/sclarea will be determined and the antioxidant and anti-inflammatory activities of the extracts of these plants and the synthesized silver nanoparticles (AgNP) will be examined in vitro. Materials and Methods: Extracts obtained from Salvia spp. (SE) collected from the Kyrenia region of Cyprus were used in the study. Methanol extracts were prepared and total phenolic, flavonoid and alkaloid contents were determined. The amount of phenolic compounds was then quantified and identified using HPLC. The antioxidant properties of Salvia aethiopis/sclarea were analyzed using various markers, including lipid peroxidation inhibition activity, 1,1-Diphenyl-2-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) scavenging capacity. Additionally, their anti-inflammatory effects were investigated by proteinase inhibitor activity, heat-induced hemolysis and albumin denaturation analyses. Results: S. aethiopis and S. sclarea were contained high levels of the phenolic compounds isoquercetin and caffeic acid or naringenin and caffeic acid, respectively. After green synthesis of silver nanoparticles, the antioxidant and anti-inflammatory properties of plant extracts increased significantly (p&lt;0.05). Conclusion: Our study found that silver nanoparticles synthesized using S. aethiopis/sclarea extract increased the antioxidant and anti-inflammatory effects of plant extracts and suggested that AgNP/SE could be used as a potential therapeutic agent in medicine.

The content of matrix metalloproteinase-1, -9 and tissue inhibitor of matrix metalloproteinase-1 in patients with bronchial asthma and obesity

Airway remodeling is a significant factor in the progression of respiratory failure in bronchial asthma (BA). Obesity can contribute to inflammation, and consequently – airway remodeling, by increased production of matrix proteinase (MMP). However, the mechanisms of remodeling in different phenotypes of BA, especially in comorbid patients, remain poorly understood. The aim of the research was to examine the levels of remodeling markers MMP-1, MMP-9 and tissue inhibitor of matrix proteinases (TIMP-1) in obese asthmatic patients, taking into account their age at the onset of BA, the severity of symptoms and the level of control. Material and methods. 195 patients with BA associated with obesity were examined. The control group consisted of 95 practically healthy people. Patients were divided into two groups: Group I included 100 patients with early onset, and Group II included 95 patients with late onset of BA. The study was approved by the Bioethics Commission of the Medical Institute of Sumy State University. Statistical analysis of the obtained results was carried out using the SPSS-17 program. Results. The content of MMP-1, -9 and TIMP-1 was higher in patients with early (129.40 ± 7.86; 177.30 ± 9.19; 142.40 ± 12.22 ng/ml, respectively) and late (196.60 ± 9.62; 289.80 ± 16.23; 187.50 ± 11.34 ng/ml, respectively) BA with obesity compared to practically healthy individuals (р &lt; 0.005). Patients with late-onset BA associated with obesity likely had higher indicators than those with early BA. In patients with early BA, the levels of MMP-1, -9, and TIMP-1 were not significantly different depending on the severity of the disease (р &gt; 0.05). However, in patients with severe late BA, the levels of these markers were higher compared to those with mild BA (р &lt; 0.05). Levels of MMP-1 and MMP-9 in patients with early-onset and late-onset BA with obesity were higher in the absence of disease control compared to those in complete and partial control, but in late-onset exceeded those in early-onset. Conclusions. Considering the obtained results, it can be assumed that the level of MMP-1, MMP-9 and TIMP-1 in obese patients with BA can serve as biomarkers of airway remodeling, namely, the severity and the level of BA control.

Bioprospecting of Meliaceae family phytomolecules for the treatment of monkeypox virus infection: a QSAR modeling and MD simulation approach

Recent monkeypox virus (MPXV) infections show the risk of MPXV transmission that persists today and the significance of surveillance and quick response methods to stop the virus’s spread. Currently, the monkeypox virus infection is not specifically treated. In this study, QSAR models were designed using known inhibitors of cysteine proteinase from the vaccinia virus, where the Random Forest model and Ridge model had showed the best correlation between predicted and observed EC50. These models were used to screen Maliaceae family phytochemicals against MPXV cysteine proteinase. The compound, IMPHY010637 was detected in top 5 from both the QSAR screening models and showed best docked score (-8.6 kcal/mol) and thus selected for further investigation. Further, the IMPHY010637 showed interaction with the catalytic residue His241 of the protein as reported in earlier studies. The ADMET analysis of the compound showed the acceptable drug-like properties of IMPHY010637. However, these properties could be improved after experimental validation of protein-ligand binding. Both docked complex and poses created in 100 ns MD simulation of the protein-ligand complex showed the presence of multiple hydrogen bonds. RMSD and conformation analysis showed stable binding of IMPHY010637 with the cysteine proteinase of MPXV at its active site. Compared to the known inhibitor, IMPHY010637 showed better binding with the protein as observed by the PCA and MM/GBSA analysis. This study concluded IMPHY010637 as a potential inhibitor for the cysteine proteinase of MPXV using computational methods that could be tested in in-vitro experiments. Communicated by Ramaswamy H. Sarma

Measuring the Concentration of Active Alpha-1 Antitrypsin in Serum.

ActiveAlpha-1 antitrypsin (AAT) circulates in blood in two isoforms in dynamic equilibrium: 1) native AAT, which binds irreversibly to neutrophil elastase, and 2) thiol-modified AAT, which binds reversibly to neutrophil elastase, either of whichcan be inactivated by oxidation, cleavage, covalent binding, and bound antibodies. Anti-AAT antibodies used for detecting AAT in plasma bind to active and inactive protein, thereby overestimating the concentration of functional protein. Active AAT can be quantitated by measuring its inhibition of active-site titrated elastase. A method for measuring active AAT in the presence of competing proteinase inhibitors in plasma has been developed and is described herein.

Novel Insights into the Link Between Myeloperoxidase Modified LDL, LOX-1, and Neuroserpin in Stroke.

Cardiovascular disease that is caused by atherosclerosis is the leading cause of death worldwide. Atherosclerosis is primarily triggered by endothelial dysfunction and the accumulation of modified low-density lipoprotein (LDL) particles in the subendothelial space of blood vessels. Early reports have associated oxidized LDL with altered fibrinolysis and atherogenesis. It has been suggested that myeloperoxidase oxidized LDL (Mox-LDL) is involved in atherosclerosis because of its significant pathophysiological role in the modification of LDL in vivo. It has been equally demonstrated that Mox-LDL binds to the lectin-like oxidized low-density lipoprotein receptor-1 (lox-1) scavenger receptor which leads to the upregulation of inflammatory mediators in endothelial cells and the progression of cardiovascular disease. It has been also shown that neuroserpin, a member of the serine proteinase inhibitor (serpin) superfamily, has an important role at the level of fibrinolysis in the nervous tissue. Since little is known about the effects of Mox-LDL on endothelial cell fibrinolytic activity and the involvement of lox-1 in this process, our study aimed at evaluating the in vitro effects of Mox-LDL on neuroserpin release from human aortic endothelial cells (HAECs) and the role of lox-1 scavenger receptor in this context by relying on lox-1 gene silencing in HAECs, culturing the cells in the presence of Mox-LDL, measuring their neuroserpin expression and release by quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively, and assessing their fibrinolytic activity using the Euglobulin Clot Lysis Time (ECLT) method. Our data show that Mox-LDL decreases endothelial cell fibrinolytic capacity by upregulating neuroserpin in lox-1 knockdown cells. Lox-1 protects the endothelial cells from a Mox-LDL-induced decrease in pro-fibrinolytic capacity, which has important consequences in the context of stroke.

Profiling the antidiabetic potential of GC–MS compounds identified from the methanolic extract of Spilanthes filicaulis: experimental and computational insight

This study examines the nutritional composition, phytochemical profiling, and antioxidant, antidiabetic, and anti-inflammatory potential of a methanolic extract of Spilanthes filicaulis leaves (MESFL) via in vitro, ex vivo, and in silico studies. In vitro antioxidant, antidiabetic, and anti-inflammatory activities were examined. In the ex vivo study, liver tissues were subjected to FeSO4-induced oxidative damage and treated with varying concentrations of MESFL. MESFL contains a reasonable amount of nitrogen-free extract, moisture, ash content, crude protein, and fat, with a lesser amount of crude fiber. According to GC–MS analysis, MESFL contains ten compounds, the most abundant of which are 13-octadecenal and Ar-tumerone. In this study, MESFL demonstrated anti-inflammatory activities via membrane stabilizing properties, proteinase inhibition, and inhibition of protein denaturation (IC50 = 72.75 ± 11.06 µg/mL). MESFL also strongly inhibited both α-amylase (IC50 = 307.02 ± 4.25 µg/mL) and α-glucosidase (IC50 = 215.51 ± 0.47 µg/mL) activities. Our findings also showed that FeSO4-induced tissue damage decreased the levels of GSH, SOD, and CAT activities while increasing the levels of MDA. In contrast, treatment with MESFL helped to restore these parameters to near-normal levels, which signifies that MESFL has great potential to address complications from oxidative stress. Furthermore, the in silico interaction of the GCMS-identified phytochemicals with the active sites of α-amylase and α-glucosidase via molecular and ensembled-based docking displayed strong binding affinities of Ar-tumerone and 4-hydroxy-3-methylacetophenone to α-amylase and α-glucosidase, respectively. Taken together, the biological activities of MESFL might be a result of the effects of these secondary metabolites. Communicated by Ramaswamy H. Sarma

Tissue inhibitor of matrix metalloproteinases 4: a novel marker associated with CAA

AbstractBackgroundWe have previously shown that matrix metalloproteinase 9 (MMP9) and tissue inhibitor of proteinases 3 (TIMP3) are specifically expressed in the vasculature of patients with cerebral amyloid angiopathy (CAA). Moreover, we have demonstrated increased MMP9 and decreased TIMP3 levels, as well as an altered MMP9:TIMP3 ratio in the vasculature of CAA patients that suffered from intracerebral haemorrhages (CAA‐ICH) compared to CAA patients without ICH (CAA‐nonICH). We now extended our studies to investigate whether TIMP4 may have a similar role in CAA, as this has not been studied before.MethodUsing immunohistochemistry, we studied occipital lobe tissue of 44 controls (mean age 78.4 years; 50% female) and 58 CAA patients (77.5 years; 55% female; 40 CAA‐nonICH and 18 CAA‐ICH). Vascular TIMP4 staining was scored in a semiquantitative manner. Using ELISA, we measured TIMP4 CSF levels of 33 CAA patients (mean age 72.2 years; 47% female) and 35 controls (mean age 71.2 years; 51% female). For a subset of CSF samples (11 CAA samples and 14 controls), the levels of MMP2, MMP9, and MMP14 have been determined previously, and now their ratios to TIMP4 were calculated. Differences between CAA patients and controls were assessed using (ordinal) linear regression analysis.ResultIn brain tissue, CAA cases had increased vascular TIMP4 expression when compared to controls (p = 0.00002, figure 1). TIMP4 expression did not differ between CAA‐nonICH and CAA‐ICH cases. In the absence of vascular TIMP4 staining, we often observed perivascular staining. Decreased levels of perivascular TIMP4 staining were observed in CAA‐ICH cases when compared to CAA‐nonICH cases (p = 0.045) and to controls (p = 0.035). CSF levels of TIMP4 were decreased in CAA patients compared to controls (p = 0.03, figure 2). In addition, the ratios of MMP2 (p = 0.038), MMP9 (p = 0.005), and MMP14 (p = 0.025) to TIMP4 were increased in CAA patients compared to controls.ConclusionWe demonstrated increased TIMP4 expression in vessel walls of CAA patients compared to controls. In contrast, TIMP4 CSF levels were decreased in CAA patients compared to controls, whereas MMP:TIMP4 ratios were increased, indicating a disbalance of MMP/TIMP4 levels in CAA pathophysiology.Funding: BIONIC project (no. 733050822, ZonMW) and CAFÉ project (no. 5R01NS104147‐02, NIH).

Biochemical markers assessing rate of reparative processes in patients with infiltrative pulmonary tuberculosis

Theeffectiveness of chemotherapy for infiltrative pulmonary tuberculosis (ITL) is determined by proper timing and adequacy of treatment. The speed of reparative changes in the pulmonary parenchyma is associated with the effectiveness of therapy. Different classes of proteinases are involved in extracellular matrix remodeling. The aim of the study was to evaluate a potential of using the markers of proteinase/inhibitors axis for predicting effectiveness of therapy in patients with ITL due to varying resistance ofMycobacterium tuberculosisto anti-tuberculosis drugs (PTP).Materials and methods.A retrospective study included 60 and 55 ITL patients with the drug sensitive (DS) and drug resistance (DR) Mtb strains, respectively. Patients were divided into two groups according to therapy effectiveness. The levels of blood serum matrix metalloproteinase MMP-1, MMP-3, MMP-8, MMP-9 and their TIMP-1 inhibitor, neutrophil elastase (NE), alpha-2 macroplobulin (MG) and proteinase inhibitor (PI) were determined. Statistica 10.0 and R software packages were used.Results. According to the model of discriminant analysis, the pattern of host inflammatory response in ITL was accounted for not only by varying pathogen resistance, but also by the different size of lung damage. Patients with a limited and widespread process with the DS of the pathogen strains were assigned to group I and II, and DR — to group III and IV group, respectively. The decisive in the speed of tissue repair was the number of neutrophils bearing various levels of proteinase MMP-8, MMP-9 (I group) or TIMP-1 and PI inhibitors (II group). Only combining biochemical data with the those on radiation methods are possible for prognosis in a group of patients with DR (III and IV).Conclusions.Combinations of the markers of the proteinase/inhibitors axis are informative in assessing a rate of reparative changes in ITL and differ not only depending on the pathogen DS strains, but also from the size of destructed pulmonary parenchyma. Differences in parameter combinations are accounted for by history of a specific disease.

Anti-inflammatory activity of lauric acid, thiocolchicoside and thiocolchicoside-lauric acid formulation.

It is of interest to develop potent and safer anti-inflammatory drugs from plants, as medicinal plants and herbs attained great attention in the medical world due to their multifunctional activities. This article studied the anti-inflammatory effects of lauric acid (LA), thiocolchicoside (TC) and thiocolchicoside-lauric acid (TC-LA) formulation. The anti-inflammatory effects of these compounds were determined by following the methods of inhibition of protein denaturation and proteinase inhibition activity. This was assessed at different concentrations to determine the 50% inhibition concentration (IC50) of the compounds. The result indicated that the activity of LA, TC, TC-LA formulation, and reference drug increased with the increase in the concentration from 10-50 µg/ml, thus proving the activity of LA, TC, and TC-LA formulation against inflammation was in a dose-dependent manner. The percentage of inhibition of protein denaturation was 59.56%, 66.94%, 86.62%, and 60.34% for LA, TC, the combination of TC-LA and standard drug, and the IC50 values were found to be 44.78 µg/mL, 37.65 µg/mL, 27.15 µg/mL and 43.42 µg/mL, respectively. The percentage of proteinase inhibition activity of LA, TC, and a combination of TC-LA and the standard drug was 66.65%, 77.49%, 94.07%, and 69.83%, and IC50 of LA, TC, a combination of TC-LA and standard drug were35.5 µg/mL, 32.12 µg/mL, 24.35 µg/mL and 37.80 µg/mL, respectively. We found out that lauric acid, thiocolchicoside, and thiocolchicoside-lauric acid formulation exhibited significant anti-inflammatory activity.

Identification of immune-responsive circular RNAs in shrimp (Litopenaeus vannamei) upon yellow head virus infection

Circular RNAs (circRNAs) are a subclass of non-coding RNAs (ncRNAs) formed through a process known as back-splicing. They play a crucial role in the genetic regulation of various biological processes. Currently, circRNAs have been identified as participants in the antiviral response within mammalian cells. However, circRNAs in shrimp infected with the yellow head virus (YHV) remain largely unexplored. Therefore, this study aims to identify circRNAs in the hemocytes of Litopenaeus vannamei during YHV infection. We discovered 358 differentially expressed circRNAs (DECs), with 177 of them being up-regulated and 181 down-regulated. Subsequently, eight DECs, including circ_alpha-1-inhibitor 3, circ_CDC42 small effector protein 2, circ_hemicentin 2, circ_integrin alpha V, circ_kazal-type proteinase inhibitor, circ_phenoloxidase 3, circ_related protein rab-8B, and circ_protein toll-like, were randomly selected for analysis of their expression patterns during YHV infection using qRT-PCR. Furthermore, the circRNAs' characteristics were confirmed through PCR, RNase R treatment, and Sanger sequencing, all of which were consistent with the features of circRNAs. These findings contribute to a better understanding of circRNAs’ involvement in the antiviral response in shrimp.

Стан локального цервікального імунітету в жінок із непліддям

Purpose - to study certain indicators of local immunity in women with infertility of various origins and early cervical lesions Materials and methods. The state of local cervical immunity was examined in 496 women, who were divided into groups depending on the cause of infertility, and each group was divided into subgroups A and B depending on the presence of cervical epithelial lesions. 128 women in the Group I had an endocrine cause, 122 patients in the Group II had a tubal-peritoneal factor of infertility, and 121 patients in the Group III had a male factor of fertility disorder. 125 healthy women formed the control group (CG). Subgroup A included women with NILM liquid cytology results: IA - 100 women, IIA - 70, IIIA - 68, GCA - 88. Subgroups B included women with ASC-US (atypical squamous cells of undetermined significance), LSIL (low-grade squamous intraepithelial lesion) and HSIL (high-grade squamous intraepithelial lesion): IB - 28 patients, IIB - 52, IIIB - 53, CGB - 37. In all patients, the content of interleukin-8 and -6 (IL-8, IL-6), lactoferrin and secretory leukocyte proteinase inhibitor in the material obtained for liquid cytology was determined by polymerase chain reaction. Results. In all patients with infertility, an increased concentration of secretory leukocyte proteinase inhibitor (from 314.1 to 398.7 pg/ml) and a decreased lactoferrin content (from 13.6 to 28.6 pg/ml) were found compared to the group of healthy women (114.8 and 64.8 pg/ml, respectively). In women with tubal and peritoneal infertility and male infertility, higher concentrations of IL-8 were found (659.9 to 878.8 pg/ml) than in the group of endocrine infertility (302.9 pg/ml) and in the group of healthy women (282.4 pg/ml). Conclusions. IL-8 content in cervical mucus is probably higher in women with tubal-peritoneal infertility (838 pg/ml) and male infertility (659.9 pg/ml) than in healthy women (289.4 pg/ml) and in women with endocrine infertility (302.3 pg/ml). The content of secretory inhibitor of leukocyte proteinase in cervical mucus was increased in all patients with infertility, regardless of its cause (from 314.1 pg/ml to 398.9 pg/ml), in the group of healthy women - 144.8 pg/ml. 3. The content of lactoferrin in the mucus of the cervical canal was reduced in all patients with impaired fertility (from 13.6 pg/ml to 28.6 pg/ml), in the group of healthy women - 64.8 pg/ml. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the author.

Carboxypeptidase inhibitors from Solanaceae as a new subclass of pathogenesis related peptide aiming biotechnological targets for plant defense.

Background: Plant protease inhibitors play a crucial role in inhibiting proteases produced by phytopathogens and exhibiting inhibitory effects on nematodes, fungi, and insects, making them promising candidates for crop protection. Specifically, carboxypeptidase inhibitors, a subset of proteinase inhibitors, have been extensively studied in potato and tomato of Solanaceae plant family. However, further research is needed to fully understand the functions and biotechnological potential of those inhibitors in plants. This work aimed to in silico characterize carboxypeptidase inhibitors from Solanaceae as potential antimicrobial and defense agents focused on biotechnological targets. Methods: The methodology employed involved search in UniProt, PDB, KNOTTIN, NCBI, and MEROPS databases for solanaceous carboxypeptidase inhibitors, phylogenetic relationships and conservation patterns analyzes using MEGA-X software and Clustal Omega/MView tools, physicochemical properties and antimicrobial potential prediction using ProtParam, ToxinPred, iAMPred, and APD3 tools, and structural features prediction using PSIPRED. Results and discussion: A systematic literature search was conducted to identify relevant studies on Solanaceae carboxypeptidase inhibitors and their activities against pathogens. The selected studies were reviewed and the main findings compiled. The characterization of Solanaceae carboxypeptidase inhibitors proposed for the first time the global sequence consensus motif CXXXCXXXXDCXXXXXCXXC, shedding light on carboxypeptidase inhibitors distribution, sequence variability, and conservation patterns. Phylogenetic analysis showed evolutionary relationships within the Solanaceae family, particularly in Capsicum, Nicotiana, and Solanum genera. Physicochemical characteristics of those peptides indicated their similarity to antimicrobial peptides. Predicted secondary structures exhibited variations, suggesting a broad spectrum of action, and studies had been demonstrated their activities against various pathogens. Conclusion: Carboxypeptidase inhibitors are being proposed here as a new subclass of PR-6 pathogenesis-related proteins, which will aid in a focused understanding of their functional roles in plant defense mechanisms. These findings confirm the Solanaceae carboxypeptidase inhibitors potential as defense agents and highlight opportunities for their biotechnological applications in pathogen control.

Charge Distribution Patterns of IA3 Impact Conformational Expansion and Hydration Diffusivity of the Disordered Ensemble.

IA3 is a 68 amino acid natural peptide/protein inhibitor of yeast aspartic proteinase A (YPRA) that is intrinsically disordered in solution with induced N-terminal helicity when in the protein complex with YPRA. Based on the intrinsically disordered protein (IDP) parameters of fractional net charge (FNC), net charge density per residue (NCPR), and charge patterning (κ), the two domains of IA3 are defined to occupy different domains within conformationally based subclasses of IDPs, thus making IA3 a bimodal domain IDP. Site-directed spin labeling (SDSL) electron paramagnetic resonance (EPR) spectroscopy and low-field Overhauser dynamic nuclear polarization (ODNP) spectroscopy results show that these two domains possess different degrees of compaction and hydration diffusivity behavior. This work suggests that SDSL EPR line shapes, analyzed in terms of their local tumbling volume (VL), provide insights into the compaction of the unstructured IDP ensemble in solution and that protein sequence and net charge distribution patterns within a conformational subclass can impact bound water hydration dynamics, thus possibly offering an alternative thermodynamic property that can encode conformational binding and behavior of IDPs and liquid-liquid phase separations.

PSIX-15 The Effects of Torula Yeast as a Protein Source on Digestibility, Inflammation, and Gut Microbiome in Labrador Retrievers with Chronically Poor Stool Quality

Abstract Four Rivers Kennel examined the effects of varying protein sources on digestibility, inflammation, and gut microbiota in Labrador Retrievers with historically poor stool quality. Thirty dogs (n = 15 males and n= 15 females; ages 3 to 10 years) with stool quality scores ≤ 2.5 on a 5 point scale were randomly assigned to 1 of 3 diets meeting AAFCO requirements with differing protein sources and similar nutrient profiles: 1) chicken meal (PC); 2) 10% brewer’s yeast (BY); and 3) 10% torula yeast (TY; SylPro, Arbiom, Durham, NC). Another 10 dogs (n = 5 males and n = 5 females) with normal stool quality (score ≥ 3) received diet 1 and served as negative control (NC). After 7 days acclimation to diet 1, dogs began treatments. Daily stool scores and weekly body weights were recorded. On days 7, 21, and 36, blood serum was analyzed for C-reactive protein (CRP), and feces for S100A12, alpha-1 proteinase inhibitor (a1PI), calprotectin, and microbiota dysbiosis index (MDI). Apparent total tract digestibility was assessed using the indicator method with 2 g titanium dioxide administered orally via gel capsules. Statistical analysis was performed by repeated measures model in SAS for stool scores, body weight, biomarkers, and microbiota or by one-way ANOVA in JMP for digestibility coefficients. Stool scores were greater in NC (P &amp;lt; 0.01), changed daily in all groups (P &amp;lt; 0.01) without noticeable overall trends, and were not affected by treatment*day interaction (P = 0.64). Body weight was greater (P = 0.01) and CRP less (P &amp;lt; 0.01) in NC dogs than poor stool dogs. Dry matter and carbohydrate digestibility did not differ between groups (P ≥ 0.14). NC dogs had greater fat digestibility compared with BY dogs (94.64 ± 1.33 % vs 91.65 ± 1.25%; P = 0.02). Protein digestibility was reduced in PC (58.91 ± 4.68%) and BY (58.26 ± 4.42%) compared with NC (75.52 ± 5.01 %), but TY diet improved protein digestibility to levels similar to NC (72.18 ± 5.01%, P = 0.03). There were no treatment effects on S100A12 or a1PI (P ≥ 0.44). Calprotectin decreased at a greater rate overtime in TY dogs (P &amp;lt; 0.01). There was a treatment*day interaction for MDI; BY and TY fluctuated less overtime (P = 0.01). Blautia (P = 0.03) and Clostridium hiranonis (P = 0.05) abundances were reduced in BY and TY. Dogs never exhibited significant dysbiosis (MDI &amp;lt; 2), and bacterial abundances remained within normal ranges. Dogs with chronically poor stool quality experience reduced body weights, increased serum CRP, and altered nutrient digestibility, but TY helped improve protein and fat digestibility, sustain microbiome, and reduce fecal calprotectin. Feeding torula yeast long-term may help improve plane of nutrition to better maintain weight and reduce inflammation in dogs with chronically poor stool quality.

Chitosan: A promising plant stimulant

Chitosan is a natural polymer produced from the cell walls of fungi and the exoskeleton of crustaceans. Chitosan has exceptional qualities, such as non-toxicity, biodegradability, biocompatibility, affordability, and its capacity to function as a proteinase inhibitor by way of the formation of phytoalexin. Furthermore, positive ionic charges on the chitosan molecule increase plant immunity and defense systems against pathogens, promote plant growth, and increase yield. Chitosan plays a significant role, particularly in reducing adverse effects of abiotic stress like salinity and drought, by affecting the formation of Reactive Oxygen Species (ROS) such as proline, antioxidant enzyme activities, and total soluble sugars, which reverse the adverse effects of stress, improve the initiation of the antioxidant system, and increase photosynthesis, consequently improving plant growth and yield. Furthermore, chitosan treatment could suppress virus infections regardless of virus types as well as plant species. There are numerous physiological responses to chitosan application to improve plant immunity and defense systems, such as the synthesis of antioxidant enzymes to enhance plant resistance against pathogens, particularly fungi and bacterial infections. There are numerous benefits to chitosan, including inducing defense genes in numerous species, improving salt tolerance, enhancing plant growth under abiotic stress, improving germination parameters, increasing seedlings’ survival, and consequently, improving plant resistance to pathogen infection, increasing tolerance to abiotic stress, and improving plant growth and productivity. Furthermore, it extends the shelf life and improves the fruit quality of numerous fruits when used as a coating film.

Transplant with MZ genotype liver: what is theclinical pulmonarypicture after 30years? a case report and review of the literature.

Alpha-1 antitrypsin, also known as alpha1 proteinase inhibitor, is a protein 90% synthesized by hepatocytes. Alpha-1 antitrypsin deficiency should be suspected if patients have unexplained emphysema or liver disease in the absence of others recognized causes. The diagnosis is based on tests that measure the amount of the enzyme in the blood and confirm by molecular analysis. We present the case of a man of Caucasian ethnicity, who started experiencing difficulty in breathing 20years after liver transplantation. After about 30years since transplantation, an intermediate alpha-1 antitrypsin deficiency is diagnosed with evidence of air trapping, pulmonary emphysema and bronchiectasis. The presence of a Z-variant synthesized from the donor liver may have contribute to the onset of respiratory disease.

Antimicrobial to anti-herbivore: Sakuranetin in rice efficiently inhibits brown planthopper by targeting their beneficial endosymbionts.

In rice, biosynthesis of specialized metabolites active against insect herbivores is elusive. The major known defense metabolites in rice against the destructive phloem-sucking herbivore brown planthoppers (BPH) (Nilaparvata lugens) are proteinase inhibitors, phenolamides and some terpenes (Xiao et al.,2012), which are induced during the invasion. Specifically, phenolamides were found to be induced upon herbivory with different feeding guild, including chewing and phloem-sucking, but could only provide defense against phloem-sucking BPH, though the clear mode of action of phenolamides has not been explored yet. Moreover, the jasmonic acid-mediated modulation of biosynthesis of these specialized metabolites in rice is not elucidated yet. However, a recent study by Liu et al.(2023) demonstrated that sakuranetin, a phytoalexin in rice, was induced upon BPH invasion and showed significant detrimental effect on herbivore's performance by targeting their beneficial endosymbionts. This is the first report on a strong bioactive anti-herbivore molecule observed in rice with an unusual mode of action. In this article, a view has been presented on this work, its impact and exceptionality.

Zymograms as a Tool to Detect PPIs in Host plants of Antheraea Assamensis

Antheraea assamensis (vernacular: ‘muga’) larvae is commercially reared on two tree species of the Lauraceae family, Persea bombycina and Litsea monopetala for its golden yellow, lustrous cocoon silk. Biochemical and molecular studies suggested that the midgut digestive enzymes and their transcripts in larvae feeding on P. bombycina differ from those found in larvae feeding on L. monopetala indicating that host plant 'choice' affects the digestive physiology of this insect. Ingestion of plant proteinase inhibitors is known to influence expression of digestive enzymes. Using reverse zymography technique trypsin and chymotrypsin inhibitors were detected in herbivore-induced leaves of L. monopetala and P. bombycina that could inhibit midgut proteinases of A. assamensis. Such interactions may affect proteolytic digestion in larvae reared on different host plant species. This work may have significance in quality of silk produced by muga silkworm, ultimately benefiting the silkworm rearers/industry.