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Abstract The bacterium Clostridium botulinum, well-known for producing botulinum neurotoxins, which cause the severe paralytic illness known as botulism, produces C2 toxin, a binary AB-toxin with ADP-ribosyltranferase activity. C2 toxin possesses two separate protein components, an enzymatically active A-component C2I and the binding and translocation B-component C2II. After proteolytic activation of C2II to C2IIa, the heptameric structure binds C2I and is taken up via receptor-mediated endocytosis into the target cells. Due to acidification of endosomes, the C2IIa/C2I complex undergoes conformational changes and consequently C2IIa forms a pore into the endosomal membrane and C2I can translocate into the cytoplasm, where it ADP-ribosylates G-actin, a key component of the cytoskeleton. This modification disrupts the actin cytoskeleton, resulting in the collapse of cytoskeleton and ultimately cell death. Here, we show that the serine-protease inhibitor α1-antitrypsin (α1AT) which we identified previously from a hemofiltrate library screen for PT from Bordetella pertussis is a multitoxin inhibitor. α1AT inhibits intoxication of cells with C2 toxin via inhibition of binding to cells and inhibition of enzyme activity of C2I. Moreover, diphtheria toxin and an anthrax fusion toxin are inhibited by α1AT. Since α1AT is commercially available as a drug for treatment of the α1AT deficiency, it could be repurposed for treatment of toxin-mediated diseases.

Objective: This study investigates the associations of α1-antitrypsin, inter-α-trypsin inhibitor heavy chain (ITIH4), and 8-isoprostane with lung function in shipyard workers exposed to occupational metal fume fine particulate matter (PM2.5), which is known to be associated with adverse respiratory outcomes. Methods: A 3-year follow-up study was conducted on 180 shipyard workers with 262 measurements. Personal exposure to welding fume PM2.5 was collected for an 8-h working day. Pre-exposure, post-exposure, and delta (∆) levels of α1-antitrypsin, ITIH4, and 8-isoprostane were determined in urine using enzyme-linked immunosorbent assays. Post-exposure urinary metals were sampled at the beginning of the next working day and analyzed by inductively coupled plasma-mass spectrometry. Lung function measurements were also conducted the next working day for post-exposure. Results: An IQR increase in PM2.5 was associated with decreases of 2.157% in FEV1, 2.806% in PEF, 4.328% in FEF25%, 5.047% in FEF50%, and 7.205% in FEF75%. An IQR increase in PM2.5 led to increases of 42.155 µg/g in ∆α1-antitrypsin and 16.273 µg/g in ∆ITIH4. Notably, IQR increases in various urinary metals were associated with increases in specific biomarkers, such as post-urinary α1-antitrypsin and ITIH4. Moreover, increases in ∆ α1-antitrypsin and ∆ITIH4 were associated with decreases in FEV1/FVC by 0.008% and 0.020%, respectively, and an increase in ∆8-isoprostane resulted in a 1.538% decline in FVC. Conclusion: Our study suggests that urinary α1-antitrypsin and ITIH4 could indicate early lung function decline in shipyard workers exposed to metal fume PM2.5, underscoring the need for better safety and health monitoring to reduce respiratory risks. [ABSTRACT FROM AUTHOR]

Objective To investigate the relationships between serum peroxisome proliferator-activated receptor γ coactivator-1α（PGC-1α) and α1-antitrypsin (A1AT) levels and type 2 diabetes mellitus (T2DM) combined with non-alcoholic fatty liver disease (NAFLD）. Methods Totally 184 patients with T2DM (T2DM group) and another 60 healthy volunteers for physical examination during the same period (control group) were selected, and the patients with T2DM were classified into the NAFLD group (95 patients) and the non-NAFLD group (89 patients) according to whether they had NAFLD or not. Serum PGC-1α and A1AT levels were measured by enzyme-linked immunosorbent assay. Factors af; fecting T2DM combined with NAFLD were analyzed using multifactorial Logistic regression, and the predictive value of se; rum PGC-1α and A1AT levels for T2DM combined with NAFLD was analyzed using receiver operating characteristic (ROC) curves. Results Compared with the control group, serum PGC-1α and A1AT levels were reduced in the T2DM group (both P<0. 05）. Compared with the non-NAFLD group, serum PGC-1α and A1AT levels were reduced in the NAFLD group (both P<0. 05）. Increased body mass index, prolonged duration of T2DM, hypertension, and elevated total of cholesterol, triacylglycerol, low-density lipoprotein cholesterol, aspartate aminotransferase, and alanine transaminase were independent risk factors for NAFLD in combination with T2DM, and elevated high-density lipoprotein cholesterol，PGC-1α, and A1AT were independent protective factors (all P<0. 05）. The area under the curve (AUC) of serum PGC1α and A1AT levels combined in predicting T2DM combined with NAFLD was 0. 885, which was greater than that of se; rum PGC-1α and A1AT levels alone, which were 0. 788 and 0. 786（both P<0. 05）. Conclusion Reduced serum PGC1α and A1AT levels are closely associated with T2DM combined with NAFLD, and the combination of serum PGC-1α and A1AT levels has a high predictive value for T2DM combined with NAFLD. [ABSTRACT FROM AUTHOR]

The neonatal period is crucial for the development of postnatal adaptation and the formation of health and quality of life. Premature birth of a child in conditions of hypoxia and morphological and functional immaturity implies a high risk of adaptation disorders in the postnatal period, causing the formation of functional and chronic diseases in the later years of life. The course of the neonatal period can be characterized both by transient states due to the formation of physiological adaptation and the development of dysfunction of organ systems, in particular the digestive system, which is a consequence of pathological labor stress in the implementation of adverse factors during pregnancy and childbirth in the mother. Timely detection and correction of early manifestations of gastrointestinal dysfunction in perinatal pathology will improve approaches to the provision of medical care to newborns, especially premature infants, should be aimed at both stabilizing the child's condition and preventing the development of its long-term consequences. Aim of the study. To study the peculiarities of clinical and laboratory indicators of intestinal dysfunction in perinatal pathology in premature infants. Materials and methods. A comprehensive clinical and laboratory examination of premature infants of gestational age 34-36 weeks who had signs of disorders of the functional state of the gastrointestinal system in perinatal pathology in the early neonatal period was carried out. Group I consisted of 55 children with clinical signs of severe perinatal pathology; group II (control) consisted of 50 conditionally healthy children of the same gestational age. Exclusion criteria were newborns with diagnosed congenital malformations. The list of laboratory indicators of the functional state of the intestine included: the level of α1-antitrypsin (A1AT), secretory IgA (sIgA), fecal elastase 1 (FE-1), PMN elastase and albumin in the coprofiltrate of newborns in the first week of life. Methods of indicators determination: using enzyme-linked immunosorbent assay (ELISA) on the basis of Ukrainian-German laboratory «BUKINTERMED» (Chernivtsi, Ukraine). The scientific research was conducted in accordance with the provisions of GCP (1996), the Convention of the Council of Europe on Human Rights and Biomedicine (April 4, 1997), the Declaration of Helsinki of the World Medical Association for the Ethical Principles of Research Involving Human Subjects (1964-2008), the Order of the Ministry of Health of Ukraine No. 690 of September 23, 2009 (as amended by the Order of the Ministry of Health of Ukraine No. 523 of July 12, 2012). Protocol of the research study of the Biomedical Ethics Committee of BSMU dated 12.09.2015. Informed written consent was obtained from the parents of the patients before the study with an explanation of the purpose, objectives and methods of laboratory research. Statistical processing of the results was performed using STATISTICA software (StatSoft Inc., USA, version 10). Comparison of quantitative indicators with normal distribution was performed using Student's t-test, the probability of differences was considered statistically significant at p<0.05. The study was conducted within the framework of the scientific theme of the Department of Pediatrics, Neonatology and Perinatal Medicine of the Bukovinian State Medical University: Research work «Improvement of directions of prognostication, diagnosis and treatment of perinatal pathology in newborns and young children, optimization of schemes of catamenial observation and rehabilitation» (0115U002768, term of execution 01.2015-12.2019); Research work «Chronobiological and adaptive aspects and features of vegetative regulation in pathological conditions in children of different age groups» (0122U002245, term of execution 01.2020-12.2024). Research results and discussion. In premature infants with clinical forms of perinatal pathology, a significant incidence of autonomic dysfunction syndrome was noted, which was accompanied by disorders of the functional state of the gastrointestinal system. Clinical signs of dysfunction were: absence or decrease of sucking reflex, regurgitation, stasis, intestinal paresis, delayed passage of meconium, flatulence, hepatomegaly. Of the total number, 43.6 % of newborns had signs of multiple organ failure. Laboratory studies of coprofiltrate showed an increase in the level of A1AT, sIgA, PMN-elastase, albumin with a decrease in the level of FE-1, which are interdependent criteria that confirm the presence and in some way explain the pathophysiological mechanisms of digestive system dysfunction. In particular, a decrease in tolerance to enteral nutrition in newborns occurs against the background of insufficient activity of pancreatic proteolytic enzymes, activation of a local allergic reaction and increased permeability of the intestinal mucosa, which, against the background of local inflammation, entails the risk of increased translocation of microorganisms and toxins into the bloodstream, resulting in deepening of endotoxemia in severe forms of perinatal pathology. Conclusions. 1. Severe diseases of the perinatal period in premature infants with clinical signs of multiorgan dysfunction require timely diagnosis of disorders of the functional state of the digestive tract. 2. Non-invasive laboratory markers using coprofiltrate (A1AT, FE-1, sIgA, PMN-elastase, albumin), together with clinical signs, provide an opportunity to clarify gastrointestinal dysfunction, considering its pathophysiological mechanisms, which is aimed at appropriate correction of treatment measures. [ABSTRACT FROM AUTHOR]

Abstract Background Current microbiological tests fail to identify the causative microorganism in more than half of all pneumonia cases. We explored biomarkers that could be used for differentiating between bacterial and viral pneumonia in patients with community-acquired pneumonia (CAP). Methods In this prospective cohort study conducted in Japan, data obtained from adult patients with bacterial pneumonia, including bacterial and viral coinfections (bacterial pneumonia [BP] group), and purely viral pneumonia (VP group) at diagnosis were analyzed using multivariate logistic regression analysis to identify predictors of bacterial pneumonia. Furthermore, a decision tree was developed using the predictors. Results A total of 210 patients were analyzed. The BP and VP groups comprised 108 and 18 patients, respectively. The other 84 patients had no identified causative microorganism. The two groups shared similar characteristics, including disease severity; however, a significant difference (p

Background: Current microbiological tests fail to identify the causative microorganism in more than half of all pneumonia cases. We explored biomarkers that could be used for differentiating between bacterial and viral pneumonia in patients with community-acquired pneumonia (CAP). Methods: In this prospective cohort study conducted in Japan, data obtained from adult patients with bacterial pneumonia, including bacterial and viral coinfections (bacterial pneumonia [BP] group), and purely viral pneumonia (VP group) at diagnosis were analyzed using multivariate logistic regression analysis to identify predictors of bacterial pneumonia. Furthermore, a decision tree was developed using the predictors. Results: A total of 210 patients were analyzed. The BP and VP groups comprised 108 and 18 patients, respectively. The other 84 patients had no identified causative microorganism. The two groups shared similar characteristics, including disease severity; however, a significant difference (p < 0.05) was observed between the two groups regarding sputum type; sputum volume score; neutrophil counts; and serum levels of interleukin (IL)-8, IL-10, and α1-antitrypsin (AAT). Sputum volume score (p < 0.001), IL-10 (p < 0.001), and AAT (p = 0.008) were ultimately identified as predictors of BP. The area under the curve for these three variables on the receiver operating characteristic (ROC) curve was 0.927 (95% confidence interval [CI]: 0.881–0.974). The ROC curve for sputum volume score and an AAT/IL-10 ratio showed a diagnostic cutoff of 1 + and 65, respectively. Logistic regression analysis using dichotomized variables at the cutoff values showed that the odds ratios for the diagnosis of BP were 10.4 (95% CI: 2.2–50.2) for sputum volume score (absence vs. presence) and 19.8 (95% CI: 4.7–83.2) for AAT/IL-10 ratio (< 65 vs. ≥ 65). Conclusions: Considering that obtaining a definitive etiologic diagnosis with the current testing methods is difficult and time consuming, a decision tree with two predictors, namely sputum volume and the AAT/IL-10 ratio, can be useful in predicting BP among patients diagnosed with CAP and facilitating the appropriate use of antibiotics. Trial registration: UMIN000034673 registered on November 29, 2018. [ABSTRACT FROM AUTHOR]

A new type of hydrophilic, biocompatible, and biodegradable polypeptide nanogel depots loaded with the natural serine protease inhibitor α1-antitrypsin (AAT) was applied for the inhibition of the inflammatory mediator trypsin. Two types of nanogels were prepared from linear synthetic polypeptides based on biocompatible and biodegradable poly[N5-(2-hydroxyethyl)-ʟ-glutamine-ran-N5-propargyl-ʟ-glutamine-ran-N5-(6-aminohexyl)-ʟ-glutamine]-ran-N5-[2-(4-hydroxyphenyl)ethyl)-ʟ-glutamine] (PHEG-Tyr) or biocompatible Nα-ʟ-lysine-grafted α,β-poly[(2-propyne)-ᴅ,ʟ-aspartamide-ran-(2-hydroxyethyl)-ᴅʟ-aspartamide-ran-(2-(4-hydroxyphenyl)ethyl)-ᴅʟ-aspartamide] (Nα-Lys-NG). Both nanogels were prepared by HRP/H2O2-mediated crosslinking in inverse miniemulsions with pH and temperature-stimuli responsive behavior confirmed by dynamic light scattering and zeta potential measurements. The loading capacity of PHEG-Tyr and Nα-Lys-NG nanogels and their release profiles were first optimized with bovine serum albumin. The nanogels were then used for loading and release of AAT. PHEG-Tyr and Nα-Lys-NG nanogels showed different loading capacities for AAT with the maximum (20%) achieved with Nα-Lys-NG nanogel. In both cases, the nanogel depots demonstrated a burst release of AAT during the first 6 h, which could be favorable for quick inhibition of trypsin. A consequent pilot in vitro inhibition study revealed that both PHEG-Tyr and Nα-Lys-NG nanogels loaded with AAT successfully inhibited the enzymatic activity of trypsin. Furthermore, the inhibitory efficiency of the AAT-loaded nanogels was higher than that of only AAT. Interestingly, also non-loaded PHEG-Tyr and Nα-Lys-NG nanogels were shown to effectively inhibit trypsin because they contain suitable amino acids in their structures that effectively block the active site of trypsin.

We have previously demonstrated that the inducible plant viral vector (CMViva) in transgenic plant cell cultures can significantly improve the productivity of extracellular functional recombinant human alpha-1-antiryspin (rAAT) compared with either a common plant constitutive promoter (Cauliflower mosaic virus (CaMV) 35S) or a chemically inducible promoter (estrogen receptor-based XVE) system. For a transgenic plant host system, however, viral or transgene-induced post-transcriptional gene silencing (PTGS) has been identified as a host response mechanism that may dramatically reduce the expression of a foreign gene. Previous studies have suggested that viral gene silencing suppressors encoded by a virus can block or interfere with the pathways of transgene-induced PTGS in plant cells. In this study, the capability of nine different viral gene silencing suppressors were evaluated for improving the production of rAAT protein in transgenic plant cell cultures (CMViva, XVE or 35S system) using an Agrobacterium-mediated transient expression co-cultivation process in which transgenic plant cells and recombinant Agrobacterium carrying the viral gene silencing suppressor were grown together in suspension cultures. Through the co-cultivation process, the impacts of gene silencing suppressors on the rAAT production were elucidated, and promising gene silencing suppressors were identified. Furthermore, the combinations of gene silencing suppressors were optimized using design of experiments methodology. The results have shown that in transgenic CMViva cell cultures, the functional rAAT as a percentage of total soluble protein is increased 5.7 fold with the expression of P19, and 17.2 fold with the co-expression of CP, P19 and P24.

The lysosomal degradation of the endoplasmic reticulum (ER), known as "reticulophagy", is important for protein quality control and organelle turnover. Here we present a noncanonical reticulophagy occurring at ER exit sites (ERESs) induced by the misfolded SERPINA1/α1-antitrypsin (AAT) mutant, Z-AAT. The accumulation of Z-AAT arrests ER-to-Golgi transport, and recruits V-ATPase and ATG16L1 to mediate LC3C decoration of ERESs. Consequently, the receptor RETREG1/FAM134B–2 is recruited by lipidated LC3C to initiate reticulophagy. Furthermore, the blockade of ER export acts as a universal signal to activate reticulophagy mediated by the V-ATPase-ATG16L1-LC3C axis. This study sheds light on the mechanism of how ERESs switch from ER export to reticulophagy for quality control. [ABSTRACT FROM AUTHOR]

α1-antitrypsin deficiency (AATD) is a genetically inherited autosomal-codominant disease with a variable clinical spectrum of lung-related diseases. Pulmonary involvement of α1-antitrypsin deficiency may also include emphysema with variable functional and radiological abnormalities, asthma, and bronchiectasis. Asthma and AATD are mutually exclusive disease entities, but the commonality of neutrophil inflammation across the diseases might suggest common underlying mechanisms of effect. The diseases share many clinical and functional features: patients with AATD commonly first present with asthma-like symptoms; functional alterations may be common to both, such as bronchial hyperresponsiveness or fixed obstruction after bronchial remodeling. It has been recognized that allergy and asthma often coexist with AATD, but the relationship between allergy, asthma and AATD is not clear. Distinguishing AATD from asthma based on presentation and clinical evaluation is not possible. The clinician must assess each of the elements in the context of the whole patient, any patient with difficult-to-manage asthma should be screened for AATD. From the clinician's point of view, improving diagnosis in this population is fundamental to optimize clinical management. Genetic studies will probably be needed in the future to unequivocally establish the causal link between AATD and asthma. [ABSTRACT FROM AUTHOR]

The COVID-19 pandemic continues, causing colossal damage to the population and the global economy. As COVID-19 is studied, new data are emerging regarding the risk of severe coronavirus infection in patients with α1-antitrypsin deficiency. α1 -Antitrypsin is the main inhibitor and key endogenous regulator of the serine leukocyte proteinase activitry released from the granules of activated neutrophils to the cell surface and into the extracellular space. It has been established that the number of cases of severe course and death of COVID-19 in the territories of 68 countries of the world correlates with the frequency of the spread of mutations in the proteinase inhibitor gene among the population of these countries, at which the concentration of α1-antitrypsin in the human blood plasma is 10 times lower than normal. All this contributes to the revision of a number of provisions of the pathogenesis and therapy of a new coronavirus infection.The review presents an analysis of the literature on the role of an inhibitor of serine leukocyte proteinases in protecting the body from COVID-19. The participation of α1-antitrypsin in the inhibition of SARS-CoV-2 penetration into the respiratory tract epithelial cells, in the protection of the vascular endothelium, blood plasma proteins and elastin of the lung tissue from the damaging effect of leukocyte elastase released during neutrophil degranulation and the formation of neutrophil extracellular traps (NETs) is considered. The role of a1-antitrypsin in suppressing inflammation by limiting the secretion of proinflammatory cytokines and neutrophil extracellular traps into the blood has been shown. The individual links in the pathogenesis of the new coronavirus infection have been detailed, which will allow revising the strategy for reducing the risks of severe course of COVID-19.

Background: More than half of the individuals diagnosed with coronavirus disease 2019 (COVID-19) have been found to have high levels of interleukin (IL)-6. A recent report showed that more elevated serum IL-6 level predicts COVID-19 disease severity and patients’ clinical outcomes. Therefore, this study aimed to compare the serum levels of α1-antitrypsin (AAT), IL-1β, and IL-6 between COVID-19 patients and healthy individuals. Methods: During the data collection phase, 90 individuals were enrolled, 45 healthy controls, and 45 patients confirmed with COVID-19 using reverse transcription-quantitative PCR (RT-qPCR) at a specialized isolation hospital in Baghdad between November 2021 and March 2022. In this cross-sectional research, venous blood samples were taken, and serum was isolated and stored for quantitative ELISA measurements of AAT, IL-1β, and IL-6 (ELISA). IBM SPSS version 24 was used to analyze the data. Results: This study revealed a significant increase in the serum levels of AAT, IL-1β, and IL-6 in the COVID-19 patients' group compared to the healthy control group with p-values < 0.001 for each of these markers. Conclusions: AAT concentrations were higher during COVID-19; this elevation is essential during infection. IL-1β and IL-6 levels were also elevated during the infection period; however, dysregulated high levels may lead to cytokine release syndrome. Therefore, these three biomarkers can be regarded as diagnostically crucial parameters.

The expression of the acute-phase reactants C-reactive protein (CRP), α1-antitrypsin (AAT), and secretory leukocyte protease inhibitor (SLPI), is induced in response to inflammation by pro-inflammatory mediators, including interleukin-1β. It is conceivable that acute-phase proteins exert protective functions, when the integrity of an organism is challenged by pathogens or trauma, which result in uncontrolled release of endogenous damage-associated molecular patterns like Toll-like receptor agonists and ATP. Acute-phase proteins can enhance or down-modulate immunity against infections or protect the host against damage caused by over-shooting effector functions of the immune system. CRP is mainly regarded as a proinflammatory opsonizing agent that binds to bacteria and damaged host cells thereby contributing to their inactivation and elimination. AAT and SLPI are well known for their anti-protease activity, which protects the lung extracellular matrix against degradation by proteases that are released by activated neutrophil granulocytes. In addition, there is growing evidence, that CRP, AAT, and SLPI can control the biosynthesis, maturation, and secretion of pro-inflammatory cytokines. The purpose of this narrative mini review is to summarize these anti-inflammatory functions with a focus on the negative control of the ATP-induced, inflammasome-dependent secretion of interleukin-1β by monocytes. CRP-, AAT- and SLPI-mediated control of interleukin-1β release involves the activation of unconventional nicotinic acetylcholine receptors that inhibits the ionotropic function of the ATP receptor P2X7. Apart from other functions, CRP, AAT, and SLPI seem to be central elements of systemic negative feedback loops that protect the host against systemic hyperinflammation, barrier dysfunction, and death by multiple organ damage. [ABSTRACT FROM AUTHOR]

Background & objectives: Type 2 diabetes mellitus (T2DM) is known to induce inflammation and activation of neutrophils causing the release of neutrophil elastase (NE), a pro-inflammatory proteinase. The activity of NE is regulated by endogenous inhibitors alpha1-antitrypsin (α1-AT) and alpha2- macroglobulin (α2-MG). Disrupted proteolytic homeostasis in T2DM patients is one of the causes for vascular complications. This study was carried out for evaluating the levels of plasma NE, α1-AT, α2-MG and NE-α1-AT complex to understand their roles in the pathophysiology of diabetic nephropathy (DN) and diabetic retinopathy (DR). Methods: A total of 240 participants (Control, n=60; T2DM, n=60; DN, n=60; and DR, n=60) were recruited after recording history, clinical examination and laboratory investigations. Retinopathy was confirmed by fundoscopy and nephropathy by urinary albumin excretion and serum creatinine levels. NE was measured using STANA. α1-AT, α2-MG and NE-α1-AT complex were estimated by ELISA. Results: Baseline clinical and laboratory findings were confirmatory to the study groups. The mean elastase activity was higher (P<0.0005) in diabetes groups (T2DM=0.73±0.31, DN=0.87±0.35, DR=0.76±0.41) than controls (0.35±0.20). The levels of α1-AT were lower in DR (8.77±2.85) than DN (26.26±6.16) and T2DM (41.13±14.06) when juxtaposed with controls (122.95±25.71). The approximate fold decrease of α1-AT levels was 15 for DR and four for DN compared to controls. The levels of α2-MG were lowered in T2DM (167.29±30.45), DN (144.66±13.72), and DR (104.67±11.47) than controls (208.87±31.16). The NE-α1-AT complex levels were: controls (215.83±13.61), T2DM (98.85±23.85), DN (129.26±20.40) and DR (153.25±17.11). Interpretation & conclusions: Homeostasis of NE, α1-AT and α2-MG is disrupted in T2DM, DN and DR. Strikingly reduced levels of α1-AT observed in DR are indicative of its possible role in the pathophysiology of retinopathy and emphasizes α1-AT as a plausible therapeutic target. [ABSTRACT FROM AUTHOR]

The expression of the acute-phase reactants C-reactive protein (CRP), α1-antitrypsin (AAT), and secretory leukocyte protease inhibitor (SLPI), is induced in response to inflammation by pro-inflammatory mediators, including interleukin-1β. It is conceivable that acute-phase proteins exert protective functions, when the integrity of an organism is challenged by pathogens or trauma, which result in uncontrolled release of endogenous damage-associated molecular patterns like Toll-like receptor agonists and ATP. Acute-phase proteins can enhance or down-modulate immunity against infections or protect the host against damage caused by over-shooting effector functions of the immune system. CRP is mainly regarded as a pro-inflammatory opsonizing agent that binds to bacteria and damaged host cells thereby contributing to their inactivation and elimination. AAT and SLPI are well known for their anti-protease activity, which protects the lung extracellular matrix against degradation by proteases that are released by activated neutrophil granulocytes. In addition, there is growing evidence, that CRP, AAT, and SLPI can control the biosynthesis, maturation, and secretion of pro-inflammatory cytokines. The purpose of this narrative mini review is to summarize these anti-inflammatory functions with a focus on the negative control of the ATP-induced, inflammasome-dependent secretion of interleukin-1β by monocytes. CRP-, AAT- and SLPI-mediated control of interleukin-1β release involves the activation of unconventional nicotinic acetylcholine receptors that inhibits the ionotropic function of the ATP receptor P2X7. Apart from other functions, CRP, AAT, and SLPI seem to be central elements of systemic negative feedback loops that protect the host against systemic hyperinflammation, barrier dysfunction, and death by multiple organ damage.

α1-antitrypsin deficiency (AATD) is a genetically inherited autosomal-codominant disease with a variable clinical spectrum of lung-related diseases. Pulmonary involvement of α1-antitrypsin deficiency may also include emphysema with variable functional and radiological abnormalities, asthma, and bronchiectasis. Asthma and AATD are mutually exclusive disease entities, but the commonality of neutrophil inflammation across the diseases might suggest common underlying mechanisms of effect. The diseases share many clinical and functional features: patients with AATD commonly first present with asthma-like symptoms; functional alterations may be common to both, such as bronchial hyperresponsiveness or fixed obstruction after bronchial remodeling. It has been recognized that allergy and asthma often coexist with AATD, but the relationship between allergy, asthma and AATD is not clear. Distinguishing AATD from asthma based on presentation and clinical evaluation is not possible. The clinician must assess each of the elements in the context of the whole patient, any patient with difficult-to-manage asthma should be screened for AATD. From the clinician’s point of view, improving diagnosis in this population is fundamental to optimize clinical management. Genetic studies will probably be needed in the future to unequivocally establish the causal link between AATD and asthma.

Convalescent plasma was proposed for passive immunization against COVID-19; but so far there are conflicting results and still open questions. However, besides antibodies, other plasma proteins may be good candidates for further research and application. Thromboinflammation frequently complicates severe COVID-19, and classical anticoagulants like heparins seem to have limited effect. The natural protease inhibitors antithrombin III (ATIII), α 1 -antitrypsin (α 1 -AT) and α 2 -macroglobulin (α 2 -M), which are found decreased in severe COVD-19, play a crucial role in prothrombotic and inflammatory pathways. While ATIII and α 1 -AT are licensed as commercially prepared therapeutic concentrates, there is no preparation of α 2 -M available. The diagnostic, prognostic, and even therapeutic potential of plasma protease inhibitors should be further explored. [ABSTRACT FROM AUTHOR]

Our previous study indicated that Clock gene could affect the thrombotic potential. In this present study, we examined the differential expression of proteins in Clock knockdown mice's plasma, including α1-antitrypsin as a potential target. The proteins were examined in AML12 cells with Clock gene being knocked down to confirm the differential expressions. RNAs of those cells were extracted every 3 h in 24 h. The transcriptional levels of α1-antitrypsin (Serpina1a) and fibronectin (Fn1) were analyzed with the least-squares fit of a 24-h cosine function by single cosinor method, but no circadian rhythm was determined in neither of these genes. The expressions of α1-antitrypsin and fibronectin in Clock knockdown cells were found upregulated in both transcriptional and translational levels. Then, we applied ELISA assay to detect the concentration of activated protein C in Clock knockdown plasma and found a risen concentration. Fibronectin was reported to play a role in inhibiting the platelet aggregation, while activated protein C was demonstrated to inhibit PAI-1. All these results indicated that downregulation of the Clock gene in the circulatory system might have effects on PAI-1 by upregulating α1-antitrypsin, and might play some roles in platelet aggregation by increased fibronectin. [ABSTRACT FROM AUTHOR]

Deficient human α1‐antitrypsin (AAT) variants are involved in pulmonary emphysema and liver cirrhosis. Especially, the Z‐type AAT (Z AAT) variant folds very slowly, and thus accumulates protein folding intermediates prone to aggregation in the endoplasmic reticulum (ER) of hepatocytes. Misfolded proteins accumulated in the ER lead to persistent ER stress and subsequent cell death. In this study, the contribution of unfolded protein response (UPR) in Z AAT‐induced cytotoxicity was investigated. Deletions of each UPR element severely hampered growth of Z AAT‐overexpressing yeast cells. Overexpression of UPR elements, except kar2, alleviated the slow growth phenotype of Z AAT‐overexpressing cells, possibly through augmentation of folding capacity. Furthermore, reinforcement of UPR elements promoted extracellular secretion of Z AAT, which may have mitigated the plasma deficiency of AAT. Our results therefore provide further information on therapeutic strategies to address protein folding diseases. [ABSTRACT FROM AUTHOR]

Summary: Circulating polymers of α1-antitrypsin (α1AT) are neutrophil chemo-attractants and contribute to inflammation, yet cellular factors affecting their secretion remain obscure. We report on a genome-wide CRISPR-Cas9 screen for genes affecting trafficking of polymerogenic α1ATH334D. A CRISPR enrichment approach based on recovery of single guide RNA (sgRNA) sequences from phenotypically selected fixed cells reveals that cells with high-polymer content are enriched in sgRNAs targeting genes involved in “cargo loading into COPII-coated vesicles,” where “COPII” is coat protein II, including the cargo receptors lectin mannose binding1 (LMAN1) and surfeit protein locus 4 (SURF4). LMAN1- and SURF4-disrupted cells display a secretion defect extending beyond α1AT monomers to polymers. Polymer secretion is especially dependent on SURF4 and correlates with a SURF4-α1ATH334D physical interaction and with their co-localization at the endoplasmic reticulum (ER). These findings indicate that ER cargo receptors co-ordinate progression of α1AT out of the ER and modulate the accumulation of polymeric α1AT not only by controlling the concentration of precursor monomers but also by promoting secretion of polymers.

Serine proteases drive important physiological processes such as coagulation, fibrinolysis, inflammation and angiogenesis. These proteases are controlled by serine protease inhibitors (SERPINs) that neutralize their activity. Currently, over 1,500 SERPINs are known in nature, but only 37 SERPINs are found in humans. Thirty of these are functional protease inhibitors. The inhibitory potential of SERPINs is in perfect balance with the proteolytic activities of its targets to enable physiological protease activity. Hence, SERPIN deficiency (either qualitative or quantitative) can lead to disease. Several SERPIN resupplementation strategies have been developed to treat SERPIN deficiencies, including concentrates derived from plasma and recombinant SERPINs. SERPINs usually inhibit multiple proteases, but only in their active state. Over the past decades, considerable insights have been acquired in the identification of SERPIN biological functions, their inhibitory mechanisms and specificity determinants. This paves the way for the development of therapeutic SERPINs. Through rational design, the inhibitory properties (selectivity and inhibitory potential) of SERPINs can be reformed and optimized. This review explores the current state of SERPIN engineering with a focus on reactive center loop modifications and backbone stabilization. We will discuss the lessons learned from these recombinant SERPINs and explore novel techniques and strategies that will be essential for the creation and application of the future generation of therapeutic SERPINs.

Objectives: Failure to produce sufficient quantities of functional α1-antitrypsin (AAT) can result in AAT deficiency (AATD) and significant comorbidities. Laboratory testing plays a vital role in AATD, with diagnosis requiring documentation of both a low AAT level and a mutated allele. This retrospective evaluation examines the efficacy of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) (proteotyping)-based algorithm for AATD detection.Methods: A 16-month retrospective data analysis was performed on two cohorts: 5,474 samples tested with the proteotype-based algorithm and 16,147 samples directly tested by isoelectric focusing (IEF) phenotyping.Results: LC-MS/MS reduced the rate of IEF testing by 97%. The 3% of cases reflexed to IEF resulted in 12 (0.2%) additional phenotype findings. Retrospectively applying the proteotype-based algorithm to the IEF cohort demonstrated a 99.9% sensitivity for the detection of deficiency-associated phenotypes. Most deficiency phenotypes missed by the proteotyping algorithm would come from heterozygous patients with an F, I, or P paired to an S or Z. In all of these cases, patient AAT levels were greater than 70 mg/dL, above the threshold for AAT augmentation therapy.Conclusions: The proteotype algorithm is a sensitive and cost-effective approach for the diagnosis of clinical AAT deficiency. [ABSTRACT FROM AUTHOR]

The formation of ordered Z (Glu342Lys) α1‐antitrypsin polymers in hepatocytes is central to liver disease in α1‐antitrypsin deficiency. In vitro experiments have identified an intermediate conformational state (M*) that precedes polymer formation, but this has yet to be identified in vivo. Moreover, the mechanism of polymer formation and their fate in cells have been incompletely characterised. We have used cell models of disease in conjunction with conformation‐selective monoclonal antibodies and a small molecule inhibitor of polymerisation to define the dynamics of polymer formation, accumulation and secretion. Pulse‐chase experiments demonstrate that Z α1‐antitrypsin accumulates as short‐chain polymers that partition with soluble cellular components and are partially secreted by cells. These precede the formation of larger, insoluble polymers with a longer half‐life (10.9 ± 1.7 h and 20.9 ± 7.4 h for soluble and insoluble polymers, respectively). The M* intermediate (or a by‐product thereof) was identified in the cells by a conformation‐specific monoclonal antibody. This was completely abrogated by treatment with the small molecule, which also blocked the formation of intracellular polymers. These data allow us to conclude that the M* conformation is central to polymerisation of Z α1‐antitrypsin in vivo; preventing its accumulation represents a tractable approach for pharmacological treatment of this condition; polymers are partially secreted; and polymers exist as two distinct populations in cells whose different dynamics have likely consequences for the aetiology of the disease. [ABSTRACT FROM AUTHOR]

BACKGROUND/AIMS: Inhibition of tumor-induced angiogenesis may restrict tumor growth and metastasis. Long-term systemic delivery of angiogenic inhibitors is associated with toxicity as well as other severe side effects. The utility of cells as vehicles for gene therapy to deliver therapeutic molecules has been suggested to be an efficient approach. Mesenchymal stem cells (MSCs) exhibit a tropism to cancer tissue, and may serve as a cellular delivery vehicle and a local producer of anti-angiogenic agents. METHODS: In the present study we attempted to assess production of the transgene, α1-antitrypsin (AAT), in lentivirus-transduced human MSCs and its cytotoxicity against human umbilical cord vein endothelial cells (HUVEC). The secreted protein from these effector cells was determined by an enzyme-linked immunosorbent assay. The cytotoxicity of hMSCs that overexpress the human AAT gene against HUVEC was evaluated with an apoptotic assay. RESULTS: Lentivirus-transduced hMSCs produced functional AAT and displayed much higher cytotoxicity against HUVEC than untransduced hMSCs. Moreover, AAT secreted from transduced hMSCs significantly inhibited HUVEC proliferation in comparison to untransduced hMSCs. Our data demonstrate for the first time that genetically-modified hMSCs released abundant and functional AAT that caused obvious cytotoxicity to HUVEC. CONCLUSION: hMSC may serve as an effective platform for the targeted delivery of therapeutic proteins to cancer sites. Copyright © 2011 John Wiley & Sons, Ltd.

The aim of the study was to assess the effect of one session of high-intensity laser therapy (HILT) on the levels of selected oxidative stress parameters, lysosomal hydrolases, and anti-inflammatory serine protease inhibitor in the peripheral blood of amateur athletes with torn or pulled tendons of the ankle or the knee joint. The group of injured athletes comprised 16 males and females aged 16.3 ± 1.3 years, while the control group of 14 healthy, noninjured amateur athletes of both sexes (controls; age 17.4 ± 4.6 years). Material for the study was peripheral blood taken at three study time points: Immediately before, 30 min after, and 24 h after HILT intervention. In plasma and erythrocytes, thiobarbituric acid reactive substances (TBARSpl and TBARSer, respectively) were determined. In erythrocytes, the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were measured. In serum, the activity of acid phosphatase (AcP), arylsulfatase (ASA), cathepsin D (CTS D), and α1-antitrypsin (AAT) were determined. Among oxidative stress parameters, only the CAT activity significantly decreased 24 h after HILT compared to measurement 30 min after the treatment in the injured individuals (P < 0.01), while the GPx activity in that group was meaningfully higher 30 min after HILT compared to controls (P < 0.05). Thirty min after the intervention, the activities of AcP and ASA were lower in the injured participants compared to the uninjured ones (P < 0.01 and P < 0.05, respectively). The CTS D activity was lower 30 min and 24 h after HILT in both groups (P < 0.001) and did not differ significantly between them (P > 0.05). Moreover, the study showed statistically significant linear relationships between the TBARSer concentration and the SOD activity before HILT in the healthy participants (r = -0.6, P = 0.021) and 24 h after HILT in the injured ones (r = 0.6, P = 0.025). In the noninjured athletes before HILT, the CTS D activity linearly correlated with the AAT activity (r = -0.70, P = 0.005), and 30 min after the treatment, with the AcP activity (r = 0.5, P = 0.041). 24 h after the HILT intervention, the CTS D and AcP activities were also correlated in the injured athletes (r = 0.8, P = 0.002). The study suggests that one HILT intervention does not significantly influence the redox equilibrium but stabilizes lysosomal membranes.

This article provides an overview of the pathophysiology of chronic obstructive pulmonary disease including the physiological mechanisms that are known precursors. The roles of environmental and genetic causes are considered. α1-Antitrypsin deficiency is also discussed as it relates to the development of airflow obstruction. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of α1-antitrypsin combined with bone marrow mesenchymal stem cells on retinopathy in diabetic rats and its mechanism. Methods: A model of diabetic retinopathy was established by intraperitoneal injection of streptozotocin. The 30 Wistar rats successfully modeled were randomly divided into a model group, a bone marrow mesenchymal stem cell group and a combined group (α1-antitrypsin combined with bone marrow Mesenchymal stem cells), the blood glucose and serum insulin levels of diabetic rats were measured 4 weeks after treatment. Enzyme-linked immunosorbent assay (ELISA) for measuring serum inflammatory factors IL-1ß, IL-6 and TNF-α in rats. Observing the pathological morphology of rat retina under hematoxylin-eosin staining (HE). TUNEL staining to observe the apoptosis of rat retinal nerve cells. Immunohistochemical method to detect the expression level of CD45 in retinal tissue. Real-time fluorescence quantitative PCR was used to detect the expression of retinal vascular endothelial growth factor (VEGF), hypoxiainducible factor-1α (HIF-1α), and angiotensin II (ANGII) mRNA. Western blot was used to detect the expression of p38 MAPK/NF-κB signaling pathway-related proteins in the retinal tissue of each group of rats. Results: Compared with the control group, the rats in the model group had increased blood glucose, decreased insulin levels, increased serum IL-1ß, IL-6, and TNF-α levels, and had obvious lesions in the retina. CD45 showed high expression in retinal tissue, VEGF, HIF-1α, ANGII mRNA expression increased, p-p38, p-p65, p-IκB α protein expression increased (P<0.05). Compared with the model group, the bone marrow mesenchymal stem cell group and the combined group have decreased blood glucose, increased insulin levels, and decreased serum IL-1ß, IL-6 and TNF-α levels. Retinopathy is improved, apoptosis of retinal nerve cells is reduced, CD45 expression in retinal tissue is reduced, VEGF, HIF-1α, ANGII mRNA expression is decreased, and p-p38, p-p65, p-IκB αprotein expression is decreased. Compared with the bone marrow mesenchymal stem cell group, the effect of the combined group was more obvious (P<0.05). Conclusion: a1-antitrypsin combined with bone marrow mesenchymal stem cell transplantation can improve the degree of retinopathy in diabetic rats. The mechanism may be related to the inhibition of p38 MAPK/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Available screening procedures for the detection of α1-antitrypsin-deficient (AATD) mutations have suboptimal cost-effectiveness ratios. The aim in this study was to evaluate and compare the viability of a composite approach, primarily based on the α1-globulin fraction, in identifying AAT genetic analysis eligible patients against standard screening procedures, based on clinically compatible profiling and circulating AAT < 1 g/L. Methods: A total of 21,094 subjects were screened for AATD and deemed eligible when meeting one of these criteria: α1-globulin ≤2.6%; α1-globulin 2.6%–2.9% and AST: >37 U/L and ALT: > 78 U/L; α1-globulin %: 2.9–4.6% and AST: >37 U/L and ALT: >78 U/L and erythrocyte sedimentation rate (ESR) >34 mm/h and C-reactive protein (CRP) >3 mg/L. Subjects were genotyped for the AAT gene mutation. Detection rates, including those of the rarest variants, were compared with results from standard clinical screenings. Siblings of mutated subjects were included in the study, and their results compared. Results: Eighty-two subjects were identified. Among these, 51.2% were found to carry some Pi*M variant versus 15.9% who were clinically screened. The detection rates of the screening, including relatives, were: 50.5% for the proposed algorithm and 18.9% for the clinically-based screening. Pi*M variant prevalence in the screened population was in line with previous studies. Interestingly, 46% of subjects with Pi*M variants had an AAT plasma level above the 1 g/L threshold. Conclusions: A composite algorithm primarily based on the α1-globulin fraction could effectively identify carriers of Pi*M gene mutation. This approach, not requiring clinical evaluation or AAT serum determination, seems suitable for clinical and epidemiological purposes. [ABSTRACT FROM AUTHOR]

Objectives: To determine the levels of endogenous α1‐antitrypsin in the perilymph of patients undergoing cochlear implant (CI), and its reverse association with the severity of hearing loss. Study Design: Retrospective study. Setting: Tertiary care university hospital. Participants: The study includes 38 patients undergoing CI surgery, 11 patients diagnosed with congenital deafness and 27 non‐congenital deafness, eight patients diagnosed with moderate hearing loss (N = 8; PTA = 70 dB), severe hearing loss (N = 11; PTA 70‐90 dB) and profound hearing loss (N = 19; PTA > 90 dB). Main outcome and measure: 1 to 12 μL perilymphatic fluids were collected by micropipette. α1‐antitrypsin levels were determined, and current and historic audiological parameters were obtained. Results: The congenital and non‐congenital group exhibited AAT concentrations of 2.5 ± 1.9 × 106 LFQ and 3.2 ± 1.2 × 106 LFQ, respectively (mean ± SD; P =.38). Mean levels of α1‐antitrypsin in the perilymph fluid within the moderate group was 3.64 × 106 ± 2.1 × 106 LFQ vs 3.5 × 106 ± 1.2 × 106 in severe hearing loss (P =.81) and 2.4 × 106 ± 1.1 × 106 LFQ in the profound hearings loss group (P =.06). The difference in levels of AAT in samples from the severe hearings loss group vs the profound hearings loss group reached statistical significance (P =.04). Conclusion: Insufficiency in α1‐antitrypsin levels in the perilymph fluid of the inner ear appears to display a relationship with the severity of hearing loss. The prospect of introducing clinical‐grade plasma‐purified α1‐antitrypsin directly onto the site of cochlear injury deserves thorough investigation. [ABSTRACT FROM AUTHOR]

We studied the effects of spiperone, a selective blocker of dopamine D2 receptors, on the model of pulmonary emphysema provoked by administration of elastase and D-galactosamine hydrochloride to female C57BL/6 mice and characterized by activation of proteases in the lungs and systemic deficiency of its inhibitor α1-antitrypsin. In this model, spiperone prevented the development of inflammatory reaction and reduced the area of emphysematous expanded alveolar tissue. The expression of angiogenic marker CD31 in the lungs increased under these conditions. Regeneration of the damaged microvascular bed under the action of spiperone resulted from recruiting of Notch1+ endothelial progenitor cells (CD45—CD31+CD34+) into the lungs and blockade of the inhibitory effect of dopamine on phosphorylation of VEGF-2 receptors in endothelial cells of different maturity. In addition, spiperone produced a protective effect on hepatocytes and restored the production and secretion of α1-antitrypsin by these cells. [ABSTRACT FROM AUTHOR]

α1‐Antitrypsin (α1‐AT) serves as an archetypal example for the serine proteinase inhibitor (serpin) protein family and has been used as a scaffold for protein engineering for >35 years. Techniques used to engineer α1‐AT include targeted mutagenesis, protein fusions, phage display, glycoengineering, and consensus protein design. The goals of engineering have also been diverse, ranging from understanding serpin structure–function relationships, to the design of more potent or more specific proteinase inhibitors with potential therapeutic relevance. Here we summarize the history of these protein engineering efforts, describing the techniques applied to engineer α1‐AT, specific mutants of interest, and providing an appended catalog of the >200 α1‐AT mutants published to date. [ABSTRACT FROM AUTHOR]

Objective: to study biomolecules associated with pathology in the respiratory system, in particular, with the development of chronic bronchitis in patients with abdominal obesity. Materials and methods: This is a pilot study. The main group consisted of 158 people with chronic bronchitis, divided into two subgroups: one with abdominal obesity, and the other without it. The control group consisted of 68 people without chronic bronchitis. We determined the blood levels of SP-A, SP-D, α1-antitrypsin, CC16, PARC, and RELM-β. Results: In the first subgroup, patients significantly more often complained of coughing, experienced shortness of breath 1.5 times more often with light physical exertion and 2.7 times more often with moderate physical exertion. In these patients, a Tiffeneau–Pinelli index (FEV1/FVC) below 70% was 1.8 times more common, more patients had FEV1 and FVC of less than 80%, and presented a statistically significant decrease in SP-A, α1-antitrypsin, CC16 levels and an increase in PARC levels than in the second subgroup. Conclusion: In patients with chronic bronchitis and abdominal obesity, there is a decrease in the levels of SP-A, α1-antitrypsin, CC16 and an increase in the level of PARC compared with patients without abdominal obesity, which is probably due to the presence of an additional source of chronic inflammation associated with adipose tissue.

Goal. To determine the blood serum level of α2-macroglobulin (α2-MG), α1-antitrypsin (α1-AT), lactoferrin (LF) and albumin in patients with IgE-mediated and IgE-independent atopic dermatitis (AD). Materials and methods. The authors tested blood serum of 60 adult patients suffering from atopic dermatitis at the exacerbation stage (30 patients with IgE-mediated and 30 patients with IgE-independent atopic dermatitis) and 20 healthy donors in order to determine the level of these proteins by rocket immunoelectrophoresis, enzyme-linked immunosorbent assay and immunoturbidimetry methods. Major results. The albumin level is reliably reduced in case of IgE-mediated AD while the LF and α1-AT levels are increased, and concentrations of LF α1-AT and a2-MG are increased as compared to healthy people. There are differences between the level of LF, α2-MG and α1-AT. There was a statistically significant correlation between the LF levels and SCORAD score in both forms, and α2-MG and α1-AT only in case of IgE-independent AD. Conclusion. These results confirm the difference in the roles of these polyfunctional proteins in the pathogenesis of IgE-mediated and IgE-independent AD.

α1-Antitrypsin (α1-AT) deficiency is a common genetic disorder characterized by low serum α1-AT levels and a clinical manifestation of pulmonary emphysema and liver disease. In addition to its classical manifestations, α1-AT deficiency frequently accompanies granulomatosis with polyangiitis (GPA); in this case the role of α1-AT deficiency in the clinical course of GPA has not been defined. Objective: to estimate the prevalence of pathological α1-AT phenotypes in GPA and other systemic vasculitides (SV) and to determinate their impact on the clinical course of GPA. Subjects and methods. The investigation enrolled 86 patients with SV, including GPA (n=47), microscopic polyangiitis (MPA) (n=16), eosinophilic granulomatosis with polyangiitis (EGPA) (n=12), and polyarteritis nodosa (PAN) (n=11). A control group included 46 healthy donors. Isoelectric focusing was used to phenotype α1-AT in blood samples and its concentrations were determined. The phenotypes of α1-AT were compared with the overall SV activity index using the Birmingham Vasculitis Activity Score (BVAS), the vasculitis damage index (VDI), the nature of an organ lesion, and the markers of immune inflammation (proteinase 3-antineutrophil cytoplasmic antibodies, total IgG, and C3 and C4 fractions of the complement system). Results and discussion. Pathological α1-AT phenotypes were detected in 17% (8/47) of the patients with GPA, 6.25% (1/16) of those with MPA and absent in EGPA and PAN. Patients with GPA had PiZZ (n=1), PiMZ (n=4), PiMF (n=2), and PiMS (n=1) phenotypes; those with MPA had a PiMS-phenotype. The detection of a pathological α1-AT phenotype in patients with GPA was characterized by the high values of BVAS and VDI (p

Heart failure (HF) is considered one of the most common diseases and one of the major causes of death. The latest studies show that HF is associated with an increase in oxidative stress. The use of antioxidants as therapy is effective in animal models, but not in humans. In this review, we analyse some emerging markers related to oxidative stress, evaluating their possible use as therapeutic targets: galectin-3, a β galactoside associated with myocardial fibrosis, α1-antitrypsin, an antiprotease and lectin-like oxidized low-density-lipoprotein receptor-1, the major receptor for ox-LDL. The up-regulation of galectin-3 appears to be associated with HF, atrial fibrillation, dilated cardiomyopathy, fibrogenesis and mortality, while in other cases it seems that galectin-3 may be protective in ischaemia–reperfusion injury. Serum α1-antitrypsin protein levels may increase in the presence of high concentrations of serum proteases, which are over expressed during reperfusion. The overexpression of α1-antitrypsin or the exogenous α1-antitrypsin treatment exhibits an anti-oxidative stress role, evaluated by increased eNOS expression and by decreased MMP9 expression, implicated in HF. The cardiac lectin-like oxidized low-density-lipoprotein receptor-1 is activated by oxidative stress in ischaemia–reperfusion injury, inducing apoptosis in cardiomyocytes through the deleterious NF-kB pathway, while the administration of anti-lectin-like oxidized low-density-lipoprotein receptor-1 antibody suppresses apoptosis and reduces the extent of myocardial infarction. In conclusion, α1-antitrypsin and lectin-like oxidized low-density-lipoprotein receptor-1 seem to represent two good markers in HF and therapeutic targets, whereas galectin-3 does not. [ABSTRACT FROM AUTHOR]

Objectives: To compare levels of α1‐antitrypsin (AT) and lactoferrin (LF) in gingival crevicular fluid (GCF) and on the tongue and determine whether MATATACORO® (MA‐T), a stabilized chlorine dioxide solution, affects GCF AT and LF levels. Methodology: The α1‐antitrypsin and lactoferrin levels in GCF and on the tongue were measured for 18 elderly patients (mean age: 83 years) and correlations were analyzed. AT and LF levels in GCF were assessed before and 1 hr after treatment with MA‐T. Results: The α1‐antitrypsin and lactoferrin levels were lower on the tongue than in GCF (median 0.6, 0.4 μg/mL vs median 2.4, 3.6 μg/mL). The respective AT and LF levels in GCF and on the tongue were positively correlated (AT: r =.43, LF: r =.69). GCF AT and LF levels were reduced following MA‐T treatment (AT: median 3.6 and 1.9 μg/mL and LF: 2.3 and 0.4 μg/mL pre‐ and post‐treatment, respectively; both P <.01). Conclusion: The α1‐antitrypsin and lactoferrin levels in GCF were higher than those on the tongue, but AT and LF were detectable for both sampling types. The reduction in GCF AT and LF levels after MA‐T treatment suggests that MA‐T could be used to control periodontal inflammation. [ABSTRACT FROM AUTHOR]

Triple-negative breast cancer (TNBC) is a subtype of breast cancer. MicroRNA (miR)-214 is closely associated with controlling the development of tumor cells; therefore, in the present study, the target gene and effects of miR-214 on TNBC cells were explored. Luciferase activity was examined by luciferase reporter assay. The viability, invasion and migration of MDA-MB-231 TNBC cells were measured using Cell Counting kit-8, Transwell and wound-healing assays, respectively. The expression levels of various factors were determined using reverse transcription-quantitative polymerase chain reaction and western blotting. The results demonstrated that the expression levels of miR-214 were higher and the levels of α1-antitrypsin (α1-AT) were lower in TNBC tissues compared with in normal tissues. Subsequently, α1-AT was revealed to be a target of miR-214. Furthermore, inhibition of miR-214 decreased cell viability, invasion and migration, enhanced the expression of E-cadherin and tissue inhibitor of metalloproteinases-2, and reduced the expression of metastatic tumour antigen 1 and matrix metalloproteinase-2. Inhibition of miR-214 also significantly downregulated the phosphorylation of protein kinase B (Akt) and mammalian target of rapamycin (mTOR), and markedly downregulated that of phosphoinositide 3-kinase (PI3K); however, the expression levels of total PI3K, Akt and mTOR remained stable in all groups. Taken together, these findings indicated that α1-AT may be a target of miR-214. Downregulation of miR-214 markedly suppressed the viability, migration and invasion of MDA-MB-231 cells, and inhibited the PI3K/Akt/mTOR pathway. These findings suggested that miR-214 targeting α1-AT may be a potential mechanism underlying TNBC development. [ABSTRACT FROM AUTHOR]

Within the 180 SPE tests that were reflexively sent for AAT phenotyping, 142 were found to have a clinically relevant AAT phenotype (Table 1), including five ZZ (severely AAT deficient) phenotypes (2.8% or 1/36). The measured AAT ranged between <=0.2 and 1.7 g/L, with a median of 1.0 g/L. In 180 cases, the AAT level was <1.0 g/L, triggering further investigation with an AAT phenotype. In addition, data were gathered for both requests for serum AAT levels and AAT phenotype results from this period that did not have an associated SPE request ("targeted testing"). [Extracted from the article]

Chronic inflammatory bowel diseases, namely ulcerative colitis and Crohn’s disease, attracted the attention of both doctors and scientists all over the world. There was a trend to an increase in the number of severe, treatment-resistant forms of diseases, complications and surgical interventions that lead to disability in the young, working-age patients. The article shows the problems in the diagnosis of chronic inflammatory bowel diseases (ulcerative colitis, Crohn’s disease).The possibilities of instrumental and laboratory methods of study are shown. Attention is paid to non-invasive diagnostic methods — determination of faecal calprotectin, lactoferrin, lysozyme, elastase, myeloperoxidase, A1-antitrypsin. A literature search was conducted using databases Pub Med, MedLine, EMBASE.

Protein-losing enteropathy (PLE) is a rare complication of intestinal diseases. Its main manifestation is hypoproteinemic edema. The diagnosis of PLE is based on the verification of protein loss into the intestinal lumen, by determining fecal α1-antitrypsin concentration and clearance. The localization of the affected colonic segment is clarified using radiologic and endoscopic techniques. The mainstay of treatment for PLE is a fat-free diet enriched with medium-chain triglycerides. Surgical resection of the affected segment of the colon may be the treatment of choice for severe hypoproteinemia resistant to drug therapy.

The Congenital Disorders of Glycosylation (CDG) are an expanding group of genetic disorders which encompass a spectrum of glycosylation defects of protein and lipids, including N- & O-linked defects and among the latter are the muscular dystroglycanopathies (MD). Initial screening of CDG is usually based on the investigation of the glycoproteins transferrin, and/or apolipoprotein CIII. These biomarkers do not always detect complex or subtle defects present in older patients, therefore there is a need to investigate additional glycoproteins in some cases. We describe a sensitive 2D-Differential Gel Electrophoresis (DIGE) method that provides a global analysis of the serum glycoproteome. Patient samples from PMM2-CDG (n = 5), CDG-II (n = 7), MD and known complex N- & O-linked glycosylation defects (n = 3) were analysed by 2D DIGE. Using this technique we demonstrated characteristic changes in mass and charge in PMM2-CDG and in charge in CDG-II for α1-antitrypsin, α1-antichymotrypsin, α2-HS-glycoprotein, ceruloplasmin, and α1-acid glycoproteins 1&2. Analysis of the samples with known N- & O-linked defects identified a lower molecular weight glycoform of C1-esterase inhibitor that was not observed in the N-linked glycosylation disorders indicating the change is likely due to affected O-glycosylation. In addition, we could identify abnormal serum glycoproteins in LARGE and B3GALNT2-deficient muscular dystrophies. The results demonstrate that the glycoform pattern is varied for some CDG patients not all glycoproteins are consistently affected and analysis of more than one protein in complex cases is warranted. 2D DIGE is an ideal method to investigate the global glycoproteome and is a potentially powerful tool and secondary test for aiding the complex diagnosis and sub classification of CDG. The technique has further potential in monitoring patients for future treatment strategies. In an era of shifting emphasis from gel- to mass-spectral based proteomics techniques, we demonstrate that 2D-DIGE remains a powerful method for studying global changes in post-translational modifications of proteins.

Objective: Preeclampsia is a major cause of mortality in pregnant women but the underlying mechanism remains unclear to date. In this study, we attempted to identify candidate proteins that might be associated with preeclampsia in pregnant women by means of proteomics tools. Materials and methods: Differentially expressed proteins in serum samples obtained from pregnant women with severe preeclampsia (n = 8) and control participants (n = 8) were identified using two-dimensional gel electrophoresis (2-DE) followed by peptide mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Additional serum samples from 50 normal and 41 pregnant women with severe preeclampsia were analyzed by immunoassay for validation. Results: Ten protein spots were found to be upregulated significantly in women with severe preeclampsia. These protein spots had the peptide mass fingerprints matched to α1-antitrypsin, α1-microglobulin, clusterin, and haptoglobin. Immunoassays in an independent series of serum samples showed that serum α1-antitrypsin, α1-microglobulin, and clusterin levels of severe preeclampsia patients (n = 41) were significantly higher than those in the normal participants (n = 50; α1-antitrypsin 295.95 ± 50.94 mg/dL vs. 259.31 ± 33.90 mg/dL, p = 0.02; α1-microglobulin 0.029 ± 0.004 mg/mL vs. 0.020 ± 0.004 mg/mL, p

Abstract Under stressful conditions, cellular heme catabolism to carbon monoxide, iron and biliverdin is mediated by the 32 kDa enzyme, heme oxygenase-1 (HO-1). A wide range of pro-oxidant and inflammatory stimuli act on diverse consensus sequences within the Hmox1 promoter to rapidly induce the gene. There is ample evidence attesting to the beneficial effects of HO-1 upregulation in brain. By converting pro-oxidant heme to the antioxidants, biliverdin and bilirubin, HO-1/biliverdin reductase may help restore a more favorable tissue redox microenvironment. Contrariwise, in some cell types and under certain circumstances, heme-derived carbon monoxide and iron may amplify intracellular oxidative stress and exacerbate the disease process. This inimical side of neural HO-1 has often been ignored in biomedical literature promulgating interventions aimed at boosting central HO-1 expression for the management of diverse CNS conditions and is the focus of the current review. A comprehensive model of astroglial stress is presented wherein sustained Hmox1 induction promotes oxidative mitochondrial membrane damage, iron sequestration and mitophagy (macroautophagy). The HO-1 mediated gliopathy renders nearby neuronal constituents vulnerable to oxidative injury and recapitulates 'core' neuropathological features of many aging-related neurodegenerative and some neurodevelopmental brain disorders. A balanced literature should acknowledge that, in a host of chronic human CNS afflictions, the glial HO-1 response may serve as a robust transducer of noxious stimuli, an important driver of relevant neuropathology and a potentially disease-modifying therapeutic target. [ABSTRACT FROM AUTHOR]

Human α1-antitrypsin (AAT) is the most abundant serine proteinase inhibitor (serpin) in the human plasma. Commercially available AAT for the medications of deficiency of α1-antitrypsin is mainly purified from human plasma. There is a high demand for a stable and low-cost supply of recombinant AAT (rAAT). In this study, the baculovirus expression vector system using silkworm larvae as host was employed and a large amount of highly active AAT was recovered from the silkworm serum (~ 15 mg/10 ml) with high purity. Both the enzymatic activity and stability of purified rAAT were comparable with those of commercial product. Our results provide an alternative method for mass production of the active rAAT in pharmaceutical use. [ABSTRACT FROM AUTHOR]