Helicobacter pylori (H. pylori) induces the production of tumor necrosis factor-alpha (TNF-α), which is closely related to a gastric epithelial injury. TNF-α gene polymorphism and TNF-α serum levels are associated with various malignant conditions. Identification of the ideal marker for gastric cancer (GC) is still the leading aim of several trials. Physio-pathological considerations of GC led us to investigate the association of two TNF-α promoter polymorphisms (-308G>A and -238G>A), and TNF-α serum levels with the susceptibility to gastric precancerous (PL) and GC. H. pylori infection , and 63 healthy controls (HC) were involved in this study. Individuals are genotyped by TNF-α gene promoter sequencing and TNF-α serum levels are measured by ELISA quantitative method. Patients suffering from gastric lesions  related to Regarding TNF-α-308 G/A locus, we noticed higher risk for GC  and PL  for individuals with AA/GA genotypes compared to GG genotype. Concerning TNF-α-238 G/A locus, we noticed higher risk for GC  and PL  for individuals with GG genotype compared to AA/GA genotypes. We noticed that TNF-α serum levels have been increased together with gastric lesions severity. Moreover, TNF-α-308 and TNF-α-238 A alleles seemed to, respectively, upregulate and downregulate TNF-α serum levels. TNF-α gene polymorphisms and TNF-α serum levels might be helpful to select those patients who are at high risk for GC. The TNF-α -308 A allele has a promotive effect for GC progression, whereas the TNF-α -238 A allele has a protective function against GC progression. High levels of TNF-α seemed to be associated with the aggressiveness of gastric lesions.  Helicobacter pylori (H. pylori)-driven inflammation, which leads to chronic gastritis which progresses to atrophy, and intestinal metaplasia. The metaplastic epithelium can undergo from other genomic and phenotypic disorganizations (dysplasia), which can then progress to invasive neoplasia  .Gastric cancer (GC) is one of the most aggressive neoplasms worldwide. It is responsible for over 1,000,000 new cases in 2018 and an estimated 783,000 deaths, making it the fifth most frequently diagnosed cancer and the third leading cause of cancer death . The process of gastric carcinogenesis is triggered by H. pylori infection induces chronic inflammation in gastric mucosa which is a critical step in gastric carcinogenesis . Tumor Necrosis Factor-alpha (TNF-α) is one of the major cytokines associated with H. pylori infection and it is closely related to gastric epithelial injury . TNF-α is a multifunctional cytokine that elicits various biological functions such as immune homeostasis, inflammatory reaction, and cell surveillance . In cancer pathologies, TNF-α plays a paradoxical role. On one hand, this cytokine can act as a tumor suppressor through vascular destruction, tumor necrosis, and immunostimulatory effects. On the other hand, TNF-α can act as a tumor promoter by inducing cellular transformation, survival, proliferation, invasion, angiogenesis, and metastasis  . TNF-α gene  contains a very large number of regulatory elements that affect the transcription of the gene . The expression of TNF-α is tightly regulated, in part, at the transcriptional level. Several polymorphisms within TNF-α promoter region  have been reported to influence the expression of TNF-α gene and diseases susceptibility  . The gene coding for TNF-α is located in the short arm of chromosome 6 (6p21) within the highly polymorphic region of the major histocompatibility complex. Its location and strict linkage disequilibrium present between some alleles of HLA-DR class II and HLA-B class I genes have allowed to hypothesize associations between TNF-α alleles and some diseases . The promoter region of the TNF-α gene encodes for a transmembrane protein (tmTNF-α) consisting of 233 amino acids (26 KDa) that is released in a soluble form (sTNF-α) of 157 amino acids (17 KDa) by TNF-converting enzyme-mediated cleavage . The different TNF-α isoforms can have different effects on inflammatory responses and tumor development. Based on several studies, it seems that tmTNF-α reduces the severity of inflammatory reactions and tumor growth, while sTNF-α stimulates those processes . The A high level of TNF-α is associated with aggressive behavior and poor prognosis in many malignant cancers . The sTNF-α is detected at low doses in the serum of healthy subjects, while high levels are detected in patients suffering from cancer and inflammatory diseases . Decreases in TNF-α serum levels in patients receiving chemotherapy have been reported by many studies . This suggests that the serum levels of sTNF-α could be an indicator of the response to chemotherapy and tumor prognosis.In most patients, GC is diagnosed in an advanced stage, which explains the poor prognosis and the high mortality rates. GC develops via several sequential lesions, which representing an opportunity for early detection and intervention to prevent the progression of precancerous lesions (PL) towards the GC. The aim of the present study is to evaluate the potential of TNF-α genotyping  and TNF-α serum levels as predictive markers in GC. Clinical and pathological characteristics of the study population H. pylori infection collected from Gastroenterology and Oncology departments at the IBN ROCHD University Hospital Center. Patients having received previous treatment for H. pylori eradication, proton pump inhibitors, anti-inflammatory medicines, chemotherapy or radiotherapy treatment, patients suffering from any other infection, inflammatory disorders, cancers other than distal gastric adenocarcinoma, were excluded from the study. Healthy asymptomatic subjects, with no history of gastrointestinal illnesses or regular use of any gastrointestinal and anti-inflammatory medicines, were recruited from the Regional Transfusion Center of Casablanca .Three biopsies  were sampled from patients admitted for endoscopy. Blood samples have been sampled from all participants. Our study focused on patients suffering from gastric pathologies related to Clinical information about the demographic characteristics of the participants including age, sex, place of birth, drug consumption , smoking and alcohol habits, were collected using a structured questionnaire. All participants were informed about their inclusion in the study and agreed to it in writing form. The study protocol has been performed in accordance with the ethical standards of Helsinki and was approved by the ethical committee of Pasteur Institute of Morocco. H. pylori infection. The biopsies were fixed in 10% formalin and taken to Histo-Cytopathology laboratory in Pasteur Institute of Morocco, where they were embedded in paraffin blocks, sectioned and stained in hematoxylin and eosin for conventional histopathology examination of gastric mucosal lesions and H. pylori infection in the healthy controls subjects, using a commercially available kit ). A cut-off value of antibody concentration ≥ 20 relative units (RU)/ml was considered as positive.The quantitative anti-H.Pylori IgG ELISA method was performed to detect DNA extraction and quantification Genomic DNA was extracted from blood taken on EDTA tube using the commercially available kit (PureLink™ Genomic DNA Mini Kit). The quantity, quality, and purity of DNA were checked by 0.8 % agarose gel electrophoresis and also by the ratios of Optical Density at 260/280 nm and 260/230 nm using a spectrophotometer (NanoVue plus). Then it was stored at −20°C until use.Genotyping of TNF-α -308 /-238 polymorphisms TNF-α gene promoter at position -308 (G/A) and -238 (G/A) by using sequencing.Individuals are genotyped for TNF-α gene (266 bp) was amplified by PCR. The reaction was carried out in a total volume of 20 µL, consisting of 200 ng of genomic DNA, 0.5 µM of primers , 1 mM de dNTPs, 3 mM MgCl2 and 0.5 unit of Taq polymerase (BIOLINE). Thermocycling conditions of PCR were as follows: initial denaturation at 95°C for 1min, followed by 35 cycles of 95°C for 15 seconds, 57°C for 15 seconds, 72°C for 30 seconds, and a final extension at 72°C for 7 min. The PCR products were analyzed by 1.5% agarose gel electrophoresis, purified using ExoSap  in a final volume of 7 µL, consisting of 2 µl ExoSAP enzyme and 5µl of PCR product, and incubated at 37°C for 4min and 90°C for 1min. Then, The DNA sequencing conditions was carried out on the basis of the method described by Essadik (2005) by using BigDyeR Terminator v3.1 Cycle Sequencing Kit (AppliedBiosystems). The DNA sequences were read by an Applied Biosystems 377 DNA sequencer and analyzed using the BIOEDITE software package. Firstly, the Measurement of TNF-α serum Levels Blood samples were collected in a serum tube from each patient before anesthesia and gastroscopy examination and conserved in an icebox during transport. The serum was separated within 1 h of blood collection after spinning for 15 min at 1,500 g. Serum samples were stored at -80 °C in aliquots of 500 µL and thawed just before testing. Serum levels of TNF-α were measured using the Human TNF-α ELISA kit from R and D Systems  and it was used according to the manufacturer’s instructions. Total TNF-α concentrations in samples were expressed as pg/ml. Statistical analysisThe results of this study were statistically analyzed using R software version 1.1.456. The descriptive data are presented in terms of frequencies. The differences between the groups were analyzed with the chi-square or Fisher’s exact test for the categorical variables, Wilcoxon test for the qualitative (2 classes) and numeric variables, and the Kruskal Wallis test for the qualitative (3 classes and more) and numeric variables. The differences were considered significant at p-value <0.05. Clinical and pathological characteristics of the study population H. pylori-positive status were selected in the present study and divided into two categories. The first category includes 63 asymptomatic healthy controls subjects (HC). The second category includes 155 patients with various gastric lesions related to H. pylori infection: 65 with chronic gastritis, 50 with PL  and 40 with gastric adenocarcinoma, were investigated. Male/Female ratio in the case group and the control group was, respectively, 73/82 and 34/29. The mean age of HC, chronic gastritis, PL and GC was, respectively, 43 ±8, 45±16, 53±14, 56±14 years. In total, 218 participants with Genotyping of TNF-α -308 /-238 polymorphisms Regarding TNF-α promoter -308 G/A locus, the frequency of A allele is 8% in HC and has shown an increase with the severity of gastric lesions, with the following order: 10.8% in chronic gastritis, 21% in PL and 27.5% in GC. Besides, the frequency of GA genotype was observed higher in patients with chronic gastritis, PL and GC  compared with the HC group (6.3%). Whereas the AA genotype was more occurred among patients with PL and GC (12% and 20% respectively) compared to patients with chronic gastritis and HC (4.6% and 4.8% respectively) . SignifiRegarding TNF-α promoter -238 G/A locus, the frequency of A allele is 17% in HC and it was observed in decline together with the increase of gastric lesions severity, in the following order: 10.7% in chronic gastritis, 4% in PL and 3.5% in GC. Besides, the genotypes GA and AA has occurred more among HC (14.3% and 9.5% respectively) and patients with chronic gastritis (12.3% and 4.6% respectively) compared to patients suffering from PL (4% and 2% respectively) and GC (2.5% and 2.5% respectively). The genotypes and alleles distribution of TNF-α -238 G/A locus were statistically different between HC and patients with PL and GC  . BesidesMeasurement of TNF-α serum Levels The relationship between serum levels of circulating TNF-α and the severity of gastric lesions was investigated. The distribution of TNF-α levels in each group of patients and controls is shown in The serum levels of circulating TNF-α were observed to be increased together with the severity of gastric lesions. TNF-α serum levels in the GC group were higher than those recorded in patients with PL and patients with chronic gastritis compared to HC. TNF-α serum levels mean were as follows 36 pg/ml in HC, 50.7 pg/ml in chronic gastritis, 55 pg/ml in PL, and 116.6 pg/ml in GC. A disparity in the TNF-α serum levels between patients and HC, and between GC and the other gastric lesions cases were noted, and it was statistically meaningful . Association between TNF-α -308 /-238 polymorphisms and TNF-α serum levels The impact of the TNF-α -308 G/A and -238 G/A polymorphisms on its serum levels were assessed. Regarding TNF-α -308 G/A locus, the TNF-α serum levels were observed to be increased with possession of the A allele. The mean level of TNF-α was 44.6 pg/ml within individuals harboring GG genotype, 75.1 pg/ml within GA, and 205.4 pg/ml within those harboring AA. The difference was statistically significant between TNF-α -308 polymorphism and TNF-α serum levels . Concerning TNF-α -238 G/A locus, the TNF-α serum levels were observed to be decreased with possession of the A allele. The mean level was 54.4 pg/ml, 36.5 pg/ml and 34.1 pg/ml within individuals harboring GG, GA and AA genotypes, respectively. The difference was statistically significant between TNF-α -238 polymorphism and TNF-α serum levels . Identification of the ideal marker for GC is still the leading aim of several trials. Physio-pathological considerations of gastric carcinogenesis led us to evaluate the potential of TNF-α genotyping  and TNF-α serum levels as predictive markers of GC.In our population, the evaluation of the association between TNF-α polymorphisms and the susceptibility to GC has revealed a noteworthy link.Regarding TNF-α -308 G/A locus, the frequency of the TNF-α -308 A allele was observed to increase with gastric lesions severity. Notable disparity in TNF-α -308 genotypes frequencies is observed between patients and HC. Indeed, the GA genotype was more often detected in patients with chronic gastritis, PL and GC compared to HC. Whereas the AA genotype was mostly detected in patients suffering from PL and GC. Our results are consolidated with several studies reported the association of TNF-α -308 G/A polymorphism with increased susceptibility to various cancers, including hepatocellular carcinoma, prostate cancer, oral cancer, lung cancer, cervical cancer, oesophageal cancer, breast cancer and colorectal cancer . In gastric diseases, TNF-α-308 G/A is associated with a high risk of chronic atrophic gastritis, intestinal metaplasia and GC . Regarding TNF-α -238 G/A polymorphism, the frequency of TNF-α -238 A allele and TNF-α-238 GA/AA genotypes were observed lower in patients with PL and GC compared to chronic gastritis and HC. According to other reports, TNF-α -238 promoter polymorphism seems to be associated with attenuated susceptibility to various cancers, including gastric carcinoma, uterine cervical carcinoma, colorectal carcinoma, renal cell carcinoma and lung cancer . The relationship between serum levels of circulating TNF-α and gastric carcinogenesis was investigated in this study. A significant disparity of TNF-α serum levels between cases and controls was noticed. We remarked that TNF-α serum levels increase among patients with chronic gastritis, with PL and mainly in those suffering from GC.H. pylori infection and in contributing to gastric inflammation . TNF-α is implicated in intestinal metaplasia and hyperplastic gastric tumors , through the stimulation of some proteins expression required in the development of intestinal cell, such as CDX1 and CDX2, intestinal mucin MUC2 and MUC4 . These findings support the elevation of TNF-α serum levels in our patients suffering from chronic gastritis, PL and GC.High levels of TNF-α have been described in several studies among patients suffering from cancer, including gastric carcinoma, colorectal carcinoma , hepatocellular carcinoma , head and neck cancer . Furthermore, a high level of TNF-α is associated with aggressive behaviour and poor prognosis in many malignant cancers . TNF-α plays a critical role in defending against The regulation of TNF-α expression is in part genetically determined. Our finding suggests that TNF-α-308 A allele may be related to an increase in TNF-α serum levels. This result agreed with previous studies reported the positive impact of this polymorphism on TNF-α production . Regarding TNF-α-238 G/A locus, we found TNF-α-238 A allele associated with a decrease in TNF-α serum levels. This finding is supported by other studies reported the downregulation of TNF-α production by TNF-α-238 polymorphism . TNF-α -308 A allele was more recorded in the GC group and was linked to high TNF-α levels. In contrast, TNF-α -238 A allele was less detected in the GC group and was associated with low TNF-α levels, we can suggest that the elevation of circulating TNF-α is implicated in the promotion of GC and not in its regression as a double act of this cytokine vis-a-vis malignant and precancerous cell. It seems that tmTNF-α reduces the severity of inflammatory reactions and tumor growth, while sTNF-α stimulates those processes .In conclusion, TNF-α -308 A and TNF-α -238 A allele seem to have a promotive and protective effect respectively on GC development. Higher TNF-α serum levels appear associated with gastric carcinogenesis. Furthermore, TNF-α -308 A and TNF-α -238 A allele seem to upregulate and downregulate TNF-α serum levels, respectively. Further studies including a large population and diverse ethnicities are needed to illumine the association of TNF-α-308 G/A and TNF-α-238 G/A polymorphisms and TNF-α serum levels with GC. QRT-PCR and Western blotting assays are recommended to confirm the relationship between TNF-α expression and TNF-α polymorphisms .


Heart disease is a leading cause of death with unmet clinical needs for targeted treatment options. Tumor necrosis factor alpha (TNF-α) represents a master pro-inflammatory cytokine that plays an important role in many immunopathogenic processes. Anti-TNF-α therapy is widely used in treating autoimmune inflammatory disorders, but in case of patients with heart disease, this treatment was unsuccessful or even harmful. The underlying reasons remain elusive until today. This review summarizes the effects of anti-TNF-α treatment in patients with and without heart disease and describes the involvement of TNF-α signaling in a number of animal models of cardiovascular diseases. We specifically focused on the role of TNF-α in specific cardiovascular conditions and in defined cardiac cell types. Although some mechanisms, mainly in disease development, are quite well known, a comprehensive understanding of TNF-α signaling in the failing heart is still incomplete. Published data identify pathogenic and cardioprotective mechanisms of TNF-α in the affected heart and highlight the differential role of two TNF-α receptors pointing to the complexity of the TNF-α signaling. In the light of these findings, it seems that targeting the TNF-α pathway in heart disease may show therapeutic benefits, but this approach must be more specific and selectively block pathogenic mechanisms. To this aim, more research is needed to better understand the molecular mechanisms of TNF-α signaling in the failing heart. Heart disease refers to a group of diseases characterized by the affected function of the heart muscle. Epidemiologic data suggest that heart disease is a leading cause of death in the world. Heart failure affects 26 million people worldwide and causes 1 million annual hospitalizations in the United States and Europe .Inflammation plays an important role in the progression of many types of cardiovascular disease. On the one hand, the systemic inflammatory condition enhances atherogenesis, leading to coronary artery disease, but it also can promote the development of diastolic heart failure. On the other hand, cardiac inflammation occurs in post-ischemic myocardial events, and in more rare cases, it develops as a response to non-ischemic cardiac injury that often causes pathogenic changes in the cardiac tissue resulting in systolic dysfunction . Thus, aTumor necrosis factor α (TNF-α) represents one of the most potent pro-inflammatory cytokines, and therefore, it was selected as the first target in the cytokine-targeted approach. Currently, TNF-α inhibitors are clinically used anti-inflammatory drugs to treat mainly patients with systemic inflammatory diseases. Roughly 1 million patients receive this type of treatment, and TNF-α antagonists are currently the most profitable class of drugs in the world, accounting for 25 billion US dollars in sales annually . In caseTNFA gene located on chromosome 6 and shares locus with major histocompatibility complex (MHC) class II genes, which plays a central role in antigen presentation [TNFA gene consists of 200 nucleotide promoters with binding sites for several transcription factors, resulting in a high plasticity of transcription and responsiveness to various types of stimuli, which also vary between cell types [In humans, TNF-α is encoded by the entation . The TNFll types .TNFA transcripts. Pro-inflammatory stimuli, such as lipopolysaccharide (LPS), regulate the activity of TTP and the translocation of HuR from the nucleus to cytoplasm, thereby enhancing TNF-α biosynthesis. TTP activity is regulated by p38 mitogen-activated protein kinases (MAPK), which controls TTP target genes at the post-transcriptional level and the binding of nuclear factor kappa B (NF-κB) to the promoter of TTP, which positively regulates its translation [At the post-transcript stage, the biosynthesis of TNF-α is controlled mainly through the competitive binding of the mRNA 3′ AU-rich untranslated region by RNA-binding proteins tristetraprolin (TTP) and stabilizing factor human antigen R (HuR). The dephosphorylated form of TTP effectively binds to mRNA and degrades it. The phosphorylation of TTP weakens its affinity to mRNA preventing its degradation. This allows the binding of HuR to mRNA and enables a more efficient translation of nslation ,10,11. Tnslation . The bionslation ,14,15.C-terminal end oriented toward cytoplasm) transmembrane protein. This membrane form of TNF-α (mTNF-α) can function as a ligand. The extracellular domain of mTNF-α can be cleaved by TNF-α cleaving enzyme  and released as soluble TNF-α (sTNF-α) [Upon translation, TNF-α is synthesized as a 17 kDa type II  . mTNF-α TNF-α represents a ligand for two types of TNF-α receptors (TNFRs), namely TNFR1 CD120a, p55) and TNFR2 . TNFRs represent single transmembrane glycoproteins with extracellular TNF-α binding domains characterized by four tandem-repeated cysteine-rich motifs . TNFRs aa, p55 anIn the presence of the ligand, TNFR1 recruits a number of adaptor proteins including TNFR-associated death domain (TRADD), TNF-α receptor associated factor 2 (TRAF2), receptor-interacting protein (RIP) kinase, inhibitors of apoptosis proteins (IAPs), Fas-associated death domain (FADD) and MAPK activating death domain (MADD) ,21. The Unlike TNFR1, TNFR2 lacks the intracellular death domain and is unable to bind TRADD and initiate caspase-mediated apoptosis. Instead, activated TNFR2 recruits adaptor proteins TRAF2 and IAPs, activating the canonical NF-κB signaling through IKK. In addition, TNFR2, TRAF2, and IAPs form a complex with NF-κB-inducing kinase (NIK). As a result, NIK is released from the complex and activated. Active NIK induces the non-canonical NF-κB pathway through IKKα that ultimately produces a transcriptionally active p52 subunit . TNFR2 cmTNF-α acts not only as a ligand for TNFRs triggering forward signaling in the target cells but also transduces a reverse signaling back to the mTNF-α expressing cell. Physiologically, the mTNF-α reverse signaling is triggered by TNFR2 expressed by the neighboring cells . FurtherIn the 20th century, TNF-α has been recognized as the key pro-inflammatory cytokine in humans. This led to the development of the first cytokine-targeted therapy with etanercept approved by the Food and Drug Administration (FDA) in 1998. Anti-TNF-α therapy has revolutionized the treatment of autoimmune inflammatory diseases by offering an alternative for non-specific immunosuppressive drugs, which cause multiple adverse effects for a long-term use . To dateTNFA gene was associated with increased risk of coronary heart disease development [The positive effects of anti-TNF-α therapy in autoimmune inflammatory diseases encouraged testing its therapeutic value in patients with systolic heart failure. These patients are characterized by elevated plasma levels of TNF-α and other pro-inflammatory cytokines ,40,41,42elopment  and in celopment . Pilot selopment ,48, as welopment . Nonetheelopment . Furtherelopment . The injelopment . In concUnlike for HFrEF patients, a long-term anti-TNF-α therapy in patients with autoimmune inflammatory diseases is generally not harmful, and it may even protect from enhanced cardiovascular complications and cardiovascular death ,54. AlthPsoriasis represents another autoimmune inflammatory disease associated with increased serum levels of TNF-α . SimilarSummarizing, in the light of published clinical data  improved cardiac functions and completely protected from cardiac death [Tnfrsf1b (gene encoding TNFR2) genetic deficiency exacerbated heart failure and increased the lethality of TNF-α overexpressing mice pointing to the cardioprotective role of TNFR2 [A gain-of-function approach is based on the overexpression of the gene of interest in a cell or in an organism and represents one strategy to study its function. A popular knock-in mice model non-specifically overexpressing human TNF-α shows high TNF-α production mainly in synovial fibroblasts and in endothelial cells (without evident effects in other cell types) and develops severe erosive arthritis ,79. Therac death . On the of TNFR2 . Interesof TNFR2 . This maAtherosclerosis is a pathogenic condition of arteries characterized by the development of atherosclerotic plaques. A rupture of atherosclerotic plaques and the subsequent blood clot formation can cause life-threatening ischemic events, such as myocardial infarction or sudden cardiac death. In a typical clinical scenario of myocardial infarction, atherosclerotic plaque rupture and the subsequent thrombosis blocks blood flow in larger coronary arteries, causing an ischemic condition in the myocardium and death of cardiomyocytes. Myocardial infarct size depends on the time and the extent of ischemia as well as the subsequent inflammatory response. In animal models, myocardial infarction is typically performed in non-atherosclerotic condition.−/−Apoe or Ldlr−/− mouse strains. In mice, TNF-α has been recognized as one of the most potent proatherogenic cytokines, and the formation of atherosclerotic lesions was preceded by an increased expression of TNF-α, TNFR1, and TNFR2, which were elevated even further during plaque growth [Tnfa gene in Apoe−/− mice fed a high-fat diet retarded the progression of plaque growth and decreased levels of pro-atherosclerotic factors without affecting cholesterol levels [APOE*3-Leiden Tnfa−/− strain fed a high-cholesterol diet also demonstrated that TNF-α promoted necrosis in plaque-infiltrating cells and enhanced advanced lesion formation [Apoe−/− mouse model of atherosclerosis [Apoe−/− mice on a high-fat, high-cholesterol diet receiving recombinant TNF-α developed an enhanced atherosclerotic phenotype, which could be reversed by NF-κB inhibitors [Experimental atherosclerosis is typically induced by feeding animals an atherogenic high-fat, high-cholesterol diet over at least 3 months. The development of atherosclerotic plaques is significantly enhanced in mice with naturally elevated plasma cholesterol levels, such as e growth . Knockoul levels ,89. Expeormation . Data obclerosis . In linehibitors .Apoe−/− model [Apoe−/− mice kept in hypoxia conditions and fed a high-fat diet [Ldlr−/− mice, monotherapy with etanercept failed to reduce the development of atherosclerotic plaques, and the atheroprotective effect of etanercept was observed only in combination with cholesterol-lowering drugs [Ldlr−/− mice [Adam17) showed enhanced macrophage adhesion and atherosclerosis in an Ldlr−/− mouse model [Tnfrsf1a−/− were initially reported to develop enhanced atherogenesis when fed an atherogenic diet [Tnfrsf1a−/− mice on a proatherogenic Apoe−/− background showed reduced atherosclerosis, and the data pointed to the key role of TNFR1 expressed in arteries [Apoe−/−, rather than Ldlr−/− or non-transgenic mice, represents the most relevant animal model to study TNF-α signaling in atherosclerosis.In contrast to data from transgenic models, the results of the pharmacological inhibition of TNF-α in mouse models were less consistent. It has been demonstrated that recombinant soluble TNFR1 successfully attenuated the formation of aortic lesions in an /− model . Beneficfat diet . Howeverng drugs . Instead−/− mice . These i−/− mice . Althoug−/− mice ,98,99. Onic diet . Howeverarteries . SummariTnfa−/− mice showed a significantly smaller infarct area, decreased expression of intercellular adhesion molecule 1 (ICAM-1), and lower numbers of heart-infiltrating neutrophils and macrophages [Tnfa−/− mice also in a myocardial ischemia–reperfusion injury model [Tnfrsf1a−/− mice were consistently reported to develop less impaired cardiac contractile functions and showed better survival rates up to several weeks after infarction [Tnfrsf1b−/− mice showed exacerbated cardiomyocyte apoptosis and fibrosis as well as worsened cardiac function and long-term survival in a permanent occlusion model [Experimental acute myocardial infarction is typically achieved by permanent or temporary mechanical ligation of the left anterior descending coronary artery. Animals surviving this procedure eventually develop fibrotic scars (that replaces necrotic myocardium), and their hearts show impaired function and hemodynamic abnormalities. Published data report elevated sTNF-α levels in the serum of post-infarcted mice and increased mTNF-α expression in the infarct and peri-infarct zones . In a perophages . Howeverrophages . A smallry model . Similarry model . Moreovery model . In anotry model . The injry model . Pathogefarction ,112,113.farction . The patfarction . Unlike on model ,111. Of on model  and cardon model . SummariNon-ischemic heart diseases refer to cardiac abnormalities occurring in the absence of coronary artery diseases. Cardiomyopathies represent the most common type of non-ischemic heart disease, in which ventricles become enlarged and stiff. Cardiomyopathies may be caused by an abnormally thick myocardium (hypertrophic cardiomyopathy) or by the dilatation of the ventricles (dilated cardiomyopathy) . HypertrTnfa−/− mice developed significantly lower inflammatory response, cardiac hypertrophy, and left ventricular remodeling, showing preserved cardiac functions in several studies [Tnfrsf1a−/− mice are partially protected from transverse aortic constriction-induced hypertrophy and are characterized by better survival rates [Tnfrsf1b showed worsened survival rates and increased cardiac hypertrophy, and the cardioprotective TNFR2 signaling has been linked to its effects in mitochondria [Experimental hypertension in rodents is commonly achieved by the partial occlusion of the aorta (transverse aortic constriction model) or by continuous infusion of the vasoconstrictor angiotensin II using osmotic minipumps. In these models, increased blood pressure in the heart induces a compensatory mechanism, by which the left ventricle becomes over time thicker and thereby less effective in muscle relaxation. On the cellular level, cardiac tissue is characterized by cardiomyocyte hypertrophy and interstitial fibrosis. Data from a mouse model of pressure overload induced by aortic banding pointed to the active role of TNF-α–TNFR1 signaling in the development of hypertensive cardiomyopathy, as initially suggested by the correlation between progressive hypertrophy and increasing myocardial levels of TNF-α, TNFR1, and TACE ,121. In  studies ,123. Thi studies , changes studies , decreas studies . The patal rates . In the al rates . This pachondria . Moreovechondria .Tnfa−/− and Tnfrsf1a−/− mice showed significantly attenuated phenotype [Tnfrsf1a−/− mice, but there was no protective effect on diastolic dysfunction in this model [Tnfrsf1b−/− mice receiving angiotensin II infusion developed fibrosis and showed only slight changes in expression of pro-fibrotic genes [Similar data were obtained in another model of hypertrophic cardiomyopathy. In an angiotensin II osmotic minipump model, henotype ,127. In-ic genes . Thus, iCardiac hypertrophy associated with diastolic dysfunction can be alternatively induced by the continuous delivery of β-adrenergic agonist isoproterenol. In this model, mice lacking TNFR1 developed reduced inflammatory response, but this was insufficient to protect them from isoproterenol-induced hypertrophy, whereas mice deficient of TNFR2 showed an increased pro-inflammatory response and exacerbated cardiac hypertrophy . Of noteMice consuming increasing doses of ethanol over longer periods of time develop a specific type of alcoholic cardiomyopathy with fibrotic and structural changes in the left ventricle. In this model, mice showed TNFR1-dependent elevated serum levels of TNF-α, left ventricle dysfunction, and increased cardiac ROS and pro-inflammatory cytokine production . IncreasSummarizing, data from hypertrophic cardiomyopathy models suggest that targeting TNF-α indeed might successfully prevent from disease development. It should be noted that as in other cardiovascular diseases, pathogenic TNF-α signaling is mainly mediated by TNFR1.+ T lymphocytes. Published data suggest that the TNF-α–TNFR1 axis plays an active role in the development of myocarditis. In a coxsackievirus B3 model, Tnfa−/− and Tnfrsf1a−/− but not Tnfrsf1b−/− mice showed strikingly reduced cardiac inflammation [Tnfrsf1a−/− mice were completely protected from the development of myocarditis induced by immunization with cardiac myosin or by an adoptive transfer of autoreactive T lymphocytes [In animal models, heart-specific inflammation is induced either by infection with cardiotropic virus (mainly coxsackievirus B3) or by the active induction of heart-specific autoimmunity . In the ammation . Interesammation . Furtherphocytes . AlthougThe heart is made up of three main cell types: cardiomyocytes, cardiac microvascular endothelial cells, and cardiac stromal cells (mainly fibroblasts). In addition, heart-resident macrophages represent a small but important cell population in the healthy heart. Furthermore, in response to injury, cardiac tissue is infiltrated by immune cells, such as inflammatory monocytes and lymphocytes. It should be noted that TNF-α can be produced by many cardiac cell types, but immune cells seem to represent a particularly important source of this cytokine in cardiac pathology. TNF-α signaling controls many biological processes ranging from pro-inflammatory and proapoptotic to regenerative and cardioprotective . The actThe activation of endothelial cells represents one of the best described pro-inflammatory mechanisms. Endothelial cells respond to TNF-α by an increased expression of adhesion molecules, which control the rolling and adhesion of inflammatory immune cells into the tissue. In this mechanism, both TNFRs are also critically engaged in the process of diapedesis . TNF-α iIn cardiomyocytes TNF-α triggers a hypertrophic response and induces apoptosis . EndogenCardiac fibroblasts represent another heart-resident cell type that can be activated by TNF-α. In response to TNF-α, cardiac fibroblasts contribute to the inflammatory cascade by secreting monocyte chemoattractant proteins (MCP)-1 and MCP-3, which control monocyte recruitment but also positively regulate TNF-α production . FurtherMost of the cardioprotective mechanisms are mediated by TNFR2. It seems that one of the most important effects of TNFR2-dependent signaling is to suppress activation of the pathogenic TNFR1 downstream pathways. It has been observed that in the absence of TNFR2, there is an increased activity of TNFR1 downstream effector molecules NF-κB  and MAPKTNF-α is undoubtedly an important pro-inflammatory cytokine playing a key role during the early inflammatory phase. Clinical studies and data from animal models confirmed that TNF-α enhanced the development of a number of cardiovascular pathologies. However, preventive anti-TNF-α therapy would not be recommended due to side effects and economic reasons. Instead, such targeted immunomodulatory therapeutic intervention was considered in heart failure patients, but the failure of anti-TNF-α therapy could be explained by the cardioprotective properties of TNF-α in the failing heart. Currently, our knowledge on the beneficial activity of TNF-α is limited; therefore, more experimental research is needed to uncover these processes and the underlying mechanisms. Available data suggest that most of the cardioprotective activity is mediated by TNFR2, whereas the activation of TNFR1 initiates pathogenic processes. So far, most of these findings have been obtained in mouse models using transgenic mice lacking one or the other TNFR. In the next step, targeting these receptors, either by blocking TNFR1 or activating TNFR2, should be performed pharmacologically. The development of drugs selectively targeting TNFRs, rather than blocking TNF-α activity, might represent a novel and more effective therapeutic concept in the treatment of heart disease.


Livestock support the livelihoods of one billion people in Africa, Asia and Latin America, but the productivity of animals remains low, reducing the potential of the sector to support higher incomes and better nutrition. Improved livestock feeding has been identified as the most important step towards higher productivity. This scoping review assessed the evidence for the uptake of improved ruminant livestock feed options, the effect of this uptake on livestock productivity and the degree to which this improves smallholder farmer livelihoods. In total, 22,981 papers were identified, of which 73 papers were included in the final analysis after a rigorous double-blind screening review. Only papers that reported farmers’ decision to use a new feed intervention were selected, thereby excluding feeding trials and participatory feed assessments. Of the 73 papers, only 6 reported combined evidence of adoption, effect on productivity and livelihood changes. A total of 58 papers looked at adoption, 19 at productivity change and 22 at livelihood change. This scoping review highlights the gap in evidence for the adoption of new livestock feeding practices and provides recommendations to support farmers’ uptake of feed interventions. Livestock support the livelihoods of a billion people in Africa, Asia and Latin America, and improved feeding could increase productivity. However, this scoping review highlights serious gaps in evidence around the adoption of new feeding practices. As well as providing income and financial security for the rural poor, livestock products are a vital source of protein and micronutrients in regions where their regular supply could reduce the currently high prevalence of childhood stunting2. Unlike in the Global North, in LMICs, urbanization and growth in incomes and populations are fuelling strong growth in the demand for livestock products. A key challenge therefore is to identify and promote solutions to increase livestock production and productivity. Currently, livestock productivity is much lower in LMICs than in high-income countries3. For example, the average cow milk yields in Western Europe are 20 times higher than in Eastern Africa4. Moreover, increasing per-animal productivity is essential for environmental sustainability as systems intensify. For example, sub-Saharan Africa is a livestock greenhouse gas emissions intensity hotspot mainly because yields are low5. Poor productivity is caused by many factors including unimproved genetic stock, inadequate veterinary provision and a general scarcity of high-quality inputs. Although access to enough high-quality feed is generally agreed to be the main constraint on better productivity7, the livestock feed challenge has proved to be intractable despite considerable research and development effort over the past three decades. This effort has included many programmes aimed at delivering high-quality feed options to smallholder livestock keepers, but the evidence for their effectiveness is sparse8. The research field is characterized by anecdotes with very little rigorous and systematic analysis of success and the factors underlying success9.One billion poor people in Low- and Middle-Income Countries (LMICs) derive part of their livelihood from livestock10, the use of multipurpose trees11, methods of increasing intake and the nutritive value of crop residues by physical or chemical treatment12, and methods of preserving fresh feed to fill seasonal feed gaps13  and the amounts offered to different animals in different seasons. Feed improvement interventions among LMIC smallholders include the introduction of improved grasses and legumesBox 1 Feed interventions and livestock production systemsBrachiaria or Pennisetum species as well as high-quality legume species such as vetches or Desmodium. If well managed, introduced grasses can greatly increase feed yields, while introduced high-protein legumes can complement basal feeds such as straws and natural pasture.Improved grasses and legumes—ruminant animals naturally consume grass, forbs or shrubby vegetation. Natural pastures, while of moderate quality, are an important feed source in low-income countries. ‘Improved’ species include naturally high-yielding tropical grasses such as Multipurpose trees—trees are important in mixed crop–livestock systems, providing multiple benefits to small-scale farmers, including livestock feed. Various (mainly leguminous) trees have been popularized in tropical regions over recent decades. If well managed, these can provide a highly digestible and high-protein livestock feed and are reasonably resilient to dry spells.Increasing the intake and nutritive value of crop residues—crop residues make up a large part of ruminant livestock feed across the tropics, particularly in semi-intensive crop–livestock systems. Residues include straws of cereals such as wheat, barley and rice; stems or leaves of cereals such as maize and sorghum; and legume straws or haulms. Crop residues are generally characterized by low nutrient density, especially cereal straws. Methods such as physical and chemical treatments as well as the selection of superior varieties have been developed to improve nutrient availability.A livestock feed intervention aims at changing practices to provide more or better feed, increasing livestock productivity. The feed interventions considered in this scoping review are of three types:Other feed interventions include preserving fresh feed, filling seasonal gaps and feeding with high-quality supplements.25 and further operationalized by Thornton et al.26. It is based on major climate and land-use categories for which data are available globally and that determine the livestock feed base: permanent grasslands support pastoral systems, which often involve seasonal movement of livestock and are focused on ruminants; land areas that are used for both cropping and grazing are home to agro-pastoral and smallholder mixed crop–livestock systems, with or without irrigation; and landless systems rely solely on purchased feeds and are typically dominated by monogastric species, such as chickens and pigs, although landless urban dairy systems and feedlot beef systems also exist. More recently, researchers have suggested expanding this categorization to also consider the potential for the intensification of production, because of its considerable implications for development, using the evolution from subsistence to market orientation as a proxy28.To describe global livestock production, various categorization approaches have been suggested. The most widely known was developed by Seré and SteinfeldBox 2 Summary of methodshttps://osf.io/5ec9k/).A comprehensive literature search for CAB Abstracts was created (see the search strategy at https://osf.io/kghtc/).The CAB Abstracts search strategy was translated to 22 additional bibliographic databases and grey literature sources  and systematic reviews (which rely on homogeneous study methodologies and address intervention and outcome scenarios), did not fit the exploratory nature of this scoping review and the available evidence base.As shown in Tables In terms of geographical representation, almost half of the papers (35) analysed sites in East Africa or in the Horn of Africa. Southeast Asia was the second most represented region 12 papers). The dominance of papers from these two regions is probably due to relevant development and research projects that have been implemented there, including those by centres of CGIAR. The two regions are also over-represented in the scoping review because unlike South America, they are dominated by small-scale livestock systems, which was an inclusion criterion. The relatively few papers from West Africa, despite our inclusion of papers in French, may be explained by the dominance of pastoral systems in West Africa, where feed interventions are more challenging. Indeed, in such systems (see Box 2 papers.Most of the papers (43) used quantitative analysis only, 19 used mixed methods and 9 papers followed qualitative research methods. The duration of the experiment was measured as the time lapse between the year the feed technology was introduced and the year the analysis was conducted. This was reported in slightly more than half of the reviewed papers (36 papers). Twenty-two papers reported a duration between 1 and 5 years, eight papers reported a duration between 6 and 10 years, and the remaining seven papers analysed cases longer than 11 years. The effects of technology adoption on ruminant productivity and the resulting impacts on households’ livelihoods can be observed only after some time, given that years are needed for the feed intervention to be implemented  and for livestock productivity to increase, especially in the case of large ruminants.In terms of agro-ecological zones and livestock systems Box , 53 papeThe majority of papers (53) dealt with planted fodder, while agroforestry was the topic of 26 papers and crop residues 7 papers Table . Most paThis scoping review analysed the impact pathway between adoption, livestock productivity and household livelihoods Table , consideAdoption is the first step along the livelihood pathway, and it was anticipated to feature in most papers, given that it is relatively easy to measure either as a yes/no decision or as the extent of adoption. We found 15 papers that did not explicitly report or analyse that first step but only reported a change in productivity and/or livelihood indicators among adopters. More frequently, papers reported livelihood changes compared to productivity changes. This may be explained by a focus on development goals, such as producers’ livelihoods, and by the technical challenges of measuring livestock productivity. Livelihood indicators such as income and diet are relatively well established, and tools are available to systematically report them. In contrast, livestock productivity indicators vary across species , require demanding measurement protocols and are calculated using different periods , making comparisons difficult.Across the three outcome sets, there were no specific differences in types of publication or years of publication. Studies in mixed systems dominated across all outcomes, with few papers covering agro-pastoral or pastoral systems. For regions, papers from East Africa were most common for adoption studies, while for livelihood outcomes, there was a relatively large number of papers from Southeast Asia (8 out of 22 papers), possibly driven by research in development projects implemented in that region. In terms of research methods, the general observation that studies were mainly quantitative applied to all three outcomes. There were more mixed-methods approaches for adoption studies, possibly reflecting the importance of not only analysing the decision-making processes quantitatively but also assessing the ‘why’ and the ‘how’ that are better captured using qualitative approaches. There was no qualitative study measuring productivity indicators.The outcome indicator results reported in the studies are shown in Table Productivity indicators included increase in milk yields, animal weight gain, improved body condition and growth in flock/herd size. The number of papers with sufficient data was very low, with only nine papers across the three feed interventions. Changes in productivity ranged from 7 to 61%, with only one paper reporting productivity change related to crop residues.Finally, for livelihood indicators, the scoping review identified 22 papers with sufficient data across the three feed types, with 14 papers quantifying the impact. Household income change (8 papers) ranged from 6 to 285%, gross margins (3 papers) increased by 58 to 519% and labour or workload change (5 papers) from −24 to −70%.To better understand the reported changes, 25 papers were identified that explicitly examined the reasons for adoption. These were further examined for underlying drivers or constraints to adoption. Of the adoption drivers, the following were mentioned most often. Farmer experience or level of education was mentioned in ten papers; these variables are commonly collected as part of the household characterization in adoption studies and tend to be associated with higher rates of adoption. Expected increased productivity or income from the livestock enterprise was mentioned in eight papers. This is of course among the primary reasons to promote improved feed technologies, so this factor is expected to be prominent. However, most of the papers did not indicate it. Eight papers mentioned access to extension or training. Many feed technologies require considerable technical skill to be successful or effective. These are often described as ‘knowledge-intensive’ technologies, such as forage seeds that require treatment or scoring to germinate and then need to be grown from seedlings. Extension and training may therefore be important to facilitate their successful implementation. Seven papers mentioned labour availability. Most improved feed technologies require the use of additional and regular labour, such as cutting and carrying planted forages to confined ruminants. Family labour may be supplemented in some seasons by casual wage labour or even full-time labour in more market-oriented enterprises. Six papers mentioned good market access; again, this factor is generally associated with higher rates of adoption and may be associated with higher livestock product prices or easier access to feed technologies such as germplasm. Other contributing factors, in descending order, were access to credit or off-farm income, market orientation of the enterprise, group membership or social pressure, and land scarcity. Only two studies indicated soil improvement as an adoption objective, although this is one of the main reasons that nitrogen-fixing leguminous forages are promoted.Of the factors that were indicated as constraining the adoption of improved feed technologies, the following were mentioned most frequently. Increased labour requirement was mentioned in six papers; just as labour availability was indicated as an important driver of adoption, the labour requirement can be a constraining factor when that labour is not easily available. Little perception of net benefit was mentioned in four papers. Feeds are an intermediate output towards livestock production, and the final benefit may not be easily perceived immediately, particularly for fattening enterprises where nutritional benefits accrue over longer periods. Four papers mentioned difficult access to the technology or inputs. For some forage species, there may be limited systematic supply of seeds or planting material, and this is often a limit to sustained use after the withdrawal of project support. Many LMICs lack functioning forage seed systems. Four papers mentioned the complexity of the technology; as indicated, some feed technologies may require specific techniques, the training in which may not be available. Finally, competition with other land uses was mentioned in four papers. In land-scarce settings, priority may be given to food crops or to short-term cash crops such as seasonal vegetables, since these may represent a more profitable use of land. Likewise, some alternative land uses may be affected by subsidies and price control and may influence the relative returns from some feeding options.The research quality assessment was conducted using three indicators for all 73 papers Table . In term15.First, it is worth noting that the scoping review identified very few studies that answer our research question on the comparative impacts of various ruminant feed interventions on the livelihoods of livestock keepers. Indeed, the exercise yielded only 73 papers from a starting population of 22,981. We found many papers that studied the technical aspects of feed supply for ruminant livestock but were excluded because they did not assess the interventions’ uptake by or usefulness to farmers. This points to a strong bias among the scientific community towards understanding the technical intricacies of ruminant feeding without paying sufficient attention to how such technologies fit into general farming practices or farmer objectives. Additionally, a number of studies were dropped as per the exclusion criteria because they focused on large-scale livestock production. Several studies from Latin America fell in this category, which is consistent with the fact that farms in Latin America and the Caribbean are generally larger than farms in other regions, including sub-Saharan Africa and AsiaSecond, among the few papers included in the final analysis, the majority only analysed the adoption of feed interventions, and only six studies additionally documented the productivity and livelihood impact pathways of the feed interventions. The funding of research to generate rigorous and relevant evidence of feed innovation outcomes and impacts has been restricted mainly to those development projects introducing such innovations. However, such development-project-linked research may not be able to analyse the whole pathway from adoption to animal and household impact given the limited lifespans of such projects, particularly since productivity gains may only translate to sustained herd growth over time, and project termination may lead to the withdrawal of needed farmer support services.16. The focus on human food production by cereal breeding research institutes leaves the residues as an ‘orphan crop’. Our scoping review points to the need to focus more research effort on improving crop residue yields and quality characteristics such as through orienting crop breeding towards improving the feed quality of underutilized residues17. Practices involving improving crop residue quality and yield may have a strong likelihood of adoption, since few changes to farming practices are required in contrast to planted forages and forage trees.Third, the literature was found to be skewed towards forages and agroforestry, yet crop residues are among the largest sources of basal feed for ruminant livestock across tropical regions . This apparent mismatch may result from the fact that forages and multipurpose trees can offer a step change in productivity, compared with the more incremental productivity benefits from the improved use of crop residues. Forages and trees also have a disciplinary home, with whole research institutes devoted to their study. Crop residues are generally seen as by-products of human food production despite the fact that the market value of straw can in some cases approach that of grain for human food18 compared with the incremental effects of introducing a new variety of an established forage species19. Second, the approaches to determine intervention impacts differ considerably between studies. Where a development project is focused on development impact , outcomes are likely to be greater than in an independent study aiming to determine how farmers benefit from a variety of interventions. An example of the former is presented by Roothaert and Kerridge20, reporting a gross margin increase of 239% among project participants, whereas a study on various fodder shrubs in central Kenya was able to detect an income improvement of only 10% (ref. 21). Third, the time horizon considered by the reviewed studies varies greatly  contain a wide variety of individual interventions with very different potentials for inducing change. For instance, introducing a new forage crop into a system without any prior forage cultivation can yield substantial improvements in productivityFinally, this scoping review has identified various factors driving or constraining the adoption of feed interventions, which can be grouped into three broad and inter-related categories. The first category refers to managing a sometimes-challenging technology, requiring certain skills on the part of the farmer as well as access to the technology, to extension and to training in its use. Second, the benefits of using a technology and its alignment with the farmer’s objectives must be perceived and valued by the farmer for adoption to occur. This is often an issue because feed is an intermediate technology in the livestock value chain, and the link between better feeding and financial benefits may not be easily perceived. Furthermore, livestock may be kept for a range of reasons other than the production of milk and meat, and the farmers’ primary objective may not be immediately obvious to well-meaning development agents. Third, the availability of the key resources of land and labour, and the trade-offs between them that the feed technology may impose, will limit or facilitate adoption, with adequate availability of both (particularly labour) generally having a positive effect. The trade-offs in certain contexts may mean that farmers can derive greater benefit from allocating land and labour to non-livestock activities, and this needs careful consideration when considering feed interventions.The consideration of these adoption drivers and constraints is helpful for considering future approaches to enhancing livestock feed supply among poor livestock keepers. Too often, technologies have been promoted without systematically considering barriers to their uptake, whether target farmers have sufficient resources  to successfully implement them and whether the technologies make economic sense given the market conditions and the competing opportunities for the use of land and labour in target communities. Box Box 3 RecommendationsFor ‘knowledge-intensive’ technologies, the capacity of local livestock keepers and the strength of the extension advice environment to support ongoing implementation should be considered. If these are limited, some re-evaluation of the technology options or a parallel effort to enhance the necessary capacity among local livestock keepers is needed.In planning development efforts for livestock feeding, the focus needs to be on small-scale, semicommercial farmers who have both the resources and the incentive to make the investments needed for feed technologies to succeed. Livestock keepers whose primary objectives for keeping livestock are not to produce milk and meat for the market need a different kind of support and are much less likely to invest in new feed technologies.The resource requirements for livestock feed options need careful consideration. If other uses for land and labour are more lucrative, livestock keepers are unlikely to invest in new feed options. This requires the whole farming system to be considered, as well as how livestock fit into overall livelihood strategies. In addition, unlike food crops, forage seed systems are underdeveloped in many regions , with unclear demand and limited supply from the private sector. Public–private partnerships and investment may be needed to develop these supply chains and can be linked to local seed-producing entrepreneurs and collective groups.Decision makers and development agents should consider these factors and constraints in deciding when and where to target investments promoting these technologies. The conditions that favour feed technology adoption go far beyond biophysical suitability, extending to the social, economic and knowledge domains.This scoping review has shown that besides technical feed efficiency characteristics, various other factors enhance or constrain the adoption of improved feeds. On the basis of our analysis, we recommend the following:https://osf.io/6ywht/.This scoping review was conducted following the PRISMA-ScR (Preferred Reporting for Items for Scoping Reviews) checklist. A protocol was registered in Open Science Framework on 5 June 2019 at A comprehensive search strategy was developed (by E.L. and E.E.) to identify all available research pertaining to the livelihoods of small-scale and agro-pastoral livestock keepers in LMICs in Africa, Asia and Latin America, in relation to the improvement of ruminant feed interventions. The search terms included variations of the key concepts in the research question: improvement or conservation of crops; small-scale producers or pastoralists; LMICs in Africa, Asia or Latin America; and innovation or adoption indicators. The comprehensiveness of the search strategy was ensured by including all known search-term synonyms and appropriate subject term searches, conducting a peer review of search strategy by expert librarians familiar with the discipline, and confirming the inclusion of eight seminal studies in the results set. See Supplementary Appendix On 5 June 2019, four bibliographic databases were searched. These included CAB Abstracts , Web of Science Core Collection , Scopus  and Dissertations and Theses Global . On the same day, 20 grey literature sources were searched, including Africa Theses and Dissertations, AgEcon Search, AGRIS, Campbell Collaboration, Cochrane Collaboration, Collaboration for Environmental Evidence, Commonwealth Scientific and Industrial Research Organization, Brazilian Agricultural Research Corporation (Embrapa), French Agricultural Research Centre for International Development, GARDIAN, International Fund for Agricultural Development, International Institute for Environment and Development, JPAL/ATAI Impact Evaluations, Overseas Development Institute, UK Department for Internal Development, United Nations Environment Programme, World Food Programme, World Health Organization, World Bank, and Cgspace.The search results were deduplicated to remove citations identified in multiple databases. The titles, abstracts and keywords of all citations were exported as RIS files.N = 20,173) were screened via this method, blindly, by author pairs , and citations that both authors excluded did not progress to the title and abstract screening phase. The remaining studies were imported into Covidence, a systematic review screening software, and were screened blindly via title and abstract by two authors with a third as a tie-breaker . Studies that were not excluded by title and abstract screening were considered at the full-text level. These were screened blindly using the same inclusion and exclusion criteria, and conflicts were resolved by a third author as a tie-breaker . Individual reasons for exclusion were recorded at the full-text screening stage , studies were excluded if they did not meet all of the following inclusion criteria: (1) the study focus includes a population of small-scale and agro-pastoral keepers of large and small ruminants; (2) the study is primary empirical research; (3) the explicit population focus is small-scale and agro-pastoral ruminant livestock keepers; (4) the study describes the adoption of ‘improved feed options’ and/or their effect on productivity, livelihoods or both; (5) the study area or focus includes target populations in LMICs; and (6) the study is in English, French, Spanish or German.In addition, studies were excluded if they met one or more of the following exclusion criteria: (1) the study is a review or a case study; (2) the study does not include small-scale or agro-pastoral as the target population; (3) the study does not take place in LMICs in Latin America, Africa or Asia; (4) the study does not consider improved feed options ; (5) the study considers only industrial by-products and/or concentrates; (6) the study is in a language other than English, French, Spanish or German; and (7) the study considers only fish, pigs, poultry, camels, wild buffaloes, yaks, alpacas, guinea pigs (or cavies), bees, equines, rabbits or any wild animal.24 and adapted to the scoping review requirements. The data extracted included the author(s), year of publication, type of paper, study location, intervention type, comparator (if any), duration of the intervention, study population and methodology; the outcome measures differentiated by adoption, effects on livestock productivity and effects on livelihoods; and important results in terms of drivers of adoption and potential for scaling. The template was tested by I.B. and A.D. before being finalized. Google forms were used to extract the data by I.B., A.D., N.T., D.C., E.J.R., S.S. and R.L. Conflicts were resolved by a third author as a tie-breaker. The data analysis tables were created and data processed by N.T., I.B. and E.J.R. Data were extracted on 51 peer-reviewed journal articles, 8 papers published in conference proceedings, 7 working papers, 6 reports and 1 book chapter. The quality assessment was conducted on the 73 included papers using three criteria. The first one considered the quality of the study methodology (low versus high), the second assessed the justification of the methodology (low versus high) and the third criterion was an overall quality assessment with three levels . Each paper was scored by two persons . The levels were first transformed into scores  and then averaged.A data extraction template was created on the basis of Barrett et al.To better understand some of the underlying drivers of adoption of improved feed technologies, a subset of the final papers was selected for more detailed examination if they mentioned analysis of factors that either facilitated adoption or constrained adoption. Of the 73 papers in the full set, 25 met this criterion. Each of these papers was then re-examined by one researcher, and a set of adoption drivers and constraints was identified; the papers were scored on whether they mentioned each adoption driver or constraint. Twelve different adoption drivers emerged, such as increased productivity and good access to markets. Nine constraining factors were also indicated across this set of papers, such as low perceived benefit of the technology and competition with other land uses.Supplementary InformationBibliography of the final list of studies, data extraction template and protocol, including all final search strategies.


In this Special Issue, we have 10 excellent papers on epitaxy. In this editorial preface, I will make comments on the following issues: (1) applications of the materials examined, (2) lattice mismatch, (3) epitaxial growth methods used, (4) characterization methods used, (5) material problems: solved and still to be solved. The “Advances in Epitaxial Materials” has a big advantage of having, in one issue, papers on different materials, but in every paper the reader should find interesting information on epitaxial growth and characterization. All ten papers deal with materials for electronics, which reflects the situation that one can rarely find other applications of epitaxial structures. Many epitaxial technologies, such as Si-Ge, are mature enough that they are very rarely mentioned in the scientific papers. The properties of others are still being intensively explored. Half of the papers in this issue deal with nitride semiconductors. This is not surprising, as the present situation in this field of science and technology is often described half-jokingly as “GaNification” because gallium nitride and its related compounds revolutionize many areas of life. White LEDs based on blue LEDs illuminating phosphor have created a market worth tens of billions of euros. Recent advances in deep-UV LEDs have created a bright future for the use of these devices in sterilization and disinfection. RGB  projectors based on LEDs or Laser Diodes (LDs) push out other projectors from the market. Lighting, displays, Li-Fi telecommunication, and many other optoelectronic applications of nitrides take advantage of the high luminosity of nitrides. Their wide band-gap and high stability make them a perfect material for high-power and high-frequency transistors already present in the market.However, one should be aware of the fact that nitrides are very difficult to grow and process.The popularity of nitride semiconductors is reflected in the Special Issue in papers of R. Czernecki et al. , Y.L. CaTwo papers describe the growth and properties of more classical III-V epitaxial layers: GaInAsSb on GaAs  and InAlPhotovoltaic applications are represented by polymorphism-based MOF  . Many el3 and LaAlO3, which are promising materials for THz devices.In the electromagnetic spectrum, there is still a gap in the THz region in both the emitters and the detectors. As THz devices are very desirable in many applications, such as cancer diagnosis, food inspection, explosive detection, electronics, and communications, there are many groups working in that field. In this Special Issue, Kazuhiro Endo et al.  report oAll epitaxial layers described in this Special Issue are highly mismatched to the substrates. Therefore, this issue is discussed in almost all of the papers. The most spectacular result in reducing the threading dislocation density for the highly mismatched GaInAsSb on GaAs was obtained by Qi Lu  who usedAn increasingly popular method of epitaxy is growing the two-dimensional object as quantum wires or stripes. In this case, the strain can be accommodated by elastic deformation. This issue is discussed in the paper of Y.L. Casallas-Moreno et al.  for InN In the Special Issue, four papers ,3,4,10 rThe second epitaxial method used mostly in academic labs is molecular beam epitaxy (MBE). This method was also used in four papers ,5,6,7. SIn almost all of the papers of the Special Issue, the High Resolution X-ray Diffraction (XRD) was used for the characterization of samples. The 2theta/omega and reciprocal lattice maps are reported. An issue not discussed in the papers is the difficulty in the interpretation of the XRD data for poor-crystallographic-quality layers. In particular, the comparison of the simulations conducted for the perfect layers and the experimental data obtained for imperfect layers must be performed with great care.The second characterization method applied is photoluminescence (PL). In this case, the interpretation of the experimental data is not easy. For example, if one examines InGaN quantum wells, the PL peak position, its half-width, and intensity depend not only on indium content and well width, but also on indium segregation, internal electric fields, and the presence of point and extended defects. All ten papers of this Special Issue significantly contribute to the knowledge on the epitaxial growth of electronic materials. Some of them propose materials which should be considered as candidates for new electronic devices. However, one should be aware of the fact that—in none of the papers—point defects were taken into account. These defects strongly influence not only optical and electrical properties but also mechanical properties (lattice mismatch relaxation). The examination of point defects is extremely difficult as very few analytical methods can give information on these defects. However, breakthroughs in electronics will happen when the nature of point defects is learnt.


Trimer stabilization effects are quantified for Adalimumab, Infliximab, Etanercept, Certolizumab, Golimumab for bivalent and monovalent binding formats. Clear differences in trimer stabilization are observed, which may provide a deeper insight into the mode-of-action of TNF-α scavengers.The cytokine tumor necrosis factor-alpha (TNF-α) readily forms homotrimers at sub-nM concentrations to promote inflammation. For the treatment of inflammatory diseases with upregulated levels of TNF-α, a number of therapeutic antibodies are currently used as scavengers to reduce the active TNF-α concentration in patients. Despite their clinical success, the mode-of-action of different antibody formats with regard to a stabilization of the trimeric state is not entirely understood. Here, we use a biosensor with dynamic nanolevers to analyze the monomeric and trimeric states of TNF-α together with the binding kinetics of therapeutic biologics. The intrinsic trimer-to-monomer decay rate k = 1.7 × 10 Monomeric subunits of TNF-α form stable non-covalent homo-trimers at sub-nanomolar concentrations in solution7. Signalling proceeds by the recognition of TNF-α trimers by endogenic TNF receptors (TNFR) 1 and 2, which form trimers themselves prior to complex formation with TNF-α8. Trimeric TNF-α is known to monomerize over time at low concentrations, which causes a loss of bioactivity9. Elevated TNF-α expression levels were found in patients suffering from a multitude of different autoimmune diseases11. Scavenging antibodies and antigen-binding fragments (Fab) against TNF-α have proven to successfully suppress TNF-α-mediated inflammation in inflammatory autoimmune diseases including rheumatoid arthritis, psoriasis, and Crohn’s disease16. Despite the significant structural differences between the therapeutic antibody formats: Infliximab, Adalimumab, Certolizumab, and Etanercept, studies have shown that all four biomolecules engage in reverse signalling by binding to membrane-bound TNF-α on human cells17, although Etanercept and Certolizumab are unable to crosslink TNF-α trimers of any type19. Structural differences in multiple binding events of Adalimumab Fab and Infliximab Fab to a single TNF-α trimer suggest a stronger stabilization of TNF trimers in complex with Adalimumab Fab, which binds in the interface between two monomeric subunits, whereas each Infliximab Fab binds to a single monomeric subunit within a TNF-α trimer21. Cross-linking appears to be irrelevant for the induction of apoptosis as in vitro assays showed no significant differences between Fab and full IgG for Infliximab22. Assays utilizing size exclusion chromatography (SEC) could show that bivalent TNF-α binders  are able to form highly stable complexes of three soluble TNF-α trimers interlinked by three IgG molecules19. Little is known about the behaviour of TNF-α scavengers with respect to the oligomeric state of TNF-α.Tumor necrosis factor-alpha (TNF-α) is a cytokine that plays a key role in mediating inflammation23. In contrast to these studies, we determined a universal monomerization rate by continuous measurements in real time using a novel Electro Switchable Biosurface technique (ESB)24. As the monomerization of TNF-α when complexed with its scavengers reflects an increase in free TNF-α, we investigated the stability of TNF-α-trimer-TNF-scavenger complexes with Adalimumab, Infliximab, Golimumab, Certolizumab, and Etanercept as well as Fab fragments derived from Adalimumab, Infliximab and Golimumab. We find that TNF-α can quickly switch between the active trimer and inactive monomer, which reveals a new, fast-acting way to regulate its bioactivity. Interestingly, there are strong differences in the trimer stabilization of the therapeutic antibody formats and their Fabs. Some do not (fully) suppress monomerization of bound TNF-α, leading to release of TNF-α monomers. These monomeric subunits of TNF-α have not shown to form clinically relevant amounts of trimeric TNF-α. However, concentration-dependent re-trimerization has been shown in vitro7.In this study we investigated the dynamics of TNF-α monomerization as well as re-trimerization in real-time. Previous studies focus on monomer exchange rates in the equilibrium state, employing Förster Resonance Energy Transfer (FRET) as well as analytical size exclusion chromatographic assays25. These studies demonstrated a loss of bioactivity over time (monomerization of TNF-α). Additional studies could even show that bioactivity can be recovered by re-addition of TNF-α to previously monomerized TNF-α (leading to trimerization of TNF-α). However, in all studies discontinuous measurements were conducted  which do not accurately reveal the dynamics of mono- and trimerization of TNF-α on a timescale of seconds and minutes.The complex nature of TNF-α being expressed as a trimeric membrane-bound, proteinogenic cytokine that undergoes a conversion to a soluble trimer (a strong trigger of inflammation) has been subject of several studies since the early 1990s27.In this study, the multimerization state of TNF-α was measured using a surface biosensor with integrated microfluidic channels Fig. . BrieflyD to avoid monomerization7. Before induction of monomerization, the size of immobilized TNF-α was determined in tetraplicates27, yielding a hydrodynamic diameter 5.6 ± 0.1 nm for the initial state  to 6.10 nm (1TNF30). Next, the monomerization of immobilized TNF-α was initiated by injection of running buffer for 2000 s with a flow rate of 1 ml/min, resulting in a gradual increase of the switching speed (decrease of hydrodynamic friction) due to dissociation of TNF-α subunits . This suggests that in the course of the experiment, complete monomerization was achieved.In order to assess the oligomeric state of TNF-α, the protein was immobilized to the sensor surface at a concentration of 500 nM, which is considerably higher than the sub-nanomolar trimerization Kate Fig. , t0. Thiits Fig. . Contrart1, Fig. . Again, 25. Here, the onset of trimer formation was found to occur at approximately 10 nM free TNF-α  of free TNF-α, full trimer can be re-gained within minutes  is monovalent. For the sake of assay simplicity, avidity (and cross-linking) effects are often avoided in literature by immobilization of the antibodyFAST = 1.42 ± 0.39 10−3 s−1 and kSLOW = 8.52 ± 0.28 10−5 s−1, respectively 35. TNF-α and antibody are mixed at different ratios in solution and allowed to bind. After equilibration, the remaining fraction of unbound TNF-α is measured by the biosensor in a concentration measurement mode. Here, we used immobilized Adalimumab to detect the remaining binding-competent TNF-α (-complexes) in pre-incubated solutions with constant concentrations of TNF-α and increasing concentrations of Adalimumab  was chosen as 100 pM and 500 pM. The solution affinity measurements confirm the dissociation constants determined in the kinetics experiments, with KD(c(TNF-α)=100pM) = 53.6 ± 29.5 pM; KD(c(TNF-α)=500pM) < 30.0 pM)  Fig. .Figure 423. However, in FRET assays a faster TNF-α monomerization in complex with Golimumab was found. This behaviour we only observed for the Fab fragment derived from Golimumab, which showed the weakest suppression of monomerization   to ensure that scavenger cannot effectively dissociate from the surface. This way, an increase in the switching speed signal indicates the decay of scavenger-bound TNF-α, but not the scavenger dissociation. The switching speed was monitored for 70 min. Full suppression of monomerization (no increase in switching speed) was observed for all avid binders of TNF-α , which can form interlinked, multiple bound complexes  or caused by binding of single Adalimumab (Fab) molecules in interfaces of TNF-α monomer subunits within a trimer19. From kinetic experiments Moreover, there are differences in the relative dissociation amplitudes: Only if ‘bridging’ of immobilized ligand molecules can be excluded Fig. , full diThe kinetic analyses and comparisons of Adalimumab full IgG and Adalimumab Fab interaction with TNF-α are – to our knowledge – the first interaction analyses of picomolar binders with consistent rate constants independent of assay orientation. In addition to that, full dissociation has not been shown for antibodies or Fabs with residence times >10,000 s before. Only the concerted employment of a two-dimensional ligand distribution, low surface density of immobilized antigen, and very high flow rates allowed for the suppression of otherwise inevitable re-binding of analyte, compromising both dissociation amplitude and rate. However, even by optimizing the experimental design, the characterization and quantitative analysis of two multivalent binders such as TNF-α and its scavenger antibodies remains a challenging task.In assays with monovalent Adalimumab Fab as the soluble analyte molecule, the formation of complexes comprising multiple TNF-α trimers can be excluded. Thus, we conclude that suppression of monomerization is not just a product of the formation of complexes of high order. Our data show that the mere binding of a TNF-α scavenger to the interfacial grooves between individual units within a TNF-α trimer is sufficient to suppress monomerization.21. The functional analysis of trimer stabilization within this study cannot detect a difference between Adalimumab Fab or Infliximab Fab, nor between the parental IgGs  analyzer  at 25 °C.Recombinant human TNF-α was obtained from Life Technologies . TNF-α was supplied as lyophilized powder and reconstituted as recommended by the supplier. All therapeutic antibody formats were bought from the original provider. All protein concentrations were determined by absorbance measurements at 280 nm using a NanoDrop 2000c spectrophotometer . All solutions for interaction measurements  were provided by Dynamic Biosensors GmbH.2HPO4/NaH2PO4 pH 7.4, 140 mM NaCl, 50 µM EGTA, 50 µM EDTA, 0.05% (v/v) polysorbate-20) after purification.For generation of Fab fragments, 2 mg of each respective antibody  were digested using the Pierce Fab Preparation Kit , following the standard protocol. Fab fragments were reconstituted in PE140 buffer  on biochip surfaces, proteins were covalently attached to 96-meric DNA oligomers  using amine coupling. Amine coupling was performed following the standard protocol (Dynamic Biosensors GmbH), whereas TNF-α and Adalimumab Fab amounts were kept at the highest recommended amount for coupling reactions (500 µg per reaction) and for Adalimumab conjugations twice the maximum amount was used (1 mg per reaction). During conjugation of TNF-α to DNA oligomers, the TNF-α-concentration was kept above 500 nM at all times, to prevent any concentration-dependent monomerization. Chromatograms of purification, following the standard protocol are shown in Figure 2HPO4/NaH2PO4 pH 7.4, 40 mM NaCl, 50 µM EGTA, 50 µM EDTA, 0.05% (v/v) polysorbate-20) and in the molecular dynamics measurement mode .To minimize mass transport limitation, the probability of re-binding, and molecular crowding effects, surface densities were kept low with fluorescence amplitudes between 80–150 kcps. In addition to the reduction of the surface density, all measurements were performed at a high flow rate of 1000 µL/min to further decrease mass transport limitation and re-binding. Size determination as well as TNF-α monomerization experiments were performed in PE40 buffer , whereas all other interaction measurements were performed in the fluorescence proximity measurement mode .Kinetic experiments of TNF-α scavengers were performed in PE140. To correct for TNF-α monomerization, interaction measurements of immobilized TNF-α with Adalimumab Fab were performed in the molecular dynamics measurement mode , it is more practical to define a switching speed parameter 27).For determination of hydrodynamic diameters of surface-tethered proteins and protein complexes, alternating potentials are applied to induce a switching motion of DNA nanolevers . After each injection, formed complexes were removed by application of regeneration solution (pH 13), followed by hybridization of fresh adalimumab-DNA conjugate.For each in-solution equilibrium titration experiment, the concentration of TNF-α was kept constant , whereas the concentration of adalimumab was titrated from 3 pM to 8 nM. All samples were incubated for 7.5 hours to reach binding equilibrium. Unbound TNF-α was determined by injection of samples at 20 µL/min for 10 minutes in the fluorescence proximity measurement mode . The hydrodynamic diameters of TNF-α were determined as described earlier28. Additionally, translational and rotational diffusion coefficients were calculated by Hydropro Version 1028, a freeware tool available here: http://leonardo.inf.um.es/macromol/programs/hydropro/hydropro.htm.The determined mean values of hydrodynamic diameters were compared to hydrodynamic diameters calculated from shell-models based on atomic coordinatesData from monomerization experiments were independently fitted using a single exponential function following Eq. OFF was determined by a global fit of the dissociation curves . krves Eq.  and subs. ON Eq. . KD was FAST rate constant of fast transition (monomerization of TNF-α), global parameter. AFAST Amplitude of fast transition, parameter without constraints. kSLOW rate constant of slow transition , global parameter. ASLOW Amplitude of slow transition, parameter without constraints. y0 y-Offset.The dissociation traces of the interaction of immobilized TNF-α with Adalimumab Fab were fitted with a bi-exponential function following Eq. D Dissociation constant of in-solution interaction of receptor and ligand. A Amplitude. y0 y-Offset.All binding curves were normalized to their initial value and fitted with individual, linear fits. Slopes of linear binding curves were plotted semi-logarithmically against the concentration of titrated Adalimumab and subsequently fitted using Eq. Supplementary Information.


Tumor necrosis factor alpha (TNF-α) was initially recognized as a factor that causes the necrosis of tumors, but it has been recently identified to have additional important functions as a pathological component of autoimmune diseases. TNF-α binds to two different receptors, which initiate signal transduction pathways. These pathways lead to various cellular responses, including cell survival, differentiation, and proliferation. However, the inappropriate or excessive activation of TNF-α signaling is associated with chronic inflammation and can eventually lead to the development of pathological complications such as autoimmune diseases. Understanding of the TNF-α signaling mechanism has been expanded and applied for the treatment of immune diseases, which has resulted in the development of effective therapeutic tools, including TNF-α inhibitors. Currently, clinically approved TNF-α inhibitors have shown noticeable potency in a variety of autoimmune diseases, and novel TNF-α signaling inhibitors are being clinically evaluated. In this review, we briefly introduce the impact of TNF-α signaling on autoimmune diseases and its inhibitors, which are used as therapeutic agents against autoimmune diseases. Tumor necrosis factor alpha (TNF-α) is a cytokine that has pleiotropic effects on various cell types. It has been identified as a major regulator of inflammatory responses and is known to be involved in the pathogenesis of some inflammatory and autoimmune diseases . StructuThe activation of TNFR1 can trigger the formation of different signaling complexes, referred to as complexes I, IIa, IIb, and IIc, which result in distinct cellular responses ,12. DuriTNFR2 is suggested to be fully activated by tmTNF-α, primarily in the context of cell-to-cell interactions. Unlike TNFR1, TNFR2 lacks a death domain and thus is unable to induce programmed cell death directly  8]. Dur. Dur8]. Physiologically, TNF-α is a crucial component for a normal immune response. TNF-α can activate the immune system to regulate; however, the inappropriate or excessive production of TNF-α can be harmful and may lead to disease. Rheumatoid arthritis (RA) , inflammDue to the involvement of TNF-α in the pathogenesis of autoimmune diseases, TNF-α inhibitors have been successfully developed and applied in the clinical treatment of autoimmune diseases such as Crohn’s disease (CD) and RA ,28. TherRheumatoid arthritis (RA) is a chronic autoimmune disorder that primarily affects the joints and typically results in redness, swelling, and arthralgia. In serious cases, it limits the range of motion . RA affePsoriatic arthritis (PsA) is a long-term autoimmune arthropathy distinct from RA . It typiInflammatory bowel disease (IBD) is a group of autoimmune diseases localized in the gastrointestinal tract. CD and ulcerative colitis (UC) are the primary types of IBD, which are distinguished by the location at which the disease occurs. CD mostly occurs in the small and large intestine, but it can also affect the mouth, esophagus, stomach, and anus, while CD commonly affects the colon and the rectum ,43. In bPsoriasis (PS) is a chronic inflammatory disease with a strong genetic diathesis and autoimmunity characterized by a specific region of abnormal skin. There are five types of PS-laque, guttate, inverse, pustular, and erythrodermic, and about 90% of PS is plaque-type PS. Plaque-type PS classically presents pruritic plaques with white scales on the top ,47. The Noninfectious uveitis (NIU), also known as autoimmune uveitis, is an autoimmune disease inside the eye that aims at the neuroretina . NIU canInfliximab is a recombinant chimeric monoclonal antibody containing a murine variable region and a human IgG1 constant region. It is specific for all forms of TNF-α in humans and effectively blocks the binding of TNF-α to its soluble and transmembrane receptors . After iEtanercept is a fusion protein that includes two identical TNFR2 extracellular regions connected to the Fc fragment of human IgG1 . There aAdalimumab is a fully human IgG1 monoclonal antibody capable of specifically blocking the binding of human TNF-α to the receptors because its function and structure are identical to that of natural human IgG1 . AdalimuCertolizumab pegol (CDP87) is a humanized monoclonal antibody that has a polyethylene glycosylated (PEG) Fab fragment and lacks the Fc region . Due to Golimumab is a completely humanized IgG1 monoclonal antibody that has functional specificity for human TNF-α . Compare®), infliximab-abda (Renflexis®), and infliximab-dyyb (Inflectra®) with infliximab as a reference product are launched and available in the US market , inflNew anti-TNF-α agents are currently being developed to overcome shortcomings such as the nonresponse of existing TNF-α inhibitors. Ozoralizumab is a humanized monoclonal antibody designed for the treatment of inflammatory diseases. In a Phase 2 study, clinical activity and safety were evaluated in patients worldwide and in Japan. When 80 mg of ozoralizumab was injected, effective and well-tolerated treatment was possible as a novel TNF-α inhibitor (NCT01007175). Due to the specific molecular characteristics of the nanobody , most patients showed remarkable improvement in disease activity, and once induced, remission can be maintained at a dose of less than 80 mg per month, which shows the desired treatment result. In addition, since Phase 3 is currently in progress, ozoralizumab is highly likely to be used as a new TNF-α inhibitor. Saddala and Huang established a new drug model, ZINC09609430, that could specifically inhibit TNF-α. It was screened based on the Zinc library using structure-based pharmacophore modeling, virtual screening, and molecular docking with in silico absorption, distribution, metabolism, excretion, and toxicity (ADMET) analysis. As a result, the identified new small compound could serve as structurally various novel inhibitors for TNF-α and could potentially be used as anti-inflammatory agents to treat related diseases. Although ZINC09609430 is safe and active in silico ADMET results, it is worthy of further evaluation for its safety and efficacy in vitro and in vivo .TNF blocking treatment for CD is safe and effective, but due to the high nonresponse rate in patients, Kwak et al. conducted an in silico study to suggest a novel drug treatment for anti-TNF refractory CD. Among them, CP-69033401 showed high drug response in the cluster of patients with anti-TNF refractory CD. These findings could provide new options for anti-TNF treatment  89]..89].Here, we introduced the association of TNF-α signaling in some autoimmune diseases and the current TNF-α inhibitors used as therapeutic drugs for these diseases. TNF-α, which is commonly known as a proinflammatory cytokine, has also been shown to apply pleiotropic effects on various cell models and has been identified as a significant factor in the pathogenesis of autoimmune diseases . The rolRegarding safety issues, there are several side effects of TNF-α inhibitors. General side effects of all TNF-α inhibitors involve headaches, rashes, anemia, transaminitis, infectious, and, most commonly, an injection site reaction through a subcutaneous route ,91. The Currently, more TNF-α signaling inhibitors for autoimmune diseases are being clinically assessed, and even biosimilars of approved therapeutic drugs are being developed by many research institutes and companies, as the market for TNF-α inhibitors shows great promise. Furthermore, TNF-α inhibitors are used in off-label indications as well as the approved therapies for autoimmune diseases. Case reports have demonstrated that infliximab, etanercept, and adalimumab are effective against diseases such as Pyoderma gangrenous, Granuloma annulare, and Pityriasis rubra pilaris . AlthougTherefore, it can be predicted that the understanding of TNF-α signaling will be much more emphasized in the near future to develop effective tools for the treatment of other autoimmune diseases as well as a wide range of diseases involving TNF.