Hereditary hemorrhagic telangiectasia is a rare autosomal dominant vascular disease defined by the presence of mucosal and cutaneous telangiectasia and visceral arterio-venous malformations. The latter are abnormal capillary-free direct communications between the pulmonary and systemic circulations with the following consequences: arterial hypoxemia caused by right-to-left shunts; paradoxical embolism with transient ischemic attack or stroke and brain abscess caused by the absence of the normally filtering capillary bed; and hemoptysis or hemothorax due to the rupture of the thin-walled arterio-venous malformations (particularly during pregnancy). It is frequently underdiagnosed, commonly presenting as complications from shunting through arterio-venous malformations: dyspnea, chronic bleeding, or embolism. Arterio-venous malformations are present not only in the lungs, but can also be found in the liver, central nervous system (mainly in the brain), nasal mucosa, or the gastrointestinal tract. The first choice of therapy is embolization of the afferent arteries of the arterio-venous malformations, a minimally invasive procedure with a high efficacy, a low morbidity, and low mortality. Other therapeutic modalities are surgery (resection) or stereotactic radiosurgery (using radiation). Routine screening for arterio-venous malformations is indicated in patients diagnosed with this condition and can prevent severe complications such as acute hemorrhages, brain abscesses, or strokes. Clinicians should provide a long-term follow-up for patients with arterio-venous malformations, in an effort to detect their growth or reperfusion in case of previously treated malformations. In spite of two experts’ consensuses, it still possesses multiple therapeutic challenges for physicians, as several aspects regarding the screening and management of arterio-venous malformations still remain controversial. Multidisciplinary teams are especially useful in complex cases. Osler–Weber–Rendu disease, known as hereditary hemorrhagic telangiectasia (HHT), is a rare autosomal dominant vascular disease defined by the presence of cutaneous and mucosal telangiectasia and visceral arterio-venous malformations (AVMs). The prevalence of HHT according to studies conducted in Europe and Asia is estimated at approximately 1:5000 persons worldwide, although an analysis using US health insurance databases detected a much lower incidence of 0.3 per 10.000 persons .The main underlying pathology in patients with HHT is a primary defect of the vascular network, characterized by a lack of intervening capillaries between arteries and veins. At a genetic level, mutations in multiple genes of the transforming growth factor-beta (TGF-β) signaling pathway are involved, normally responsible for regulating cell proliferation, migration, differentiation, and apoptosis . Some stArterial hypoxemia caused by right-to-left shunts;A paradoxical embolism with transient ischemic attack or stroke, and brain abscess caused by the absence of normally filtering capillary bed;Chronic bleeding like hemoptysis or hemothorax due to the rupture of the thin wall of the AVMs.Visceral AVMs are abnormal capillary-free direct communications between the pulmonary and systemic circulations with the following consequences :ArterialENG gene mutation, encoding endoglin, and involves a higher number of pulmonary and central nervous system vascular malformations.Type 1 is associated with an ACVRL1) gene mutations and with a higher incidence of primary pulmonary hypertension and hepatic manifestations of the disease. These two types are found in approximately 85% of cases referred to genetic testing for clinical suspicion of HHT [Type 2 is associated with activin A receptor type II—like 1 , and ADAM 17 are associated with pulmonary AVMs. Mutations in the SMAD4 gene, which is normally an essential effector of the aforementioned TGF-β signaling pathway, are detected in less than 2% of cases with clinical suspicion of HHT and determine an increased risk of juvenile polyposis syndrome (juvenile polyposis/HHT overlap syndrome) [GDF2 mutations, encoding the activin receptor-like kinase-1-ligand BMP9, which have only been reported in a few cases so far [RASA1 gene and suggest that their inclusion in molecular screening for HHT patients without ENG, ACVRL1, or SMAD4 mutations could aid in a correct diagnosis [Additionally, other gene mutations that can cause HHT have been studied: protein tyrosine phosphatase, non-receptor type 14 (yndrome) . Anotheriagnosis .A distinct genetic disorder associated with AVMs formation is the capillary malformation-arteriovenous malformation syndrome, which is defined by multiple oval capillary malformations with a surrounding white halo and synchronous AVMs.simple  andcomplex .According to the anatomic structure, pulmonary AVMs are classified in:Even if it is an autosomal dominant inherited disease, its manifestations are not typically present from birth, but more commonly develop with increasing age. The most frequent manifestation, epistaxis, is present in approximately 50% of affected individuals from the age of 10 and in 80–90% by the age of 21 . After tA clinical diagnosis of hereditary hemorrhagic telangiectasia is established based on the Curaçao criteria, family history  ,12 or byCuraçao criteria are represented by:Telangiectasia: these are pink to red lesions, usually 0.5–1.0 mm in diameter, which may appear at any site, especially on the face, nose, fingertips, lips, tongue, oral, and gastrointestinal mucosa. These lesions blanch when pressure is applied and refill immediately after release. Given their thin walls and close situation to the surface of the skin and mucosa, they are predisposed to rupture and bleeding [bleeding .Epistaxis: the most prevalent symptom in HHT is epistaxis related to nasal telangiectasia, which can be spontaneous or recurrent and in severe cases can lead to anemia.Visceral lesions: represented by AMVs that may arise in various organs, with characteristic clinical presentations. Gastrointestinal lesions are usually evaluated by upper or lower digestive endoscopies and may become symptomatic through massive hemorrhages or anemia [r anemia . Pulmonar anemia . Neurolor anemia . Lesionsr anemia . In the r anemia . These lFamily history: first degree relatives affected by the disease.The Pulmonary AVMs are thin-walled abnormal vessels, usually resulting in sac-like structures that replace normal capillaries between the pulmonary arterial and venous circulations. These are frequently multifocal, bilateral, and located predominantly at the bases of the pulmonary lobes, providing direct communication between pulmonary and systemic circulations. They tend to gradually increase in size over time; puberty, pregnancy, and pulmonary arterial hypertension may be contributing factors to this phenomenon . A recenA recent multi-center study has found that the presence of symptomatic liver AVMs and gastrointestinal bleeding are predictors of mortality in HHT patients, irrespective of age. Intriguingly, brain and pulmonary AVMs were not significantly associated with mortality rates in the study population .Stage I (latency or inactivity): localized hyperemia. Pediatric patients can remain stable for long time in this stage if AVMs are small.Stage II (expansion): increasing of arterio-venous shunting.Stage III (progression): onset of symptoms and signs, such as pain, bleeding, or ulceration.Stage IV: large AVMs with a risk of cardiac involvement and decompensation.The risk and severity of AVMs might be assessed by Schobinger’s clinical staging :Stage I Current recommendations state that in asymptomatic children of a parent affected by the disease, a HHT diagnosis should be suspected, unless clearly excluded by genetic testing .At the time of stroke and brain abscess, pulmonary AVMs can be clinically silent, so it is very important for these to be detected before becoming symptomatic. As chest X-ray and blood oxygen levels are not sensitive enough to detect the pulmonary AVMs, in this this regard, one must use a chest CT scan , magnetiTransthoracic contrast echocardiography should be used as the initial screening test for pulmonary AVMs . In intrIn adult patients with a definite or suspected HHT diagnosis, screening for liver AVMs should be performed . In the in the absence of symptoms, pulmonary AVMs screening should be performed initially, either by an agitated saline transthoracic contrast echocardiography or by a low-dose chest CT without contrast, according to local availability and expertise. When opting for a chest CT examination, it should ideally be performed early in the second trimester.in cases presenting with symptoms indicative of pulmonary AVMs, diagnostic testing by a low-dose non-contrast chest CT scan should be performed. This examination can be carried out at any gestational age, as clinically indicated.treatment of pulmonary AVMs is recommended starting with the second trimester, in the absence of other clinical contraindications.In pregnant women, the management of pulmonary AVMs is recommended as follows :in the aNovel evidence regarding issues previously addressed in the first International HHT Guidelines for Diagnosis and Management  have beeManagement of liver AVMs:Screening for liver AVMs should be performed in adults with definite or suspected HHT.In HHT patients with signs or symptoms suggestive of complicated liver AVMs, such as abnormal liver function tests, abdominal pain, portal hypertension, encephalopathy, pulmonary hypertension, heart failure, or abnormal levels of cardiac biomarkers, diagnostic testing should be performed using a Doppler ultrasound, a multiphase contrast CT scan, or contrast abdominal magnetic resonance imaging.An intensive first-line approach should only be instated in cases presenting with complicated or symptomatic liver AVMs, adapted to the specific type of complication.HHT cases that exhibit high-output cardiac failure and pulmonary hypertension should be managed multidisciplinarily by the Center of Excellence for HHT and a HHT cardiologist or a pulmonary hypertension specialty clinic.Clinicians should evaluate liver AVMs prognosis using available predictors in an effort to select patients in which closer monitoring may be advised.Intravenous administration of bevacizumab should be taken into consideration in cases with symptomatic high-output cardiac failure caused by liver AVMs with an insufficient response to first-line treatment.A referral to evaluation for liver transplantation is recommended in cases of refractory high-output cardiac failure, biliary ischemia, or complicated portal hypertension that arise as complications of liver AVMs.In patients with a proven or suspected HHT diagnosis, liver biopsy should be avoided.Given the fact that hepatic artery embolization is a temporising intervention with significant morbidity and mortality, it should not be performed in cases with liver AVMs.Management of pulmonary AVMs:Clinicians should screen all patients with a suspected or confirmed HHT diagnosis for pulmonary AVMs.Clinicians should use transthoracic contrast echocardiography as the initial screening test for pulmonary AVMs.Clinicians should treat pulmonary AVMs with transcatheter embolotherapy.In the event of procedures with risk of bacteremia, antibiotic prophylaxis should be done;When intravenous access is in place, special care should be taken in order to avoid intravenous air;SCUBA diving should be avoided.Clinicians should provide the following long-term recommendations to patients with diagnosed pulmonary AVMs (irrespective of previous treatment):Long-term follow-up should be provided in cases with pulmonary AVMs, in an effort to diagnose the possible growth of untreated AVMs or the reperfusion of AVMs that have undegone treatment.Management of brain AVMs:Magnetic resonance imaging as a screening method for brain AVMs in patients with suspected or definite HHT should be performed using a protocol both with and without contrast administration and implementing sequences that detect blood products, for sensitivity optimization.In cases of acute hemorrhage arrising as a complication of brain AVMs, definitive treatment should be recommended in a specialized center with neurovascular expertise.In all other patients diagnosed with brain AVMs, invasive testing and individualized management should be recommended in a center with neurovascular expertise.In pregnant women with possible or confirmed HHT with concurrent asymptomatic brain AVMs, definitive treatment of the brain AVMs should ideally be postponed until after delivery .Management of epistaxis:In HHT-related epistaxis, moisturizing topical therapies that humidify the nasal mucosa should be used.Oral administration of tranexamic acid should be considered in cases of epistaxis that do not resolve with the use of moisturizing topical therapies.Ablative therapies such as laser treatment, radiofrequency ablation, electrosurgery, and sclerotherapy should be considered for the treatment for nasal telangiectasias, in cases that are non-responsive to moisturizing topical therapies.Systemic antiangiogenic agents should be considered for the management of epistaxis that has failed to resolve with any of the aforementioned therapies.Septodermoplasty or nasal closure should be taken into consideration in cases of epistaxis with insufficient improvement after topical application of moisturising therapies, ablative procedures and/or administration of tranexamic acid.Physicians should recommend the topical use of nasal mucosa humidifying agents for HHT-related epistaxis prophylaxis.Patients with HHT-related epistaxis who are willing to undergo treatment should be referred to otorhinolaryngologists with HHT expertise.If nasal surgery is considered for reasons other than epistaxis, an evaluation should be performed by an otorhinolaryngologist with expertise in HHT-related epistaxis.Interventions performed for acute epistaxis should involve packing with material or products that are associated with a low likelihood of rebleeding upon removal, such as lubricated low-pressure pneumatic packing.Management of gastrointestinal bleeding:Esophagogastroduodenoscopy should be used as the first-line diagnostic investigation in patients with possible HHT-related bleeding. If colorectal cancer screening criteria are fullfilled and in cases with either genetically proven or suspected SMAD4-HHT syndrome, a colonoscopy should also be performed.If the esophagogastroduodenoscopy does not identify significant HHT-related telangiectasia, a videocapsule endoscopy should be taken into consideration.Mild: oral iron replacement is sufficient to achieve the target hemoglobin level;Moderate: hemoglobin goals are met with intravenous iron treatment;Severe: target hemoglobin levels are not met in spite of adequate iron replacement or blood transfusions are necessary.HHT-related gastrointestinal bleeding severity should be assessed using the following framework:The use of endoscopic argon plasma coagulation during endoscopy should be limited.In mild HHT-related gastrointestinal bleeding, the administration of oral antifibrinolytics should be taken into consideration.In moderate to severe HHT-related gastrointestinal bleeding, the administration of intravenous systemic antiangiogenic therapies, such as bevacizumab, should be taken into consideration.Management of AVMs in children:Diagnostic genetic testing should be performed in asymptomatic children of an affected parent.Asymptomatic children with HHT or at risk for HHT at the time of presentation should be screened for brain and pulmonary AVMs.Large pulmonary AVMs and those associated with decreased oxygen saturation levels should undergo treatment in an effort to avoid major complications.Pulmonary AVM screening in asymptomatic children with definite HHT or at risk for HHT should typically be repeated at 5-year intervals.Brain AVMs that harbour high-risk features should be treated.Management of anemia and anticoagulation:Adults, irrespective of symptoms;Children presenting with symptoms of anemia and/or recurrent bleeding episodes.Iron deficiency and anemia should be investigated in the following HHT patients:Oral iron administration constitutes the first therapeutic step;In cases in which oral administration of iron proves ineffective, cannot be absorbed or tolerated, and in patients presenting with severe anemia, intravenous iron replacement should be performed.Iron replacement therapy is recommended as follows:Hemodynamic instability or shock;Comorbidities in which a higher hemoglobin target is needed;Clinical settings in which an urgent increase in hemoglobin level is necessary ;Inadequate hemoglobin levels in spite of frequent intravenous iron infusions.Red blood cell transfusion is recommended as follows:Additional causes of anemia should be investigated in case of an inadequate response to iron replacement.Prophylactic or therapeutic anticoagulation or antiplatelets should be administered in the presence of an appropriate indication, taking into account the individualized bleeding risks of HHT patients; HHT-related bleeding does not constitute an absolute contraindication for these medications.Dual antiplatelet therapy and/or a combination of antiplatelet medication and anticoagulation should be avoided in HHT when possible.Management of AVMs in pregnacy and delivery:Clinicians should address preconception and prenatal diagnostic options in patients with HHT.Testing by unenhanced magnetic resonance imaging should be done in pregnant women presenting with symptoms indicative of brain AVMs.In the absence of symptoms, pulmonary AVMs screening should be performed initially, either by an agitated saline transthoracic contrast echocardiography or by a low-dose chest CT without contrast, according to local availability and expertise. When opting for a chest CT examination, it should ideally be performed early in the second trimester.In cases presenting with symptoms indicative of pulmonary AVMs, diagnostic testing by a low-dose non-contrast chest CT scan should be performed. This examination can be carried out at any gestational age, as clinically indicated.The treatment of pulmonary AVMs is recommended starting with the second trimester, in the absence of other clinical contraindications.HHT-affected pregnant women without recent screening and/or treatment for pulmonary AVMs should be managed as follows:HHT-affected pregnant women with previously untreated pulmonary and/or brain AVMs or those who have not undergone pulmonary AVMs screening recently should be managed by a multidisciplinary team, ideally in a tertiary care center.Epidural anesthesia should not be withheld on the basis of a HHT diagnosis and spinal vascular malformations screening is not necessary.Labor and vaginal delivery are not contraindicated in pregnant HHT women who harbour objectively defined, non-high-risk brain AVMs. An assisted second stage may be necessary on a case-by-case basis.Treatment of AVMs  is focusEmbolization therapy is the preferred method as it avoids the risks associated with major surgery, general anesthesia, and loss of pulmonary parenchyma. It has a good success rate, and the most commonly encountered complication is pleuritic chest pain, usually self-limited. Other complications are the migration of the device used in embolization, pulmonary infarction, air embolism in the coronary arteries with consequent angina, and bradycardia . For thaAn association between oral microorganisms and brain abscesses was documented. A very good oral hygiene is indicated along with antibiotic prophylaxis prior to dental procedures associated with risk of bacteremia for patients with pulmonary AVMs. In addition to that, given their high risk of paradoxical embolic stroke and brain abscess, pulmonary AVMs should be treated irrespective of their diameters.The intravenous administration of bevacizumab should be taken into consideration in cases with symptomatic high-output cardiac failure arrising as a complication of liver AVMs with an insufficient response to first-line treatment .Taking into account the fact that hepatic artery embolization is a temporising intervention with significant morbidity and mortality, it should not be performed in cases with liver AVMs . A referHHT cases that exhibit high-output cardiac failure and pulmonary hypertension should be managed multidisciplinarily by the Center of Excellence for HHT and a HHT cardiologist or a pulmonary hypertension specialty clinic .In the long-term, patients with documented pulmonary AVMs should avoid SCUBA diving in order to avoid their rupture, irrespective of previous treatment. In addition to that, antibiotic prophylaxis should be recommended before procedures associated with the risk of bacteremia and special care should be taken in order to avoid intravenous air when intravenous access is in place . PulmonaACVRL1, ENG, or SMAD4 mutation) have higher epistaxis severity scores. It seems that anti-fibrinolytics are effective therapies in decreasing epistaxis severity and their long-term use in HHT patients has been proven safe [Recurrent, spontaneous bleeding is associated with the development of iron deficiency anemia. Epistaxis severity scores correlate with age, hemoglobin, and ferritin levels. HHT patients with genetic mutations  for complex AVMs is recommended, with recent studies observing a significantly more sustained response in patients who have undergone both medical treatment with sirolimus and embolization .High-dose radiation therapy of an AVM has been shown to induce its obliteration. Radiation beam doses seem to be one of the essential factors that influence the AVMs angiogenesis process following radiosurgery or gamma knife .There is increasing interest in recent literature regarding potential novel therapies for HHT-related bleeding non-responsive to currently recommended treatments. One such example is tamoxifen, a selective estrogen receptor modulator, which has been administered in a transfusion-dependent patient with HHT-related epistaxis and gastrointestinal hemorrhage with favorable outcome and no significant adverse effects . In addiHereditary hemorrhagic telangiectasia is a rare condition, that is usually underdiagnosed. The clinical presentation is typically the result of complications arising from shunting through arterio-venous malformations, such as dyspnea, chronic bleeding, or embolism. It is associated with multiple therapeutic challenges for specialists. Routine screening for AVMs in patients diagnosed with this condition is recommended and can prevent severe complications, such as brain abscesses and strokes. A long-term follow-up should be provided in cases with AVMs, in an effort to diagnose the possible growth of untreated AVMs or the reperfusion of those that have undergone treatment.In spite of two consensuses, several aspects of AVM screening and management remain controversial even among experts. Multidisciplinary teams are useful in complex cases.


In recent years, host–microbiome interactions in both animals and plants has emerged as a novel research area for studying the relationship between host organisms and their commensal microbial communities. The fitness advantages of this mutualistic interaction can be found in both plant hosts and their associated microbiome, however, the driving forces mediating this beneficial interaction are poorly understood. Alternative splicing (AS), a pivotal post-transcriptional mechanism, has been demonstrated to play a crucial role in plant development and stress responses among diverse plant ecotypes. This natural variation of plants also has an impact on their commensal microbiome. In this article, we review the current progress of plant natural variation on their microbiome community, and discuss knowledge gaps between AS regulation of plants in response to their intimately related microbiota. Through the impact of this article, an avenue could be established to study the biological mechanism of naturally varied splicing isoforms on plant-associated microbiome assembly. The interaction between host plants and their commensal microbiome has become a research hotspot in recent years. As sessile organisms, plants have evolved a series of mechanisms to cope with changing environments ,2. In faInfluencing factors that affect this mutualistic interaction have been extensively reported, including internal and external factors in both plants and microorganisms . SpecifiIn addition, plants can translate their genetic differences by adjusting the repertoire of proteins they express in response to surrounding abiotic and biotic factors. One post-transcriptional regulatory mechanism, notably alternative splicing (AS), has emerged as a pivotal process for plants, to adapt to changing environments by significantly increasing proteome diversity or fine-tuning transcript abundance . To datePseudomonas spp., Bacillus spp., Azospirillum spp., and Streptomyce spp. [Bacillus thuringiensis is renowned as a good bioinsecticidal bacterium, and acyl homoserine lactone lactonase created by B. thuringiensis can open the lactone ring of N-acyl homoserine lactone, a signature molecule within the bacterial quorum-sensing system [F. graminearum, as suppression of fungal growth, virulence, and mycotoxin biogenesis [Chitinophaga spp., Chryseobacterium spp., Flavobacterium spp., Microbacterium spp., Pseudomonas spp., Sphingomonas spp., Stenotrophomonas spp., and Xanthomonas spp. are typically found to complement plant responses to totally different pathogen/pest attacks. However, the benefits of microorganisms on plants may occur via two modes. For example, a change in the structure of the inter-root microbiome would give the common tomato the property of resistance to wilt. The model dicot plant Arabidopsis thaliana specifically promotes three bacterial species within the rhizosphere upon foliar defence activation by the mildew pathogen Hyaloperonospora arabidopsidis [Pseudomonas on roots of wheat grown in a soil suppressive to take-all of wheat. Phl-producing fluorescent Pseudomonas are key elements of the natural biological control that operates in take-all-suppressive soils [The microbial community is a crucial environmental component, as approximately 10 billion microorganisms are present in every gram of soil . The intyce spp. ,36,37. Fg system . The capg system ,40,41. Fdopsidis . 2,4-diave soils . Recruitve soils ,45,46. TThe plant microbiome is not static: its structure and therefore the provided host functions in response to stresses and environmental stimuli ,48,49,50Pseudomonas spp., Bacillus spp., streptomyces spp., and plant growth-promoting fungi (PGPF), as well as Trichoderma spp. and Serendipita indica (formerly Piriformospora indica) [Pseudomonas piscium ZJU60 strain, which was isolated from infected wheat head, antagonizes the flora Fusarium graminearum by inhibiting its simple protein acetyltransferases [Arabidopsis leaf microbiome [Emerging evidence has indicated that plant-associated microbiomes are closely related to plant health  and the  indica) ,57,58,59sferases . The abicrobiome .Plant–pathogen interactions are mediated by the interplay of multifaceted processes, which are expedited by pathogen- and plant-oriented molecules ,63. PathArabidopsis leaves is in response to the associated degree of activation of the SA signalling pathway [The vital next step, currently, is to check these molecular mechanisms and unravel how commensal microbes move with plant immunity under various plant conditions. Such interactions between the system and microbiome are likely critical for plant defence against diverse stresses and influence host microbiota associations. Evidently, internal secretion signalling pathways of jasmonic acid (JA), salicylic acid (SA), and ethylene (ET) and their interactions are crucial in regulating plant defences and their associated microbiota ,70. BiotTaken together, unravelling the interactions between plant defence systems, microbiomes, and environmental factors will be an essential next step for understanding plant selection of microbes and immune modulation shifts of plant microbiota of various environmental conditions.Arabidopsis thaliana demonstrates that host cultivars (genotypes) mediate a weak but measurable impact on the root-associated microbiota [Lolium perenne cultivars (AberDove and S23), and microbial communities with differences were discovered [In light of growing concern over the threat of water and nutrient stress facing terrestrial ecosystems, particularly those used for agricultural production, increased emphasis has been placed on understanding how abiotic stress conditions influence the composition and functioning of the root microbiome and its ultimate consequences for plant health. However, under abiotic stress conditions, the composition of root microorganisms will change accordingly, which will be reflected in changes in root secretions. Although there are many conclusions about abiotic stresses on microbial community alteration, in recent years, attention has gradually shifted to the genotype differences of plant hosts. Following the appearance of next-generation sequencing, many studies have characterized the rhizospheric wheat microbiome and investigated the influence of the compartment (rhizoplane vs. endosphere), crop management, or wheat genotypes on the diversity and structure of those complicated microbial communities ,72,73,74crobiota . Plant gcrobiota . The effcrobiota . Lamit icrobiota . Jiang ecrobiota . Microbiscovered . Hou et scovered . A signiscovered . Huang escovered . By analscovered . Zhou etscovered . A compascovered , winter scovered , and maiPhaseolus vulgaris) cultivars with different levels of resistance to the fungal root pathogen Fusarium (Fox), differences in microbial composition between susceptible and nonsusceptible cultivars and higher network complexity of resistant cultivars were found. Pseudomonadaceae and Bacillaceae had a high abundance in the rhizosphere of the Fox-resistant cultivar. Breeding for Fox resistance in common bean could have co-selected other unknown plant traits that support the next abundance of specific helpful microorganism families in the rhizosphere with functional traits that reinforce the primary line of defence [flavobacterial genome that was much more abundant in the resistant plant, which could suppress R. solanacearum-disease development in a susceptible plant [All of this evidence suggests that microbial communities change in response to changes in hosts, so a hypothesis has been proposed that plants will recruit different microorganisms by releasing different root secretions based on their own demand recruitment to relieve stress on their own growth or when they encounter stressful environments. Specifically, a mechanism employed by plants is the ‘cry for help’ strategy , a pheno defence . Microbile plant . Thus, oArabidopsis root metabolic patterns and their variability between plants and naturally occurring germplasm were analysed, and for some secondary metabolites, the ratio of total plant-to-plant variability was high and significant [Arabidopsis plants exhibit hyperbolic coumarin and oligolignol exudation [Populus tremula) iatrogenic organic acid exudation, and zinc-deficient wheat hyperbolic phytosiderophore exudation [It is not surprising that various active genes of the soil microbiota play important roles in competition or cooperation with other microbes. Microorganisms synthesize completely different products that affect microbe–microbe interactions. Distinct and numerous gene clusters for biosynthesis of natural products have been identified in plant-associated microorganisms ,92. Addinificant . Exudatinificant . Additioxudation , and couxudation , seriousxudation . DiffereMost eukaryotic genes are interrupted by introns. Therefore, an important step in gene expression is the removal of introns through the splicing of precursor mRNA transcripts (premRNAs). Emerging evidence has indicated that plant-associated microbiomes are engaged with plant health and the helpful features of plant-associated microbes will boost the immune responses in plants against biotic/abiotic environmental constraints. Recent progress in high outturn sequencing of ribonucleic acid and bioinformatics tools to research AS events ,106,107 Arabidopsis). Examples include XA21 from rice [Arabidopsis [Platanus × acerifolia: phyletic and structural, we see that some RLKs have indeed been involved in the expression of phenotypic plasticity and are therefore a decent candidate for investigations into pathogen resistance [Plant immune receptors belonging to the receptor-like kinase (RLK) family play important roles in the recognition of microbial pathogens and activation of downstream defence responses. The function of RLKs in plant immune responses involves specific phosphorylation events within and outside the structural domain of the kinase, which leads to altered kinase activity and consequently to immune signalling. The function of several RLKs in pathogen resistance has been extensively studied in model plants with simpler genomes  is a classical PRR receptor protein that detects conserved bacterial flagellin 2 (FLS2) in plants, sensing external bacterial infestation. Flagellin perception in Arabidopsis works through recognition of its extremely conserved N-terminal epitope (flg22) by flagellin-sensitive 2 (FLS2) to initiate a series of immune responses. In nine families of dicotyledons, FLS2 was shown to undergo AS of its first exon. Point mutations and gene swaps indicated that the position and potency of exitron splicing primarily relied on the nucleotide sequences of FLS2 genes. The position and efficiency of splicing depend mainly on the FLS2 nucleotide sequence of FLS2; exon of FLS2 is spliced via an intron-mediated enhancement (IME), which regulates the accumulation of transcriptional products. Some ATs have the potential to encode suppressors for the FLS2 pathway, and transformed transcripts can encode FLS2 pathway repressors that affect FLS2-mediated reactive oxygen species production. This study reveals that alternative splicing finely regulates the pattern of receptor recognition and thus influences the downstream disease resistance response . The recArabidopsis thaliana NPR1 loss-of-function mutant PR gene expression is reduced, and the ability to develop SAR loses the capacity to develop SAR. The backfill npr1 mutant to obtain the SNC  series of mutants can restore both abilities to some extent [Arabidopsis thaliana mutant SNC4 1D contains an RLK SNC4  gain-of-function mutation, which leads to constitutive activation of the defence response. Identification of two conserved shearing factors, SUA (ABI3-5 repressor gene) and RSN2 , are necessary to construct a defence response by mutating the snc4 1D mutant.The e extent . The AraIn SUA and RSN2 mutants, SNC4 shearing is altered, and further analysis showed that SUA and RSN2 are important receptors for shearing CERK1 . The precursor mRNA is sheared in the receptor-like kinase protein kinase SNC4 and CERK1-mediated regulation of plant immunity . All thePaulownia tomentosa [Populus throughout salt stress, suggesting a completely different tree species produce different AS types to cope with constant stress. Apparently, an alternative acceptor is the major AS pattern when camellia sinensis is treated with drought and heat stress [Interestingly, we found that AS events seem to have specificity across various plant species or in response to a variety of stressful environments. For resistance to different pathogenic bacteria, independent splice variants are also formed. IR was the most common AS pattern observed in omentosa . Exon skt stress .Arabidopsis RPS4 senses and is resistant to the Avr Rps4-expressing strain Pst.DC3000, and RPS4 undergoes variable splicing to produce six different transcripts. After infestation, RPS4 expression was upregulated in intact transcripts, whereas clonally incomplete transcripts could not backfill the resistance of the deficient RPS4 plants to Pst. DC3000, suggesting that only intact RPS4 transcripts are resistant to Pst [C. sinensis is treated with drought and heat stress [t to Pst . In tobat to Pst ,128. Dift stress . Comparit stress . A totalt stress .There are fewer reports related to alternative splicing, but it is easy to see that conserved AS events seem to be consistently present, including the feature that alternative splicing is specific under different stresses as well as under different species. We have also previously summarized the changes in microbial communities under different plants and different stresses, and it is unclear how the release of phytohormones relates to AS, including the recruitment of their preferred microbes by plants, so we speculate that AS may have its own unique splicing.Arabidopsis thaliana to spot genomic regions (splicing quantitative trait loci (sQTLs)) that regulated differential AS, and observed enrichment for trans-sQTLs (trans-sQTL hotspots) on chromosome regions. Many sQTLs were enriched, including the circadian clock, flowering, and stress-responsive genes, suggesting the potential role of differential isoform usage in controlling these necessary processes among diverse ecotypes of Arabidopsis [A. thaliana ecotypes [Although AS is pervasive, the genetic basis for differential isoform usage in plants is increasing, widespread natural variation in AS has been observed in plants, and how AS is regulated and contributes to phenotypic variation is poorly understood. One study performed genome-wide analysis in 666 geographically distributed diverse ecotypes of bidopsis . The preecotypes . Compariecotypes   identified from these plant ecotypes. For instance, a deep understanding of the genetic linkage between AS and secretion of root exudates for microbiome recruitment will be the key to mastering changes in plant ecology and plant breeding. However, further investigations are required to validate this hypothesis and proof of its value to form the new research direction in this field.To this end, studies of both host and microbiome aspects are necessary to unravel the underlying mechanism between plant–microbiome interactions. In the host aspect particularly, the propensity for AS varies under different stress conditions, and unique splice variants are developed to fight against different pathogens. AS triggers intracellular defence signals and mass expression of defence genes, yet the interactions between different plant-associated symbionts and the plant immune system are unknown. Plants recruit microbes in a ‘call for help’ strategy, with different species recruiting different microbes and susceptible versus resistant species recruiting different microbial communities. One major challenge is going to be to research root exudation in natural settings. Due to the chemical complexity of soil, exudation is historically analysed in aquacultural culture, an environment distant from many natural settings of plant microbiome studies. In addition, new technologies are needed for high-throughput screening of functional microorganisms that can be targeted to reveal the impact of core microorganisms on the inter-root biota and, in turn, on plant health. With further understanding of root morphology and secretions, the core strains involved may alter the engineering or breeding of plants with altered talent to act against pathogens. This must be complemented with an improved understanding of the substrate preferences of plant-associated microbes, their interactions, and also the mechanisms which profit the plant. Although research on alternative splicing in plant disease resistance has been ongoing, there are still many unanswered questions about the role of alternative splicing in plant immunity.Regarding commensal microbes, with the evolution of host–microbe interactions, microorganisms have evolved evolutionary strategies for manipulating host AS machinery to disrupt host immunity, and alternative splicing is involved in the mechanism of R-Avr interactions. Whether this mechanism is widespread in multiple species and in disease-resistance systems in which different microorganisms interact with plants has not been fully explained. The mechanism of alternative splicing in the R–Avr interaction, and whether it is widespread in multiple species and in different microbe–plant interactions in the disease resistance system, is also not fully understood; nor are the functional mechanisms of R proteins and subcellular localization. In the future, we need to investigate the functional mechanism, subcellular localization, and interaction with the target protein Avr to further expand our understanding of the molecular mechanism of plant resistance.


It is well-known that some microRNAs (miRNAs) are more important than the others for life, hinting the wide range of miRNA in essentiality or importance. Functional enrichment analysis is a quite pervasive method to dig out the underlying biological pathway for a given gene list and several tools of miRNA set enrichment analysis have been developed. However, all those tools treat each miRNA equally and neglect the importance score of miRNA itself, which could be an obstacle to seek more insightful biological processes for researchers. Here, we developed wTAM, a tool for annotation of weighted human miRNAs, introducing the miRNA importance scores into enrichment analysis. In addition, the annotation repository has been enlarged comparing to TAM. Finally, the case study demonstrated the availability and flexibility of wTAM.http://www.cuilab.cn/wtam/.wTAM is freely available at Bioinformatics Advances online. MicroRNAs (miRNAs) as a class of noncoding RNAs, regulating the expression or translation of messenger RNAs, play crucial roles in various biological processes and human diseases. The strategy to infer the potential pathways or functions that miRNAs involve in after screening differentially expressed miRNAs (DEMs) in a transcriptomic dataset is common and useful. Currently, there are several available tools for annotation of a given miRNA list , has beeFirstly, we obtained 1239 well-annotated miRNA sets from TAM ; PKU-Baidu Fund (2019BD014); the Natural Science Foundation of China (62025102/81970440/81921001); and Peking University Basic Research Program (BMU2020JC001).The data underlying this article are available in the article and in its online supplementary material.Conflict of Interest: There is no conflict.vbab040_Supplementary_DataClick here for additional data file.


Cellular biomechanics underlie a diversity of cellular processes and behaviours, simultaneously being a fundamental determinant of tissue patterning. Core cellular activities, from differentiation and proliferation to migration and apoptosis, are influenced by the cell’s mechanical properties and the ability to discern mechanical signals from their environment. The intrinsic cell machinery internalizes exogenous physical signals from their environment, and a high-precision regulation from a molecular to multicellular level ensures each cell interpret such signals and meets its designated fate. Thus, cells within tissues constantly acquire sensory inputs from their neighbours and the extracellular matrix—detecting, transducing and processing mechanochemical signals to modulate processes and coordinate responses. In this issue, we highlight recent developments in three areas of cell biomechanics in these chapters: 1) Cell and Developmental Biology, 2) Cancer and 3) Bones & Connective Tissues.Alvarez and Smutny reviewed the recent advancements on how actomyosin-contractile forces affect tissue morphogenesis and regulate cell fate in mouse and zebrafish embryonic development. Next, Procès et al. developed a comprehensive review of the heterogenous mechanobiology of the brain, from a molecular to organ level, to help engineer early interventional therapies or treatments for traumatic brain injury, neurodegenerative diseases, and glioblastoma.In the first chapter, Khataee et al. put forward a general model based on the Cellular Potts Model to encapsulate and analyze the interplay between cellular mechanical properties and dynamical transitions in epithelial cell shapes and structures. Interestingly, cell extrusion promotes monolayer-to-multilayer transition based on the mechanical properties of cells and the orientation of cell division.As the complexity of our understanding of mechanobiology grows, theoretical modelling can help interweave existing experimental data and consolidate various parameters into a singular, quantitative system. These computational models can help to form new hypotheses or to unify concepts. Afzal et al. studied how the interaction of placenta and the endometrium eventually results in the deep invasion of placental extravillous trophoblasts into the maternal stroma. They demonstrated that paracrine HB-EGF signalling reduces activated decidual fibroblasts’ enhanced contractility and energetic state by rebalancing the SRF-MRTF-TCF transcriptional axis.While mechanochemical signalling pathways can control both cellular dynamics  and gene expression , their co-regulation is critical for the self-organization of cells into tissues. Dobrokhotov et al. found that actomyosin activity is impaired in cutaneous squamous cell carcinoma (cSCC). External application of tensile loads to adherens junctions by sustained mechanical stretch attenuates the proliferation of cSCC cells. Force-dependent activation of actomyosin in cSCC cells also inhibits their proliferation in a cell-cell contact-dependent manner. Taken together, the malignancy of cSCC cells may be reduced by applying tensile loads to adherens junctions.Further, misregulation of the aforementioned mechanobiological mechanisms can lead to pathological consequences such as cancer and suppressed immunity. In Chapter 2, we focus on cancer mechanobiology. Vahala and Choi review current platforms and biomaterials engineered to mimic the micro and nano-properties of the tumour microenvironment, and subsequent understanding of mechanically regulated pathways in cancer. In essence, cancer cells morphologically adapt to survive in altered environments. Thus, changing tumour ECM properties, including stiffness and ligand chemistry and spacing, are factors that should be considered and incorporated when designing future tools.Besides the adherens junctions, cancer progression also involves remodelling the extracellular matrix (ECM). Wong et al.Next, how contractile forces regulate non-adherent natural killer (NK) cells during cancer surveillance was addressed by They found that lung cancer cells can provoke NK cells and enhance their actomyosin-mediated contractility as a potential early phase activation mechanism. This action shuttles Eomes, an evolutionarily conserved NK cell transcription factor, into the nucleus. NK cells responded to the presumed immunosuppressive TGFβ in the NK-lung cancer coculture medium to sustain its intracellular contractility through myosin light chain phosphorylation, thereby promoting the nuclear localization of Eomes, which likely responds downstream to mechanical stimuli for increased NK cytotoxicity.Choi et al. reviewing the current knowledge of the osteocyte’s role in maintaining bone health and the key regulatory pathways of these mechanosensitive cells. Subsequently, they highlighted the therapeutic opportunities offered by existing treatments and the potential for targeting osteocyte-directed signalling.In the final chapter, we focus on the study of bones and connective tissues, which lies precisely at the interface of biomechanics and mechanobiology. The chapter commences with Boos et al. reviewed the different cartilages and chondrocyte mechanosensing types, then moved on to the multiscale strain transfer through cartilage tissue the involvement of individual ECM components before finally outlining insights to understand multiscale strain transfer in cartilage further. Essentially, the heterogeneity in the spatial variation of ECM molecules leads to a non-uniform, depth-dependent strain transfer and alters the magnitude of forces sensed by cells in articular cartilage and fibrocartilage, influencing chondrocyte metabolism and biochemical response.Delving into the cartilage, Royer and Han discussed the potential mechanobiological pathways involved in the two most popular types of EDS: classical and hypermobile, and their respective, associated comorbidities: mast cell activation syndrome and impaired wound healing—finding that altered mechanosensitive proteins and the lack of collagen V to be main contributors respectively.We end the issue with a review of Ehlers-Danlos Syndromes (EDSs), a group of connective tissue disorders characterized by skin stretchability, joint hypermobility and instability. Though EDS patients typically exhibit lowered elasticity, recent evidence suggests that comorbidities of EDS could also be associated with reduced tissue stiffness. Extending the traditions of our first issue (Forces in Biology) , we hope


Nature, Levo et al. reported that paralogous genes separated by long distances are regulated by specialized DNA stretches called tethering elements, which enable their physical associations and co-dependent transcriptional coupling during Drosophila embryogenesis.1In a recent study published in 3 Accumulating evidence suggests that the dynamic regulation of higher-order chromatin organization is mechanistically interesting and, more importantly, biologically significant. For instance, loop-like structure generated by cohesin complex facilitates the tethering of separated DNA elements into physical proximity for long-range gene regulation. Functionally, the proper organization of chromatin architecture plays an essential role in various biological processes such as lineage specification and cell differentiation, and disruption of these structures can lead to developmental disorders and human diseases.3 While the domain formation by loop extrusion mechanism has been extensively investigated to delineate the key events underlying promoter-enhancer communication and provide a foundation for understanding how distant regulatory elements act on corresponding individual genes to fine-tune their expression, other modes of regulatory interactions have been relatively understudied in the past. In particular, genes with long-range connectivity have been reported to be preferentially transcribed in a co-regulated manner.4 Yet, the causal relationship between spatial engagement of co-regulated genes and their transcriptional coupling remains debatable.The eukaryotic genome is spatially organized via a hierarchy of multi-scale structures.5 for mapping chromatin folding in early fly embryos. This analysis revealed a large number of long-range focal contacts, most of which correspond to paralogues that are co-regulated by shared enhancers. These results therefore hinted that the conventional view of metazoan gene regulation, which claims that individual genes are independently controlled via their own regulatory elements, needs to be revisited to elucidate the co-dependent activation of these interconnected paralogues.Paralogues are duplicated genes that reside at different genomic locations but play interconnected roles in a common biological process. Taking advantage of their overlapping expression pattern during embryonic development, Levo et al. explored the mechanistic principles underlying the coordinated expression of these genes. They first employed Micro-C, a state-of-art 3C-based method that provides finer-scale chromosome organization at nucleosome-resolution,knrl/kni and scyl/chrb, for an in-depth characterization of their coordinated expression. They used CRISPR/Cas9-based targeted genome editing to insert structurally unique stem loops into the intronic region of respective genes, enabling the corresponding nascent transcript to be visualized by fluorescent coat proteins. This tagging strategy allowed the group to spatiotemporally evaluate the transcriptional activity in live embryos. Time-lapse quantitative analysis of nascent transcripts revealed that these distantly located paralogues became physically associated when transcription was turned on. They also exhibited higher co-initiation frequencies than uncoupled genes, supporting their transcriptional coupling during development.The authors then focused on two sets of paralogous genes, Finally, the researchers sought to investigate the molecular underpinnings of synchronized gene expression. Owing to the enhanced resolution of Micro-C, the authors could unambiguously pinpoint the direct attachment sites between long-range connected gene pairs. The prominent feature of these promoter-proximal tethering elements is that they are physically segregated from the respective shared enhancers and are not bound by CTCF, indicating that they are likely formed in a CTCF-independent manner. Genetic perturbation in the tethering elements not only disrupted the spatial contacts between these genes, but also altered their co-dependent transcriptional bursting dynamics, presumably reflecting the destabilization of enhancer-promoter communications. Therefore, the authors concluded that promoter-proximal tethering elements support the promoter-anchored interactions between distant paralogues and contribute to their coordinated expression Fig. .Fig. 1ToAnalogous to bacterial operons that provide a common control mechanism for simultaneous expression of functionally related genes to quickly adapt to environmental changes, eukaryotic paralogous genes involved in the same pathway are also co-regulated to ensure their synchronized activation in response to developmental cues. However, whereas bacterial genes falling under the same operon are physically close in the genome, eukaryotic paralogous genes might be separated by long linear distances. Reflecting this difference, the authors proposed the term “topological operon” to highlight the importance of higher-order chromatin organization for coordinated gene expression. In such eukaryotic operons, functionally related genes are not always in physical proximity, allowing the tethering elements to play a switch-like function to determine when and where the operons become activated. Moreover, the grouping of interconnected genes is no longer constrained by linear genomic distances in topological operons. This gives rise to the possibility that a given enhancer might be shared by different sets of genes because of the changes in 3D organization when cells are shifted from one developmental stage to another. Another possibility is that multiple tethering elements within the topological operon might have different degrees of tethering effect on gene-gene associations, thereby enabling distinct modes of co-regulation under different conditions. These intriguing prospects remain to be rigorously tested in the future.1 They may act in trans and function together with tethering elements to establish the long-range integration for transcriptional co-regulation. Nevertheless, the factors involved in the formation of topological operons remain to be identified. Finally, given the much larger size of mammalian genome than that of Drosophila, whether and to what extent the current findings can provide novel insights into the coordinated expression of functionally-related genes in mammalian cells is currently unclear. Thus, further studies on these crucial issues will be extremely informative in illuminating the detailed mechanisms of topological operon regulation.This study also raises several other interesting questions. For example, what are the upstream signals that instruct the formation of topological operons? Are topological operons exclusively involved in developmental processes? Or can topological operons be reprogrammed in disease settings to promote aberrant gene expression? More importantly, what is the molecular grammar underlying the coordinated regulation of topological operons? Transcription factors such as GAF, CLAMP and Ph appear to be enriched at the promoter-proximal tethering elements.In summary, this study sheds new light on the critical role of genome topology in the co-dependent expression of distant regulatory genes. It raises an interesting question: could this topological operon be the “top secret” underlying long-range transcriptional coupling? As technologies to map genome organization are constantly evolving, the answer to this query will undoubtedly be unveiled soon.


Herbivore-induced plant volatiles (HIPVs) are released by plants upon damaged or disturbance by phytophagous insects. Plants emit HIPV signals not merely in reaction to tissue damage, but also in response to herbivore salivary secretions, oviposition, and excrement. Although certain volatile chemicals are retained in plant tissues and released rapidly upon damaged, others are synthesized de novo in response to herbivore feeding and emitted not only from damaged tissue but also from nearby by undamaged leaves. HIPVs can be used by predators and parasitoids to locate herbivores at different spatial scales. The HIPV-emitting spatial pattern is dynamic and heterogeneous in nature and influenced by the concentration, chemical makeup, breakdown of the emitted mixes and environmental elements  which affect the foraging of biocontrol agents. In addition, sensory capability to detect volatiles and the physical ability to move towards the source were also different between natural enemy individuals. The impacts of HIPVs on arthropod natural enemies have been partially studied at spatial scales, that is why the functions of HIPVs is still subject under much debate. In this review, we summarized the current knowledge and loopholes regarding the role of HIPVs in tritrophic interactions at multiple scale levels. Therefore, we contend that closing these loopholes will make it much easier to use HIPVs for sustainable pest management in agriculture. Volatile organic compounds (VOCs) are released by the majority of vascular plants on a constant basis; however, under biotic and abiotic stress, emissions may significantly increase and change . HerbivoTortrixvirid ian L. (Lepidoptera: Tortricidae) avoided English oak tree, Quercus robur L. , with an herbivore resistant phenotype because they released HIPVs that contained the sesquiterpenes α-farnesene and germacrene-D . Stu. StuHeliInformation transfer within and between plants is one of the many hypothesized functions of HIPVs . NumerouPlant interactions lead to associational resistance. Biological control agents in the immediate environment can detect and process the volatile blend’s composition, which provides precise information about the physiological status of the plant . These oMacrosiphum euphorbiae Thomas (Hemiptera: Aphididae) into potato, Solanum tuberosum L. (Solanum: Solanaceae) was significantly reduced by intercropping with the following three sequences, only potato plant: highest damage, potato with garlic, Allium sativum L. (Amaryllis: Amaryllidaceae) intercropping: lower damaged, potato with onion, Allium cepa L.  intercropping: lowest damaged (Coccinella septempunctata Linnaeus (Coleoptera: Coccinellidae) on potato exposed to onion volatiles: TMTT  was an attractant, whereas (E)-nerolidol acted as a repellent  based HIPVs-lures have been significantly used to attract insect predators  and parasitoids (Braconidae and Ichneumonidae)  in fieldC. carnea, became less attracted to synthetic caryophyllene, and thus detection of the lure thereby waned. However, C. carnea was not attracted to caryophyllene in wheat plants [Field HIPV-based lures are generally applied in crops which also emit HIPV of interest, and this may improve interference with the successful location of lures by natural enemies . Flint et plants  for whict plants . In thest plants ,165.C. carnea than un-baited traps when deployed in peach and cherry orchards [C. carnea would be attracted to phenylacetaldehyde on cotton and wheat of which the headspace volatiles lack this compound [An alternative strategy for reducing the interaction between kairomone-based lure and volatile emission by target field crops could be to use an attractant HIPV in crop fields where HIPV is not emitted or only emitted in minor amounts to improve lure detection within the crop background odor. Phenylacetaldehyde baited field traps attracted 10–100-fold more predator orchards . Interesorchards ,168,169.compound ,171,172.The integration of many aspects that have been covered in this review is necessary to comprehend the interactions between arthropod natural enemies-herbivores-plants in a volatile-mosaic framework. The impression of the volatile mosaic may alter greatly based on the biocontrol agent size and style of movement. For instance, predator/parasitoids with low mobility may view volatile mosaics as fragmented, whereas with high dispersion capacity may not. Further research regarding the variables and mechanisms behind these natural enemies’ mobility and host hunting at the landscape scale needs to be better understood.The discovery that HIPVs are major drivers of trophic interactions sparked a thriving field of highly multidisciplinary study. Since then, a lot of information has been developed, and it is becoming abundantly evident that inducible volatiles support a wide range of functions and ecological roles. Yet, many uncertainties exist, particularly about the mechanisms that are involved in the induction, evolution, release, and perception of the volatiles. An improved understanding of these mechanisms will also help us to better understand the ecological significance and evolution of HIPVs, as well as how we might better utilize them for crop protection. Technological advancements, such as molecular engineering and HIPV detection in real time, will enable precise modification, monitoring, and early identification of agricultural pests and diseases. In the long term, we believe that capitalizing on nature’s own innovations will pave the way for more sustainable, cost-effective agricultural production.The HIPVs play a significant role in host plant–herbivore–natural enemy interactions and have the capacity to improve the effectiveness of host plant resistance and biological control for integrated pest management. However, there is a need to comprehend the ecological importance of HIPVs by integrating molecular and biochemical mechanisms in the production and recognize their ecological functions. Understanding such linkages will offer new options for future research on primary signaling pathways and their ecological repercussions in diverse natural and man-made ecosystems.Longer-term studies of pest management considering HIPVs via promoting natural enemies are required because most research have been short in duration and consequently unable to disclose the effects of HIPVs concerning the changes in environment. In particular, it is yet unclear the impacts of genetically modified crops and pesticide use, shifts in land use in the surrounding landscape, and global warming. Moreover, agri environmental initiatives that pay farmers for stewardship activities provide chances to promote biological control by using HIPVs to encourage pest suppression. However, further study is needed to determine the impact of different HIPVs from plant taxa in a specific agri-ecosystem.Another challenge is to attract and retain adequate natural enemies in crop fields, which are frequently suboptimal environments. To achieve this goal, the “Attract-and-Reward approach” could be achieved by incorporating attractive synthetically manufactured HIPVs with companion non-crop plants, which provide alternative resources to the targeted natural enemies ,173. AltLast but not least, further research needs to be carried out to spot the volatile compounds that command the olfaction-directed behavior of insect pests and their natural enemies. Manipulation of such volatiles would attract the natural enemies of the crop pests for enhancing the effectiveness of bio-control agents for pest management. It will also be uttermost helpful to formulate strategies for developing pest resistant crop varieties with constitutive and induced resistance to insect pests.


Extracellular vesicles (EVs) are small packages that contain proteins, lipids and nucleic acids and are released by various cell types ,2. With EVs are known to mirror the host cells by carrying a variety of molecular cargo such as nucleic acids, lipids, proteins and metabolites . Upon thAmong the many molecules that are incorporated into EVs that are highly implicated in cancer progression, the epidermal growth factor receptor (EGFR) has taken center stage in recent years . This reThe cargo content of EVs often differs with the pathophysiological conditions of the host cells. For instance, the EVs that are isolated from cigarette smoke condensate-treated human small airway epithelial (SAE) cells have a diverse RNA signature compared to the control human airway epithelial cell (AEC)-derived EVs. Corsello et al. were able to identify novel miRNAs that could play a role as potential biomarkers for the diagnosis of cigarette smoke-related diseases . HoweverThe tumour microenvironment (TME) consists of a heterogeneous population of cells that secretes rich cargo-containing EVs, which in turn, can mediate phenotypical changes in the cells within TME and distant sites ,5. This EVs also have recently gained interest as modulators of inflammation and cell death during conditions such as sepsis. The comprehensive review by Sanwlani et al. has summarised types of EVs known to have roles in mediating immune responses leading to cell death and the role of EVs in lung inflammatory disorders . Due to EVs also play an important role in cross-species communication. In recent years, there is an exponential increase in studies that have exploited the anti-cancer properties of bovine milk-derived extracellular vesicles (MEVs) . Fonseka


Bipolar disorder (BD) is a severe mental illness with a strong genetic component. Genetic variations have been involved in the risk of this disorder, including those mediating brain function and neurodevelopment. Early neurodevelopment and neuroprogression processes could be reflected in brain gyrification patterns and help optimize the prediction and diagnosis of such disorders that is often delayed. Previous neuroimaging studies using this measure in patients with bipolar disorder revealed controversial results. This systematic review aimed to summarize available neuroimaging investigations on gyrification in BD compared to healthy controls (HC) and/or other psychiatric groups. Fourteen studies including 733 patients with BD, 585 patients with schizophrenia (SCZ), 90 with schizoaffective disorder (SZA), and 1380 healthy subjects were identified. Overall, a heterogeneous pattern of gyrification emerged between patients with BD and HC. Interestingly, increased gyrification or no differences were also observed in patients with BD compared to those with the schizophrenia-spectrum disorders. Furthermore, relatives of patients with BD showed lower or no differences in gyrification compared to healthy subjects without a family history of affective illness. Differences in the design and in methodological approaches could have contributed to the heterogeneity of the findings. The current review supports an altered brain gyrification pattern that underlies the pathophysiology of BD spanning large anatomical and functional neural networks, associated with altered cognitive functioning, difficulties in processing and affective regulation, and clinical symptoms. Longitudinal studies are needed to test different bipolar phenotypes and pharmacological effects on gyrification.The online version contains supplementary material available at 10.1007/s11682-022-00713-x. Bipolar disorder (BD) is a clinically severe, episodic, lifelong mood disorder  images , defined as the ratio between the perimeters of the pial and outer hull from structural Unfortunately, previous studies employing this measure in BD revealed controversial results and the literature related to gyrification measures in this disorder has not been systematically evaluated so far. Therefore, this systematic review aimed to summarize available published MRI research reporting GI in patients with BD compared to healthy controls (HC) and/or other psychiatric groups.https://osf.io/54xcf/), and adhered to the procedures of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement  AND (bipolar disorder or Bipolar*)".This systematic review followed a pre-defined protocol available online  reporting original findings; c) patients formally diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders (DSM) or the International Classification of Diseases (ICD); d) using high-resolution MRI T1 weighted images; e) including at least a sample of patients with BD; f) evaluated gyrification as part of the analysis and reported the GI values of patients with BD.Experimental, case–control, cross-sectional, and prospective or cohort study designs were considered eligible. Articles meeting the following criteria were included in this review:The authors independently screened the existing literature and extracted the data , with a senior author solving disagreements and making the final decision (F.S.). Each article was required to pass a first- and second-level inspection to be included in this systematic review. This involved a title and abstract search and a full-text search, respectively. The following variables were extracted from a predefined excel spreadsheet: author, year, study design, country, sample size, age group, sex, diagnosis, patient care (inpatient vs. outpatient), duration of illness, diagnostic criteria, and diagnostic tools.The quality of the included studies was assessed with the Imaging Methodology Quality Assessment Checklist , 29 relatives of patients with mixed BD-major depressive disorder , and 1380 healthy controls  were included in this systematic review  approach and ten performed a whole-brain analysis. In general, the results were heterogeneous due to different study protocols and methodological approaches, with seven investigations that did not reveal differences in cortical gyrification between samples  was performed in affected and unaffected offspring of BD and HC. Although volumetric and surface area differences were found, gyrification did not differ between groups Below is the link to the electronic supplementary material.


Non-alcoholic fatty liver disease (NAFLD) is a spectrum of liver disease, ranging from simple steatosis to hepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). Liver fibrosis, which portends a poor prognosis in NAFLD, is characterized by the excessive accumulation of extracellular matrix (ECM) proteins resulting from abnormal wound repair response and metabolic disorders. Various metabolic factors play crucial roles in the progression of NAFLD, including abnormal lipid, bile acid, and endotoxin metabolism, leading to chronic inflammation and hepatic stellate cell (HSC) activation. Autophagy is a conserved process within cells that removes unnecessary or dysfunctional components through a lysosome-dependent regulated mechanism. Accumulating evidence has shown the importance of autophagy in NAFLD and its close relation to NAFLD progression. Thus, regulation of autophagy appears to be beneficial in treating NAFLD and could become an important therapeutic target. NAFLD represents a spectrum of liver disease that can lead to progressive nonalcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma . It is cThe liver is the central organ for fatty acid metabolism. Dietary intake of carbohydrates and lipids, fatty acids (FAs) derived from adipose tissue, can be used for gluconeogenesis, lipogenesis, and ketogenesis in the liver . An imbaAutophagy was first described in the early 1960s, and it has already been proven that certain conditions, such as starvation, lead to its activation . MicroauMacroautophagy is the most prevalent form of autophagy and is considered the dominant activity for selectively trafficking LDs to the lysosome in hepatocytes. Macroautophagy (hereafter referred to as autophagy) involves the de novo formation of a phagophore to engulf cytoplasmic material to form double-membraned vesicles called autophagosomes. The subsequent fusion of autophagosomes with lysosomes forming autolysosomes degrades and recycles cargo to maintain cellular homeostasis .Saccharomyces cerevisiae and Magnaporthe oryzae, autophagy is initiated in the pre-autophagosomal structure (PAS), which is localized in the vicinity of the vacuole [Autophagy is regulated by a set of conserved genes called autophagy-related genes (ATGs). To date, more than 40 ATGs have been identified . Autopha vacuole . In mamm vacuole .The autophagic process is initiated by nutrient starvation, followed by discontinuing TORC1 stimulation and subsequent activation of the Atg1 kinase complex . The ULKThe subsequent expansion and closure of the phagophore are mediated by two ubiquitin (Ubl)-like molecules, Atg12 and LC3/Atg8 . Atg12 iIn eukaryotic cells, autophagy occurs at a basal level and is responsible for the clearance of abnormal proteins, organelles, and infectious agents . DysfuncThe liver is the central organ that regulates lipid metabolism by fatty acid β-oxidation, lipogenesis, and lipoprotein uptake and secretion. However, the liver is not a storage depot for fat. Under normal conditions, the liver processes large quantities of fatty acids with less than 5% steady-state triglyceride (TG) storage . HepatocDietary fat enters the liver as intestinally derived chylomicron remnants or spillover fatty acids . These lIn the fasted state, most fatty acids enter the liver from lipolysis in adipose tissue  . The livExcessive hepatic accumulation of NEFAs leads to cytotoxicity, resulting in cell injuries and cell death in hepatocytes . To prevVLDL assembly is divided into two steps: the first involves the transfer of a small amount of lipid by the microsomal triglyceride transfer protein (MTP) to apolipoprotein; the second requires the fusion of apolipoprotein B-containing precursor particles with TG droplets to form mature VLDLs . An elevHepatic lipid homeostasis is supported by the coordinated regulation of lipid uptake, de novo synthesis, and elimination. The overflow of any metabolite pool can lead to organ dysfunction and subsequent pathologies.The regulatory effect of autophagy on lipid metabolism was first demonstrated by Singh et al. . The stuIn the liver, two major pathways mediate the breakdown of TAGs stored in LDs for subsequent oxidation: lipolysis and lipophagy . LipolysLipophagy involves the formation of autophagosomes, which pinch part of the LD and then fuse with lysosomes to provide the liver with FFAs that can be used as an energy source . Though Various factors can regulate the process of lipophagy through different pathways. Understanding these pathways may provide an attractive approach to preventing NAFLD. As autophagy is usually suppressed by amino acids and insulin through the mTOR- or/and Akt-dependent pathways, suppressed lipophagy is observed in the liver of mice with insulin resistance and hyperinsulinemia induced by a high-fat diet (HFD) ,47. On tLipophagy is also regulated by transcriptional regulators. TFEB, a basic helix-loop-helix leucine zipper protein, is the most studied transcriptional regulator of lipophagy . TFEB reThe nuclear receptor PPARα belongs to the peroxisome proliferator-activated receptors (PPARs) family, which plays an essential role in lipid metabolism. PPARα is highly expressed in the liver, and its activation lowers lipid levels . PharmacFXR is activated by increased bile acid levels after feeding. As bile acids regulate postprandial hepatic transition from a catabolic to an anabolic state, they inhibit hepatic autophagic activity ,58. A prWhile lipophagy appears beneficial in attenuating steatosis in hepatocytes, lipophagy in HSCs is thought to exacerbate its activation and lead to liver fibrosis . Liver fInhibition of autophagy in HSCs has shown the effect of fibrosis attenuation in vivo and in vitro . ConsistIn recent years, a microarray analysis of hepatic miRNAs identified 19 upregulated and 18 downregulated miRNAs in mouse livers with fibrosis . One of However, those results are controversial since some of the studies demonstrated the beneficial role of autophagy in liver fibrosis. For example, the study by Zhang et al.  showed tThe gut and the liver are anatomically and physiologically connected, and their relationship is called the “gut–liver axis” . PreviouPioneering studies showed mutations in autophagy-associated genes as susceptibility factors for Crohn’s disease . SubsequS. typhimurine infection in mice [S. typhimurine both in intestinal epithelial cells and extraintestinal tissues, indicating that autophagy plays a role in limiting bacterial dissemination. A similar study described the role of the autophagy-related gene Atg16l1 in bacteria clearance [Intestinal epithelial autophagy is an essential mechanism for eliminating invading bacteria. A previous study showed that autophagosomes form within 24 h in the small intestinal epithelium after oral  in mice . Immunoflearance . In addilearance . Anotherlearance . Intestilearance ,87, indi−/− mice, indicating that autophagy-deficient mice may be susceptible to pathogenic bacteria [Intracellular tight junctions (TJs) are essential to the gut barrier; they function as gatekeepers for the paracellular pathway. Disruption of the intestinal TJ barrier leads to the permeation of pro-inflammatory molecules from the gut lumen into the circulating system, which is considered a risk factor for NAFLD development . Autophabacteria . A recenbacteria . The aboThe current first-line treatment of NAFLD is lifestyle intervention targeting weight loss through exercise and a hypocaloric diet . BariatrAlthough autophagy is an old concept, its research in liver disease is still at a relatively early stage. In recent years, the role of autophagy in NAFLD has attracted the attention of the academic community and has been extensively studied . AutophaCalorie restriction is the most robust modifiable inducer of autophagy and is a natural dietary therapy that improves health and extends longevity . AutophaEven though current data on the therapeutic effects and mechanism of autophagy in liver diseases are mostly based on in vitro and animal experiments, there is accumulating evidence that abnormal autophagy in the liver is closely related to NAFLD progression. Thus, the regulation of autophagy appears to be beneficial in treating NAFLD and could become an important therapeutic target.


Editorial on the Research TopicImproving voice outcomes after thyroid surgery and ultrasound-guided ablation proceduresThyroid nodules are a common clinical problem, and thyroid surgery remains one of the more common head and neck procedures . In recePost-thyroid procedures dysphonia (PTD) is a common complaint of patients that not only affects the performance of professional voice users but also causes a decline in the quality of life of nonprofessional voice users . PossiblWu et al. reported combined models based on machine learning incorporated with CT radiomics features and the clinicoradiological risk factor and proved the model could be an effective, non-invasive, and safe tool for the preoperative prediction of TCI in PTC. In the study by Liu et al., the authors investigated the impact of Hashimoto's Thyroiditis (HT) on the predictive risk factors of CLNM in PTC and suggested that different predictive systems should be used for HT and non-HT patients to have a more accurate evaluation of central lymph node and determine the appropriate scope of lymph node dissection. In addition, Wang et al. proposed a risk assessment system for CLNM in papillary thyroid microcarcinoma (PTMC) of stage cN0 and explored its application value in clinical practice.Papillary thyroid carcinoma (PTC) is the most common histological subtype of thyroid cancer, and surgery remains the mainstay of treatment. In PTC with thyroid capsule invasion (TCI), extrathyroid invasion (ETE), and central lymph node metastasis (CLNM), the risk of RLN injury is increased during surgery. Therefore, accurate preoperative evaluation TCI, ETE, or CLNM for PTC is important to determine surgical strategies and improve voice outcomes. Yan et al. constructed a nomogram to predict regrowth in patients with benign thyroid nodules after RFA with good discrimination and calibration capabilities. Patients with a high score had an increased probability of nodule regrowth and were potential candidates for additional ablation or surgical treatment. This reliable prognostic nomogram can guide the physician in stratifying patients and provide precise guidance for individualized treatment protocols and improve voice outcomes.Compared to surgery for thyroid cancer, there has been a growing interest in developing the minimally invasive US-guided ablation treatment for benign thyroid nodules. Pace-Asciak et al., surgical and nonsurgical techniques for minimizing and treating PTD caused by an open approach or remote access thyroidectomy and RFA were introduced. Among the techniques for improving voice outcomes, intraoperative neural monitoring (IONM) has now gained widespread acceptance in the international community as an adjunct to visual nerve identification of the RLN and a useful tool for the external branch of superior laryngeal nerve (EBSLN) mapping during thyroid surgery  consensus statement, which outlines general and specific considerations as well as recommended criteria for informed consent for the use of IONM. This consensus statement can assist surgeons and patients in the processes of informed consent and shared decision-making before thyroid surgery.In the review article by  surgery . Wu et aLu et al. proposed a useful clinical surgeon-centered sugammadex protocol according to NMB degree (0.5 mg/kg for deep block and 0.25 mg/kg for others) that provided high IONM quality and adequate surgical relaxation. Recently, Lu et al. also explored the feasibility of neostigmine (0.04 mg/kg) timely reversing NMB by both cisatracurium (0.2 mg/kg) and rocuronium (0.6 mg/kg) in a porcine model. These clinical and experimental studies can expand the options for precision NMB management during monitored thyroidectomy to improve vocal outcomes.Teamwork between surgeons and anesthesiologists plays an important role in successful IONM. The utilization of neuromuscular blocking (NMB) agents facilitates tracheal intubation for general anesthesia. Once tracheal intubation is complete, the degree of NMB turns into a key factor for EMG signaling during IONM. Proper NMB management through the timing and dosage of reversal agents such as sugammadex and neostigmine are undergoing increasing amounts of investigations . Lu et aLiu et al. reviewed the major recent developments of newly emerging transcartilage, percutaneous, and transcutaneous anterior laryngeal recording techniques used in IONM and highlighted their contribution to improved voice outcomes in modern thyroid surgery.During IONM, the major limitation of the EMG tube recording system is the difficulty in maintaining stable contact between tube electrodes and vocal folds during surgical manipulation. Aygun et al. analyzed the clinical and anatomical factors that affect RLN injury and reported that the RLN–inferior thyroid artery (ITA) relationship, extralaryngeal branches, and entrapment of the RLN at the Berry ligament are important factors affecting the development of postoperative VFP. This study concluded that revealing anatomical features with IONM and careful dissection can contribute to the risk reduction of PTD. Chiu et al. analyzed the intraoperative EMG recovery patterns and outcomes after RLN traction injury during IONM. The result shows different recovery patterns have different vocal cord function outcomes, and elucidating the recovery patterns can assist surgeons in intraoperative decision-making and postoperative management.With the application of IONM, Wang et al. reviewed relevant medical literature and compares the safety parameters, such as safe activation distance and cooling time, between different types of EBDs used for thyroid surgery When using EBDs near the RLN in thyroid surgery, surgeons can adopt these safety parameters and follow the standard procedures to avoid laryngeal nerves or soft tissue injuries to improve the postoperative voice outcomes.In addition to the IONM, another important technological advance in thyroid surgery in recent years is the introduction and development of energy-based devices (EBDs). The use of EBDs has many advantages such as reduced blood loss, lower rate of post-operative hypocalcemia, and shorter operation time. However, EBDs generate high temperatures that can cause iatrogenic thermal injury to the RLN by direct or indirect thermal spread. Huang et al. investigated the correlations between IONM findings and voice outcomes in patients with impaired VFM after thyroid surgery. The result showed that severe type RLN injury  raises the risk of unrecovered VFM and moderate/severe long-term postoperative subjective voice impairment. This study suggested that objective voice parameters  can be used as prognostic indicators not only to enable surgeons to earlier identify patients with low voice satisfaction after surgery, but also to enable the implementation of interventions sufficiently early to maintain quality of life.Unrecovered VFP and subjective voice impairment after thyroid surgery causes extreme distress in patients. Huang et al. evaluated the correlation between subjective and objective HPVI in patients after thyroid surgery. The result showed factors that affect a patient's subjective HPVI are complex, and voice stability (Jitter and Shimmer) is no less important than the Fmax level. Therefore, the authors suggested that maximum frequency (Fmax) and Index of voice and swallowing handicap of thyroidectomy (IVST) scores should be interpreted comprehensively, and surgeons and speech-language pathologists (SLPs) should work together to identify patients with HPVI early and arrange speech therapy for them.High-pitched voice impairment (HPVI) is not uncommon in patients without RLN or EBSLN injury after thyroidectomy, which is commonly caused by fibrosis and limited movement of the strap, lateral extralaryngeal, or cricothyroid muscles. Wen et al. reported a retrospective case series in a tertiary teaching hospital to compare the clinical outcomes between different treatment options, including voice therapy (VT), hyaluronic acid (HA) injection, autologous fat injection (FI), and medialization thyroplasty (MT). The results revealed that VT, HA, FI, and MT can all improve the voice outcomes of patients and suggested that the optimal treatment approach should be individualized according to the patient's preference, vocal demand, and the interval between thyroidectomy and intervention. Finally, Liao et al., performed a literature review using the PubMed, Medline, and EMBASE databases to determine the timing, materials, methods, and outcomes of injection laryngoplasty (IL) for unilateral VFP after thyroid surgery. This state of art review supported that IL could improve the voice outcome for unilateral VFP after thyroid surgery, and the autologous fat remains a good augmentation material with a potential longer lasting effect.Concerning the treatment in patients with unilateral VFP after thyroidectomy, In conclusion, this special issue comprises highly qualified papers, which provide novel and comprehensive information for clinicians involved in managing thyroid tumor patients, which includes surgeons, otolaryngologists, endocrinologists, radiologists, internists, speech-language pathologists, family physicians, and other primary care providers, anesthesiologists, nurses, and others. The increasing interest in prevention rather than treatment of PTD may lead to a greater increase in the cost-effectiveness of thyroid tumor treatment, a reduction in procedure-related morbidity, and better health-related quality-of-life .


Angiopathy is a common complication of diabetes mellitus. Vascular endothelium is among the first targets to experience blood-borne metabolic alterations, such as hyperglycemia and hyperlipidemia, the hallmarks of type 2 diabetes. To explore mechanisms of vascular dysfunction and eventual damage brought by these pathologic conditions and to find ways to protect vasculature in diabetic patients, various research approaches are used including in vitro endothelial cell-based models. We present an analysis of the data available from these models that identifies early endothelial cell apoptosis associated with oxidative stress as the major outcome of mimicking hyperglycemia and hyperlipidemia in vitro. However, the fate of endothelial cells observed in these studies does not closely follow it in vivo where massive endothelial damage occurs mainly in the terminal stages of diabetes and in conjunction with comorbidities. We propose that the discrepancy is likely in missing essentials that should be available to cultured endothelial cells to adjust the metabolic state and withstand the immediate apoptosis. We discuss the role of carnitine, creatine, and AMP-activated protein kinase (AMPK) in suiting the endothelial metabolism for long-term function in diabetic type milieu in vitro. Engagement of these essentials is anticipated to expand diabetes research options when using endothelial cell-based models. Vascular endothelium is the primary target of hyperlipidemia, hyperglycemia, and insulin in metabolic disorders and diabetes, leading to endothelial dysfunction and cardiovascular complications at various stages of pathological changes ,2,3. BeiThe critical issue that arises in such an approach is whether the culture conditions adequately match those in situ . ConsideIn particular, metabolic aberrations typical for type 2 diabetes (T2D) have been reproduced in endothelial cell culture in multiple studies in order to reveal how these pathologic conditions affect vascular endothelium. They included exposure of endothelial cells to high environmental glucose or lipid levels typical for T2D progression and implicated in endothelial dysfunction. High content of fatty acids with low carbon number and few/no double bonds in plasma triglycerides and phospholipids are associated with increased risk of diabetes . PalmitiIn vitro studies promptly identified endothelial apoptosis and cell death as the most common and immediate cell response to high glucose (HG) or palmitate . OverallBelow, we review evidence that some essential substances might be missing from the endothelial cell growth media, especially those substances, which are normally produced by other tissues, carried by the blood, and uptaken by vascular endothelium to sustain viability. We suggest that endothelial cells possess an intrinsic mechanism that acts in concert with these essential substances to sense and respond to stressful metabolic conditions in order to tune up metabolism and withstand apoptosis. We discuss metabolic aspects and the key molecules, deteriorated activity of which may lead to vascular endothelium dysfunction.+ and inhibits a key glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase, by its polyADP-ribosylation. This causes upstream accumulation of early glycolytic intermediates, which are diverted to activate sorbitol, hexosamine, diacylglycerol/PKC, and advanced glycation end-products (AGE) pathways leading to apoptosis thymidine incorporation, DNA laddering, TUNEL or annexin-V assays, expression of pro-apoptotic proteins, such as Bax, Bcl-2, or Bak, and caspase-3 activation [Measurement of apoptosis in endothelial cell studies requires caution while interpreting the results. Apoptosis is usually assessed indirectly using [tivation ,25,26,27tivation . AdditioVascular endothelium is exposed to the blood-borne FFA, including the albumin-bound FFA and those liberated by the lipoprotein lipase (LPL) from triacylglycerol (TAG)-enriched lipoproteins at the endothelial surface. About a third of LPL-generated FFA is released into circulation in the process called “spillover” . The lipWhat makes the external FFA detrimental to vascular endothelium in vitro? Low solubility of long-chain FFA in aqueous solutions is the pivotal factor . In blooNormally, the bulk of FFA traverses the vascular endothelium for ultimate destination in underlying tissues such as skeletal muscle or adipose tissue. A portion of FFA is retained in endothelial cells and may enter various pathways.The ability of carnitine to enhance oxidation of fatty acids by vascular endothelium in vitro was demonstrated more than three decades ago . The samCarnitine supplementation appears important for all standard growth media  used to culture endothelial cells in stressful conditions that may critically involve oxidation of fatty acids . As emphThe key observation made by Ido et al.  indicateAdditionally, carnitine may participate in maintaining an equilibrium between the pools of the long-chain acyl-CoA (LCACoA) and free HS-CoA in cells. The intracellular concentrations of LCACoA are normally in the 5–160 μM range but vary considerably in different metabolic conditions . The excCarnitine is synthesized mainly in the liver and distributed through the blood to other tissues including vascular endothelium. With carnitine concentration in the blood plasma at ~50 μM, it is expected to be about the same in endothelium that is constantly exposed to blood. This is in the range of free and acyl-bound CoA. Thus, due to intrinsic ability to reversibly exchange the acyl groups with many acyl-CoA species ranging from simple acetyl-CoA to LCACoA, carnitine may be essential to maintain the availability of the HS-CoA pool for metabolic needs and to sequester excess LCACoA  in metabThe original discovery that, in addition to being a precursor for the de novo synthesis of fatty acids, malonyl-CoA serves as an acyl-carnitine transferase inhibitor  phagy. This appears to occur in cells treated with exogenous fatty acids ,58,59 orWhether and to what extent the above-mentioned mechanisms are realized in the presence of carnitine and sufficient AMPK activity and lead to endothelial cell dysfunction and damage needs to be re-evaluated.H+) across the inner mitochondrial membrane and reduces ATP synthase activity [Mitochondria are considered the major source of superoxide anion and other ROS, at least in hyperglycemia ,69,70. Ractivity . In mammactivity . Protectactivity . High glactivity ,78, likeactivity ,80. In tactivity ,79. By tactivity . All theactivity .H+), increases oxidation of electron carrier pools, and reduces local oxygen to cease superoxide anion radical production in mitochondria [There is also another potential mechanism that may rescue the mitochondrial electron transport chain from overloading. Inability of a cell to spend ATP increases the mitochondrial ROS; restoring the ATP generation lowers the proton electrochemical potential gradient  shares A number of studies have elaborated the antioxidant potential of creatine , howeverSumming up, the in vitro endothelial models require a thoughtful approach with regard to essential natural components of the culture medium to be available for endothelial cells in order to cope with external metabolic interrogations, such as those occurring in diabetic states. Both high glucose and increased FFA load alter the cellular metabolism to increase intracellular malonyl-CoA and LCACoA and thrust them into the cytosolic lipogenesis flux and/or into mitochondria for the β-Ox/FAO and OxPhos pathways . The choUnder high glucose conditions, an increased glycolytic flux triggers three major pathways involved in endothelial cell damage: through polyol/sorbitol or G3P-initiated AGE formation, or F1,6PP-initiated hexosamine pathway . It alsoInvolvement of ROS in endothelial dysfunction in diabetic conditions is hardly disputed and seems to be a unifying pathogenic mechanism ,20. Yet,Is there a safeguard regulatory circuit present in cultured/diabetic endothelial cells to protect from detrimental ROS generation? AMPK appears to be an important molecular component of such a circuit. AMPK is an intrinsic double-faced regulator of the cytoplasmic malonyl-CoA and LCACoA levels and mitochondrial ROS . AMPK exIn conclusion, the endothelial cell-based models appear indispensable to explore cellular mechanisms of vascular dysfunction in diabetes-related conditions such as hyperglycemia and/or dyslipidemia. For metabolic studies, the cells need to be constantly supplied with essential factors. Some of them may not yet be fully elaborated, likely because the harsh metabolic conditions had not been envisioned when common cell media were being developed. As evidence accumulates, these essentials may be needed to revive the intrinsic safeguard mechanisms, such as those involving AMPK, to sustain mitochondrial function, combat oxidative stress and prevent artificially high apoptosis. This allows maintaining the cultures for up to several weeks to monitor the long-term progression of cell metabolic dysfunctions. Still, endothelial cell-based models retain limitations with regard to systemic responses imposed by hemodynamic forces or communication with other cells, including blood cells or vascular wall neighbors. These aspects can be also addressed in advanced coculture systems. Clearly, more studies are needed, under the controlled cell culture conditions, to reap the harvest.