2,4,6-trinitrobenzene sulfonic acid-induced chronic colitis with fibrosis and modulation of TGF-β1 signaling

AIM:To investigate whether targeting proteasome might reverse intestinal fibrosis in rats.METHODS:Chronic colitis was induced in rats by repeated administration of increasing dose of2,4,6-trinitrobenzene sulfonic acid(TNBS,15,30,45,60,60,60 mg)by rectal injection for 6 wk(from day0 to day 35),while control rats received the vehicle.TNBS+bortezomib(BTZ)rats received intraperitoneal injections of BTZ twice weekly(from day 37 to day44)at a dose of 25 mg/kg,whereas the control and TNBS groups received the same amount of the vehicle.Histologic scoring of inflammation and fibrosis was performed.Colonic production of transforming growth factor(TGF)-βwas measured by ELISA.Colon fibrosisrelated proteins such as phospho-p38,phosphoSMAD2/3,Akt and peroxisome proliferator activated receptorγ(PPARγ)were studied by western blot.Expression of the tight junction proteins,occludin and claudin-1,were assessed by Western blot.Colon proteasome activities(chymotrypsin-like and trypsinlike activities)were assessed.RESULTS:TNBS-treated rats had a higher colon weight/length ratio compared to control rats(P<0.01).Furthermore,fibrosis and inflammation scores were higher in TNBS-treated rats compared to control rats(P<0.01 for both).Colonic production of TGF-βproduction tended to be higher in TNBS-treated rats(P<0.06).Fibrosis-related proteins such as phospho-p38,phospho-SMAD2/3,and PPARγwere significantly higher in TNBS-treated rats compared to control rats(all P<0.05).TNBS rats had a higher expression of Akt compared to control rats(P<0.01).Tight junction proteins were modified by repeated TNBS challenge:colon occludin expression rose significantly(P<0.01),whereas claudin-1 expression fell(P<0.01).Bortezomib inhibition significantly decreased chymotrypsin-like activity(P<0.05),but had no significant effect on trypsin-like activity(P>0.05).In contrast,bortezomib had no effect on other studied parameters such as fibrosis score,TGF-βsignaling,or tight junction expression(P>0.05 for all).CONCLUSION:Rats with TNBS-induced chronic colitis exhibited colon fibrosis associated with higher TGF-βsignaling.Proteasome inhibition by bortezomib had no effect on fibrosis in our experimental conditions.

Acellular Mineralized Exoskeleton Shrimp (MES): A Natural Source of Bioactive Factors for Tissue Regeneration—Preliminary Results

Current challenges in the development of scaffolds for bone regeneration include the engineering of biomaterials that can withstand a natural dynamic physiology on the bone that provides a matrix capable of supporting cell migration and tissue ingrowth. The objective of the present work was to develop and characterize a new biomembrane—Mineralized Exoskeleton Shrimp (MES) developed from the exoskeleton of paleomonetes. The integration of MES as a biomaterial for tissue regeneration relies on the growing evidence that the shrimp is characterized by a hierarchically arranged chitin fiber structure, mineralized predominately by calcium carbonate and/or calcium phosphate, bringing beneficial effects in bone regeneration. Additionally, the tridimensional MES structure, can act as a “tent” for Guided Bone Regeneration (GBR). Recently, our team has characterized the MES biomaterial by in vitro (human osteoblastic cellular cultures and immersion of the membrane in modified synthetic plasma) and in vivo (soft tissue in lab mice and hard tissue in rabbit model). The cellular growth in the MES membrane was very exuberant in cellular culture with osteoblastic colonization on its surface (histophilic and biocompatible). After the immersion in modified synthetic plasma for one week, a mass mineralization occurred throughout the membrane’s surface (bioactive). The analysis of histological samples from experimental surgery in lab mice showed that the MES membrane wasn’t toxic to soft tissues and that it caused a moderate inflammatory response (first reabsorption signs at 8 weeks). The MES could act as a cell-guiding template that contains the necessary cues and adequate three-dimensional set to facilitate cell adhesion and promote tissue regeneration upon implantation and subsequent biodegradation.

Crohn’s disease: Why the ileum?

Crohn’s disease(CD)is an inflammatory bowel disease characterized by immunemediated flares affecting any region of the intestine alternating with remission periods.In CD,the ileum is frequently affected and about one third of patients presents with a pure ileal type.Moreover,the ileal type of CD presents epidemiological specificities like a younger age at onset and often a strong link with smoking and genetic susceptibility genes.Most of these genes are associated with Paneth cell dysfunction,a cell type found in the intestinal crypts of the ileum.Besides,a Western-type diet is associated in epidemiological studies with CD onset and increasing evidence shows that diet can modulate the composition of bile acids and gut microbiota,which in turn modulates the susceptibility of the ileum to inflammation.Thus,the interplay between environmental factors and the histological and anatomical features of the ileum is thought to explain the specific transcriptome profile observed in CD ileitis.Indeed,both immune response and cellular healing processes harbour differences between ileal and non-ileal CD.Taken together,these findings advocate for a dedicated therapeutic approach to managing ileal CD.Currently,interventional pharmacological studies have failed to clearly demonstrate distinct response profiles according to disease site.However,the high rate of stricturing disease in ileal CD requires the identification of new therapeutic targets to significantly change the natural history of this debilitating disease.

Longitudinal decrease in platelet counts as a surrogate marker of liver fibrosis

BACKGROUND Liver cirrhosis is a significant source of morbidity and mortality worldwide.The disease is usually indolent and asymptomatic early in its course while many cirrhotic patients are diagnosed late when severe complications occur.A major challenge is to diagnose advanced fibrosis as early as possible,using simple and non-invasive diagnostics tools.Thrombocytopenia represents advanced fibrosis and portal hypertension(HTN)and most non-invasive scores that predict liver fibrosis incorporate platelets as a strong risk factor.However,little is known about the association between longitudinal changes in platelet counts(PTC),when still within the normal range,and the risk of cirrhosis.AIM To explore whether platelet counts trajectories over time,can predict advanced liver fibrosis across the different etiologies of liver diseases.METHODS A nested case-control study utilizing a large computerized database.Cirrhosis cases(n=5258)were compared to controls(n=15744)matched for age and sex at a ratio of 1:3.All participants had multiple laboratory measurements prior to enrollment.We calculated the trends of PTC,liver enzymes,bilirubin,international normalized ratio,albumin and fibrosis scores(fibrosis-4 and aspartate transaminase-to-platelet ratio index)throughout the preceding 20 years prior to cirrhosis diagnosis compared to healthy controls.The association between PTC,cirrhosis complications and fibrosis scores prior to cirrhosis diagnosis was investigated.RESULTS The mean age in both groups was 56(SD 15.8).Cirrhotic patients were more likely to be smokers,diabetic with chronic kidney disease and had a higher prevalence of HTN.The leading cirrhosis etiologies were viral,alcoholic and fatty liver disease.The mean PTC decreased from 240000/μL to 190000/μL up to 15 years prior to cirrhosis diagnosis compared to controls who’s PTC remained stable around the values of 240000/μL.This trend was consistent regardless of sex,cirrhosis etiology and was more pronounced in patients who developed varices and ascites.Compared to controls whose values remained in the normal range,in the cirrhosis group aspartate aminotransferase and alanine aminotransferase,increased from 40 U/L to 75 U/L and FIB-4 increased gradually from 1.3 to 3 prior to cirrhosis diagnosis.In multivariable regression analysis,a decrease of 50 units in PTC was associated with 1.3 times odds of cirrhosis(95%CI 1.25-1.35).CONCLUSION In the preceding years before the diagnosis of cirrhosis,there is a progressive decline in PTC,within the normal range,matched to a gradual increase in fibrosis scores.

Spinal cord injury:can we repair spinal cord non-invasively by using magnetic stimulation?

Spinal cord injury(SCI)is current l y an incurable condition which induces sensorimotor impairments below the injury level.Mainly,SCI are the consequence of physical damages which occur on spinal cord due to traffic accidents or sports and recreation injuries.To date,nor treatment of therapy could be proposed to patients with SCI(Wilson et al.,2012).

纤维与纺织品的阻燃新进展与挑战（英文）

20世纪50～60年代，各种织物、纤维的耐久阻燃整理技术飞速发展，这些产品和工艺现已被大量应用于织物和纤维的阻燃耐久整理。但随着市场对环境、化学品毒性、成本及性能等方面的要求愈加严苛，其中多数产品的使用受到了质疑。“可否找到使用中释放甲醛类阻燃剂的替代品用于纤维素织物的耐久阻燃”已成为常被提及的问题。而关于“能否生产出成炭性能良好的聚酯阻燃剂”和“能否制备出高效无卤阻燃剂用于织物涂层和背涂”也备受关注。相关研究回顾表明，人们从20世纪下半叶就在寻求这些问题的解决方法，而这些工作也为目前最新的商业化阻燃纤维和纺织品发展奠定了基础；其次，本世纪前十年的研究主要侧重于解决环境可持续发展问题、寻找可替代的阻燃配方、满足合成纤维提高成炭的需求以及纳米技术的应用。另外，文章还对纤维的微表面改性和纳米表面改性的作用以及整理过程中等离子体、层层自组装、溶胶一凝胶等技术在织物阻燃整理中的应用进行了综述。

Bladder-colon chronic cross-sensitization involves neuro-glial pathways in male mice

BACKGROUND Irritable bowel syndrome and bladder pain syndrome often overlap and are both characterized by visceral hypersensitivity.Since pelvic organs share common sensory pathways,it is likely that those syndromes involve a cross-sensitization of the bladder and the colon.The precise pathophysiology remains poorly understood.AIM To develop a model of chronic bladder-colon cross-sensitization and to investigate the mechanisms involved.METHODS Chronic cross-organ visceral sensitization was obtained in C57BL/6 mice using ultrasound-guided intravesical injections of acetic acid under brief isoflurane anesthesia.Colorectal sensitivity was assessed in conscious mice by measuring intracolonic pressure during isobaric colorectal distensions.Myeloperoxidase,used as a marker of colorectal inflammation,was measured in the colon,and colorectal permeability was measured using chambers.c-Fos protein expression,used as a marker of neuronal activation,was assessed in the spinal cord(L6-S1 level)using immunohistochemistry.Green fluorescent protein on the fractalkine receptor-positive mice were used to identify and count microglia cells in the L6-S1 dorsal horn of the spinal cord.The expression of NK1 receptors and MAPK-p38 were quantified in the spinal cord using western blot.RESULTS Visceral hypersensitivity to colorectal distension was observed after the intravesical injection of acetic acid vs saline(P<0.0001).This effect started 1 h post-injection and lasted up to 7 d postinjection.No increased permeability or inflammation was shown in the bladder or colon 7 d postinjection.Visceral hypersensitivity was associated with the increased expression of c-Fos protein in the spinal cord(P<0.0001).In green fluorescent protein on the fractalkine receptor-positive mice,intravesical acetic acid injection resulted in an increased number of microglia cells in the L6-S1 dorsal horn of the spinal cord(P<0.0001).NK1 receptor and MAPK-p38 levels were increased in the spinal cord up to 7 d after injection(P=0.007 and 0.023 respectively).Colorectal sensitization was prevented by intrathecal or intracerebroventricular injections of minocycline,a microglia inhibitor,by intracerebroventricular injection of CP-99994 dihydrochloride,a NK1 antagonist,and by intracerebroventricular injection of SB203580,a MAPK-p38 inhibitor.CONCLUSION We describe a new model of cross-organ visceral sensitization between the bladder and the colon in mice.Intravesical injections of acetic acid induced a long-lasting colorectal hypersensitivity to distension,mediated by neuroglial interactions,MAPK-p38 phosphorylation and the NK1 receptor.

Dental pulp stem cells and BonelikeVR for bone regeneration in ovine model

Development of synthetic bone substitutes has arisen as a major research interest in the need to find an alternative to autologous bone grafts.Using an ovine model,the present pre-clinical study presents a synthetic bone graft(BonelikeVR)in combination with a cellular system as an alternative for the regeneration of non-critical defects.The association of biomaterials and cell-based therapies is a promising strategy for bone tissue engineering.Mesenchymal stem cells(MSCs)from human dental pulp have demonstrated both in vitro and in vivo to interact with diverse biomaterial systems and promote mineral deposition,aiming at the reconstruction of osseous defects.Moreover,these cells can be found and isolated from many species.Non-critical bone defects were treated with BonelikeVR with or without MSCs obtained from the human dental pulp.Results showed that BonelikeVR and MSCs treated defects showed improved bone regeneration compared with the defects treated with BonelikeVR alone.Also,it was observed that the biomaterial matrix was reabsorbed and gradually replaced by new bone during the healing process.We therefore propose this combination as an efficient binomial strategy that promotes bone growth and vascularization in non-critical bone defects.

Immune regulatory networks coordinated by glycans and glycan-binding proteins in autoimmunity and infection

The immune system is coordinated by an intricate network of stimulatory and inhibitory circuits that regulate host responses against endogenous and exogenous insults.Disruption of these safeguard and homeostatic mechanisms can lead to unpredictable inflammatory and autoimmune responses,whereas deficiency of immune stimulatory pathways may orchestrate immunosuppressive programs that contribute to perpetuate chronic infections,but also influence cancer development and progression.Glycans have emerged as essential components of homeostatic circuits,acting as fine-tuners of immunological responses and potential molecular targets for manipulation of immune tolerance and activation in a wide range of pathologic settings.Cell surface glycans,present in cells,tissues and the extracellular matrix,have been proposed to serve as“self-associated molecular patterns”that store structurally relevant biological data.The responsibility of deciphering this information relies on different families of glycan-binding proteins(including galectins,siglecs and C-type lectins)which,upon recognition of specific carbohydrate structures,can recalibrate the magnitude,nature and fate of immune responses.This process is tightly regulated by the diversity of glycan structures and the establishment of multivalent interactions on cell surface receptors and the extracellular matrix.Here we review the spatiotemporal regulation of selected glycan-modifying processes including mannosylation,complex N-glycan branching,core 2 O-glycan elongation,LacNAc extension,as well as terminal sialylation and fucosylation.Moreover,we illustrate examples that highlight the contribution of these processes to the control of immune responses and their integration with canonical tolerogenic pathways.Finally,we discuss the power of glycans and glycan-binding proteins as a source of immunomodulatory signals that could be leveraged for the treatment of autoimmune inflammation and chronic infection.

Mannosylated glycans impair normal T-cell development by reprogramming commitment and repertoire diversity

T-cell development ensures the formation of diverse repertoires of T-cell receptors(TCRs)that recognize a variety of antigens.Glycosylation is a major posttranslational modification present in virtually all cells,including T-lymphocytes,that regulates activity/functions.Although these structures are known to be involved in TCR-selection in DP thymocytes,it is unclear how glycans regulate other thymic development processes and how they influence susceptibility to disease.Here,we discovered stage-specific glycome compositions during T-cell development in human and murine thymocytes,as well as dynamic alterations.After restricting the N-glycosylation profile of thymocytes to high-mannose structures,using specific glycoengineered mice(Rag1CreMgat1fl/fl),we showed remarkable defects in key developmental checkpoints,includingß-selection,regulatory T-cell generation andγδT-cell development,associated with increased susceptibility to colon and kidney inflammation and infection.We further demonstrated that a single N-glycan antenna(modeled in Rag1CreMgat2fl/fl mice)is the sine-qua-non condition to ensure normal development.In conclusion,we revealed that mannosylated thymocytes lead to a dysregulation in T-cell development that is associated with inflammation susceptibility.

The impact of the design of learning spaces on attention and memory from a neuroarchitectural approach:A systematic review

Enriched environments in animal models have demonstrated that exposure to an optimal stimulus improves behavior,cognition,and genomics.However,the evidence base for the neurophysiological influence of human environment enrichment has not been extensively studied.This systematic review compiles indicators about the effect of built,indoor environments on the cognitive processes of memory and attention in humans.This work pursues two main objectives:(1)to define current knowledge and the methods that are useful and identify whether previously published studies indicate consistencies and(2)to report the approaches and strategies that can be used in evaluating cognitive processes affected by environment response.Results of this systematic review show that(1)form and geometry,(2)space distribution and context,(3)color and texture,(4)height,width,and enclosure,(5)transition and circulation,and(6)light,sound,and temperature have an impact on memory and/or attention,and they can be assessed objectively.Despite all the advances in this field,methodological limitations and a lack of cross-validated standard protocols are found.Therefore,future research is necessary to provide a deep insight into how human cognition can be heightened by the environment to which it is exposed.

Impact of phytoconstituents on oral health practices:a post COVID-19 observation

Appropriate oral hygiene significantly reduces the possibility of oral infections.However,dental caries and periodontal diseases are major oral health issues causing chronic diseases due to poor oral health.Recently,herbal compounds have gained interest in maintaining oral health.Extracts of burdock root(Arctium),noni fruit(Morinda citrifolia),and neem leaf(Azadirachta indica)are now used as intracanal medicaments in endodontics and periodontics.Plectranthus amboinicus species and other plants produces essential oil likeβ-caryophyllene,p-cymene,andγ-terpinene can exhibit antibacterial activity;highlighting phytoconstituents plays a vital role in oral health.The COVID-19 pandemic highlighted the importance of hygiene and sanitization,to curb SARS-CoV-2.Oral cavity is among the gateways for virus entry into saliva.Saliva is a potential reservoir of SARS-CoV-2,and there is an increased risk of infection if there is any fissure in the mouth.This enables entry of virus into the vascular system through gingival or periodontal pocket,possibly reaching lung periphery then to lung vessels by interacting with endothelial surface receptors triggering pulmonary vasoconstriction and lung damage due to endothelial dysfunction.This review aims to draw attention to the possible route of SARS-CoV-2 infection via the oral cavity and the importance of oral hygiene against COVID-19.

Sustained activation of P2X7 induces MMP-2- evoked cleavage and functional purinoceptor inhibition

P2X7 purinoceptor 支持幸存或 cytotoxicity 取决于细胞外的腺苷 triphosphate (ATP ) 刺激紧张控制它的离子隧道或 P2X7 依赖的大毛孔(LP ) 工作。管理这运作的分叉和功能的特质的机制是听说病的。在 P2X7 的持续激活触发活跃矩阵 metalloproteinase 的版本的地方，我们发现了一个反馈环 2 (MMP-2 ) ，它经由 MMP-2-dependent 受体劈开停止离子隧道和 LP 回答。这机制在象巨噬细胞， dystrophic myoblasts， P2X7-transfected HEK293，和人的瘤房间一样多样的房间操作。给也堵住的那项浆液出生的 MMP-2 活动受体功能，在 vivo 的 P2X7 回答可以与可渗透的 capillaries 在机关减少。因此，这机制代表 P2X7 功能的重要调整，依赖房间自治、体外的因素。确实，它与高 P2X7 表示层次在巨噬细胞和人的癌症房间从导致 ATP 的 cytotoxicity 允许避免。最后，我们证明那 P2X7 脱离在 vivo 在煽动的 dystrophic 肌肉消除了 gelatinase 活动。因此， P2X7 对手能在煽动性的疾病和癌症的治疗被用作高度有毒的 MMP 禁止者的一种选择。

Use of Intraoral Three-dimensional Images for the Identification of Dental Morphological Traits Related to Ancestry Estimation

Victim identification through dental features is one of the main objectives of forensic dentistry.In circumstances where information regarding antemortem dental records is missing,reconstruction of a biological profile can be useful as a first step toward personal identification.This reconstructive method provides valuable information,namely regarding the individual’s ancestry,through the detection and degree of expression of dental morphological traits,which may help to restrict the number of candidates for identification.Technological advances allowed the development of alternative methods for dental evaluation,that complement or substitute those already in use in clinical practice.Among these,intraoral three‑dimensional(3D)images are increasingly used in dentistry,as they have a high level of accuracy and are easy to obtain and store.However,a fundamental question regarding forensic dentistry is whether they allow recognition and analysis of dental morphological traits in detail,namely those related to ancestry.In this study,we evaluated 20 teeth morphological features using intraoral 3D imaging from 77 individuals from Northern Portugal.Our results showed that it was possible to identify and classify a large part of the main morphological traits used in the estimation of ancestry.As these 3D images present sufficient morphological detail to be classified,we believe that future applications of this technique can be expected in forensic dentistry.

Salicylic Acid Regulates Pollen Tip Growth ：hrough an NPR3/NPR4-1ndependent Pathway

Tip growth is a common strategy for the rapid elongation of cells to forage the environment and/or to target to long-distance destinations. In the model tip growth system of Arabidopsis pollen tubes, several small- molecule hormones regulate their elongation, but how these rapidly diffusing molecules control extremely localized growth remains mysterious. Here we show that the interconvertible salicylic acid （SA） and meth- ylated SA （MESA）, well characterized for their roles in plant defense, oppositely regulate Arabidopsis pollen tip growth with SA being inhibitory and MeSA stimulatory. The effect of SA and MeSA was independent of known NPR3/NPR4 SA receptor-mediated signaling pathways. SA inhibited clathrin-mediated endocytosis in pollen tubes associated with an increased accumulation of less stretchable demethylated pectin in the apical wall, whereas MeSA did the opposite. Furthermore, SA and MeSA alter the apical activation of ROP1 GTPase, a key regulator of tip growth in pollen tubes, in an opposite manner. Interestingly, both MeSA methylesterase and SA methyltransferase, which catalyze the interconversion between SA and MESA, are localized at the apical region of pollen tubes, indicating of the tip-localized production of SA and MeSA and consistent with their effects on the apical cellular activities. These findings suggest that local generation of a highly diffusible signal can regulate polarized cell growth, providing a novel mechanism of cell polarity control apart from the one involving protein and mRNA polarization.