Diagnosis of neurodegenerative disease (NDD) is complex, therefore simpler, less invasive, more accurate biomarkers are needed. small non-coding RNA (sncRNA) dysregulates in NDDs and sncRNA signatures have been explored for the diagnosis of NDDs, however, the performance of previous biomarkers is still better. Astrocyte dysfunction promotes neurodegeneration and thus derived scnRNA signatures could provide a more precise way to identify of changes related to NDD course and pathogenesis, and it could be useful for the dissection of mechanistic insights operating in NDD. Often sncRNA are transported outside the cell by the action of secreted particles such as extracellular vesicles (EV), which protect sncRNA from degradation. Furthermore, EV associated sncRNA can cross the BBB to be found in easier to obtain peripheral samples, EVs also inherit cell-specific surface markers that can be used for the identification of Astrocyte Derived Extracellular Vesicles (ADEVs) in a peripheral sample. By the study of the sncRNA transported in ADEVs it is possible to identify astrocyte specific sncRNA signatures that could show astrocyte dysfunction in a more simpler manner than previous methods. However, sncRNA signatures in ADEV are not a copy of intracellular transcriptome and methodological aspects such as the yield of sncRNA produced in ADEV or the variable amount of ADEV captured after separation protocols must be considered. Here we review the role as signaling molecules of ADEV derived sncRNA dysregulated in conditions associated with risk of neurodegeneration, providing an explanation of why to choose ADEV for the identification of astrocyte-specific transcriptome. Finally, we discuss possible limitations of this approach and the need to improve the detection limits of sncRNA for the use of ADEV derived sncRNA signatures. Neurodegenerative diseases (NDDs) are caused by a general dysfunction of the Central Nervous System (CNS), characterized by progressive loss of neuron structure and function . In receAnother characteristic of NDD diagnosis is the complexity associated with paraclinical assessment which troubles timely diagnosis. Detection of distinct hallmarks of NDDs has been possible with neuroimaging and CSF evaluation up to very early time points before symptoms onset (~30 years) . HoweverAstrocytes regulate central processes for neuronal microenvironment maintenance, protection, and repair ,22, inclAnother family of biomarkers deserving interest for their discriminative potential are small non-coding RNAs (sncRNAs) signatures. It is well known that micro-RNA (miRNA) dysregulation is associated with NDD and neurodegeneration ,30. AddiFurthermore, studying sncRNA signatures in a cell specific manner would improve the precision of peripheral signatures of sncRNA. This could be accomplished by isolating cell-specific EVs—which overpass the BBB—and studying their sncRNA content. Recently, flow cytometry separation of Astrocyte Derived Extracellular Vesicles (ADEVs) with GFAP revealed upregulation in the symptomatic phase of a MS mouse model ; this reJovicic & Gitler, (2017)  showed t11C-deuterium-L-deprenyl (11C-DED) and 11C-BU99008, both of which recognize reactive astrocytes [11C-DED recognize monoamine oxidase B (MAO-B) and have also been found in MS [Neurodegenerative disease (ND) etiology is particularly difficult to define, for instance Aβ peptides, which are the most representative hallmark of Alzheimer’s disease (AD), have been observed to reduce oxidative stress in some circumstances, blurring the limits between pathological and physiological changes , therefotrocytes ,50 and htrocytes , and symnd in MS . TherefoAstrocytes perform multifold functions some of them essential such as regulating electrochemical balance, distributing the energy uptake (meeting the high energy requirement of neurons), and accelerating detoxification which protect neurons from oxidative damage . SeveralFor instance, in Alzheimer Disease (AD) abnormal astrocyte exposure to saturated fatty acids (SFF) such as stearic, linoleic, oleic and palmitic acid (PA) has been linked higher risk ,57. HighAnimal models and human also support in vitro observations situating astrocyte dysfunction as an early pathological event in AD. In animal models, High Fat Diet (HFD) robustly induced cognitive impairment in healthy rat  and Tg25Parkinson’s Disease (PD) development is partly due to astrocyte dysfunction with evidence building up to suggest early astroglial participation in neuroinflammation and in the disruption of several neuroprotective mechanisms as partly initiator events . SeveralEfforts in Vascular Dementia (VD) early detection center in the identification of subclinical symptoms of cerebrovascular conditions that precede it since a stroke constitute a risk marker by itself. Most VD cases arise after vascular damage caused by smaller and less evident pathophysiological lesions such as lacunar infarct . In mostIn Multiple Sclerosis (MS), astrocyte dysfunction takes place earlier than previously believed . Evidence shows that, before lesion demyelination, astrocytes contribute to the inflammatory response that results in the recruitment of lymphocytes and regulate BBB and BSCB permeability to favor lesion . EvidencOther alterations associated with increased risk of NDD cause alterations in astrocyte function. A proinflammatory microenvironment, oxygen and glucose deprivation, intracerebral hemorrhage, traumatic brain injury (TBI), Postoperative Cognitive Dysfunction (PCD) and ethanol exposure, represent conditions inducing increased risk of NDD ,77,78,79Biomarkers can be classified according to their application in risk, screening, diagnostic, prognostic, monitoring and predictive/therapeutic ,166,167,Misdiagnosis is also found in NDD increasing in early phases ,169,170,RNA signatures consist of the validation of an expression profile in a subcellular location, cell, tissue, or organism for the recognition of pathological or physiological states, and the activated cellular pathways . These sIdentification of cell-specific signatures would increase accuracy of sncRNA measures, by blocking the influence of confounding factors affecting circulating sncRNA expression such as changes in cell proportion and secretion  and can An RNA carrier is any cell derived structure known to carry extracellular RNA with a protective role of its RNA. Extracellular vesicles (EVs) participate in a safe cell-to-cell RNA delivery and according to their biogenesis can be subtyped in at least three categories: exosomes, microvesicles (MVs) and apoptotic bodies (ApoBDs) . ExosomeFor a widespread diagnosis of neurodegenerative disease (ND), cheaper, painless, and less invasive, simpler biomarkers should be designed, especially for preclinical and prodromal sceneries. Several reports evidence that EVs originated from astrocytes (ADEV) appear in peripheral fluids such as mouse  or humanNon vesicular carriers for sncRNA include ribonucleoprotein (RNPs) and lipoprotein (LPPs) complexes. RNPs can employ nucleophosmin or argonaute proteins, while LPPs are lipid-protein particles of varying density that subdivide into; chylomicrons (75–1200 nm), very low-density lipoproteins (VLDLs) (30–80 nm), low-density lipoproteins (LDLs) (18–25 nm), and high-density lipoproteins (HDLs) (5–12 nm) ,186. ExtBesides having the ability to cross BBB to be found in peripheral samples, sncRNA carriers should be sortable to assign a cell-specific sncRNA signature to astrocytes. This depends on both the availability of cell markers for the cell in question and how often the carrier integrates the marker within their structure. EV formation involves budding of different membranes of the cell of origin from which they inherit cell surface markers that can be exploited  using anNon-vesicular carriers have more limitations regard to recognition of cell specific signatures. RNPs express ubiquitously with a 2% showing strict tissue specificity representing about 20 families of RNPs, whether isoform tissue specificity occurs and to which extent is yet to be discovered . ConditiThe sncRNA carriers also differ in the type of sncRNA transported. While MV sncRNA resembles expression in the cell of origin, EV and RNPs sncRNA show strong differential expression ,187. SevMicroglia are the resident immune cells in CNS which also contribute the most to internalize vesicles via micropinocytosis mechanisms under physiological conditions with in vitro and mouse model supporting this uptake role . ADEV-deNeuronal internalization of ADEV has been reported in physiological states and multiple studies demonstrate assimilation resulting in expression changes ,85,198. On the other hand, the same stimulus causing the secretion of neuroprotective sncRNA on astrocyte can stimulate the secretion of miR-16-5p, which reduces neuronal function after inflammatory stimulus . MiR-146Ischemic preconditioning elevates miR-92b-3p favoring neurite growth, viability and mitochondrial function on neuron . HoweverInterestingly ADEV uptake by astrocytes is expected to occur in pathological conditions such as NDD . RegardiThe sncRNA effects on astrocytes can be quite dynamic. For example, miR-21 exacerbates astrocyte activation worsening optic nerve lesion  but caus2+ levels in mSOD1 mice, while in NPCs and OPCs promotes myelin production [Some stimuli associated with early neurodegeneration conditions such as proinflammatory cytokine secretion, lesions, and toxicity, in the “right cell” cause the secretion of context dependent neuroprotector signaling ,146 encloduction ,146. On oduction ,145. Addoduction ,145,146 oduction  but beneoduction ,146, it Astrocytes are one of the most numerous glial cells in CNS and are key mediators in the neuroinflammatory process which recruits peripheral immune cells to the CNS , hence aTBI and fractures are a frequent occurrence in clinical routine (polytraumatic injuries), with TBI patients with concomitant fractures reporting and improved fracture healing process with faster and more robust regeneration, which later was supported by studies. In the finding of candidate healing molecules to improve fracture healing process various cytokines, hormones, correlated with improved healing and more importantly, serum derived of TBI patients accelerated healing by increasing cell proliferation . TBI cauTissue-specific markers such as ADEV-derived sncRNA signatures would improve signal-to-noise ratio and decrease variability of extracellular sncRNA signatures enriching for disease-specific biomarkers . It woul6 particles/mL for human CSF [10 particles/mL for mouse plasma [9 particles/mL for mesenchymal stem cell derived EVs in culture [Although bodily fluids are expected to have a higher amount of EV this is not always the case. EVs counts as measured by nanoparticle tracking analysis (NTA) reveals yields of 10uman CSF , 1010 pae plasma , and 6 × culture . Variabi culture  but, con culture , which h culture  and it iIn addition, current separation protocols seem to fail in the recovery of the total EV load produced by the cell, with variable results depending on the method representing no consensus in a gold standard separation method ,203. MetBiomarkers for neurodegenerative disease are a necessity but their design is challenging. We briefly reviewed the central role of astrocyte dysfunction in the course and development of neurodegenerative disease (NDD) and how sncRNA changes in response to astrocyte function could regulate function on target cells. Different biomarker applications such as the diagnosis in early stages of neurodegeneration would be possible by employing astrocyte specific signatures. Such developments could take part in future molecular diagnostics whose development is expected to improve treatments supporting true neuroprotective/neurorestorative therapies through early diagnosis.However, several methodological aspects should be considered to make ADEV-derived sncRNA a reliable biomarker. New ADEV separation protocols have to increase EV yields without increase contamination with both other particles such as lipoprotein complexes or non-astrocytic EVs. Furthermore, if contamination remains deconvolution techniques may be explored to diminish noisy signal of circulating RNA in complex with RNPs and lipoprotein. Small nuclear (snRNA) is the most abundant class of sncRNA in EVs samples but this should be confirmed in astrocytes and small quantities of miRNA have been shown to influence target cell function and we should employ sensitive enough techniques to detect these small increases. Furthermore, pathological conditions and stress tend to increase EV production and it could generate a dose dependent signal of sncRNA that responds to an early neurodegeneration condition.Astrocyte-derived extracellular RNA binding protein (RBP) role in the functional delivery of sncRNA should be explored in near future. A total of 2% of RBP genes in human genome with cell expression specificity represent enough specificity to develop astrocyte specific detection against epitopes expressed in astrocyte expressed RNPs due to the large quantity of RBP genes. Further studies in RBP biology and function in relation with secretion are necessary to determine basic question such as whether secretion of tissue exclusive RNPs occurs. The same techniques employed for the isolation of EV should be useful for the enrichment of astrocyte derived RNPs due to their shared physicochemical properties. Because of these similarities it should be easier for research groups working in vesicular area shifting to explore RNPs role in sncRNA delivery.sncRNA has a functional role as extracellular vesicle cargo that we are beginning to understand, thus we must improve sncRNA detection capabilities considering the relatively lows amount of ADEV derived sncRNA reported by different studies in human body fluids and the EV lost in methodological procedures before a cell specific signal can be generated.


Injectable and biodegradable hydrogels have been regarded as encouraging materials in the biomedical field because they had a similar three-dimensional microenvironment with native tissues and cells that could be effortlessly encapsulated within these hydrogels dispersedly. During the period of tissue regeneration, biodegradable hydrogels could be degraded with the deposition of extracellular matrix secreted by cells, thus serving as a promising “temporary extracellular matrix” in remodeling. However, due to the lack of precise microarchitecture control and the limited ability to reconstruct living structures containing complex anisotropic microenvironment, hydrogels with precise topographic cues remains an ongoing challenge. Considering these shortcomings, nanocomposite hydrogels which consisted of nanomaterials and hydrogels enable improve directed structure formation at the multiscale. The integration of nanomaterials, such as nanofibers, nanoparticles, nanoclay and nanogel, and hydrogel endow new mechanical and functional properties, which will be highly beneficial for various biomedical applications, especially tissue engineering and regeneration.Here, this Research Topic collected a total of four original research papers, which described different kinds of injectable hydrogels from the design, preparation and evaluation. The papers on this Research Topic are briefly shared below.Haghighattalab et al. added magnetic nanoparticles (MNPs) into silk fibroin hydrogel and loaded with doxorubicin hydrochloride (DOX) in the meantime using a straightforward blending method to determine the capacity of controlled drug release under the diversified external magnetic field (EMF). With a higher DOX concentration, the Wilms’ tumor cells showed drug resistance which revealed the importance of controlled drug delivery. This study demonstrated that the magnetic SF hydrogel provided a new strategy for smart drug delivery with non-invasive injection and remote control capability.Silk fibroin (SF) materials have been applied as an appealing biomaterial in the biomedical and pharmaceutical area over the past few decades. Wang et al. designed an injectable hydrogel based on Bionic Self-Assembling Peptide (BSAP) with hemostatic and bactericidal function for wound healing. BSAP could co-assemble swiftly into a hydrogel network structure driven by Ca2+in situ. This hydrogel displayed a lower blood clotting index and reduced blood clotting time and bleeding volume dramatically in the foot trauma model and tail amputation model. Besides, by fighting bacterial infection, this hydrogel could reduce inflammatory responses and enhance wound healing.It is urgent to develop a potent coagulation and anti-infection strategy save patients suffering from wounds or trauma. Hydrogels are distinguished as the most suitable biomaterials for accelerating wound healing. Wang et al. prepared a new organic-inorganic composite hydrogel (FPIGP@BGN-Sr hydrogel) comprising diacrylated PF-127, β-GP-modified polyitaconate and BGN-Sr via radical polymerization and coordination interactions. The composite hydrogel could effectively enhance the mouse embryonic mesenchymal stem cells’ osteogenic differentiation Therefore, the appropriate concentration of BGN-Sr, PF-127 and β-GP-modified polyitaconate exhibited good synergistic effects on promoting osteogenic differentiation of MSCs, therefore offered a promising approach for stem cell-based bone regeneration.To achieve better mechanical match and stronger biological properties of bone biomaterials, Chen et al. fabricated an injectable mesh using N-isopropylacrylamide (NIPAAm), N-hydroxyethyl acrylamide (HMAAm) and Polyvinyl alcohol (PVA) based on the electrospinning method. The temperature-responsive mesh could be injected at the tumor site to modulate and control anticancer effects by loading magnetic nanoparticles (MNPs) and chemotherapeutic drugs such as paclitaxel (PTX). This study revealed a new therapeutic platform for cancer treatment using hyperthermia combined with chemotherapy.Many advances have been made in the application of injectable and biodegradable hydrogels in the treatment of various diseases, tissue regeneration and controlled release of drugs. However, the application of injectable biodegradable hydrogels was still restricted by some practical problems. For example, the needle loaded with temperature-responsive hydrogels may be blocked when injected subcutaneously due to the temperature increase, thus causing a failed injection. Moreover, the long-term storage and stability of injectable and biodegradable hydrogels still needed to be improved. To solve these dilemmas, developments of injectable and biodegradable hydrogels that respond to other stimuli, such as hypoxia, light, or enzymes, may be alternative strategies. Most current studies focus on improving the performance of biodegradable and injectable hydrogels. However, the degradation of products’ toxicity is an overlooked problem. Therefore, more work needs to be done to evaluate the safety of biodegradable hydrogels after the practical application tasks are completed. Future work should also focus on the fabrication of biodegradable and injectable hydrogels using more convenient, cost-effective, and environmentally friendly methods. Additionally, the development of biodegradable and injectable hydrogels with more advanced properties should be further explored to provide more options in various application scenarios. Through these efforts, biodegradable and injectable hydrogels will benefit an increasing number of patients in the near future.


Diffuse gastric mucosal injury is a chronic injury with altered cell differentiation, including spasmolytic polypeptide expression metaplasia (SPEM) and intestinal metaplasia (IM), which are considered precancerous lesions of gastric cancer (GC). Previously, most studies have focused on how parietal cell loss causes SPEM through transdifferentiation of chief cells. In theory, alteration or loss of chief cells seems to be a secondary phenomenon due to initial partial cell loss. However, whether initial chief cell loss causes SPEM needs to be further investigated. Currently, increasing evidence shows that initial chief cell loss is sufficient to induce gastric mucosal injury, including SPEM and IM, and ultimately lead to GC. Therefore, we summarized the two main types of models that explain the development of gastric mucosal injury due to initial chief cell loss. We hope to provide a novel perspective for the prevention and treatment of diffuse gastric mucosal injury. Gastric mucosal injury can be divided into two types: focal and diffuse. Generally, the common causes of focal injury are toxin ingestion, bile reflux, and certain infectious agents, which result in focal erosions or full-thickness ulcerations that are rapidly repaired by increased proliferation in neighboring units and migration of surface cells; these processes eventually reestablish normal differentiation in damaged units. Diffuse gastric mucosal injury is a chronic injury with altered cell differentiation, including spasmolytic polypeptide expression metaplasia (SPEM) and intestinal metaplasia (IM), which are considered precancerous lesions of gastric cancer (GC) –6.Helicobacter pylori (H. pylori) infection [Previously, most studies focused on parietal cell loss as the initial factor inducing diffuse gastric mucosal injury, with alteration or loss of chief cells following. These mouse models of metaplasia have been generated by gene knockout (KO) and treatment with various drugs ), as well as nfection . Howevernfection –17, suggChief cells reside at the base of oxyntic glands in the gastric corpus. These cells not only secrete pepsinogen and participate in digestive functions in normal gastric physiology  but alsoIn most previous studies, after injury to the gastric epithelium, especially after the loss of parietal cells, chief cells undergo metaplasia that causes them to transition into mucous cells, a process called SPEM. Thus, chief cells have been considered the origin of metaplasia after gastric mucosal injury. These cells usually undergo metaplasia by transdifferentiation or dedifferentiation , 22. How+ chief cells [In the chief cells of the stomach, multiple markers with various functions have been identified. Basic helix-loop-helix family member a15  is a marker of chief cell health and maturation and is mainly expressed within chief cells. Its expression is lost in metaplasia, atypical hyperplasia and various carcinomas, providing evidence that metaplasia originates from mature chief cells . Leucineef cells . Acidic ef cells  and is sef cells . FurtherChief cells responding to mucosal injury are subject to many regulatory mechanisms, such as maturation and aging , 28, swiThe loss of chief cells results in various types of gastric diffuse mucosal injury, epithelial structural disorders and types of gastric metaplasia, classic precancerous lesions –17, inclThese data suggest that the initial loss of chief cells is sufficient to cause gastric mucosal injury, providing promising research directions for understanding the mechanisms of mucosal injury. Currently, two types of mouse models with initial chief cell loss have been established. The first involves KO of genes that are highly expressed in chief cells –14. The RUNX3 is considered a tumor suppressor in GC  and is s+/K+-ATPase, Slc26a9 and Sonic hedgehog. These changes were observed only at the HGIN stage, not in the SPEM phase  [CHIA is also predominantly expressed in chief cells of the stomach . Accordied data) , 14.H. pylori should be considered.In conclusion, the loss of chief cells should be noted in diffuse gastric mucosal injury. An independent origin of malignant events in the gastric epithelium unrelated to the absence of parietal cells and infection with + chief cells and Mist1+ chief cells, and both agents induced short-term SPEM without affecting parietal cells, accompanied by rapid, short-term expansion of GSII+ GIF+ cells [Drug-induced initial chief cell loss also causes gastric mucosal injury. Researchers administered diphtheria toxin and small doses of tamoxifen to ablate Lgr5F+ cells . MoreoveF+ cells .+ chief cells, and mouse glands also showed metaplasia. These metaplastic glands showed strong expression of Tff2 and Muc5ac and the absence of parietal cells, as well as macrophage infiltration, consistent with SPEM features [+ chief cells with Kras mutations, since they found that Gpr30 is widely and specifically expressed in chief cells and can be used as a chief cell-specific marker. After prolonged Kras induction by doxycycline, the chief cells were lost as expected, while the parietal cells remained viable. These effects were accompanied by the appearance of a GIF+GSII+ SPEM lineage, which resulted from regenerative expansion of Kitl+ isthmus progenitors in response to the loss of chief cells rather than transdifferentiation or dedifferentiation of Gpr30+ chief cells. Chief cells are subject to Gpr30 regulation by rapid ablation from epithelial cells through PDK-dependent cell competition. These effects may occur via induction of the Warburg effect caused by enhanced PDH phosphorylation in Gpr30+ chief cells [Activation of Kras has been suggested to be a key signal in the development of GC –39, and features . Moreoveef cells . It is aef cells .These findings suggest that both gene deletion- and drug-induced initial chief cell loss are also origins of gastric mucosal injury and not just effects of parietal cell loss.The gastric mucosal barrier provides protection for the stomach, and an imbalance between invasive and protective factors can lead to the development of gastric mucosal disease. Most of the previous studies on gastric mucosal diseases have focused on parietal cells. However, in-depth research on mucosal diseases has revealed that chief cells should also be emphasized in the prevention and treatment of gastric mucosal injury. The initial loss of chief cells is an independent key event in gastric mucosal injury and in the development of gastric precancerous lesions and gastric tumors. This review provides a new perspective on gastric mucosal diseases, furthering the understanding of the role of chief cells in these diseases and providing new targets for prevention and treatment.


COPA syndrome is a rare hereditary inflammatory disease caused by mutations in the gene encoding the coatomer protein subunit alpha, causing excessive production of type I interferon.This case is a reminder for the general paediatrician, highlighting the relevance of the association between arthritis and lung involvement in toddlers.We report the case of a 2-year-old girl with intermittent limping and joint pain. Her family history was relevant for a Still disease with lung involvement in the mother. Physical examination showed moderate wrist swelling. Laboratory findings on admission showed an increase in inflammatory markers, positive rheumatoid factor, antibodies antinuclear antibody (ANA) and cyclic citrullinated peptide (anti-CCP). Wrists’ ultrasound documented synovial thickening, and chest X-rays showed an unexpected severe interstitial pneumopathy. Genetic testing confirmed the diagnosis of a heterozygous mutation of the COPA gene in c.841C > T (p.R281W). Janus kinase treatment was started  with a remarkable improvement in limping and joint pain after two weeks.In cases of recurrent arthritis with family history and multiple involvement organs, a genetic disorder should be suspected and genetic testing should be performed. Furthermore, this case suggests that therapy with jak inhibitors may be effective and safe in interferonopathies. COPA syndrome is a rare hereditary inflammatory disease caused by mutations in the gene encoding the coatomer protein subunit alpha, causing excessive production of type I interferon , 2. ThesA 2-year-old girl was seen for a 6-month history of intermittent limping and joint pain, previously treated with acetaminophen or nonsteroidal anti-inflammatory drugs (NSAIDs) with little benefit. The child was born preterm (27 weeks gestation), she suffered from mild bronchopulmonary dysplasia and bilateral renal dysplasia, and her mother reported a Still disease with lung involvement. In the previous months, an ultrasound of lower limbs’ joints had been performed, documenting a mild right hip effusion.; objective chest evaluation, saturation values and a slit lamp eye examination were unremarkable.Physical examination showed moderate wrist swelling.Blood sampling showed average blood count, increased ESR, positive antibodies ANA and anti-CCP, hypergammaglobulinemia, hypercomplementemia and highly positive rheumatoid factor Table . Kidney Due to the discrepancy between the mild articular involvement and the marked increase of rheumatoid factor and acute inflammation indexes, the girl was admitted in order to perform further testing.. Remarkably, the toddler did not show any sign of respiratory distress. A pulmonary CT highlighted severe interstitial pneumopathy with diffuse fibrosis, patchy ground-glass opacities, cystic areas, bronchiectasis and mediastinal lymphadenopathy .Janus kinase inhibitor treatment was started  with a remarkable improvement in limping and joint pain after two weeks.In children with arthritis and lung involvement various diagnoses should be considered, such as systemic lupus erythematosus, vasculitis (e.g. granulomatosis with polyangiitis), systemic juvenile idiopathic arthritis, dermatomyositis. The association of arthritis and renal involvement also evokes systemic lupus erythematosus and vasculitis.In this case, the young age at onset of the disease and the absence of other specific clinical and laboratory features suggestive of Lupus , or dermatomyositis , prompted us to rule out a genetic inflammatory disorder.. Generally, in the presence of significant lung involvement, the clinical presentation is characterized by chronic cough, tachypnoea, and rarely haemoptysis. Lung examination may highlight retractions, crackles and signs of hypoxia such as cyanosis or clubbing.The association of lung and/or renal involvement with arthritis should suggest a COPA syndrome among differential diagnoses. In children with COPA, pneumopathy can be initially asymptomatic and eventually progressive, with the development of alveolar bleeding, severe fibrosis and end-stage respiratory failureThe glomerular disease has been documented in half of the patients with COPA. The main histologic features are crescentic glomerulonephritis and focal mesangial hypercellularity with immune complex deposits, ranging from isolated IgA deposits to “full-house” immunofluorescence. Polyarticular chronic inflammatory arthritis can be severe with involvement of the cervical spine with vertebrae fusion.In this patient, besides rheumatological symptoms, physical chest examination, saturation values and respiratory rate were standard. A chest X-ray performed to rule out other pathologies unexpectedly showed a severe picture of interstitial pneumopathy, confirmed by pulmonary CT. The genetic test confirmed COPA Syndrome’s diagnosis revealing heterozygous mutation of the COPA gene in c.841C > T (p.R281W). Disease transmission is autosomal dominant, but mutations de novo are also possible. The genetic test performed on the mother was negative while the genetic test performed on the father and siblings is still ongoing.Remarkably, an NSE moderate increase has been found in inflammatory diseases  and neurCOPA syndrome does poorly respond to steroids or other immune-suppressive treatments such as methotrexate and TNF-alpha inhibitors. In the literature, two cases of COPA syndrome have been reported responding to Janus Kinase inhibitor, one to baricitinib and one to ruxolitinib . RegardiJanus kinase inhibitors are supposed to act by hampering Type I interferon and IL6. Although not yet approved in children in Europe, they are approved in other countries for use in other childhood inflammatory diseases, allowing to demonstrate and describe their pharmacokinetics and safety profile . The expThe limit of this report is the short follow-up after starting treatment with baricitinib; the point of strength is the take-home message highlighting the association between arthritis, family history and lung involvement.Paediatric practitioners should be aware of the risk of a genetic disorder in front of recurrent arthritis with family history and multiple organ involvement. Even in the absence of respiratory or renal symptoms genetic testing should be considered, being the gold standard of diagnosis.


Although cardiovascular diseases (CVD) are the leading cause of non-communicable diseases-dependent death worldwide, their effects are still largely underestimated in women . The exiSince, for many decades, CVD research has been performed primarily on men, this Special Issue aims to encourage a translational approach involving contributions from basic and clinic researchers that would lead to a sex/gender-tailored medicine with the identification of sex specific biomolecules profiling, with clinical relevance for predictive, diagnostic or prognostic purposes for CVD prevention in both sexes.


Brain tissue temperature is a dynamic balance between heat generation from metabolism, passive loss of energy to the environment, and thermoregulatory processes such as perfusion. Perinatal brain injuries, particularly neonatal encephalopathy, and seizures, have a significant impact on the metabolic and haemodynamic state of the developing brain, and thereby likely induce changes in brain temperature. In healthy newborn brains, brain temperature is higher than the core temperature. Magnetic resonance spectroscopy (MRS) has been used as a viable, non-invasive tool to measure temperature in the newborn brain with a reported accuracy of up to 0.2 degrees Celcius and a precision of 0.3 degrees Celcius. This measurement is based on the separation of chemical shifts between the temperature-sensitive water peaks and temperature-insensitive singlet metabolite peaks. MRS thermometry requires transport to an MRI scanner and a lengthy single-point measurement. Optical monitoring, using near infrared spectroscopy (NIRS), offers an alternative which overcomes this limitation in its ability to monitor newborn brain tissue temperature continuously at the cot side in real-time. Near infrared spectroscopy uses linear temperature-dependent changes in water absorption spectra in the near infrared range to estimate the tissue temperature. This review focuses on the currently available methodologies and their viability for accurate measurement, the potential benefits of monitoring newborn brain temperature in the neonatal intensive care unit, and the important challenges that still need to be addressed. In term infants, neonatal encephalopathy (NE) is the most common form of perinatal brain injury in both high-income countries (HIC) as well as low- and middle-income countries (LMIC). Hypoxic-ischemic encephalopathy (HIE) affects 1–6 per 1,000 live births and remains a major cause of morbidity and mortality in term infants . SeizureBrain temperature is determined by the balance of heat production and heat removal, and it is influenced by multiple factors including cerebral metabolism, brain tissue injury, cerebral blood flow (CBF), body-brain temperature difference, various medications and infection –9. The b31P) magnetic resonance spectroscopy (MRS) described the evolution of primary and secondary energy failure with a reduction in high energy phosphates and a rise in cerebral lactate following injury ] and deoxygenated haemoglobin [Δ(HHb)] which can then be used to derive changes in total haemoglobin [Δ(HbT)] and haemoglobin difference [Δ(HbD)]. Changes in [Δ(HbT)] and [Δ(HbD)] represent changes in cerebral blood volume and oxygenation respectively. In addition, cerebral oximeters can also measure absolute brain tissue saturation  as an absolute percentage measurement of the mixed arterial and venous saturation. A seizure typically increases both cerebral blood flow and cerebral metabolic rate (Previous studies with near-infrared spectroscopy (NIRS) during neonatal seizures have also described changes in cerebral hemodynamics and oxygenation –36. NIRSlic rate . Any inclic rate  which calic rate , while alic rate , 41 to blic rate . NeuronaSeveral different methodologies have been evaluated so far for brain temperature measurements . Proton MRS thermometry has been used most for non-invasive and in-vivo brain temperature measurement in the neonatal population , 59, 60.in vivo for many applications. The chemical shift of water is approximately linearly dependent on temperature in the physiological range (Hindman). So, by measuring the chemical shift separation between water and one or more reference metabolites, the absolute temperature can be inferred. Non-invasive local temperature measurement using proton MRS has been demonstrated in vivo of about 0.05 ppm. Despite this challenge, accuracies of 0.2–0.5 °C have been reported , the major endogenous tissue chromophores, responsible for the light’s absorption of the tissue, are water and oxy- and deoxy- haemoglobin. The aim of NIRS is to quantify the concentrations, or change in concentrations related to a specific event, of these chromophores in tissue and its focus is to quantify the oxygenation of the tissues. However, the linear temperature-dependent changes in NIR water absorption spectra make the measurement of tissue temperature possible with NIRS, as the tissues are mostly composed of water (more than 70% of the brain tissue composition for example).Indeed, the water absorption spectra and its temperature dependency has been well established , 75. TheOther studies using broadband diffuse optical spectroscopy (DOS) reported the measurement of the adult breas  and foreFinally, the brain temperature of piglets was measured using a time-domain NIRS (TD-NIRS) . TD-NIRSLooking to the future, broadband TD-NIRS systems could further benefit brain temperature monitoring by NIRS techniques, as in general, the accuracy of the temperature prediction can be improved by acquiring a continuous absorption spectrum. Indeed, it allows a more accurate determination of chromophore concentrations compared to discrete wavelengths, as more chromophores can be quantified, and a more refined data analysis technique can be used. Such systems have been reported in the literature , 90, howThe recent technological developments, notably in terms of electronics, enabled to reduce the footprint of TD-NIRS, which facilitated its use for clinical applications  and compBrain temperature is determined by the balance of heat production and heat removal, and it is influenced by multiple factors including cerebral metabolism, brain tissue injury, blood flow, body/brain temperature, drugs, sedation, seizures, and infection –9. The bHypothermia decreases the metabolic demand for glucose and oxygen and attenuates secondary energy failure and neuroapoptosis , 93. TheThe current hypothermic strategy for NE uses rectal temperature for servo-controlled feedback to maintain a steady temperature profile. An in-vivo assessment of regional brain temperature using proton MRS during whole-body TH revealed heterogenicity of the brain temperature profile. Hypothermia effectively lower deep grey matter structures, whereas temperatures of more superficial structures in the grey matter and white matter are significantly greater than rectal temperatures . FindingAn animal study suggested that focal seizures produce an increase in neuronal activity and led to an elevation of local blood flow, cerebral metabolism and a significant rise in brain temperature . GeneralIn animal studies focal cooling rapidly terminates experimental neocortical seizures and histological examination of the cortex after cooling has shown no evidence of acute or delayed neuronal injury, and blood pressure and temperature remained stable . Animal Confirming brain injury at the bedside and determining the type and severity remains a challenge, as does bedside identification and monitoring of injuries likely involving ongoing processes of oxidative stress, excitotoxicity, inflammation, repair, and cell death, evolving over hours to weeks. As novel treatment strategies for neonatal brain injuries and seizures become available, the need for non-invasive and continuous bedside monitoring of disease severity and response to treatment becomes increasingly apparent. Non-invasive and continuous measurements of brain temperature in the neonatal neurocritical care set-up may permit the selection of neonatal candidates who may benefit from an adjustment in their hypothermia therapy or for additional neuroprotective therapies. Further studies are now urgently needed to establish whether optical brain monitoring can be a useful neuromonitoring tool in neonatal neurocritical care set-up. In view of the heterogeneous profile of brain temperature during TH, an option of continuous non-invasive monitoring of brain temperature at the bedside might also provide an opportunity to review and improve the current cooling methodologies in neonatal neurocritical care following NE.


Patients suffering from musculoskeletal diseases must cope with a diminished quality of life and an increased burden on medical expenses. The interaction of immune cells and mesenchymal stromal cells during bone regeneration is one of the key requirements for the restoration of skeletal integrity. While stromal cells of the osteo-chondral lineage support bone regeneration, an excessive accumulation of cells of the adipogenic lineage is thought to promote low-grade inflammation and impair bone regeneration. Increasing evidence indicates that pro-inflammatory signaling from adipocytes is responsible for various chronic musculoskeletal diseases. This review aims to summarize the features of bone marrow adipocytes by phenotype, function, secretory features, metabolic properties and their impact on bone formation. In detail, the master regulator of adipogenesis and prominent diabetes drug target, peroxisome proliferator-activated receptor γ (PPARG), will be debated as a potential therapeutic approach to enhance bone regeneration. We will explore the possibilities of using clinically established PPARG agonists, the thiazolidinediones (TZDs), as a treatment strategy to guide the induction of a pro-regenerative, metabolically active bone marrow adipose tissue. The impact of this PPARG induced bone marrow adipose tissue type on providing the necessary metabolites to sustain osteogenic-as well as beneficial immune cells during bone fracture healing will be highlighted. The osteogenic differentiation trajectory of mesenchymal stromal cells (MSCs) is crucial for bone tissue function, as they likely contribute to the formation of adult skeletal stem cells. For example, there are bone-resident populations of cells specialized to undergo the typical osteogenic maturation stages, including osteoblasts and osteocytes. Moreover, MSCs probably contribute significantly to chondrogenic lineages as a key component of endochondral ossification . The conin vitro and in vivo experiments, there is no clear definition of MSCs in bone marrow , which shows tri-lineage  differentiation potential in > 80% of individual cells  exhibit similar sensitivity towards PPARG agonists than white adipocytes . For insIn general, BMAds differentiate from MSCs within the bone marrow, which arise from a mesenchymal-osteogenic lineage and undergo lineage specialization involving a decision-making step between osteogenic and adipogenic lineage commitment .Interestingly, the potential role of adipogenic cells in bone has been further emphasized by using porous scaffolds seeded with human adipose tissue-derived mesenchymal stromal cells. In these scaffolds, adipocytes have been shown to contribute to osteoblast formation directly and to vascularization of bone defects as well as the induction of bone mineralization by the secretion of paracrine factors . Thus, wRecent studies have emphasized that BMAds are also clearly distinct from brown adipocytes and show very little propensity for expression of Ucp1 or cold-induced browning. In addition, their response to many physiological stimuli, including glucose homeostasis and insulin responses, is markedly different to brown adipocytes .Regarding differences between BMAds, subtypes have previously been defined by their anatomical distribution within long-bones . The iniOf note, cBMAT starts to appear during embryogenesis and/or in early childhood whereas the development of rBMAT is thought to occur later in life and during aging in particular . The travia indirect or direct cell-cell contacts  of AMP-activated kinase (AMPK) signaling pathways. Here, controversial data on the bioactivity of adiponectin in the bone has been observed in animal and human studies .A complete lack of adipocyte derived endocrine factors is shown to exert detrimental effects on bone integrity . LiteratTo date, adiponectin is the most prominent systemically active endocrine factor for which BMAT has been described as a source . AdiponeDuring caloric restriction (anorexia nervosa), BMAT which paradoxically increases under these conditions, is a main source of adiponectin. Thus BMAT is able to exert systemic endocrine effects by contributing to increased serum adiponectin. . Systemiin vitro and bone formation in vivo. Although further analysis is required, these observations suggest that modulation of adipocyte phenotypes by appropriate signals may promote a beneficial or at least non-pathological profile of local adipocytes  and cathepsin K, a protease involved in bone resorption, differs between both BMAT types, where PAI-1 is described to prevent bone loss by increasing bone formation and cathepsin K is indispensable for osteoclast-mediated bone resorption and remodeling . The sitvia cAMP signaling  has been shown to correlate with higher bone mineral density (BMD) and a reduced fracture risk . It shouignaling . The proignaling . Omega-3ignaling . Bone maignaling   and TNFa. Subsequently, additional cells of the adaptive immune system, as well as CD34+ endothelial progenitors and MSCs are actively attracted towards the vicinity of the fracture gap, further driving the healing process. Thus, the adaptive immune response results in a high production of pro-inflammatory cytokines, activating and attracting MSCs for the facilitation of tissue regeneration , to a pro-inflammatory, activated state with increased energy demand . As the iewed in ]. M1-likneration . During neration . Endotheneration .In bone and many other tissues, a pro-regenerative switch subsequently requires a transition from the pro-inflammatory milieu towards an anti-inflammatory signaling . This chInterestingly, PPARG is shown to promote glycolysis, glucose uptake and triglyceride synthesis by adiponectin induction . During Most diabetes drugs induce a beneficial metabolic and anti-inflammatory switch within adipocytes. This metabolic switch is linked to diverse effects on bone integrity, depending on the specific drug treatment. Aside from recombinant insulin-treatments, the most commonly used anti-diabetic drugs to date include sodium-glucose linked transporter 2 (SGLT2) inhibitors, TZDs, metformin, and regulators of the incretin effect, i.e., DPP4-inhibtors and incretin analogues .The traditional antidiabetic drugs frequently display undesired side effects such as hypoglycemia, weight gain, cardiovascular complications and, in relation to bone health, increased fracture risk . HoweverTo influence the accumulation of adipocytes in bone marrow, distinct approaches, including interventions targeting energy metabolism as well as certain drugs can be applied . Since tAmong the know antidiabetic drug treatments, TZDs are known to most effectively influence BMAT accumulation and metabolism, as well as bone integrity . NotablyTZDs, such as rosiglitazone, act as PPARG-agonists and induce systemic insulin sensitivity, partially by the release of insulin-sensitizing adipokines from adipose tissue, and thereby improving systemic glucose homeostasis. PPARG-agonists also can decrease lipotoxicity by enhancing deposition of fatty acids into adipose tissue as triglycerides and thereby sequestering fatty acids away from other non-adipose tissues. In effect, the activation of PPARG as a key regulator of lipogenesis results in the reduction of fatty acids in circulation. Hereby, PPARG activation is regulating metabolic flexibility of adipose tissue under challenging metabolic conditions . DiffereWhile previous reviews on the clinical aspects of bone health and TZD treatment generally agree that treatment is detrimental to bone health in diabetics, the extent has also been considered modest . Some stIn humans, the detrimental or beneficial impact of PPARG agonists, such as rosiglitazone, on bone seems to be highly dependent on the duration of therapy, with the adverse effects of rosiglitazone on bone structure only becoming apparent after approximately one year . It is, On a cellular level, the sensitivity of osteoblast progenitors towards rosiglitazone decreases with age while the adipogenic lineage appears to become more sensitive to TZDs. This change in responsiveness coincides with increased levels of PPARG in the aged MSC pool which steer the rosiglitazone-driven increase in osteoclast progenitor development, leading to rosiglitazone-stimulated bone loss . RosigliSince the incidence of patients suffering from T2DM is steadily increasing, even in younger individuals, TZDs are more frequently prescribed. Thus, it is important to analyze TZD-based treatment and bone loss driven by anti-diabetic drugs such as rosiglitazone. Decreased osteoblast function or fatty acids that inhibit osteogenesis could also be a factor reducing bone quality .A previous study tested the effects of rosiglitazone on osteogenic differentiation and reported an enhanced adipogenic as well as osteogenic differentiation. Specifically, human mesenchymal stromal cells and other cell lines were either exposed to adipogenic or osteogenic differentiation conditions with and without addition of rosiglitazone. An RNAi-mediated knockdown of Pparg gene expression yielded similar results . Under tAlso, BMAT contains high levels of essential polyunsaturated fatty acids (PUFAs) in the form of triglycerides. TZDs can also bind these bioactive lipids, initiating processes relevant to osteogenesis, such as inflammation and marrow adipocyte formation . In addiProminent lipid mediators in bone, such as prostaglandins and leukotrienes, are well-known to regulate bone formation and inflammation . These mOnly a limited number of studies have shed light on the process of lipogenesis in BMAT. However, the fatty acid composition of rBMAT, which contains higher levels of saturated fatty acids closely resembles the saturation profile of white adipocytes in other, localized depots of WAT  Griffit. ConversIn BMAT, lipid unsaturation ranges from lower amounts in proximal rBMAT sites towards increased amounts in the distally located areas of cBMAT  Schelle. Fatty aIn summary, the association of bone function and health with PPARG-activation, and the related interactions within a physiological context, are highly complex and remain only partially understood. There is some experimental evidence suggesting that TZDs, in their role as PPARG agonists, may not only promote formation of BMAT but additionally drive a bone-resident adipocyte phenotype that could exert beneficial physiological effects. For instance, the well-established immunomodulatory activity of adipocytes could be a potential mechanism of interaction between BMAT and immune cells during bone tissue repair and maintenance. However, there is a clear need for further research on this immune-BMAT interaction as a component of bone healing.Without a balanced immune reaction during the initial bone healing phase, the regeneration cascade is disrupted. This in turn leads to detrimental effects which cause a delay in bone tissue repair as well as long-term consequences, such as non-union or sustained mechanical instability . Other fPro-inflammatory conditions inhibit PPARG activity at the gene expression level and pro-inflammatory cytokines, such as TNFa, IFNg and IL1b, negatively regulate PPARG expression in the bone marrow, targeting e.g., macrophages . Takada Converse+ T cells with PPARG-expressing Tregs resulted in a downregulation of IFNg production, which is beneficial for bone healing  and naïve adaptive immune cells that balance pro-inflammatory signals . Consist healing . Additio healing . CD4+ Trsitivity . Phenotysitivity . Thus, weoclasts .+ memory T helper cells, especially CD4+ memory T helper 2 (TH2) cells, are associated with regulatory and anti-inflammatory features during bone repair. The induction of PPARG is required for the activation and proliferation of CD4+ naïve T cells and their development into CD4+ memory TH2 cells  niche remains only partially understood. Recently published evidence stresses that marrow adipocytes are distinct from other types of white adipocytes and have little, if any, resemblance to thermogenic brown adipocytes. However, the recognition of different types of bone marrow-resident adipocytes, cBMAT and rBMAT, in combination with the early-onset of marrow adipogenesis at a relatively young age, offers the intriguing possibility that this type of adipocyte is a key element of bone physiology, involved in the tissues energy homeostasis and a regulator of skeletal regeneration processes. Marrow adipocyte may target bone healing through different processes, including their insulin-sensitivity and metabolic activity. To this end, we review published evidence to explore the hypothesis that induction of marrow adipogenesis, for instance mediated by the insulin-sensitizing drug class of thiazolidinediones (TZD), in the context of skeletal repair may create a metabolically active type of marrow adipocytes as a pro-regenerative niche component .An important second aspect of this hypothesis is the timing of induction of marrow adipogenesis, which is clearly distinct from traditional inducers that promote marrow adipogenesis over a prolonged period. Accordingly, we review evidence to highlight a potential involvement of TZDs as agonist of the nuclear receptor PPARG and immune-modulation. To develop a strategy to enhance bone regeneration through targeting of PPARG-sensitive signaling and metabolic pathways, studying the combined effects of PPARG on bone and BMAT while also considering the immune reaction is of great importance. To this end, we have reviewed T cell-specific actions of PPARG as potential mechanisms relevant during bone regeneration. These processes have at present mainly received attention in the context of extra-skeletal adipose depots, i.e., the traditional depots of brown and white fat. However, the adaptive immune system, comprising distinct T cell subsets, has independently been shown to be essential for endogenous bone regeneration.The presence and impact of metabolically distinct BMAT subtypes during bone regeneration is unknown and remains to be investigated in future studies. In summary, the characterization of BMAT subtypes within the bone compartment and their specific impact on bone injury repair and regeneration may require a more nuanced definition of BMAT functions, highlighting site-specific differences of BMAT as well as the impact of different metabolic and pathological conditions that BMAT is exposed to. The concept of adaptive changes in BMAT characteristics resembles the immune response during healing of bone, where the plasticity of cell types and inflammatory cues is highly dependent on the surrounding environmental milieu and the stage of bone healing.


Differences in speech prosody are a widely observed feature of Autism Spectrum Disorder (ASD). However, it is unclear how prosodic differences in ASD manifest across different languages that demonstrate cross-linguistic variability in prosody. Using a supervised machine-learning analytic approach, we examined acoustic features relevant to rhythmic and intonational aspects of prosody derived from narrative samples elicited in English and Cantonese, two typologically and prosodically distinct languages. Our models revealed successful classification of ASD diagnosis using rhythm-relative features within and across both languages. Classification with intonation-relevant features was significant for English but not Cantonese. Results highlight differences in rhythm as a key prosodic feature impacted in ASD, and also demonstrate important variability in other prosodic properties that appear to be modulated by language-specific differences, such as intonation. Differences in prosody are observed in many individuals with autism spectrum disorder (ASD) , 2. AlthAlthough a number of studies have provided important insights into the characteristics of speech prosody in individuals with ASD, little is known concerning the underlying causes of prosodic differences in ASD. Moreover, most studies have concentrated on studying prosody in English-speaking groups , 6. LangSince the earliest delineations of ASD , prosodilexical meaning in tone languages such as Cantonese. For example, in Cantonese, a syllable /ji/ means ‘to cure’ when produced with a high-level pitch pattern but means ‘two’ when produced with a low-level pitch pattern.Traditionally, speech prosody is categorized into affective and linguistic categories . AffectiProsodic expression in individuals with ASD speaking a tone language has been scarcely investigated. One study examined the f0 and duration of the five Thai lexical tones produced by native speakers with ASD . A separTaking a cross-linguistic approach, the present study compared prosodic characteristics of speech from individuals with and without ASD across English and Cantonese, two typologically and prosodically distinct languages. English is an Indo-European Germanic non-tone language that belongs to the rhythmic group of stress-timed languages , whereasrhythm, and 2) intonation, considered to be the two core elements of speech prosody [rhythm and/or intonation) derived from larger temporal windows  could reliably classify individuals with ASD vs. individuals with TD, both within and across English and Cantonese.We implemented a series of novel machine learning (ML)-based analytics to examine prosodic characteristics of ASD both within and across the two languages. ML capitalizes on the multivariate nature of acoustics features  that richly represents the dynamicity of speech prosody, and have been proven to be able to delineate the acoustic characteristics of the prosodic profile of ASD , 42–44.  prosody . A serie prosody –47. ML c prosody  for a re prosody –51. In pThis study included participants from two languages groups, consisting of native speakers of American English  and Hong Kong Cantonese . The English group included 55 individuals with ASD (English ASD group) and 39 individuals with TD (English TD group). The Cantonese group included 28 individuals with ASD (Cantonese ASD group) and 24 individuals with TD (Cantonese TD group).We acknowledge the variability in preferences related to person-first and identity-first terminology . We do not endorse one style over the other. For the purpose of this manuscript, to ensure that language is parallel when referencing both participant groups, we remain consistent with the following terminology: individuals with autism spectrum disorder (ASD) and individuals with typical development (TD).Participants from the English group were recruited in the United States through a larger family-genetic study of ASD, which included individuals with ASD, their parents, and respective controls. Participants from the Cantonese group were recruited in Hong Kong using advertisements posted on social media platforms  and directly sent to schools and organizations with existing populations of individuals with ASD, as well as from employment programs particularly designed for adults previously diagnosed with Asperger syndrome or high-functioning Autism. Informed assent/consent was obtained from all participants and guardians (as applicable), and procedures were approved by the respective institution’s ethics committee. All procedures were in accordance with ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.Only participants having no reported history of brain injury, major psychiatric disorder, known genetic syndrome, or neurodevelopmental disorder (other than ASD) were included. Participants with TD in both language groups were screened for personal or family history of ASD or related genetic disorders . ASD status was confirmed with research reliable administration and scoring of the Autism Diagnostic Observation Schedule 2nd Edition (ADOS-2). ADOS-2 Overall, Social Affect, and Restricted and Repetitive Behaviors (RRB) calibrated severity scores were used to determine ASD severity .To assess IQ, the Wechsler Abbreviated Scale of Intelligence , Wechsler Adult Intelligence Scale-Third or Fourth Editions , or the Wechsler Intelligence Scale for Children-Fourth Edition  were administered to participants in the English group. The Test of Nonverbal Intelligence, Fourth Edition (TONI-4) was administered to participants in the Cantonese group.Frog, Where Are You? [Participants were asked to narrate (in their respective languages) the 24-page wordless picture book, Are You? . The booAre You? , 54–58 aAre You? . Particiutterance level. In contrast to syllabic-level units, the utterance is associated with the intonational phrase, the most fundamental unit of prosody argued to be universal across all languages [The narrative data elicited from the English group were the focus of a prior published study , examinianguages –63. The Speech Rhythm: Speech rhythm is traditionally considered as durational variations across syllables in an utterance that signal linguistic and affective properties [operties . The temoperties . Such rhoperties . Three moperties –47 were Intonation: Speech intonation, by definition [finition , refers finition –21, 44. finition –21, oftefinition , 61, 66.finition , as it eFurther technical descriptions of the acoustic feature extraction procedures are presented in §1.2 in Two classes of multivariate acoustic features were extracted from utterances in the narrative samples utilizing the MATLAB Audio Toolbox and MATLAB scripts provided by prior studies –47, 64.In two sets of ML models, we trained a series of linear support vector machine (SVM) classifiers to classify participants with ASD from those with TD using principal components derived from the multivariate acoustic features extracted from participants’ narrative samples. The SVM performed classifications by finding a hyperplane from a multidimensional space that divided data points from the principal components according to participants’ diagnosis.The SVM was chosen because it is well suited to handle data of high-dimension, and the types of speech acoustic features implicated in ASD  in particular . The linSeveral procedures were performed in the classification pipeline to further avoid overfitting and optimistic bias due to limited sample size and the high-dimensionality of data   or intonation-relevant (f0 contours) features, on English and Cantonese samples respectively. Model statistics of all classifications are presented in rhythm-relevant features were significant for both English , and Cantonese . In contrast, classifications with intonation-relevant features were significant only for English  but not Cantonese . A post-hoc analysis , whereas the classification using intonation-relevant features was near chance level .Model 2 further examined the classifications between ASD and TD diagnosis using Confusion matrices of all classifications are presented in The present study applied machine learning (ML)-based analyses to acoustic features of speech in ASD, in an attempt to identify prosodic differences in ASD that span two typologically distinct languages, as well as those that may be shaped by specific linguistic and cultural influences. Capitalizing on the power of ML algorithms for performing classifications using high dimensional data , 42–44, Specifically, results of Model 1 indicated that prosodic features in the rhythm of speech contained crucial information captured by the ML algorithm to differentiate individuals with ASD from controls. Such findings complement the many rich, descriptive studies of prosody conducted previously, that described differences in stress pattern, speech rate, and loudness that could be attributed to ASD , 71–75. Perhaps a more revealing aspect of Model 1 is its cross-linguistic design, examining both English and Cantonese utterance samples. Relatively few studies , 34–36 hSuch cross-linguistic commonalities in rhythmic characteristics of prosody associated with ASD may partly stem from the crucial role of prosody in social communication that is partly invariant across cultures. Speech prosody, in general, demonstrates strong similarities across cultures and languages in terms of the ability to recognize emotion from prosody . Such inIn contrast with strong performance for classifications using rhythm-relevant acoustic features in both languages in Model 1, classifications using intonation-relevant acoustic features  were significant for English, but not Cantonese. Unlike non-tone languages where pitch serves linguistic and para-linguistic functions at word, phrase, and utterance levels, in tone languages such as Cantonese, pitch is also used to convey meaning at the syllabic and lexical levels. Therefore, differences in intonation may manifest differently across languages due to cross-linguistic variability . Indeedisorders  that conSeveral potential limitations should be considered in interpreting findings. First, there was a wide age range across our Cantonese and English groups and a small number of females included in the current study. With exhaustive efforts made to avoid model overfitting , robust classification results were found, potentially representing aspects of prosodic profiles of ASD which are relatively age and gender invariant. Moreover, neither age nor gender appeared to influence classification results, as per our post-hoc analysis §2 in . NeverthUsing brief utterance samples from a structured narrative task, ML models applied in this study were able to robustly classify individuals with ASD from those with TD , based on rhythm-relevant features in both English- and Cantonese-speaking populations. The success of our ML approach yields implications for its future clinical utility, such as developing automatic detection of ASD to augment diagnosis  for persTogether, findings from this study provide some of the first evidence for cross-linguistic commonalities in rhythmic characteristics in ASD across two typologically and prosodically distinct languages, while suggesting intonational characteristics in ASD are different across the two languages. Future studies examining languages rhythmically similar to English , as well as those distant from both English and Cantonese , could provide additional crucial insights into establishing prosodic profiles of ASD  which could potentially be invariant across languages, while highlighting those  potentially more malleable to be shaped by language-specific linguistic properties.S1 File(PDF)Click here for additional data file.


The T-OPLL natural course has not been extensively reported, and evidence to support the timing of surgery is also lacking. Owing to the correlation between symptom duration and a low postoperative improvement rate, which has been described, the choice to perform surgery should be proactively examined in patients with progressing myelopathy .In theory, direct decompression induced by OPLL appears plausible. However, operations near the spinal cord that cause extensive injury increase the risk of exacerbating neurological disorders. Therefore, techniques for posterior surgery for T-OPLL without ossification excision are frequently selected. Posterior procedures are divided into posterior decompression alone (laminectomy and laminoplasty) and posterior decompression and fusion (including the method combining fusion with laminectomy and laminoplasty and a kyphosis-reducing procedure (dekyphosis)). Both increased instability induced by posterior invasion into the tissue supporting the spinal column and advanced kyphosis due to age-related intervertebral degeneration and the concurrent compression fracture of the spine exacerbate neurological impairment in posterior decompression alone. Except in individuals with continuous ossification, decompression alone for the mid-thoracic spine is rarely used. Fusion is unnecessary when performing posterior surgery for T-OPLL. Notably, surgical outcomes are good with posterior decompression alone for OPLL when the severity is at a level at which alignment indicates mild lordosis and kyphosis, such as in the upper thoracic spine .The recent increase in improvement rate has been affected by simultaneous fusion via the posterior approach, and various procedures, including posterior decompression and fusion, anterior decompression and fusion, and anterior decompression via the posterior approach, are useful surgical procedures . HoweverAdditionally, factors related to inadequate long-term improvement after surgery include symptom duration, multilevel decompression, cervical OPLL concurrent with the ossification of the ligamentum flavum (OLF), maximum ossification located at the T1–T4 levels, length of the long axis of the OPLL, ossification occupancy rate within the spinal canal, decompression of five or more vertebral bodies, insufficient kyphosis correction during posterior decompression and fixation, intraoperative neurophysiological monitoring (IONM) aggravation, simultaneous surgery of the cervical and lumbar spine, instrumentation utilization, and mid-thoracic spine position .According to the evaluation of symptoms such as numbness or spasticity in a persistent prone or supine posture, changes in spinal alignment while in bed worsened the symptoms. This test was deemed negative in cases where symptoms did not progress after five minutes because these symptoms worsen within one minute in most cases. The authors referred to this test as a prone and supine position test (PST), and situations when symptoms worsened were considered positive .Preoperative physical examinations of PST results are significant and independent elements that affect the success of surgery. Although the PST is positive, the result might not be favorable, even if T-OPLL patients can walk with assistance. Because these patients’ symptoms worsen while in bed and their spinal cord alignment changes somewhat, PST can identify fragile spinal cord damage before surgery. Preoperative PST evaluation is recommended in all T-OPLL cases to identify any serious spinal cord damage and to allow the planning of surgery with appropriate timing to improve the likelihood of a successful procedure .Imagama S et al. prospectively determined the surgical strategy for beak-type T-OPLL, with the additional resection of T-OPLL from a posterior approach planned for three weeks after the first surgery if the patient shows no improvement or has worsening symptoms . The iniThe perioperative complications of T-OPLL surgery were assessed in a prospective, multicenter, and national study . The incThe previous study’s findings support IONM utilization in preventing postoperative motor palsy . After IIn a prospective multicenter study by the monitoring committee of the Japanese Society for Spine Surgery and Related Research, patients with a transcranial motor-evoked potential (Tc-MEP) alert during posterior decompression and fusion surgery for T-OPLL were examined . Tc-MEP Although T-OLF is a relatively uncommon spinal condition globally, it is more prevalent in Asian nations. Due to the progressive disease and the poor response of conservative therapy, patients with T-OLF myelopathy generally require surgery. Notably, thermal nociception, improvement of vibration sensation disorder, history of trauma, and other concurrent ossifications of the spinal ligaments are associated factors regarding outcome-related aspects . The prePreoperative Japanese Orthopedic Association for Myelopathy scores (JOA score) of less than 6 points, fused-type OLF, and beak-type OLF are factors related to surgical outcomes. Other factors include symptom duration, changes in intramedullary signal intensity on magnetic resonance imaging, longer duration of preoperative symptoms, and anteroposterior compression degree .Whether decompression and fusion improve surgical results for OLF compared to decompression alone cannot be determined owing to data insufficiency. In the systematic review, randomized controlled trial (RCT), which directly compared the postoperative results of decompression alone with those of decompression and fusion for T-OLF, were not found. Notably, owing to the need for reoperation and poor improvement rate in symptom, postoperative kyphosis development can become problematic in some individuals when only laminectomy is used to treat T-OLF. Extensive laminectomy without fusion can minimize postoperative kyphosis progression without associated worsening symptoms, with noticeable postoperative alterations absent in thoracic kyphosis .Patients who received laminectomy to remove the ossified lesion concomitant with lateral fusion utilizing bone graft alone experienced a considerably higher rate of JOA score improvement than those without simultaneous fusion. However, regardless of whether fusion was performed, no statistically significant difference was found between the preoperative and postoperative local kyphotic angles. Only a few studies in the same report compared decompression alone with decompression with fusion; however, the patients who underwent internal fusion were limited to those in whom the cervicothoracic junction or thoracolumbar junction was affected and those with osteoporosis .The use of instrumentation can result in higher expenses and decreased spinal mobility due to decompression and fusion. However, some benefits include a reduction in the local kyphotic angle’s postoperative progression and an improvement in neurological symptoms. Therefore, it is not yet possible to conclude that simultaneous fusion considerably improves postoperative results. In addition, patients may find it challenging to select a treatment course given the hazy cost-effectiveness .For patients who have instability before surgery and for those who need to have the facet joint removed for decompression and who are at risk for postoperative instability, simultaneous fusion may be the best option. However, additional studies are required to support these conclusions. It should be highlighted that the pathophysiology of OLF positioned posterior to the spinal cord fundamentally differs from that of OPLL located anterior to the spinal cord, and direct decompression can be performed for OLF via posterior decompression .A multi-institutional retrospective study of patients receiving T-OLF surgery was conducted. The duration of symptoms and the presence of an ossified dura mater were significant predictors of surgical success . A prospOLF occurs frequently at lower thoracic levels due to certain factors: increased mechanical stress caused by the thoracic spine’s position at the intersection of the lumbar spine’s elasticity and rib cage’s rigidity, a direct link between increased spinal mobility and recurrent mild trauma, and the posterior column’s high tensile force. Three cases of thoracic spinal cord injury without major bone injury associated with OLF were reported . FurtherWhile the prospective evaluation of surgical treatment outcomes is made possible by focusing on perioperative complications and risk variables, the RCT of surgical procedures remains uniquely challenging. Spinal surgeons must continue to study these cases prospectively for an extended period to assess the surgical results over time and select the best surgical approach based on patient needs in the future.


Cancer is a major health concern and accounts for one of the main causes of death worldwide. Innovative strategies are needed to aid in the diagnosis and treatment of different types of cancers. Recently, there has been an evolving interest in utilizing nanobodies of camel origin as therapeutic tools against cancer. Nanotechnology uses nanobodies an emerging attractive field that provides promises to researchers in advancing different scientific sectors including medicine and oncology. Nanobodies are characteristically small-sized biologics featured with the ability for deep tissue penetration and dissemination and harbour high stability at high pH and temperatures. The current review highlights the potential use of nanobodies that are naturally secreted in camels’ biological fluids, both milk and urine, in the development of nanotechnology-based therapy for treating different typesQuery of cancers and other diseases. Moreover, the role of nano proteomics in the invention of novel therapeutic agents specifically used for cancer intervention is also illustrated. Cancer denotes one of the leading causes of death globally albeit with the rapid innovations in the molecular biology . AccordiSmall molecules, or nanoparticles, have a diameter of roughly 200 nm. Many pharmaceutical companies have aspired to create efficient drug delivery methods, and as a result, a variety of nanoparticle-based delivery modalities, including magnetic, polymeric, and inorganic nanoparticles, have been developed . An attaNanobodies exemplify unique structural and functional features credited to their small size 15–74 kDa) and large surface area, high stability and solubility, high binding affinity and detection of different epitopes, fast tissue internalization, ease of production and manipulation, and low immunogenic reactions . Consequ5–74 kDa Targeted delivery is a major milestone in cancer therapy as it mitigates the damage caused by non-selective drugs. Nanoparticles allow a controlled release of drugs inside the cancer cells using multi-functional nanocarriers. Consequently, the therapeutic potential of nanobodies as tools to deliver drug-loaded nanoparticles toward tumours has recently attracted the attention of many researchers , 28. TheThere are two surviving species of camels: Camelus dromedarius and Camelus bactrianus with each having distinguishing features. Camelus dromedarius (C. dromedarius) also called the dromedary, one-humped camel, or Arabian camel mainly lives in the hot deserts of North African and Middle Eastern countries, including Saudi Arabia (SA) , 33. WheThe genome of the C. dromedarius camel has been completely sequenced as a result of the joint efforts of researchers from the Beijing Genomics Institute (BGI), China and the National Center for Genomic Technology (NCGT) at King Abdulaziz City for Science and Technology (KACST), Saudi Arabia . SequencThe fact that all camelids naturally produce different antibodies that circulate in their blood should be kept in mind. Two identical heavy chains and two identical light chains joined by disulphide bonds through non-covalent interactions make up the usual structure for conventional antibodies . On the Development of a therapeutic tool targeting selectively the tumour cells and leaving the normal cells unaffected is a challenging aim. Cancer cells are featured by overexpression of specific receptors or biomarkers that can act as an attractive spot to increase the delivery and the accumulation of the drug into the affected tissues . FollowiThe employment of the camelid-derived nanobodies in diverse areas of nanomedicine and nano therapy has contributed to the advancement of disease screening and prevention, diagnosis, treatment, and follow-up. Indeed, many commercial companies such as Ablynx, Camel-IDS, and Hybrigenics were reported to generate nanobodies . In addiCamels’ milk has a high nutritional value  as it coThe highest milk production is found in the Majaheem breed (of C. dromedarius species) however there is a significant difference in the milk components among the C. dromedarius dairy camel breeds. For example, milk produced by the Sofor camel is higher in protein, fat, and lactose than that produced by other camel breeds in SA . On the The milk proteins, particularly lactoferrin, deliver potent probiotics, antioxidant, anti-microbial along with anti-inflammatory effects  as shownThe therapeutic ability of camel milk can be delivered by its gifted biological components such as vitamin E and C , casein,Moreover, camel milk expressed distinctive immunoglobulins characterized as VHH antibodies or nanobodies . These ihttp://kmplot.com/analysis/) [To complete the picture, we investigated the correlation between the expression of the camel milk protein components combined and the overall survival (OS) of cancer patients with different tumour types using the online KM plotter software  . This toDespite being a waste product, camel urine has been utilized as a foundation for several therapeutic mediators. In alignment with the previous reports, a study conducted by Abdul Qader et al. showed that camel urine consists of more than 32 proteins including alpha-1B-glycoprotein, alpha-1-acid glycoprotein, serotransferrin, vitamin D-binding protein, serum albumin, and thyroxine-binding globulin. These proteins could explain the anti-inflammatory and anti-infectious effects delivered by camel urine . In vitrMoreover, camels’ secretions, including milk and urine, revealed favourable results as a therapeutic modality in the hepatoma , breast Likewise, mitigation of tumour angiogenesis that is delivered by the camel urine was recently confirmed by Gader and Alhaider . AdditioThe precise function of the anticancer components in camel urine and milk remains poorly understood. The tumorigenic suppression capacity depicted by the camel urine was attributed mostly to the abundance of iron-binding lactoferrin component , 90–92. Furthermore, several forms of camel urine have existed such as Prophet Medicine Fraction (PMF) and PM701 found to be enriched in crystals, nanorods, nanoparticles, and different organic and inorganic compounds. PMF crystals constitute numerous elements such as calcium oxalate, cystine, uric acid crystals, ammonium urate, calcium phosphate, benzoic acids, glycine, alanine, and arginine. Additionally, PMF is highly concentrated in ions including Cs, Rb, K, Ca, Cd, Y, Eu, Th, and Zn . Importa18F were successfully used in the detection of HER2 expression in breast cancer cases using PET/CT imaging technique [131I IB-Mal-D-GEEEK were produced for determining HER2 expression in breast cancer patients before trastuzumab administration. This conjugated nanobody revealed encouraging results in targeting HER2 tumours promoting receptor internalization and subsequent signals inactivation both in vitro and in mice models, . They studied the consequence of adding PMF nanoparticles to a lung cancer cells line (A549) using various microscopic and spectroscopic techniques, including scanning electron microscopy. A549 cells treated with PMF showed that the PMF exhibited two modes of action against the target cells. First, these nanobodies attacked the membrane of A549 cells and changed the packing and polarity of the membrane. Second, these nanobodies selectively targeted cancer cells and transferred the PMF into the nucleus and other organelles of abnormal cells through endocytosis. In this regard, this nanobody-based drug delivery system (DDS) achieved the purpose of the smart drug treatment . SimilarThe insinuation of nanobodies in medical determinations has immensely progressed as numerous nanobodies have been tested in the preclinical setting, while many of them are being examined in clinical trials as demonstrated in Table As previously described the camels’ biological secretions have shown to exert therapeutic benefits in treating numerous health conditions. However, the precise subcellular molecular mechanisms that are regulated by these secretions necessitate further investigations. Camel milk was reported to induce a hypoglycemic effect in both human and animal models and was suggested as a complementary therapy in the type 1 diabetes –132. To The anti-inflammatory and antioxidant effects exhibited by camel milk components were also validated in a rats model with acute respiratory distress syndrome (ARDS). ARDS is defined as the formation of significant inflammatory background causing damagethe to alveolar epithelial and endothelial barriers ending in respiratory failure , 136. ThCyclosporin is an immune suppressant agent that is widely used for several autoimmune diseases. The most harmful side effects of this drug are kidney injury that can progress to renal failure. Camel milk, as an anti-inflammatory/antioxidant mediator, was proposed as a natural agent to ameliorate the side effects associated with cyclosporine administration. In vivo Administration of camel milk for 3 weeks reduced dramatically the levels of the biomarkers related to renal damage such as serum creatinine, BUN, and KIM-1. Furthermore, camel milk treatment is associated with a marked lowering in numerous inflammatory cytokines and degradation signals such as MCP-1, IL-8, TNF, MMP-2, and MMP-9. Besides the ability of the camel milk to inhibit p38/ERK/JNK MAPK signalling, camel milk blocked NF-κB pathway activation by suppressing the expression of NF-κBp65, p-NF-κBp65, and p-IκBα proteins. Curtailing of pathways together with enhancement of glutathione and antioxidant activities resulted in alleviating of the oxidative stress on the kidneys. Therefore, camel milk can be used as a natural protecting remedy against renal damage triggered by cytotoxic drugs .Camel urine, the other camel’s biological secretion, was also reported to exert a tumour suppression effect and augment the doxorubicin treatment for breast cancer cells. This anticancer impact was obtained by inducing cancer cells apoptosis and DNA damage, reversing the EMT markers, and retrieving the expression of the E-cadherin epithelial marker. The author suggested the potential involvement of the camel urine component in attenuation of NF-κB-Snail signalling and its downstream inflammatory mediators thus abrogating breast cancer cells' aggressive phenotype .The camel genome sequencing project was completed in 2003, and the data it produced gave researchers the chance to use proteomics to analyze and gain insight into all of the cellular processes that could not be explained by genomics alone. A proteomics study offers helpful information on the post-translational modifications, localizations, structures, and interactions of all proteins expressed by an organism in addition to merely studying the entire proteome . Also, pThe immense challenge of finding minute quantities of biomarkers from natural fluids such as urine, saliva and blood, compared to the simpler detection of high-abundance proteins, has led to the emergence of a new field called nano proteomics, which integrates proteomics and nano techniques –150. NanThe exceptional pharmacokinetic and physicochemical properties of camel nanobodies are consistent with those needed for cancer therapy and provide added benefits over conventional antibody technology in drug delivery, immunotherapy, and diagnostics. The affinities of camel-heavy antibodies are higher than those of conventional antibodies and can reach up to 100 pM affinity constants , 159. ThAdvancements in computational molecular biology, such as the formation of large genomic and proteomic databases and the development of various tools and bioinformatic software, facilitated the achievement of various benchmarks that can be used for therapeutic purposes. Bioinformatics is playing a vital role in understanding complex metabolic pathways and discovering various vital components of living systems . SeveralResearch has also been carried out to utilize and explore the combined potential of nanoparticles and bioinformatic tools for the treatment of cancer . In 2012Exosomes are nano-sized extracellular membrane vesicles that range in size from 50 to 200 nm and are unique nanoparticle drug carriers with important nanoparticle features , 175. ExPreliminary studies have shown that camel milk exosomes could play an important role in inhibiting the growth of breast cancer cells . They coFurthermore, EL-Kattawy et al. found that camel colostrum-derived exosomes, that are enriched in milk protein contents, showed remarkable apoptotic effects on liver cancer cells exclusively sparing the normal cells unaffected. These carcinogenic suppression effects are attained by elevation in the expression level of both Bax and caspase3 and reduction in Bcl2 level. This was accompanied by a reduced expression level of the inflammatory mediators such as TNFα, NFkB, TGFβ1, and Cox2 as well as the angiogenic-related factors VEGF .In this review, we discussed the therapeutic value of the camelid-derived nanobodies along with the therapeutic effects provided by the ingredients of camels’ biological secretion both milk and urine in combatting different human diseases mostly cancers. Many research areas such as medical, biotechnology, engineering, and economics have been cultivated after the discovery of the naturally occurring camelids nanobodies. Nanobodies have recently emerged as attractive robust strategies for diagnostic and therapeutic aims, particularly in the cancer field. Providentially, nanobodies exhibit several biophysical and biochemical proprieties besides the ability to cross the biological membranes such as the blood-brain barrier. Furthermore, they show the capacity to enter smoothly into solid organs including the brain, lymph nodes, lungs, and liver. Because nanobodies presented flexible manufacturing and assembly they have been incorporating into multipotent constructs or conjugating with chemotherapeutic drugs to produce highly specific and efficient compounds. These features make nanobody-based therapies useful and powerful tools to deliver the drug into the targeted tissues exclusively. Nevertheless, nanobodies still displayed some pitfalls due to their minute sizes including rapid renal clearance and a possibility of renal toxicity. This drawback can be modified by coupling nanobodies with the serum albumin to increase their retention time in the circulation, yet such an approach will inexorably reduce their diffusion and penetration benefits. Further investigation is pivotal to elucidate their exact molecular mechanisms in modulating oncogenic signalling pathways and to assess their safety profile in preclinical experimental studies before the implication in any clinical trials.