Currarino triad is a rare syndrome that may be occasionally encountered during managing cases of anorectal anomalies. The triad consists of anorectal anomaly, sacral bony defect, and a presacral mass. It may be familial or sporadic, with a reported female predominance. Identification of the characteristic notched sacrum  in plain X-ray (anteroposterior view) is considered the key for the diagnosis; however, not infrequently, this radiological sign is overlooked, especially with a small sacral defect.Excision of the presacral cyst is usually performed concomitantly during anorectoplasty. The prone position is the standard approach for posterior sagittal anorectoplasty (PSARP) in males; however, in females, the supine position can be used as an alternative . In this case report, excision of the presacral cyst took place in two steps: the first excision during the PSARP procedure in the prone position, and a second operation in the supine lithotomy position to remove a residual component of the lesion that was missed during the primary operation. It was clear that the supine lithotomy position provided better access to explore the presacral space than the prone position, especially with a deeply located cyst as in our case. The role of magnetic resonance imaging (MRI) in the identification of the exact nature and extent of the lesion before surgery is crucial and should be performed in all cases. Currarino triad is a rare congenital disorder characterized by the presence of three components: anorectal anomaly, sacral vertebral defect, and a presacral mass .Early reports in the literature were described by KennedyA male patient presented at birth with anorectal malformation ; there At the age of 3 months, a high-pressure distal colostogram was performed as a preparatory step for a PSARP  procedure.At the time of operation, the mother revealed that her elder daughter (now 6 years old) underwent a similar operation. Revising the medical records of her daughter revealed a case of Currarino triad: anal stenosis (funnel anus), vertebral defect, and presacral cyst. Because of the reported autosomal dominant inheritance in cases of Currarino triad,With the patient in the prone position, a generous posterior sagittal incision was made for mobilization of the anorectum and exploration of the presacral space. We started as usual by dissection of the fascia around the posterior and the lateral aspects of the rectal pouch. A small midline incision was made in the posterior wall of the rectum to inspect for any missed communication with the urinary tract and to guide the dissection during separation of the anterior rectal wall from the intimately adherent urinary tract. After mobilization of the anorectum, a retrorectal cyst (∼2 cm in diameter) was found and excised. Then, the steps of anorectoplasty were completed as usual by reconstruction of the levator ani and striated muscle complex around the neorectum.Postoperative recovery was uneventful. Histopathological examination of excised specimen revealed fibromuscular tissue showing multiple clefts lined by stratified squamous epithelium and entrapped transitional epithelial islands. These histopathological findings go with the diagnosis of a developmental cyst associating with Currarino triad.Later, a postoperative pelvic MRI was ordered that revealed residual presacral cystic lesion , with tThe patient was seen at follow-up 3 months after the closure of colostomy. He defecates spontaneously once every 1 to 2 days. He is still only 19 months old and therefore below the age of voluntary bowel control. However, the mother was informed about the possibility of constipation, problems with fecal continence, and the need for long-term follow-up. The mother has had a similar experience with her elder daughter who is now 6 years old and has voluntary bowel and urinary control. The parents (being the first-degree relatives of our case) were advised to perform screening by plain X-ray sacrum.Currarino triad is a rare syndrome that may be occasionally encountered during managing cases of anorectal anomalies. The triad consists of anorectal anomaly, sacral bony defect, and a presacral mass.MNX1(previouslyHLXB9) gene on chromosome 7q36. The disorder is an autosomal dominant genetic trait that has variable expressivity.MNX1gene have been identified in nearly all cases of familial Currarino's syndrome and in approximately 30% of patients with sporadic Currarino's syndrome.Currarino triad is caused by heterozygous mutation in the-ray due to a highly variable phenotype within most families.-ray should be referred to a surgeon for further investigations, including pelvic MRI for an occult presacral mass due to significant inter- and intrafamilial variability in expression.All affected families should be offered genetic counseling, as awareness of the hereditary nature of the disorder allows identification of asymptomatic heterozygotes and patients at risk, which leads to better planning of pregnancies and appropriate management of affected patients. All first-degree relatives of patients with Currarino triad should be screened by pelvic XThe presacral mass in Currarino triad may be a lipoma, lipomyelomeningocele, or some sort of developmental cyst.Excision of the presacral cyst is usually performed concomitantly during anorectoplasty. The prone position is the standard approach for PSARP in males;There was some delay initially in completing the diagnosis for this case; however, the management was completed with a satisfactory outcome. Lastly, the prognosis for continence may be generally good for this case owing to the absence of associated spinal cord anomalies.


This study aimed to assess the feasibility and acceptability of the first known implementation study of these standards in a low resource setting.Perinatal and newborn complications are major risk factors for unfavorable fetal and neonatal outcomes. Gestational dating and growth monitoring can be instrumental in the identification and management of high-risk pregnancies and births. The INTERGROWTH-21The study was performed in two 12-month phases from March 2016 to March 2018 at Jacaranda Health, a private maternity hospital in peri-urban Nairobi, Kenya. In-depth interviews, focus group discussions and a provider survey were utilized to evaluate providers’ experiences during implementation. Client chart data, for pregnant women attending antenatal care and/or delivering at Jacaranda Health along with their newborns, were captured to assess uptake and effect of the standards on clinical decision-making.Facility-level support and provider buy-in proved to be critical factors driving the success of implementing the standards. However, additional support was needed to strengthen capacity to conduct and interpret ultrasounds and maintain motivation among providers. We observed a significant increase in the uptake of obstetric ultrasounds, particularly gestational dating, during the implementation of the standards. Although no significant changes were detected in the identification of high-risk pregnancies, referrals and deliveries by Cesarean section during implementation, we did observe a significant reduction in inductions for post-date. No significant barriers were reported regarding the use of the newborn standards. Over 80% of providers advocated for the standards to remain in place with some enhancements related mainly to training, advocacy and procurement.The findings are timely with increasing global adoption of the standards and the challenging and multi-faceted nature of translating new, evidence-based guidelines into routine clinical practice. Despite progress made in reducing maternal and newborn mortality, large gaps remain. Currently, 2.6 million neonates die yearly worldwide, most from preventable causes . Fetal gst Project developed the first global standards for gestational dating, fetal growth monitoring, and newborn size at birth, supplying a toolkit for clinicians working to reduce the burden of preterm birth as well as growth faltering and its complications  because skill is something that is now in me and I feel empowered and so am grateful and we would like to have the practical skill for the ultrasound.”Anthropometry Nurse-Midwife (Implementation Phase– 12 months)This improvement in knowledge and skills was also reflected in the enhanced quality of ultrasound images over time. Analysis of variance and subsequent pairwise comparisons indicated that the quality scores improved significantly over time for head circumference and femur length. For both, we found that there was continuous and significant improvement from batch one to five which then continued to batch six.st standards had become part of routine practice at Jacaranda Health. More than half of respondents (17 of 29) reported that the effort to integrate the tools into the clinical flow was worthwhile. The main reported facilitator to this integration was the change in the clinical workflow model to one dedicated nurse-midwife performing gestational dating ultrasounds. Reported barriers included long waiting times that turned away clients or resulted in complaints to already overwhelmed providers, especially in the first half of implementation; the time required to conduct procedures on top of regular clinical duties; and the lack of full-time availability of the nurse-midwife providing gestational dating services in the second half of the implementation phase.Over 86% of providers (25 of 29) surveyed at eight months into the implementation phase agreed or strongly agreed that the INTERGROWTH-21st gestational dating and newborn size at birth standards had an overall positive effect on their workload. The expert sonographer at Jacaranda Health also expressed that the process of implementing the standards reduced her workload and allowed her to concentrate on fetal growth monitoring and other advanced obstetric scans. Similarly, other nurse-midwives reported feeling relieved that a dedicated provider was assigned to carry out all gestational dating ultrasounds; this adaptation eased the burden of combining gestational dating scans with their regular ANC duties, as originally planned and implemented.At four months into the implementation phase, providers reported an initial increase in workload, but it was later normalized as they began to see the benefit of the standards and flow issues were addressed. At the end of the implementation phase, the majority of respondents (10 of 14) noted that the INTERGROWTH-21“Initially, I can say it [the ultrasound scans] came as an extra tool without really knowing why I have to do this. But, through getting used to the tools and doing it regularly, I came to get used to it and think right now I can say it is something we feel like we cannot do without.”Anthropometry Nurse-Midwife (Implementation Phase– 12 months)Providers were already used to assessing newborns at birth, and reported that the standards increased their efficiency and delivery of accurate information.st standards, documentation of the ultrasound form included six extra indicators related to the determination of the EDD via ultrasound and the LMP. Significantly more ultrasounds had complete documentation in the pre-implementation phase (96%) than the implementation phase (52%) (p<0.0001).We observed a significant increase in the percentage of ANC clients receiving any obstetric ultrasounds , from 919 (53%) in the pre-implementation phase to 1156 (64%) in the implementation phase (p<0.0001). In total, 1217 obstetric ultrasound scans were performed in the pre-implementation phase and 1653 in the implementation phase. There was also a significant increase in the uptake of gestational dating ultrasounds in the implementation phase compared to the pre-implementation phase; from 373 scans representing 34% of eligible ANC clients to 580 scans representing 55% of eligible ANC clients (p<0.0001). The difference in uptake was further intensified when we changed from a multi-nurse model (40% of eligible ANC clients) to a dedicated ultrasound nurse model (73% of eligible ANC clients). Information on clients who chose to opt out of scans was only recorded during the implementation period so we were not able to compare opt-outs between the two study phases. With the implementation of the INTERGROWTH-21According to providers, the promotion of the gestational dating scan as free of charge during the implementation phase served as an incentive for eligible pregnant women to opt in and helped providers to advocate for the scan’s importance. Providers felt that the novelty and perception of the accuracy of the gestational dating services also facilitated their promotion and uptake.“Because it has been free so there are no negative reactions but if it was coming with a cost then they [clients] would have said that am not prepared for it today and let me do it next time. Such things would crop up.”Ultrasound Nurse-Midwife (Implementation Phase– 12 months)At the same time, providers shared a number of issues that they felt needed to be tackled to further promote the uptake of the ultrasound standards. Reported demand side obstacles included long wait times; lack of print outs accompanying scan results to show family members; first ANC visits late in the pregnancy; and client misconceptions such as the fear that scans would harm the fetus. Client perceptions and experiences of care during the implementation of the standards are described in detail in another paper.Perceived supply side barriers at four months into the implementation phase included lack of initial provider motivation to take on additional work; added requirements for documentation; long wait times that caused bottlenecks in providers’ schedules; longer time needed to conduct scans limiting the number of clients who could be seen; and an insufficient number of nurse-midwives trained in ultrasounds resulting in staff coverage gaps. At the end of implementation, providers noted that improvements were made as a result of the one-nurse model, but expressed concerns about the times when that designated nurse-midwife was off duty.st calculator utilized by data entry clerks, accuracy of diagnoses by nurse-midwives were found to be 76%, 80% and 77% for AGA, SGA and LGA respectively. The discrepancies were mainly due to the diagnosis falling between two diagnostic categories and incomplete documentation of the forms. Similar to the ultrasound form, there was a significantly lower (p < 0.0001) completeness of documentation of the newborn assessment form in the implementation phase (63%) compared to the pre-implementation phase (80%).All documented deliveries at Jacaranda Health, both live births and stillbirths, were enumerated for the study. Out of the 1092 newborns whose assessment forms indicated a newborn size at birth diagnosis in the implementation phase, 1037 (95%) were identified as being appropriate for gestational age (AGA), 25 (2%) as SGA and 30 (3%) as large for gestational age (LGA) using the new growth charts. When compared with the results generated by an online INTERGROWTH-21Generally, no major barriers were reported in the use of the newborn size at birth standards apart from some additional documentation needs. Reported facilitators for uptake of the newborn size at birth standards included enhanced infection control because the new scale was easy to clean and the head circumference tapes were disposable; decreased workload because weight and height could be measured on one apparatus; and increased accuracy and standardization. Finally, providers shared that the implementation of these standards built upon existing newborn anthropometry practices which facilitated their routine integration and helped to promptly identify newborn size at birth diagnoses.st standards improved overall growth monitoring (90%), high-risk identification and referral (76%) and quality of care (72%). While the proportion of ANC clients identified to have high-risk pregnancies and those referred  increased slightly from the pre-implementation to the implementation phase, these increases were small and not statistically significant  inductions of labor due to post-date pregnancies . This wa“[We are] getting almost accurate EDD’s to [help us] avoid unnecessary post-datism and thus it has helped us to act when necessary.”Ultrasound Nurse-Midwife (Implementation Phase– 12 months)When evaluating the association between the implementation of the standards and Cesarean section rates in one study phase versus the other, we did not find a statistically significant difference . HoweverThe table below summarizes the main facilitators and barriers of implementing the standards as experienced by providers .The majority of providers (13 of 16) expressed that the standards should continue to be used, highlighting how instrumental they have been in enhancing the quality of care as well as their potential to improve the health and survival of women and newborns.st standards at Jacaranda Health were assessed through the examination of two broad dimensions: (1) facility-level support to introduce and integrate the standards into practice; and (2) use and integration of the standards to influence uptake of ultrasound and newborn size at birth standards, and clinical decision-making around pregnancy and delivery care. Facility-level support and provider buy-in proved to be critical factors driving the success of implementing the standards. Training and job aids were instrumental in the introduction and reinforcement of the standards. A literature review of clinical guideline implementation in a number of low resource settings also highlights the importance of introducing checklists and other job aids and making context-specific adjustments to foster integration [st standards, Uauy and colleagues [The feasibility and acceptability of implementing the INTERGROWTH-21egration . In theilleagues , 22–24. lleagues .Our implementation study sheds light on an innovative model of ultrasound delivery by nurse-midwives for whom the optimal training regimen has yet to be determined . Nurse-lWe observed an increase in the uptake of obstetric ultrasounds, particularly gestational dating scans. However, only about 60% of ANC clients presented for their initial visit in the first two trimesters with most of these visits occurring in the second trimester. First ANC visits in the second and third trimesters are common in sub-Saharan Africa and lead to missed opportunities, during the first trimester, for the delivery of crucial components of ANC and management of obstetric complications , 30–32. st standards.The use of the newborn size at birth standards was deemed feasible and acceptable by the nurse-midwives administering them; no significant barriers were reported. Unlike gestational dating ultrasounds, administration of these standards required minimal adaptations to existing newborn anthropometry practices while offering additional support for decision-making. Additionally, uptake was guaranteed to be universal since newborn anthropometry was a part of essential newborn care. Data on clinical decision-making indicators, when considered together with provider inputs on the standardization of post-date cut-offs and indications for Cesarean section during the study period, do not suggest a significant association between the implementation of the standards and clinical decision-making outcomes. Similar to our results, health workers in rural Kenya and Rwanda also reported that ultrasounds were instrumental in helping them to make clinical decisions, manage patients and improve the quality of ANC , 31. Givst standards. Providers’ and clients’ experiences were equally prioritized and captured; this paper describes the former. Additional strengths include pre-implementation versus implementation comparisons, the use of both quantitative and qualitative methods and the setting of the study in a low resource context. This study also had a number of limitations. Our findings related to the ultrasound-based standards focus largely on gestational dating scans, a service that was not routinely provided before the study. Conversely, fetal growth monitoring was already part of routine care at Jacaranda Health; thus, we did not expect the implementation of the standards to influence a change but rather focused on overall uptake in obstetric ultrasounds. Results on the effect of the implementation of the standards on clinical decision-making outcomes were limited. We recommend that further time and financial investment would enable the standards to become engrained in clinical practice and lead to changes in maternal, perinatal and newborn outcomes, in particular at a low-risk health facility such as Jacaranda Health. The implementation process and its findings may not be generalizable to all health facilities in Kenya and beyond, but we have gleaned important insights that can facilitate adaptation and implementation of the full or partial INTERGROWTH-21st package in other settings.This is the first implementation study evaluating the feasibility and acceptability of the INTERGROWTH-21st standards are being adopted in a number of countries around the world and are proving to be an instrumental tool for the improvement of the quality of routine maternal and perinatal care and emergency situations that affect pregnancy and perinatal outcomes, such as the zika epidemic [st standards built into them, thus making it more feasible to implement the standards on a larger scale. We hope that providers’ experiences and insights will be beneficial to facility managers who are considering, preparing for or in the process of implementing the INTERGROWTH-21st standards or other similar clinical initiatives. This paper highlights the need to engage health providers in understanding and addressing their needs; the time and resource investment required for training and capacity building; the flexibility and monitoring necessary to facilitate adaptations to clinical flow; and the recognition that changes in clinical practice take time, especially when they deviate greatly from existing care. More research is needed to evaluate the financial, political and health implications of introducing routine gestational dating in low resource settings and community-based approaches to increase demand for early initiation of ANC.This implementation study is timely as the INTERGROWTH-21epidemic . The staepidemic . AdditioS1 File(PDF)Click here for additional data file.S2 File(PDF)Click here for additional data file.S3 File(PDF)Click here for additional data file.S4 File(PDF)Click here for additional data file.


CSF leak in penetrating skull base injury is relatively rare compared to close head injury involving skull base fracture. We report a 5-year-old boy presented with epistaxis and impacted pencil into the left nostril. The child was hemodynamically stable without any neurological deficit. Intraoperatively, there was a nasal septal defect posteriorly with anterior skull base fracture associated with CSF leak. The pencil was removed from the left nostril and the CSF leak was repaired using harvested abdominal fat under the same setting. Computed Tomography (CT) of the brain showed right cribriform plate fracture with small pneumocranium. Postoperatively, a prophylactic antibiotic was given for seven days and he was discharged well. Subsequent clinic visits up to one-year postoperative period showed no recurrence of the CSF leak. History taking, physical examination and CT imaging give valuable diagnostic values in managing the penetrating skull base injury. Early intervention for removal of the foreign body and repair of the CSF leak is advocated to prevent catastrophic complication.  Presence of fistulous tract in between intracranium and nasal cavity may result in cerebrospinal fluid (CSF) rhinorrhea. Craniofacial trauma contributed 80 % of the etiologies of CSF rhinorrhea. Traumatic CSF rhinorrhea is more commonly seen in skull base fractures. Majority of the skull base fractures (90 %) are caused by closed head injuries, whereas penetrating trauma constitutes the remaining 10 % of the etiology.Most of the penetrating head injuries involved oro-cranial or transorbital intracranial penetration of the sharp object with a significant external injury. Foreign bodies such as glass, air gun projectiles, metal splinters and plastic chair glides have been reported. Traumatic transnasal penetrating injury to the skull base is rare in literature. Removal of the penetrating object followed by repair of the CSF leakage depicts more challenging management.In this case report, we described an unusual case of penetrating injury of the anterior skull base by a pencil via the left nostril, without external, orbital and paranasal sinuses involvement. The importance of radiological assessment prior to surgery was emphasized in this report. We also discussed the approaches of removal of the penetrating object and endoscopic CSF leakage repair in the same setting.A 5-year-old boy presented to the emergency room with epistaxis following an impacted pencil over the left nostril. He admitted to inserting the blunted end into his left nostril and was pushed by his sibling from behind. He fell forward on his face causing impaction of the pencil into his left nostril. His mother claimed that they have successfully pulled out the pencil and the epistaxis subsequently resolved. Upon review, he was hemodynamically stable, alert and conscious. Anterior rhinoscopy showed part of the pencil shaft embedded in the left nasal cavity. The child was however uncooperative for further assessment of the extent of the injury. Otherwise, there was no external wound and deformity over the craniofacial region, bilateral eye examination and neurological examinations were unremarkable. Radiograph of the skull Figure 1 showed fHe was brought into the operating theatre for examination under anesthesia. Intraoperatively, the pencil was impacted firmly in the left nostril and nasal septum Figure 2. The penAfter removal of the foreign body, the CSF leakage was repaired with autologous graft by placing the septal cartilage and abdominal fat on to the defect until the CSF leakage ceased. The septal mucosal graft was repositioned on top of the fat graft and covered with Gelfoam and tissue glue.Post-operatively, he was ventilated for 24 hours and kept under bed rest with intravenous ceftriaxone administered for one week in view of overwhelming intraoperative findings. Computed Tomography (CT) scan of the brain, skull base and paranasal sinuses post-operatively showed right cribriform plate fracture with small pneumocranium Figure 4. No evidNasal foreign bodies are common encounter in ENT setting. It ranges from simple foreign bodies to catastrophic scenarios with skull base fractures and intracranial complications.Transnasal septal penetrating injury to the anterior skull base without an external wound and orbital involvement makes this a unique case. Furthermore, management of penetrating injury of the head and neck region in a pediatric patient can be very challenging as history may be inaccurate with limited cooperation during an examination. Additionally, clinical examination showed no external wound over the face and orbital region with intact neurological function masquerade the extent of the injury made..When the history is uncertain, the evaluation of suspected penetrating injury of the nose lies on the radiological assessment. A plain radiograph is easily available with low radiation, it offers an initial assessment of the bony framework of the facial and skull bone to detect any fracture lines. The panoramic view of plain radiograph helps in locating the foreign body. However, the visibility of the foreign body in plain radiograph depends on the types of object. Metallic foreign body is radio-opaque on radiograph whereas organic foreign body like peanut or wood would be less radio-opaque hence easily missed .CT scan gives a good osseous delineation which is important in detecting subtle fractures. This fine thin cut of the CT scan with the image in axial, coronal and sagittal plane provides an accurate assessment of the location of the foreign body. With the three-dimensional (3D) images reconstructed from the fine cut CT scan, the extent of the injury can be accurately localized and essential in planning the surgical approach. For cases suspected with vasculature or intracranial injury, a CT angiography and Magnetic Resonance Imaging (MRI) brain would be warranted .rd generation of cephalosporin with the property of crossing blood-brain barrier for prophylaxis against intracranial infection.Early removal of the penetrating foreign body is the definitive treatment in regards to avoid the unwanted sequelae . Early There are two suggested methods in the removal of the penetrating foreign body into the maxillofacial region, namely the endonasal approach and external approach . The shCSF leak was demonstrated immediately after removal of the foreign body. Most of the traumatic CSF leak will be resolved with conservative management or CSF diversion with lumbar drain. However, the incidence of ascending intracranial infection was relatively high in the group of CSF leak patient when conservative treatment was given. Thus, repair of the CSF leak was advocated whereby it significantly reduces the risk of ascending intracranial infection . Presence of fistulous tract in between intracranial and nasal cavity predisposes the patient to CSF leaks, pneumocephalus and intracranial infection such as ascending meningitis and abscess. The goals of skull base reconstruction include separation of the cranial cavity from the sinonasal tract, protection of neurovascular structures, preservation of functions and avoidance of dead spaces.As stated by Wormald and McDonough in 1997).Indications for endoscopic repair include post-traumatic CSF leak, iatrogenic and spontaneous CSF rhinorrhea associated with either malformation of the skull base, meningoencephaloceles, empty sella and increased intracranial pressure. With the improvement of skills and instrumentation, amazingly high success rates with primary closure of the defect between 88 % and 94 % were achieved in the early 1990s .The key to successful management of CSF leak is to precisely identify the site of the dural tear. Preoperative imaging such as high-resolution CT scan of the skull base and paranasal sinuses can be performed to locate the fracture site. Intrathecal application of sodium fluorescein in combination with the use of blue light and a blocking filter over the endoscope's eyepiece will precisely guide the endoscopic surgeon to the site of the lesions. Laboratory techniques such as Beta-2-transferrin and beta-trace protein testing are the standard tests that help to differentiate non-CSF-related nasal secretion from the true CSF in an unclear situation .In this case, we harvested the fat graft from the periumbilical region as the autologous graft for repair. The indentation mark over the anterior skull base was covered by the lacerated septal cartilage with septal mucosa. For defect closure, it is recommended that no mucosa should be placed under any graft or fat to avoid mucocele formation. Hence the septal cartilage with mucosa was taken out temporary, the area of skull base defect was denuded of its mucosal layer. Adequate size of the harvested fat graft was gently placed over the skull base defect with pediatric Blakesley forcep. With the use of Freers elevator, the fat graft was gently placed in between the bone and surrounding mucosa, which is described as overlay technique. Subsequently, the septal cartilage with mucosa was repositioned back on top of the fat graft. The grafted area was reinforced with Gelfoam and tissue glue. The end result of the repair was excellent without intracranial complication and further CSF leak up to date.Management of transnasal penetrating injury of the anterior skull base can be very challenging in terms of diagnosis and treatment. When the history and physical examination are in doubts, CT imaging is mandatory to rule out suspecting skull base injury. Having said that accurate diagnosis and early intervention are important in order to prevent intracranial complications, continuous follow up is advocated to detect the delayed complication.The authors declare no conflict of interest.


Acropora millepora), a sea urchin (Tripneustes gratilla), and a reef fish (Epinephelus sp). Connectivity metrics and reef clusters were computed from the settlement probability matrices. Fitted power law functions derived from the dispersal kernels provided relative probabilities of connection given only the distance between reefs, and demonstrated that 95% of the larvae across organisms settled within a third of their maximum settlement distances. The magnitude of the connectivity metric values of reef cells were sensitive to differences both in the type of organism and temporal variability. Seasonal variability of connections was more dominant than interannual variability. However, despite these differences, the moderate to high correlation of metrics between organisms and seasonal matrices suggest that the spatial patterns are relatively similar between reefs. A cluster analysis based on the Bray-Curtis Dissimilarity of sink and source connections synthesized the inherent variability of these multiple large connectivity matrices. Through this, similarities in regional connectivity patterns were determined at various cluster sizes depending on the scale of interest. The validity of the model is supported by 1) the simulated dispersal kernels being within the range of reported parentage analysis estimates; and, 2) the clusters that emerged reflect the dispersal barriers implied by previously published population genetics studies. The tools presented here  can be used to include regional patterns of connectivity into the spatial management of coral reefs.Coral reefs of the North Indo-West Pacific provide important ecosystem services to the region but are subjected to multiple local and global threats. Strengthening management measures necessitate understanding the variability of larval connectivity and bridging global connectivity models to local scales. An individual-based Lagrangian biophysical model was used to simulate connectivity between coral reefs for three organisms with different early life history characteristics: a coral ( The North Indo-West Pacific (NIWP) is an archipelagic region composed of several marginal seas, narrow straits, and shallow bays that host the highest levels of biodiversity and a significant portion of the global coral reef area ,2. TheseLarval connectivity, henceforth referred to as connectivity, is the exchange of individuals between populations which results from processes involving larval production, transport by currents, larval behaviors, and post-settlement conditions. Population growth, regulation, and recovery from disturbances is dependent on the supply of larvae including both self-recruitment and immigration from other reefs –22. LarvAcropora millepora was also the source of variability in connectivity patterns for the SCS [Connectivity has been found to have high spatiotemporal variability due to a variety of physical and biological factors –21,26,27 the SCS . The ree the SCS ,31.The variability in connectivity patterns in the highly diverse and complex area of the NIWP need to be better understood to gain a more detailed picture of between-basin and within-basin larval transport. This would contribute to possible management strategies from the regional NIWP scale to the level of local natural parks and community-based marine protected areas (MPAs). Given that different organisms may produce different connectivity patterns, the management of coral reef ecosystems should consider multiple species towards a metacommunity approach to connectivity . Thus, sThis study provides a mesoscale analysis of connectivity variability in the NIWP that could be considered in marine resource conservation and management decisions at both regional and more local scales. In particular, we aimed to (1) characterize how connectivity in the NIWP varies between three functionally different coral reef organisms representing a range of early life history conditions, (2) assess the sensitivity of the results to possible seasonal and interannual circulation variabilities, and (3) present a way to identify reef clusters which could serve as management units that integrate the connectivity features based on between-reef similarities in sources and sinks across the three model organisms.Acropora millepora, a reef-associated keystone herbivore sea urchin Tripneustes gratilla, and a predatory coral trout Epinephelus sp. These organisms were chosen for their key ecological roles in the reef ecosystem and for their economic value in the NIWP. The temporal span of the study covered three representative years to capture interannual variations of circulation [The connectivity model simulating larval dispersal was written in Java, utilizing the MASON simulation toolkit . The modculation  likely dculation –40. SimuThis study covered all coral reef areas  of the NThe hydrodynamic input used was the surface circulation of daily global Hybrid Coordinate Ocean Model (HYCOM) ,50 + NavCoral reef locations were identified based on the UNEP World Conservation Monitoring Center (UNEP-WCMC) coral reef database . IndividA. millepora was regarded as a short-distance spawner. Both T. gratilla and Epinephelus sp. were considered long-distance spawners and were simulated to produce passively dispersing and actively swimming larvae, respectively. Acropora millepora was observed to spawn during the summer months in the Philippines [T. gratilla spawns year-round [Epinephelus sp. may also spawn during different periods of the year, but has peak spawning during the summer months [Various biological factors affect larval dispersal, but the most influential ones considered in this study  were morlippines  while T.ar-round . Epinephr months . This stA. millepora, T. gratilla, and Epinephelus sp., respectively. Based on a calibration exercise  to a sink site (j) for each simulation. The rows of the matrix constituted the sources while the columns constituted the sinks. The raw connectivity matrix was then converted into a settlement probability matrix according to:Pset was the settlement probability representing the proportion of larvae exported from a source i to a sink j relative to the number of larvae released from the source. A total of 36 matrices were generated in this study based on 3 organisms, 4 seasons, and 3 years.The model outputs were recorded as raw connectivity matrices of the total number of simulated larvae that settled from a source site , was a measure of local retention [Seven connectivity metrics were used to analyze connectivity patterns: local retention, export and import probabilities, out- and in-degrees, and the average export and import distances. The diagonal of the matrix, etention  or the petention , in whicThe mean direction of exports was also computed for temporal analysis. This was based on the mean Euclidean source-to-sink directions weighted by the settlement probabilities. For this, only long-distance connections beyond 13 km  were considered in the averaging to remove local retention and adjacent-cell connections.The Kruskal-Wallis one-way analysis of variance and the post-hoc Dunn’s test were used to compare the distributions of connectivity metrics testing for significant differences in the distribution between model organisms and between seasonal matrices. Pearson correlation coefficient was used to explore the relationships of connectivity metrics between model organisms and between seasonal matrices.Circular analysis was employed to compare the distributions of the mean direction of exports of each reef cell between seasons and years for each organism. A multi-sample test for equal median directions, a circular analogue of the Kruskal-Wallis test , was usei and j was:Cij was the sum of the smaller magnitudes of connections shared between both cells, and S was the sum of all connections in each cell. A BCDij value close to zero meant that reef cells had similar connections and variability across the matrices used in the analysis while a value close to one meant that cells had dissimilar connections. The BCD matrix used here was based on the known spawning periods of each organism  to reflect how managing coral reef ecosystems could cut across multiple key organisms.The settlement probability matrices were subjected to agglomerative cluster analysis to explore possible clusters of reefs which covary in connectivity patterns based on similarities of the sources and sinks of each reef cell. The distance metric of the cluster analysis was based on the Bray-Curtis dissimilarity (BCD) equation  in whichThe resulting BCD matrix was subjected to a dynamic tree cut algorithm implemented in the R programming language  using deA. millepora larvae settled within 18.5 km or around two model cells away and most settled less than 171 km from their natal reefs. Tripneustes gratilla and Epinephelus sp. had similar settlement distances despite the latter having a longer larval duration. Relative to the median, the settlement probability at the 95th percentile was more than half and the probabilities at the farthest settlement distances were an order of magnitude lower for all organisms.Fitted power law functions for the dispersal kernels provided the highest coefficient of determination and the lowest mean standard error . These dth percentile; beyond which, there were spikes of higher export probabilities at distances of around 800 km and 1,300 km . On the contrary, A. millepora scored lowest in terms of the range of external connections for out-degree, in-degree, mean distance of export, and mean distance of imports. Tripneustes gratilla and Epinephelus sp. performed similarly for these metrics. Most of the between-organism pairs for all metrics were highly correlated  using Kruskal-Wallis one-way analysis of variance showed that distributions of all metrics  were sigrrelated  except fA. millepora. Comparing the annual and seasonal connectivity matrices, the metrics derived from the annually-averaged matrices of each model organism were highly correlated with the metrics from the seasonal matrices   decreased and fewer cells were excluded from clustering . The cluA total of 13 clusters emerged at MCS = 100 . The NorThe South China Sea was subdivided into six clusters which differentiated near-coast reefs from offshore reefs. The cluster composed of Vietnam and Paracel Islands (cluster 8) highlighted the reefs which were strong larval sources during the southwest monsoon and conversely, the main larval sinks of the northern South China Sea and northeastern Luzon during the northeast monsoon. The Western Luzon cluster (9) similarly was the sink of the Vietnam-Paracel Island cluster during the southwest monsoon and the source of larvae to the South China Sea during the rest of the year. The western Palawan shelf was subdivided into the northern reefs (cluster 10), connecting mostly with the northern South China Sea and the Mindoro Strait, and the southern reefs (cluster 11), connecting with the southern South China Sea and the Balabac strait. The Spratly Islands were also subdivided to the northern cluster (12) connecting with the northern South China Sea and the southern cluster (13) including Western Borneo mostly connected with the southern South China Sea and the Celebes Sea.The Karimata Strait cluster emerged at MCS = 75  . These rThe level of clustering used would need to be informed by the spatial scale of interest. For example, in the Philippines, management of coral reefs is usually implemented at the level of local municipalities through community-based MPAs . While tClustering at a coarse scale pointed out some general connectivity patterns between countries of the NIWP. The global connectivity model by Wood et al.  found thTridacna crocea [Chanos chanos [Siganus fuscescens [The location of inferred barriers to gene flow summarized in Von der Heyden et al.  appears a crocea , Chanos s chanos , and Sigscescens . Observescescens  can be escescens  reflectsscescens  is concoscescens  differenscescens  resultinscescens  was likeThe spatial match between the modelled regional connectivity patterns with inferred genetic barriers provides a coarse validation of large-scale exchange of larvae in the NIWP. Validation with population structure is still insufficient for finer-scale spatial patterns especially since genetic and ecological interconnectedness can be mutually exclusive, i.e., populations that are evolutionarily linked may be demographically separated . The cluC. vagabundus [P. maculatus and P. leopardus [Recent studies on parentage analyses of various reef organisms provide optimistic empirical demonstrations of demographic connectivity at both local and regional scales. Parent-juvenile pairings have shown both greater local retention at the natal reef or reef region and more importantly, long-distance connections representing the tail of dispersal up to distances of 48 km to 400 km for different species ,100–106.gabundus , and 480eopardus .The model was only able to account for surface flow and near-surface reefs. Subsurface pathways that likely retained more larvae near the natal reef would best be resolved with three-dimensions. Such a modelling approach could potentially capture depth variability of spawning and settlement, especially for reef slopes and mesophotic reefs ,107, togImproving the model results would primarily entail (1) increasing the resolution which would resolve the narrow straits and complex coastlines and (2) coupling larval dispersal with a population growth model using realistic estimates of larval supply and settlement habitat quality. Applying the connectivity results to matrix projections  and seasOur simulation of the larval connectivity of three coral reef organisms demonstrated that connectivity is inherently variable in a dynamic region like the NIWP. Both early-life history characteristics and temporal differences in circulation resulted in variations in the magnitudes of the settlement probabilities and connectivity metrics. However, based on the generally moderate to high correlations of these metrics between organisms and between time periods, connectivity in the NIWP seems more sensitive to the geographic configuration of reefs compared to differences in dispersal potential of larvae or circulation. Concordance of the modeled ranges of dispersal with the range of the empirical estimates from parentage analyses, and the identification of clusters of co-varying connectivity patterns with population genetic studies, provide support for the validity of the model results.Apart from illustrating the mesoscale patterns in larval connectivity for the NIWP, we offer three tools which could be used to bridge connectivity from the global to local levels for the NIWP: (1) fitted dispersal kernel functions providing estimates of the relative probabilities of connection given only the distances between reefs; (2) maps on the relative sensitivity of different reefs to temporal connectivity variability; and, (3) the clustering of reefs using the BCD algorithm, which produced clusters defined by the geographical position of reefs relative to dominant circulation patterns and features. These clusters of co-varying connectivity patterns can serve as inputs into efforts for the conservation and management of NIWP coral reefs from regional to local levels.S1 Appendix(DOCX)Click here for additional data file.S2 Appendix(DOCX)Click here for additional data file.S3 Appendix(DOCX)Click here for additional data file.S4 Appendix(DOCX)Click here for additional data file.S5 Appendix(DOCX)Click here for additional data file.


The aim of this study was to discover the functional impact of non-synonymous single nucleotide polymorphisms (nsSNPs) that were found in selective sweep regions of the Landrace genomeWhole-genome re-sequencing data were obtained from 40 pigs, including 14 Landrace, 16 Yorkshire, and 10 wild boars, which were generated with the Illumina HiSeq 2000 platform. The nsSNPs in the selective sweep regions of the Landrace genome were identified, and the impacts of these variations on protein function were predicted to reveal their potential association with traits of the Landrace breed, such as reproductive capacity.Total of 53,998 nsSNPs in the mapped regions of pigs were identified, and among them, 345 nsSNPs were found in the selective sweep regions of the Landrace genome which were reported previously. The genes featuring these nsSNPs fell into various functional categories, such as reproductive capacity or growth and development during the perinatal period. The impacts of amino acid sequence changes by nsSNPs on protein function were predicted using two in silico SNP prediction algorithms, i.e., sorting intolerant from tolerant and polymorphism phenotyping v2, to reveal their potential roles in biological processes that might be associated with the reproductive capacity of the Landrace breed.The findings elucidated the domestication history of the Landrace breed and illustrated how Landrace domestication led to patterns of genetic variation related to superior reproductive capacity. Our novel findings will help understand the process of Landrace domestication at the genome level and provide SNPs that are informative for breeding. The recently developed high-throughput and cost-effective genotyping techniques allow the thorough exploration of genetic variation in domestic animals. In particular, whole-genome sequencing is a powerful approach for detecting massive amounts of single nucleotide polymorphisms (SNPs) in genome-wide sequence data. One of the strategies for studying genetic variation is to detect the selective sweep signatures based on patterns of linkage disequilibrium (LD) , which wDiverse types of variants, e.g. copy number variations, insertion/deletion (InDel) and structural variations, have been identified in the selective sweep regions of the Landrace genome . Unlike in silico bioinformatic tools.In this study, we aimed to identify nsSNPs in the selective sweep regions of the Landrace genome that might be related to superior reproductive capacity or growth and development during the perinatal period, and gene networks that were enriched in Landrace genome. Finally, impact of amino acid changes by nsSNPs on protein function was also investigated using Sscrofa 10.2.75) from the Ensembl database using Bowtie2 , CCL1, kinesin family member 20B, plasminogen, and PTPRC) were shown to be involved in the positive regulation of locomotion, and its network consisted of four GO terms . This cellular movement is a central process in the development and maintenance of multicellular organisms. In addition, tissue formation during embryonic development requires the orchestrated movement of cells in a particular direction. It is reasonable to assume that several genes of these four significant GO terms in the selective sweep regions of the Landrace genome might be related to the superior growth and development of Landrace during the perinatal period. Ten genes  were associated with exocytosis, and five genes  were classified under the secretory granule membrane term in the GO network. The acrosome contains a single secretory granule and is located in the head of mammalian sperm; secretion from this granule is an absolute requirement for fertilization [Among 90 genes, the functions of 64 genes were predicted, and we performed GO network analysis of these 64 genes using ClueGO  to draw e sweeps . Metal ie sweeps . Five gelization . Acrosomlization . Therefolization .in silico SNP prediction algorithms, SIFT [Two ms, SIFT  and Polyms, SIFT , were apMOGAT2) gene was shown to have the greatest number of nsSNPs with a strong effect among the 90 genes. Five SNPs of the total of 11 nsSNPs in the MOGAT2 gene had strong effects on protein function in this study. Although our GO network analysis did not reveal any particularly important network of MOGAT2, this gene has been reported to be important in porcine backfat adipose tissue, which is related to the concentration of lipid and lipid synthesis, as revealed by a transcriptome analysis comparing Landrace and other breeds [DMBT1 gene were considered to have strong effects on protein function, as revealed by the SIFT and Polyphen-2 results.According to the SIFT analysis, 75 of 345 nsSNPs were classified as being deleterious . PolyPhen-2 calculates the true-positive rate as a fraction of predicted mutations; its results showed that 82 amino acid variants involving nsSNPs in the selective sweep regions of the Landrace genome were likely to exert deleterious functional effects. In addition, 46 of these nsSNPs overlapped with the SIFT results. From the results of the two bioinformatics tools, we reasoned that 46 of the 345 nsSNPs might have strong effects on biological mechanisms during the process of Landrace domestication . Forty-sr breeds . In addiGiven the interest of the meat production industry in improving the meat quality or piglet number, a genetic investigation focusing on the selective sweep regions of the Landrace genome was previously performed . This stOur results strongly suggested that Landrace genetic variants, which could give rise to changes in amino acid sequences, might be important factors for the superior reproductive capacity of this breed. We aimed to perform analyses of the Landrace genome using nsSNPs in selective sweep regions. Our results showed that most of the genes affected by nsSNPs in the selective sweep regions may be closely related to the superior reproductive capacity or growth and development of the Landrace breed during the perinatal period. Furthermore, there were indications that nsSNPs in selection had impacted in Landrace breed establishment. This study will provide insights into the impact of the process of domestication on the Landrace genome.


Pain and depression affect one another, and this bidirectional interaction implies the existence of common or interacting neural pathways. Among the neural circuits relevant to negative affection, the medial prefrontal cortex (mPFC) is known to be involved in both pain and depression. Persistent stress from physical pain and mental distress can evoke maladaptive changes in mPFC circuits to induce depression. Conversely, the unpleasant mood condition alters mPFC circuits to distort the appraisal of aversion and make individuals vulnerable to pain. In this article, recent findings regarding mPFC in chronic pain and/or depression are reviewed, with particular focus on the metabotropic glutamate receptor 5 (mGluR5). Although the involvement of mGluR5 within the mPFC in both pain and depressive disorders has been extensively studied, there are controversies regarding changes in the activity of the mPFC during chronic pain and depression, and the functional roles of mGluR5 on altered mPFC activity. We discuss alterations in the availability of mGluR5 in the mPFC in these disorders, its role in behavioral manifestations, and its possible influence on cellular subpopulations that mediate dysfunction in the mPFC. We also propose molecular mechanisms that may cause expressional changes in mGluR5 within the mPFC circuitry. Chronic pain patients often develop negative mood symptoms such as depression . ConversThe neural circuits within the mPFC undergo various changes during the development of chronic pain and/or depression, and these alterations play a key role in the persistence of the disorder . The molHowever, there have been discrepancies in the previous reports regarding the direction of altered mPFC activity, the expressional changes of mGluR5 within the mPFC (mPFC-mGluR5), and their functional consequences. In this article, we discuss chronic pain-induced hypo- or hyper-excitability of mPFC pyramidal neurons, and their roles in pain and depression. Furthermore, we offer our perspective on the issue of contrasting reports, the specific mPFC neuronal subpopulation that may be affected by mGluR5 alteration, and possible underlying molecular mechanisms.Numerous animal model studies have found that the activity of mPFC neurons is altered in the pain state, and that the altered activity of pyramidal and/or GABAergic neurons is associated with increased pain perception. In rodents, the mPFC consists of three subregions: anterior cingulate cortex (ACC); prelimbic cortex; and infralimbic cortex. Although the role of hyperactive ACC activity on abnormal pain is well established , controvMany researchers have associated hypoactive prelimbic pyramidal neurons with increased pain perception and attenuation of pain modulatory function. Previous studies have demonstrated that pyramidal neuronal activity in the prelimbic cortex is decreased in several animal models of chronic pain, and that increasing its activity ameliorates pain. According to electrophysiological analyses, reduction of prelimbic pyramidal neuronal activity was associated with decreased intrinsic excitability of pyramidal neurons , reducedAmong these mechanisms, the increased influence of GABAergic neurons on pyramidal neurons has been actively studied. The increase in inhibitory inputs to pyramidal neurons is due to the loss of available endocannabinoid in pyramidal neurons , and thein vivo. On the other hand, some studies have found increased mPFC neuronal activity in the pain state. These investigations showed that pharmacological deactivation of activity reduces chronic pain symptoms. In a study by The inconsistencies indicate that the changes in synaptic transmission and intrinsic excitability are highly specific to mPFC subregion, cell type, cortical layer, the cause and the duration of the pain. In neuropathic pain models, excitability of layer 5 pyramidal neurons was reduced in the prelimbic cortex , but notAlthough further studies are needed to clarify apparent contradictions between a hyperactive and hypoactive mPFC in the chronic pain state, research to date has commonly observed alterations in excitatory and/or inhibitory influences on pyramidal neurons, and these disturbances are critically involved in the pain itself and the ensuing affective and cognitive disorders. Interestingly, multiple studies have described mGluR5 as a molecular mediator of altered mPFC pyramidal neuronal activity in the chronic pain state, but with inconsistent descriptions of its functional roles.There are conflicting reports regarding the effect of mGluR5 blockade in the mPFC on pain modulation . A previIn the field of depression research, previous studies investigating non-pain depression have reported inconsistent results regarding the alteration of prefrontal mGluR5 expression. A few studies have reported lower prefrontal mGluR5 levels in patients with major depressive disorder (MDD) compared with healthy controls , whereasin vivo resulted in an increase in mPFC glutamatergic neuronal activity  agonists to the mPFC activated the endogenous analgesic action of the PAG . AlternaOne possible explanation is that increased prelimbic mGluR5 levels eventually result in a reduction in the firing of excitatory pyramidal mPFC neurons, which project to the PAG or the NAc. Previous studies have reported that the neuronal excitability of mPFC-PAG projection neurons is decreased in chronic pain states, and the increased activity of inhibitory interneurons is responsible for the reduced activity of pyramidal neurons . InteresFigure 1A). Second, apart from this explanation, mGluR5 may be upregulated in excitatory neurons that preferentially excite inhibitory interneurons within the local circuits . In the mPFC, the activity of local GABAergic neurons is influenced by the glutamatergic excitatory inputs they receive, and the frequency – but not the amplitude – of the excitatory transmission to the GABAergic neurons were increased in the neuropathic pain state . For example, a recent study found that coactivation of mGluR5 and the M1 muscarinic acetylcholine receptor in the mPFC exerted a long-lasting decrease in excitatory transmission and concurrent enhancement of GABAergic inhibitory tone . Glial mGluR5 serves to evoke complex and bi-directional effects on the modulation of neuronal activity, and upregulated mGluR5 in cortical astrocytes contributes to synaptic remodeling during the chronic neuropathic pain state -mediated control of mGluR5 expression. From the perspective of a reinforcement learning paradigm, dopamine signaling plays a key role in motivated approach or avoidance behavior following aversion. Because D2Rs have a strong binding affinity compared with the dopamine 1 receptor (D1R), persistent dopamine would be mainly occupied by D2R in the nervous system. When negative prediction error occurs by salient event (aversion), dopaminergic neurons would cease firing, resulting in transient dopamine depletion in the relevant brain regions . TheoretExperiencing unavoidable stress, such as somatic pain or psychological distress, alters mPFC circuitry and distorts the perception of the self-state and ensuing decisions regarding coping strategies. We propose that mGluR5 in the mPFC may be a common mediator for both pain and depression. mGluR5 levels would regulate the gain for the assessment of internal state, guide the appraisal of the external state, and be involved in updating the expectancies of behavioral outcome in the mPFC. Increased mPFC-mGluR5 levels during the pathological state would inhibit the activity of pyramidal neurons and reduce the flexibility of the system, resulting in the loss of control of the appropriate processing of the information.GC conceived the idea, performed the bibliographical research, wrote the draft, and revised the manuscript. SJK and SKK supervised and revised the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Mechanochemical transformation of the powders of lactose monohydrate and sodium chloride as models of nutrients and components of dosage forms was investigated by the complex of physicochemical and biological methods. The solubility and kinetic activity of substances dispersed in various ways showed a positive correlation with the solvent isotope profile. Substances dissolved in heavy water were more active than solutes in natural water. Differential IR spectroscopy confirmed the modification of substituents in the sample of lactose monohydrate, demonstrating physicochemical changes during mechanical intervention. The biological activity of the compounds was determined by the method of Spirotox. The activation energy was determined by Arrhenius. Compared with the native compound, dispersed lactose monohydrate showed lower activation energy and, therefore, greater efficiency. In conclusion, proposed data confirm the statement that mechanical changes in compounds can lead to physicochemical changes that affect chemical and biological profiles.This study demonstrates the link between the modification of the solid-phase pharmaceutical substances mechanical structure and their activity in waters with different molar ratio  To predict expected bioavailability characteristics for drug substances the pharmaceutical scientists give their significant attention to investigate the dependence of the pharmaceutical ingredient's activity on the degree of dispersion , 2.There is direct relationship between the pharmaceutical substance dissolution rate in biological fluids and its bioavailability –6. The i in vitro , 3433, 3 S. ambiguum is linearized in Arrhenius coordinates: lg(1/t)-1000/T. The values of the apparent activation energy (obsEa) were found by the tangent of the lines slope in semilogarithmic coordinates  in the waters of different deuterium/protium ratio (kinetic isotope effect), IR spectral properties in the middle and terahertz ranges, and differences in ligand-receptor interactions as a function of the dispersity of pharmaceutical substances samples for the biological model .


Although current desalination technologies are mature enough and advanced, the shortage of freshwater is still considered as one of the most pressing global issues. Therefore, there is a strong incentive to explore and investigate new potential methods with low energy consumption. We have previously reported that reversible thermally induced sorption/desorption process using polymeric hydrogels hold promise for water desalination with further development. In order to develop a more effective hydrogels architecture, polyelectrolyte moieties were introduced in this work as pendent chains and a thermally responsive polymer as network backbone using reversible addition-fragmentation chain transfer (RAFT) polymerisation. The ability of the comb-type polymeric hydrogels to desalinate water was evaluated. These hydrogels were proved to absorb water with low salinity from brine solution of 2 g L Loosely cross-linked hydrophilic polymers that swell in the presence of water are often used to form jelly-like material known as hydrogels ,2. TheseN-isopropylacrylamide) (PNIPAAm), which demonstrates a sharp phase transition at a lower critical solution temperature (LCST) around 32 One of the most frequently used and studied temperature-responsive polymers and hydrogels is poly were obtained after a second RAFT polymerisation using NIPAAm, PAAc-TTC, and cross-linkers with varying composition.In the present study, we will discuss the preparation of comb-type polymer hydrogels composed of NIPAAm and a copolymer of polyacrylic acid (PAAc) via a two-step RAFT polymerisation, and verify their viability for water desalination. The influence of the length and the fraction of the grafted PAAc chains on the response rate and the equilibrium-swelling ratio will be investigated. Towards this end, three trithiocarbonate macro-RAFT agents (PAAc-TTC) differing in the average molecular weight  was purified by recrystallisation in hexane and dried in vacuo. Acrylic acid  was vacuum-distilled to remove the inhibitor MEHQ (hydroquinone monomethyl ether) and possible oligomer content. 4,4N,N1H-NMR spectroscopy in The poly(acrylic acid)-based macro-RAFT agent was prepared similarly as described elsewhere . In a tyTo synthesise the comb-type grafted gels (GG), a mixture of PAAc-TTC, NIPAAm, MBAm, ACPA were dissolved in 3 mL ethanol and the solution was charged to a glass vial (10 mm in diameter) equipped with a magnetic stirrer. The solution was purged over a period of 20 min with nitrogen and then degassed by a freeze-thaw cycle and sealed under nitrogen. The polymerisation was conducted for at least 72 h at 70 A comprehensive account on the samples used and their respective composition as well as the degree of monomer conversions are given in The morphology of hydrogels were examined using scanning electron microscopy . The hydrogel specimens were firstly immersed in deionised water at room temperature (23 FTIR spectra of the macro-RAFT agents (PAAc-TTC) and hydrogels were recorded on a 3100 Fourier-Transform infrared spectrometer  in the range of 600–4000 cmThe water content was estimated using differential scanning calorimetry (DSC)  from −30 to 30  Malcolm . The amoThe hydrogel samples were placed in excess water with a typical mass ratio of 0.05 The dewatering experiments were carried out at 50 +. The salt rejection was determined as follows:The salt rejection was obtained by measuring the concentrations of sodium ions in the water recovered from the hydrogels by ion chromatography (Metrohm 833 Basic IC plus). Silica gel with carboxyl groups column (particle size 5 mm) was used at 25 The mechanical properties of the fully swollen hydrogels were investigated via the frequency sweep experiments using a rheometer . Solvent trap was used to minimize evaporation during the measurments. The experiments were performed in the linear range of R is the ideal gas constant L=ξ=1L=ξ=1(8)G1H-NMR and FTIR  as macro-RAFT agent and MBAm as cross-linker agent . The reaThe polymer hydrogels were characterised with FTIR . All speThe morphological features of the swollen freeze-dried hydrogels were evaluated via SEM studies. The micrographs shown in The swelling behaviour of the comb-type grafted PAAc gels was investigated in both brine and deionised (DI) water. It is well known that the swelling and osmotic pressure within the hydrogel increases with increasing the ionic component and thus the equilibrium-swelling ratio of the hydrogel will be increased. The strong hydration of the gels is related to the fact that longer chains are structurally separated from the backbone of the cross-linked network. The hydrogels with different PAAc-grafted chain lengths may have different cross-linking density. The effect of a change in crosslink density on the equilibrium-swelling ratios was not studied. No effect of the chain length on the equilibrium-swelling ratio in brine was found indicating that the contribution of charges to the osmotic pressure is the same in all systems. We therefore expect samples containing the same total amount of PAAc to show a similar salt depletion capacity.The dynamic of the swelling process of the dry materials was also quantified via the half-swelling time  . All gelHowever, the change in modulus arise from entropy changes in the rubbery range. Therefore, the change in toughness or stiffness of the hydrogel networks can be correlated with their composition. onfirmed . Gels wiThe mesh size of hydrogels, as a measure of their microstructure, was estimated from the rheological data using Equation . As expedashed line). Generally, for all studied gels, the endothermic peaks are broad and structured.In order to get an insight into the dewatering of gel matrices, the type of water binding inside the fully swollen hydrogels was determined by means of DSC analysis. Generally, water inside hydrogel networks can be thermodynamically classified as one of three different states : (1) freThe different fraction of freezable and non-freezable water for the studied hydrogels can be correlated with their swelling ratio at equilibrium, which is also associated with both grafted PAAc content and chain length. The grafted PAAc with freely mobile ends can hydrate sufficiently and contain a large amount of freezable water. As we stated in the Introduction, the solution taken up by gels should not only be depleted from salt, but also can swiftly be released under thermal stimulus. Therefore, the phase transition behaviour of hydrogels was explored by measuring the changes of the swelling ratios at equilibrium as a function of external temperature in deionised water as shown in gels cf. .The LCST at which the volume phase transition of the gels start is obtained by a fit of a sigmoidal function to the data . Gels wiThe water release from the polymer hydrogels  was achiIn addition, gels with longer PAAc chain length have a slightly better dewatering performance, resulting probably from the increase in void volume within the polymer network upon the dehydration of PNIPAAm chains  to determine the number of consecutive steps required to achieve a complete salt rejection . An initFrom an application point of view, it is also important to study the reversibility of the desalination process. We evaluated the reversibility of the swelling ratio and the salt rejection on temperature cycles between 20 and 50 k–20 gel . After eThe total world water demand is predicted to be a global interdisciplinary research challenge. This need is primarily driven by rapid growth of population and industries, which leads to high water consumption. Despite the vast abundance of water, a lot of effort has to be done for human consumption or other beneficial purpose. Advanced technologies for water purification and desalination are currently in use. Most of these technologies such as reverse osmosis and depend on nonrenewable traditional fossil fuels, which are subject to a rapid decrease with increasing world population. Therefore, the search for a viable alternative technology is becoming important from an economic standpoint. In this research, we reported on the development of polymer-based hydrogels for salty water desalination. These hydrogels were synthesised by a combination of grafted polyelectrolyte and a thermally responsive polymer with different grafted chain content and chain length. We have shown that the efficiency of this desalination process strongly depends on their composition. In particular, altering the grafted PAAc content has a major influence on swelling/dewatering behaviour as well as the salt expelling ability of the hydrogels. The recovered water in the dewatering step decreases and meanwhile the salt rejection increases as the PAAc content increases. In contrast, hydrogels with the same chemical composition but different PAAc-grafted chain length reveal a higher swelling capacity as the chain length increases, while the percentage of released water increases only slightly as a function of the PAAc-grafted chain length. It was found that the performance of a material with 30 wt % of grafted PAAc and a chain length of 10 kD is the best trade-off with respect to salt removal and and water recovery for the proposed desalination experiment. Using this material, the total salt depletion of 2 g L


Species delineation based on bacterial genomes is an essential part of the research of prokaryotes. In silico genome-to-genome comparison methods are computationally demanding, but much less tedious and error prone than the wet-lab methods. In this paper, we present a novel method for the delineation of bacterial genomes based on genomic signal processing. The proposed method uses numerical representations of whole bacterial genomes, phase signal and cumulated phase signal, from which four parameters are derived for each genome. The parameters characterize a genome and their calculation is independent of the other genomes comprising a delineation dataset. The delineation itself is processed as a calculation of the parameters' average similarity. The method was statistically verified on 1826 bacterial genomes. A similarity threshold of 96% was set based on the receiver operating characteristic curve that featured sensitivity of 99.78% and specificity of 97.25%. Additionally, comparative analysis on another 33 bacterial genomes was conducted using standard delineation tools as these tools were not able to process the dataset of 1826 genomes using desktop computer. The proposed method achieved comparable or better delineation results in comparison with the standard tools. Besides the excellent delineation results, another great advantage of the method is its small computational demands, which enables the delineation of thousands of genomes on a desktop computer. The calculation of the parameters takes tens of minutes for thousands of genomes. Moreover, they can be calculated in advance by creating a database, meaning the delineation itself is then completed in a matter of seconds.


Observations such as the presence of tau in the cerebrospinal fluid (CSF) from control individuals and in the CSF of transgenic mice overexpressing human tau before the detection of neuronal death indicate that tau can be secreted by neurons. The increase of tau in the CSF in pathological conditions such as AD suggests that tau secretion is enhanced and/or other secretory pathways take place when neuronal function is compromised. In physiological conditions, extracellular tau could exert beneficial effects as observed for other cytosolic proteins also released in the extracellular space. In such a case, blocking tau secretion could have negative effects on neurons unless the mechanism of tau secretion are different in physiological and pathological conditions allowing the prevention of pathological tau secretion without affecting the secretion of physiological tau. Furthermore, distinct extracellular tau species could be secreted in physiological and pathological conditions, species having the capacity to induce tau pathology being only secreted in the latter condition. In the present review, we will focus on the mechanisms and function of tau secretion in both physiological and pathological conditions and how this information can help to elaborate an efficient therapeutic strategy to prevent tau pathology and its propagation.In Alzheimer’s disease (AD), neurofibrillary tangles (NFTs), lesions composed of hyperphosphorylated and aggregated tau, spread from the transentorhinal cortex to the hippocampal formation and neocortex. Growing evidence indicates that tau pathology propagates  This melocation . PIecretion . Such faecretion . The proecretion . Interesecretion .The contribution of membranous organelles to tau secretion was revealed by demonstrating that SNAREs and Rabs, two classes of proteins implicated in membrane trafficking are involved in tau secretion. In a recent study, it was observed that the suppression of SNAP-23, a SNARE protein implicated in neurotransmitter exocytosis, resulted in a significant reduction of tau secretion . The co-We recently investigated tau secretory pathways  and showWe also reported that Rab1A was involved in tau secretion . Rab1A iExosomes and ectosomes are the two main types of extracellular vesicles involved in the secretion of cytosolic proteins . In the Although several studies indicate that tau found in CSF most likely results from its secretion by neurons, this remains to be experimentally demonstrated. Observations performed in lamprey where human tau was overexpressed in anterior bulbar neurons indicate that tau released by neurons could reach the CSF by periventricular and perimeningeal routes . The facThe PTMs of exosomal tau were also examined. Most studies reported that both FL-tau and C- and N-terminal truncated tau was found in exosomes, truncated forms being more abundant than FL-tau as noted for free-tau . In someIn adult human brain, six tau isoforms  are expressed . Very spAltogether, the above data reveal that several forms of tau co-exist in both the culture medium and CSF and strongly suggest that CSF-tau corresponds to secreted tau.The fact that tau is released by neurons in physiological conditions strongly suggests that it has a role in the extracellular space. This was demonstrated for other cytosolic proteins. For example, annexin A4 and A5, calcium and phospholipids-binding proteins, act as anticoagulant factors via their Ca2+-regulated binding to phospholipids in the extracellular space . Heat shUnconventional secretion of cytosolic proteins could also take place to prevent PTMs that can inactivate them. For example, the unconventional secretion of FGF2 prevents its O-glycosylation that renders its biologically inactive . The PTMtrans-synaptically by the release of tau at the pre-synaptic terminal and its uptake by the post-synaptic neurons  compartment as noted for TDP-43. In either case, the elucidation of tau secretory pathways is central for the elaboration of a therapeutic strategy to prevent the development of tau pathology and its propagation. An efficient therapeutic strategy should prevent the intracellular and extracellular accumulation of pathological tau. To achieve this, an increase of pathological tau secretion for prevention of its intracellular accumulation and its sequestration in the extracellular space by an antibody to prevent its propagation and synaptic detrimental effects are necessary . FurtherFrom recent studies, one could conclude that combined therapeutic strategies with tau secretion would be the most efficient way for preventing the propagation of tau pathology in the brain. Three combined strategies could be elaborated. Firstly, a recent study reported that impaired dural lymphatic function resulted in an increased retention of extracellular tau in the brain as well as a delayed clearance of tau to the periphery . A theraLastly, a recent study reported that END-binding proteins found at the tip of microtubules could modulate tau secretion in non-neuronal cells indicating that microtubules could play a determinant role in the release of tau by neurons . AlthougThe non-vesicular extracellular tau, the main target for immunotherapy, was shown to be able to transfer from neuron to neuron . SeveralThe isoform composition of extracellular tau remains to be well characterized for most tauopathies. This point is important since it was recently reported that tau isoforms do not have the same propensity for spreading in the brain. Indeed, the 4R isoforms were found to propagate farther than 3R tau . In the It remains to be demonstrated whether CSF-tau could be used for the differential diagnosis of tauopathies at early stages of the disease and/or for following the progression of a tauopathy. The amount of total and phosphorylated tau is increased in the CSF of AD patients . More reIn conclusion, the elucidation of tau secretory pathways is central to further investigate the physiological function of extracellular tau and its contribution to tau pathology. Extracellular tau is the main target of therapies involving the administration of an anti-tau antibody. A better characterization of all tau forms released in the extracellular space and their function is needed for the design of more efficient antibodies that will solely sequester toxic tau species. This characterization will require the use of more sensitive techniques for the detection of extracellular tau PTMs and aggregation state in the CSF.CP, AD, and NL wrote the manuscript. CP prepared the figures.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Cholangiocarcinoma (CCA), or cancer of the biliary epithelium is a relatively rare but aggressive form of biliary duct cancer which has a 5-year survival rate post metastasis of 2%. Although a number of risk factors are established for CCA growth and progression, a careful evaluation of the existing literature on CCA reveals that an inflammatory environment near the biliary tree is the most common causal link between the risk factors and the development of CCA. The fact that inflammation predisposes affected individuals to CCA is further bolstered by multiple observations where the presence and maintenance of an inflammatory microenvironment at the site of the primary tumor plays a significant role in the development and metastasis of CCA. In addition, mechanisms activating the tumor vasculature and enhancing angiogenesis and lymphangiogenesis significantly contribute to CCA aggressiveness and metastasis. This review aims to address the role of an inflammatory microenvironment-CCA crosstalk and will present the basic concepts, observations, and current perspectives from recent research studies in the field of tumor stroma of CCA. Cholangiocarcinoma (CCA) is a term used to define a group of different biliary epithelial cancers and is the second most common type of liver cancer. This group of primary biliary malignancy represents three different classically recognized kinds of biliary tree cancers, classified on the basis of anatomical point of origin in the bile duct, intrahepatic CCA (iCCA), perihilar CCA (pCCA), and distal CCA (dCCA) . Among tThe number of people afflicted with CCA differs geographically primarily because of the difference in the presence of the risk factors that predispose an individual toward CCA. The number of CCA cases is higher in Asian countries . CCA alsInflammation and inflammatory mediators form a key underlying basis for several risk factors significantly associated with CCA . An inflLiver cirrhosis develops as a consequence of liver diseases and/or conditions such as alcoholism and hepatitis. As a result, the liver parenchyma is dominated by fibrosis/scarring of liver tissue resulting in disruption and eventual loss of normal liver function. Cirrhosis is an important risk factor for iCCA  and showClonorchis sinensis, Opisthorchis viverrini) infections have been identified as critical risk factors for CCA especially in eastern Asian countries where these infections are deemed to be endemic  is due to its role in the development of CCA in affected individuals. These infections are associated with a rise in inflammation, generation of fibrotic nodules, obstruction of bile ducts and/or cholestasis. Chronic inflammation in the biliary tree in O. viverrini infected patients  leads to CCA development , serve as important risk factors for CCA . While HCholedolithiasis and cholelithiasis are conditions that involve the presence of stones in the gall bladder and common bile ducts. The presence of these gall stones causes biliary obstruction resulting in cholestasis and serve especially as a risk factor for extrahepatic CCA . The preChronic pancreatitis is a strong risk factor for extrahepatic CCA with an odds ratio of 6.61 (95% CI 5.21–8.40) in comparison to the 2.66 odds ratio of iCCA . In chronic pancreatitis too, cholestasis and inflammation may arise leading to CCA . Also, tAs CCAs originate from cholangiocytes from different anatomical locations of the biliary tree, they also exhibit considerable tumor heterogeneity that points to the possibility of diverse cellular origins , 26. In The undeniable role of stem cells in CCA development and origin is proven by the fact that human hepatic stem cells (hHPSCs) are the progenitor cells giving rise to cholangiocytes and hepatocytes that mutate to give rise to CCA . The PBGCCA is one of the most desmoplastic tumors and the tumor microenvironment of CCA is characterized by a dense bed of connective tissue intertwining the tumor cells. This dense stroma is composed of a contiguously activated subset of fibroblasts called CAFs that play key roles in modulating several aspects of CCA progression . FurtherCAFs release a number of molecules functioning as extracellular matrix proteins (ECM) such as collagen I and fibronectin . In CCA,Alpha-smooth muscle actin-positive (α-SMA) fibroblasts promote biliary cell proliferation and correlate with poor survival in CCA. CCA fibroblasts have proliferative effects that enhance tumor promotion and progression of CCA . CCA patCAFs in CCA show unique characteristics and gene signatures . Gene exEGFA/EGFR binding has been shown to promote tumorigenesis and metastasis in CCA, another important EGFR ligand, HB-EGF was found to be highly expressed in myofibroblasts. HB-EGF activated EGF signaling promotes proliferation of CCA cells and also induces epithelial-mesenchymal (EMT) changes as well as invasion. HB-EGF secretion from fibroblasts is also activated by the pro-tumorigenic growth factor TGF-β secreted by tumor cells that in turn favors CCA growth .In vitro studies indicate that when SDF-1 is expressed by HSCs, a number of pro-tumorigenic responses are induced such Bcl-2, and activation of PI3K/Akt pathway. These responses initiate increased CCA cell invasion and prolonged survival in addition to inducing epithelial-mesenchymal transition  play an important role by regulating angiogenesis, lymphangiogenesis, tumor proliferation and also modulating matrix related changes , 60. In ogenesis . In a cod VEGF-A . A high d VEGF-A .The association between chronic inflammation and the development and progression of malignancy is significantly pronounced in onset and development of CCA . InflammTNF-α is one of the most well-known mediators of inflammatory stimuli in the tumor microenvironment . AlthougClassified as one of the most important pro-inflammatory cytokines, IL-1β has been shown to be highly expressed from HSCs. The autocrine signaling mediated by CCA cells also becomes prominent in this regard as CCA cells have been shown to produce high levels of IL-1β that further enhances the CXCL5/CXCR2 pathway that in turn activates AKT/PI3K or ERK1/2 pathways. In fact, heightened CXCL5 expression has been seen to indicate poor rates of survival in CCA patients , 72.WAF/CIP1 a cyclin dependent kinase inhibitor, involved in cell cycle regulation (Bone marrow derived MSCs (BM-MSC) when exposed to tumor conditioned medium can transform into CAFs and stimulate tumor growth via secretion of inflammatory cytokine IL-6 in the tumor stroma. In CCA, this IL-6 overexpression was found to decrease the methylation of the EGFR promoter and enhance EGFR expression that in turn is associated with poor prognosis and overall survival , 73. IL-gulation . IL-6 algulation . In addigulation .TGF-β plays dual roles in cancer progression and inhibits cell proliferation, regulates anti-inflammatory, and pro-apoptotic effects in cells under normal physiological conditions . It alsoOverall, inflammatory cytokines set the stage for CCA growth by enhancing proliferation, activation of tumor promoting mechanisms such as EMT, activation of signaling pathways that promote tumor growth and loss of cell cycle checkpoints. However, the major cause for the high mortality associated with these cancers is its ability to metastasize, that is aided by the activation of lymphangiogenic (growth of new lymphatic vessels) and angiogenic (growth of new blood vessels). The various growth factors secreted by CCA cells into their stroma and other components of the tumor microenvironment foster the development of new lymphatic and blood vessels that in turn promote tumor growth and dissemination to distant organs.Tumor cells employ several mechanisms to establish a functional and integrated vascular system comprised of both blood and lymphatic vessels to promote cellular growth and metabolism. Expansion of these vascular networks is key to migration of the tumor cells to distant sites where they establish tumor niches. A surge of recent data has implicated the roles of both lymphatic and the blood vascular in promoting CCA metastasis.Tumor-associated lymphangiogenesis, or the sprouting of new lymphatic vessels in the tumor microenvironment is a form of tumor-associated neovascularization that has been the focus of studies concerning the metastatic spread of highly aggressive form of cancers . Lymphatin vivo however, significantly reduced lymphatic vascularization and reduced lymph node metastases  results in the inhibition of angiogenesis inhibition. In tumor cells however, hypoxia or a low oxygen environment in the region of the tumor induces expression of VEGF hormones. Cholangiocytes, accordingly, have been found to secrete high levels of both VEGF-A in the tumor stroma and VEGFR-2 during cholangiocyte hyperplasia , endogenous small non-coding RNAs (19–24 nucleotides) regulate various aspects of cholangiopathies including CCA and has been extensively reviewed elsewhere , 117. HoThe epidemiology of CCA varies across different regions owing to the differences in the number and intensity of the risk factors present in each place, the malignancy also varies in terms of the epidemiology of its types , however based on the data above it can be postulated that inflammation of the tumor microenvironment and its associated players have a crucial role in shaping the response of the CCA cells to therapeutic strategies, their growth and progression. To this end, the early metastatic events of CCA is an area that can be pursued in the future to look for new therapeutic targets as well as to unravel the intricacies of the inflammatory tumor microenvironment-CCA crosstalk. While therapies targeting specific molecules and signaling pathways have shown promise, combinatorial therapies as a whole have come up to be effective in different cancer types. Hence, a better understanding of the different components of the tumor stroma that the CCA cells modulate and exploit in order to give rise to a pro-inflammatory and pro-tumorigenic environment can lead to a holistic understanding and approach toward treating CCA. Some of these key mechanisms that interact and promote the onset and progression and subsequent metastasis of CCA is shown in SR, SG, and SC planned and wrote the manuscript and approved the final submitted version. SR made the figure.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.