People typically rely heavily on visual information when finding their way to unfamiliar locations. For individuals with reduced vision, there are a variety of navigational tools available to assist with this task if needed. However, for wayfinding in unfamiliar indoor environments the applicability of existing tools is limited. One potential approach to assist with this task is to enhance visual information about the location and content of existing signage in the environment. With this aim, we developed a prototype software application, which runs on a consumer head-mounted augmented reality (AR) device, to assist visually impaired users with sign-reading. The sign-reading assistant identifies real-world text  on command, highlights the text location, converts it to high contrast AR lettering, and optionally reads the content aloud via text-to-speech. We assessed the usability of this application in a behavioral experiment. Participants with simulated visual impairment were asked to locate a particular office within a hallway, either with or without AR assistance . Subjective assessments indicated that participants in the AR group found the application helpful for this task, and an analysis of walking paths indicated that these participants took more direct routes compared to the control group. However, participants in the AR group also walked more slowly and took more time to complete the task than the control group. The results point to several specific future goals for usability and system performance in AR-based assistive tools. Navigation is a multifaceted task, for which people typically rely heavily on vision ,2. Two bWhile existing aids such as canes and GPS technology –7 can asFor individuals who have limited vision but are still able to read magnified text, another possibility for assisting indoor wayfinding is to provide the user with enhanced visual information about the location and content of existing signage in the environment. For this purpose, systems that utilize head-mounted displays (HMDs) may offer unique features: the user can naturally rotate their head to scan for signs, and the enhanced information may be integrated directly with their remaining vision. Existing HMD devices for people with low vision can perform overall contrast enhancement and magnification via real-time near-eye video displays ,19. OtheMoving forward, tools that allow for visual enhancement via off-the-shelf hardware hold promise to increase the utility of HMDs for users with reduced vision. In particular, augmented reality (AR) via optical see-through HMDs may allow for task-specific information to be provided in a way that does not completely occlude the user’s natural vision –23. ReceHere, we describe the design and implementation of an application for a consumer AR platform (Microsoft HoloLens) that provides indoor wayfinding assistance directly in the user’s natural field of view. The system leverages recent advances in computer vision and HMDs to allow visually impaired users to interact with existing wayfinding infrastructure (text-based signs), without modification to the physical environment. Specifically, the system uses optical character recognition (OCR) to identify signs, which are then localized in 3D and visually enhanced in AR. In a behavioral experiment, participants with simulated visual impairment were asked to use the system while locating a particular office within a hallway. The results suggest improvements in subjective task assessment and route efficiency – but overall negative effects on task speed – as compared to a control group. The conclusions drawn from the current study can be used to inform future work on AR visual assistance.Microsoft HoloLens is an untethered optical see-through HMD system designed to support AR for consumer and enterprise applications . The senSoftware development was performed using Microsoft’s HoloToolkit and Unity (Unity Technologies). When looking for a sign, the user initiates OCR either by pressing the clicker or by saying “What’s here?”. On this command, the PhotoCapture API in Unity captures an image with the HoloLens’ front-facing camera . The application constructs a JavaScript Object Notation (JSON) request containing this image and sends it to the Google Cloud Vision API, a cloud-based computer vision engine, over Wi-Fi. The response from the API contains a series of entries, each with a description and pixel bounding box for a region of text detected in the image. This process takes 3–4 sec on average.If the response is empty, the user is notified with synthetic speech: “No text was detected.” If the response is not empty, there is no guarantee that separate response entries from the API correspond to physically separate signs. For example, a sign for Room 323 might in some instances return two entries (“Room” and “323”), which should ideally not result in two separate AR signs. Thus, if the distance between multiple response entries within a given image capture falls below a specified threshold (in pixels), the text is combined and allocated to the same AR sign. In the current implementation, this threshold is set to 30 pixels.To determine the 3D location of each sign in physical space, a virtual camera is placed at the location and orientation that the picture was captured from, using the on-board motion tracking. A ray is cast from this camera through the center of the pixel bounding box, and the intersection with the 3D map of the surroundings is recorded. Note that this 3D map is constructed in real-time via the depth sensor, not loaded from an external source. Importantly, each time a new image is captured, any new AR signs must be compared to the existing signs to avoid generation of duplicates. Signs are likely to be duplicates when a new sign has both similar text and similar 3D coordinates to an existing sign. To prevent duplicates, locality hashing is performed on all sign texts, and the application discards any new signs with similar text and similar 3D coordinates to an existing sign. In the current implementation, a distance threshold of 13.5 cm is used, such that signs with similar text that are less than 13.5 cm from an existing sign are considered duplicates. This threshold was determined during pilot testing. This method accounts for minor errors in the OCR, as well as offsets that can occur due to errors in motion tracking as the user moves around an environment.The next step is to create highly visible AR sign indicators. This is done by placing a virtual sphere at the location of the text. As the user continues to move, the world coordinates of the sphere stay the same, creating an AR sign in the same location as the real sign. The diameter of the sphere matches the maximum dimension of the text in the world, and each sphere flashes at 3 Hz to improve visibility. Indicators are also colored based on the confidence that the associated text has been accurately recognized, using the height of the bounding box in pixels. For the current implementation, signs with text height >20 pixels, 11–20 pixels, and ≤ 10 pixels were colored green, orange, and red, respectively. When the user sets the cursor on a sign, the indicator stops flashing and turns bright magenta. Pilot tests confirmed that the spheres, colors, and cursor were all visible with visual acuity of ~1.30 logMAR (~20/400), although color vision deficits could impede users from distinguishing between confidence colors.http://github.com/eacooper/HoloLensSignARApp.The user has the option to either read all AR signs at once or select a single sign. To read all of the AR signs, the user says “Read all here,” which reveals the enhanced text of every sign in the current field of view. Alternatively, the user can select an individual sign with the cursor, and say “Show me” or press the clicker. When a sign is being read, the sphere is hidden and replaced by bright green Arial font subtending 2° vertically . All participants wore swim goggles modified with Bangerter occlusion foils (two layers of LT 0.1 foils) that reduced their vision to 1.28 logMAR on average . ParticiParticipants started at one end of an unfamiliar hallway (2.1 m × 27 m) that contained 12 doors, including 10 offices with name and room number signs, and two restrooms also with signs. They were asked to find a particular professor’s office in the hallway, located approximately 16.7 m from the starting position. They were given the professor’s name, and were unaware of the office location prior to performing the task. The AR group practiced using the system briefly in a separate room before performing the task. After completing the task, participants were asked to consider times when they have had to locate a room in an unknown building, and rate their agreement with three statements from 1(low) to 5(high) based on their experience in the study: “This task would be easy,” “I would feel comfortable doing this task in a typical indoor environment,” and “I would feel confident in my ability to perform the task.”The HoloLens motion tracking recorded the participant’s position during the task at approximately 50 Hz. To determine average walking speed, for each participant we first calculated the distance travelled between each pair of consecutive position measurements. Data points were excluded in which participants were stationary (speed < 0.005 m/s). We also excluded data points with high speeds that were likely due to recording artifacts (speed > 5 m/s). The average speed was then computed for each participant from the remaining measurements.While our primary focus was to compare the control and AR groups, we also wanted to examine whether individuals preferred doing this task with or without the application. Thus, after giving their responses, the control group repeated the task using the sign-reading assistant, and the AR group repeated the task without it. The second trial was conducted in a different hallway. Afterwards, participants reported which trial was easier, and on which trial they were more comfortable and confident. Raw motion tracking and response data for each participant, as well as plotting and analysis code, are included in W = 151, p < 0.001), and felt more comfortable , and confident  than the control group . On average, participants in the AR group initiated the OCR 9.8 times during the task, with a minimum of 3 and a maximum of 22. Based on the average processing time see , this suThe results of the behavioral study suggest that an AR-based sign-reading assistant may provide a useful tool for increasing ease, comfort and confidence when visual wayfinding is impaired. However, there are several limitations in the current system that can be addressed with future work.First, the application relies on a cloud-based service for OCR; as such, it requires a wireless internet connection and takes several seconds to obtain results. Second, while the 3D localization of the signs was relatively accurate, there were clear cases in which the AR sign did not overlap with the physical sign. While these errors were not substantial enough to reduce the users’ ratings of the task relative to the control group, localization could be improved with higher resolution  3D mapping and/or motion tracking. In addition, the current system does not have an efficient solution for areas with a lot of signage, which could lead to an obtrusive number of blinking indicators, or enhanced text that is too long to comfortably read on the screen. Third, the current application only searches for signs when the user manually initiates it, and as such requires visual guidance . Future In conclusion, consumer AR systems hold the potential to augment visual information for everyone. In the process of developing these technologies, it is essential to consider the needs of and opportunities presented for users with a range of different levels of vision. While consumer AR systems are not yet common place in day-to-day life, our work suggests that AR provides a promising and flexible future platform to address gaps in common assistive tools.S1 Appendix(ZIP)Click here for additional data file.


Gastric cancer (GC) is a leading global health problem as it is the fifth most common cancer type and the third most common cause of cancer-related deaths worldwide. In most areas of the world, the incidence rate of GC is 1.5- to 3-fold higher in males than in females. The androgen receptor (AR) is an independent adverse prognostic factor in patients with GC. However, the mechanism by which AR regulates the progression of GC remains unclear. In this study, we found that AR expression was upregulated in 6/8 GC cell lines, and this expression was higher than that in immortalized gastric cells. AR expression was also higher in GC tissues than in adjacent tissues. Moreover, the ectopic expression of AR promoted the colony formation ability, migration and invasion of GC cells. In contrast, AR knockdown had the opposite effects on GC cell lines. Remarkably, we found that AR regulated cell cycle-related kinase (CCRK) expression through transcriptional mechanisms. The AR-CCRK axis promoted GC development through the phosphorylation of GSK3β and β-catenin. Furthermore, TCGA data revealed that high expression of AR or CCRK was related to poor prognosis in GC patients. The prognosis was significantly worse in patients with concurrent high AR and CCRK expression than in patients with low AR and CCRK expression. In conclusion, our study demonstrated that AR and CCRK acted as oncogenes in GC progression. However, their clinical roles require further exploration. Globally, the incidence of GC was 1,000,000 cases in 2018 et al. found that AR could indirectly suppress NFκB to inhibit invasion in HCC The androgen/AR signaling pathway is involved in the development of male-specific phenotypes during embryonic development, spermatogenesis, sexual behavior and adult reproduction The aim of this study is to determine the effect of AR on the progression of GC and the mechanism underlying these effects, which will provide novel ideas for the treatment of GC.2 at 37 °C until they reached 80% confluency. The cells were then digested with 0.25% trypsin and inoculated into a new flat.In our research, 7 human GC cell lines  and 1 normal immortalized gastric mucosal cell line GES-1 were used for the assays. AGS and MGC803 cells were obtained from the ATCC. MKN45, MKN28, HGC27, BGC-823 and SGC7901 cells were purchased from the Cancer Hospital of the Chinese Academy of Medical Sciences . MKN45, MKN28, HGC27 and SGC7901 cells were cultured in DMEM  with 10% FBS . AGS, MGC-803, BGC-823 and SGC-7901 cells were cultured in RPMI-1640 medium  supplemented with 10% FBS . Cells were cultured with 100 μg/mL penicillin and 100 μg/mL streptomycin in a humid atmosphere containing 5% COFour pairs of clinical tissue samples from primary tumors and adjacent nontumorous sites were collected from the Inner Mongolia People's Hospital . The project was approved by the Ethics Committee of Clinical Research of the Inner Mongolia People's Hospital.We downloaded the gene expression data of 440 clinical gastric adenocarcinoma samples  from the cBioPortal database. Then, we used Rstudio software to develop a program to simulate the high or low expression of AR by selecting 30 samples randomly from 440 clinical samples that had the 25% highest or 25% lowest AR expression values and then used this gene set for GSEA. GSEA software analysis included the following four important factors: enrichment score (ES), normalized enrichment score (NES), false discovery rate (FDR) and P value.Total RNA was extracted from GC cells and tumor tissues using triazole reagent . cDNA was synthesized using a GoScript reverse transcription kit . The relative expression of AR and CCRK was detected by semiquantitative RT-PCR and real-time quantitative PCR. Primers: AR RT-sense: 5'-AAGCCAGAGCTGTGCAGATGA-3'; AR-RT-anti-sense: 5'-TGTCCTGCAGCCACTGGTTC-3'; CCRK-RT-sense: 5'-AGACTGGCGAGATAGTTGCC-3'; CCRK-RT-anti-sense: 5'-GTGGGAACACAGCCTTCAGT-3'.5 cells/mL with serum-free medium. Then, 200-μl cell suspensions were placed in a transwell upper chamber (for the migration assay) or a Matrigel invasion chamber (for the invasion assay). The lower chamber contained 10% fetal bovine serum culture medium and was maintained at 37 °C with 5% CO2 for cultivation for six to eight hours (for the migration assay) or for 18 to 24 hours (for the invasive assay). Adherent cells were fixed with 4% paraformaldehyde and stained with crystal violet. The cells were counted with an optical microscope.GC cells were digested to produce a single-cell suspension, which was then diluted to a concentration of 1X104 cells/mL. Then, 1000 cells were plated on a 6-well plate. Cell culture conditions were 37 °C for 7-14 days. The colony number was counted after staining with crystal violet.AR cells were digested with 0.25% trypsin, and the cell suspension was diluted to 1X10ChIP analysis was performed using anti-AR antibody (Abcam: #5153) and IgG (Abcam: #3900) according to the manufacturer's protocol. PCR was performed using the following DNA promoter-specific primers: CCRK-RT-promoter-sense: 5'-CGCAACGGCCCAAAGTAG-3'; CCRK-RT-promoter-anti-sense: 5'-CGCAACGGCCCAAAGTAG-3'.7 cells/mL. We assigned five 6-week-old BALB/c nude mice to each group and then injected 200 μL of the cell suspension into their right dorsal sides. We collected data, including mouse weight and tumor volume, every week. Following observation for 6 weeks, the nude mice were sacrificed; the stomach tumors of the mice were collected, and photos were taken. Animal care and all laboratory procedures were approved by the Animal Ethics Committee of the Inner Mongolia People's Hospital.CCRK was transformed with or without vector into AGS cells. BGC823 cells were transfected with shCCRK, shVector or no vector. The cell concentration was adjusted to 1 x 10We purchased viruses  from Shanghai GenePharma Company . Transfections were performed according to the manufacturer's instructions.We used RIPA lysates to cleave cellular proteins and then combined them for use with a BCA protein quantification kit . Protein was isolated from each sample and separated using 10% SDS-PAGE prior to being transferred onto polyvinylidene fluoride (PVDF) membranes. The PVDF membranes were then incubated with 8% milk, followed by incubation with primary antibodies  and secondary antibodies. The signals were detected by enhanced chemiluminescence.AR and CCRK mRNA expression data (GDC TCGA Stomach Cancer) were downloaded from the UCSC Xena database. The clinical data were obtained from the cBioPortal database website. Data analysis of the correlation between AR and CCRK was performed with GraphPad Prism 5.0 . We found the genes related to AR in GC tissues from cBioPortal. ChIP-Seq data for AR were downloaded from GEO (GSM2219852). We used the intersection of the top fifty percent of AR-related genes and the top fifteen percent of putative AR targets to analyze their functional categories with the DAVID website. We used X-tile software to find AR and CCRK cut-off values  to maximize the Kaplan-Meier survival curve difference. A student's t-test was used for comparisons between groups. Pearson's correlation analysis was performed to analyze the correlation between AR and CCRK mRNA expression. Kaplan-Meier survival curves and log-rank tests were used to evaluate the overall survival (OS) corresponding to AR and CCRK mRNA expression. When the P value was less than 0.05, the difference was considered statistically significant.It has been reported that the expression of AR is higher in GC tissues than in adjacent tissues Based on the GSEA results, we explored whether AR affected the migration and invasion of GC cell lines. Transwell experiments showed that when AR was overexpressed in AGS cells, the migration ability of the cells was increased Figure A, B. A Met al. found that in HCC, AR could promote the expression of CCRK through transcriptional regulation We next aimed to explore the underlying mechanisms of how AR promotes GC. We used the intersection of AR-related genes and putative AR targets to analyze their functional categories in DAVID. We focused on 22 genes of cancer-related cell cycle functional categories Table  and analin vitro.Since AR was found to regulate the expression of CCRK transcriptionally, we wondered whether CCRK might also affect the progression of GC. Therefore, we conducted a series of functional experiments on CCRK in GC cell lines. Cloning formation experiments showed that in the AGS and GES-1 cell lines, CCRK overexpression could enhance the clonal formation ability of the cells  and glutamine synthase (GS) We examined the mechanism of how AR promoted GC. Feng H. Finally, we investigated the clinical application prospects of AR and CCRK. We analyzed the prognostic ability of AR and CCRK in GC patients with TCGA data. As shown by the Kaplan-Meier survival curve, GC patients with lower AR mRNA expression had significant survival advantages over those with higher AR expression  is well-known as a target of prostate cancer which is a malignancy in the male reproductive system et al. tested six commercially available inhibitors  and one preqin enzyme development inhibitor (MER151). The authors found moderate inhibitory effects in vitro because these compounds were not completely specific for CCRKOur data demonstrated that AR could promote CCRK expression in GC through transcriptional regulation. Transwell experiments showed that CCRK increased invasion and metastasis in GC cells. Our results also showed that the high expression of CCRK increased the ability of cells to clone; subcutaneous tumorigenesis experiments in nude mice showed that CCRK knockdown reduced the tumor volume, but CCRK overexpression increased the tumor volume. Our experimental results indicate that CCRK is an oncogene in GC. Therefore, CCRK antagonists may also play a role in the progression of GC. CDK inhibitors have also been used to inhibit the expression of CCRK. Mueller In summary, we show that the expression of CCRK is increased by AR in GC. Low expression levels of AR and CCRK are related to a better prognosis in GC patients. AR and CCRK can promote the development of GC, suggesting that we should investigate the effects of AR or CCRK inhibitors. In addition, these molecules are strong candidates to be prognostic indicators of GC.


Allergic Rhinitis (AR) is a common inflammatory disease of the nasal mucosa. The CD14 is a receptor for lipopolysaccharide and inhaled endotoxin which can stimulate the production of interleukins by antigen presenting cells. Accordingly, CD14 plays an important role in allergic and atopic diseases, which can be one of the etiological factors for allergic diseases. The present study investigated the association between the CD14 gene polymorphism C-159T and AR and aimed to detect the correlation between serum levels of CD14 and AR.This study was conducted on two groups of participants. The experimental group consisted of 125 patients with AR referring to Ghaem Hospital, Mashhad University of Medical Sciences in Mashhad, Iran, and the control group included 125 healthy subjects from Mashhad National Blood Center, Iran. Serum CD14 levels were measured by enzyme-linked immunosorbent assay. Polymerase chain reaction-restriction fragment length polymorphism was employed to detect C-159T gene polymorphism in the CD14 promoter region.There was a significant association between CD14 C-159T gene polymorphism and AR (P<0.001). The results of the statistical analysis showed that the TT genotype could significantly increase the risk of AR (P<0.001). Additionally, a significant association was observed between C-159T gene polymorphism and the serum level of CD14 (P<0.001). Regardless of the genotypes, the serum CD14 levels were significantly higher in AR patients than in those of the participants in the controls (P=0.007).According to the obtained results of this study, CD14 in serum might be a potential marker for the diagnosis of AR, and in genetic levels it might be a predictive factor for the disease. Allergic rhinitis (AR) is a type of inflammation of the mucous membrane in the nose, which affects 10-40% of people around the world . When seThis case-control study was conducted on 125 patients with AR referring to Ghaem Hospital, Mashhad University of Medical Sciences in 2014-2015. For the sample size, each of the case and control groups was supposed to have 105 individuals according to Yazdani et al. and the comparison of two population proportions formula . On the Patients signed a written consent form and participated in the study when their AR was confirmed by allergists. Inclusion criteria of the study were a diagnosis of AR and cutaneous sensitivity to local allergens. The exclusion criteria were a history of antihistamine usage, dermographism disease, and reluctance to participate due to personal reasons.After patients were diagnosed with AR through clinical history-taking and skin prick tests, they completed 22-item Sinonasal Outcome Test . On the Skin prick testthe skin prick test was performed using allergens, such as Salsola kali, grass mix, orchard grass, ash, Amaranthus retroflexus, lamb’s quarters, tree mix, Timothy, D. Farinae, D. Pteronyssinus, Bermuda grass, Kentucky grass, mugwort, willow, Alternaria, and Aspergillus mix allergens. Allergens were purchased from Greer Company . Genomic DNA extraction and CD14 genotypingThe PCR-RFLP technique was employed for the detection of CD14 gene polymorphism C-159T based on a previously published study . The proMeasurement of the serum levels of CD14Serum levels of CD14 were measured by ELISA according to the manufacturer’s instructions . Statistical analysisStatistical analysis was performed using SPSS (version 16) through t-test and logistic regression tests. Hardy–Weinberg equilibrium was performed using the Chi-square test. The frequency of genotypes and allelotypes of C-159T gene polymorphism was calculated by direct counting. A P-value<0.05 was considered statistically significant. Study populationPatients with AR included 61 men and 64 women with a mean age of 34.67±10.04 years. On the other hand, the controls consisted of 61 men and 64 women with a mean age of 35.01±8.01 years. No significant difference was observed between patients with AR and the controls in terms of age (P=0.77) and gender (P=0.899). As can be seen in Prick test The results of the prick test demonstrated that the most frequent allergens were Salsola kali, lamb’s quarters, pigweed mix with the allergenicity of 90.4%, 82.4%, and 80.8%, respectively. Moreover, willow with 16.8% allergenicity was the least frequent allergen among patients in this study. The highest and lowest rates of skin inflammation were related to the Salsola kali (10.88±6.66 mm) and the willow (0.57±1.38 mm), respectively. Serum levels of CD14The mean serum levels of CD14 was 6237.2±3618.25 ng/ml in patients with AR and 4589.38±2907.22 ng/ml in controls. As shown in The genotype frequency for CD14 gene polymorphism C-159T in the control group was 42.4% (n=53) for the CC, 51.2% (n=64) for the CT, and 6.4% (n=8) for the TT genotypes. In patients with AR, it was 14.4% (n=18) for the CC, 65.6% (n=82) for the CT, and 20% (n=25) for the TT genotypes . The genRunny nose and sneeze were the most common clinical symptoms in 84% of people with the TT genotype. A significant difference was observed between patients with AR and clinical symptoms, including nasal congestion (P=0.034), sneezing (P=0.013), and wheezing (P=0.012). According to The etiology of AR and other allergic diseases is related to factors that cause an excessive increase in IgE-dependent responses ,3,4. AltSeveral studies investigated the relationship between a polymorphism named C-159T in the CD14 gene promoter and allergic diseases . The aboTo the best knowledge of the researchers, this research is the first study which investigated the relationship between the functional polymorphism C-159T in the CD14 gene promoter region and serum CD14 levels in Iranian patients with AR. The results showed that C-159T polymorphism in the CD14 gene is highly associated with AR since AR was a significantly associated with TT and CT genotypes (P<0.001). The risk of AR in people with the TT and the CT genotypes is 9.2 and 3.8 times higher, respectively, compared to the individuals with the CC genotype. In other words, the risk of AR is higher in carriers of the T allele. Several previous studies showed a positive association between CD14 gene polymorphism C-159T and allergic diseases, as well as serum CD14 levels. Studies conducted by Gao et al. in England and Buckova et al. in the U.S. showed that the TT genotype was associated with the increase in the serum levels of CD14 and decrease in the serum levels of IgE in patients with allergic diseases such as asthma, allergic rhinitis ,21, eczeIn the studies performed by Baldini et al. and Demin Han et al. on patients with allergic diseases ,22, the As Koppelman et al. reported CC genotype was associated with a decrease in positive prick test cases in Dutch people. In addition, they observed an association between carriers of the C allele in CD14 polymorphism C-159T and atopy in a Czechoslovakian population . The stuThese controversies might be due to differences in the pathophysiologies of asthma and AR. Additionally, the influence of gene-to-gene interactions and ethnic diversity might be other causative factors for this discrepancy. Differences in environmental conditions can also be considered as highly important factors in shaping such contraindications, as well. The interaction between CD14 gene polymorphism C-159T and environmental factors, such as smoking, pets and exposure to endotoxin, were reported in several studies ,29-31. MThe results of the current study also showed that the most frequent allergens in Mashhad were Salsola kali (90.4%) and lamb’s quarters 82.4%). Moreover, the maximum levels of inflammation in the skin prick test were observed for the above-mentioned allergens. Studies conducted by Barber et al. in Spain and Pereira et al. in Portugal reported similar findings,34. The .4%. MoreThis is the first time an association between sensitivity to Salsola kali, ash, and pigweed mix in patients with AR and CD14 polymorphism C-159T is being reported. Patients who had CT and TT genotypes showed significantly higher sensitivity to the above-mentioned allergens than those patients with the CC genotype. The results of the present study showed that CD14 in both protein levels and genetics might influence AR. Therefore, it is suggested that CD14 might be a serum immune biomarker for the diagnosis of AR. Additionally, it is suggested that CD14 gene polymorphism C-159T is a potential genetic predictive factor for AR. As a matter of fact, toll-like receptor (TLR) family, can recognize microbial products and be responsible for induction of both innate and adaptive immune responses. The TLR-4 is essential for the recognition of bacterial LPS and activation of signaling pathway. The signaling pathway is mediated by adaptor proteins, namely myeloid differentiation primary response 88, TIRAP, translocating chain-associated membrane protein, and TIR-domain-containing adapter-inducing interferon-β. These adaptor proteins are mediators between TLR-4 and transcription factors, such as NF-κB, AP-1 and Interferon Regulatory Factor-3, in order to produce proinflammatory cytokines ,40. We suggest to evaluate the expression of TLR-4 as well as serum levels of DC14 and total level of IgE in patients with AR for future studies. This might help us to find out the influence of CD14 on Th1 and Th2 balance, more precisely. Additionally, it might be helpful for future studies to divide patients with AR into atopic and non-atopic.


Development of novel engineering solutions that incorporate robotic systems offers huge potential, as both assistive devices and therapeutic aids, for patients with reduced motor and/or cognitive abilities. Indeed, such technologies are capable of outperforming existing therapeutic systems for improving patients' achievable functional recovery. As a result, advanced engineering solutions for rehabilitation robotics have received a considerable amount of interest in recent years, from both the academic community and the industrial sector. Emphasis is typically focused on the improvement of sensor systems, control engineering, computer vision, robotic mechanics, human-machine interfaces, modelling and simulation, in-built intelligence, and informatics, to meet the broad range of challenges posed during patient rehabilitation.This special issue is aimed at exhibiting the latest research achievements, findings, and ideas in the field of rehabilitation robotics and systems. Researchers were invited to contribute with their original research articles, as well as review articles, that summarize the most recent developments in the field of rehabilitation robotics and systems.Plasticity and motor-learning robotic devicesAssistive and therapeutic robotic aidsUpper and lower limb rehabilitation roboticsHuman-machine and brain-machine interfacesMobile, wearable, and prosthetic robotic devicesSensors, intelligent sensors, and sensor networksArtificial intelligence-based rehabilitation systemsComplex, nonlinear, and intelligent systemsVision, awareness, perception, and signal processingSensor networks for precise motion control and visual servoingAdaptive control, robust control, and active disturbance rejection controlModelling and simulation for robotic rehabilitation systemsNetworked rehabilitation roboticsIntelligent automation of rehabilitation roboticsInformational monitoring, control, and data fusion for rehabilitation systemsPotential topics included but were not limited to the following:A total of 22 papers were submitted to this special issue. After peer review, finally 13 of them were accepted and published, covering a wide range of the topics proposed in the call for papers.Three interesting papers are based on reviews of literature papers on rehabilitation robotics and systems. Firstly, a work by G. Carpino et al. includes a meta-analysis of robot-mediated lower limbs rehabilitation for stroke-affected patients; it is aimed at evaluating the effectiveness of the robotic approach using wearable robots or operational machines with respect to the conventional approach. The primary assessed outcome is the patient's ability to regain walking independence, whereas the secondary outcome is the average walking speed. Also, E. D. Oña et al. conduct a systematic literature review to identify the contribution of robotics to upper limb neurorehabilitation, highlighting its relationship with the rehabilitation cycle, and to clarify the prospective research directions in the development of more autonomous rehabilitation processes. In addition, a series of technical requirements that should be considered when designing and implementing autonomous robotic systems for rehabilitation are presented and discussed. Lastly, T. Eiammanussakul and V. Sangveraphunsiri review the training activities that were realized by rehabilitation robots in the literature to offer insights into developing a novel robot suitable for stroke rehabilitation. The control system of the lower limb rehabilitation robot in sitting position of the authors' previous work is discussed in detail to demonstrate the behavior of the robot while training a subject. A preliminary experiment is conducted on a healthy subject to show that the robot can perform active assistive exercises with various training activities and assist the subject to complete the training with a desired level of assistance.Next, other six papers are more directed towards the experimental evaluation of rehabilitation mechanics. Thus, W. Liang and Y. Yu propose the so-called manipulability inclusive principle (MIP) to evaluate the assistive mechanism's assistive feasibility and assistive effect through the manipulability comparison between the assisted limb and slave-active-assistive mechanism. The optimization based on MIP can make the assistive mechanism realize better kinematical performance and assistance. Another paper by B. D. M. Chaparro-Rico et al. presents an experimental characterization of NURSE, a device for arm motion guidance. The laboratory setup and testing modes are presented. Two exercises for the upper limb are used to test the NURSE behavior. Trajectories and linear accelerations are tested when the device performs the exercises with and without load. Z. Ray and E. D. Engeberg face the problem of autonomously preventing grasped objects from slipping out of prosthetic hands for limb-absent people. Their paper explores a human-inspired grasp reflex controller for prosthetic hands to prevent slip of objects when they are rotated. This human-inspired grasped object slip prevention controller is evaluated with six different objects in benchtop tests and by 12 able-bodied subjects during realistic tasks of daily life. Moreover, in another article, M. Wu, M. R. Haque, and X. Shen present a new sit-to-stand (STS) control approach for powered lower limb prostheses, which can regulate the power delivery of the prosthetic knee joint to obtain natural STS motion like that displayed by healthy subjects. Mimicking the dynamic behavior of the knee in both phases of the STS, a unified control structure provides the desired control actions by combining an impedance function with a time-based ramp-up function. In addition, H. Guang et al. present an interactive upper limb rehabilitation robot with a parallel mechanism and an isometric screen embedded in the platform to display trajectories. In the dynamic modeling for impedance control, the effects of friction and inertia are reduced by introducing the principle of virtual work and derivative of the Jacobian matrix. To achieve the assist-as-needed impedance control for arbitrary trajectories, the strategy based on orthogonal deviations is proposed. Simulations and experiments were performed to validate the dynamic modeling and impedance control. Lastly, in a study by M. R. Afzal et al., a wearable system based on the reaction wheel is used to deliver light-touch-based balance biofeedback on the subject's back. The system can sense torso tilt and, using reaction wheels, generates light-touch. A group of 7 healthy young individuals performed balance tasks under 12 trial combinations based on two conditions each of standing stance and surface types and three of biofeedback device status.A last group of four papers is directly focused demonstrating implementations for rehabilitation. A paper by J. Cantillo-Negrete et al. presents the implementation of a brain-computer interface system, coupled to a robotic hand orthosis, driven by hand motor imagery of healthy subjects and the paralyzed hand of stroke patients. A novel processing stage is designed using a bank of temporal filters, the common spatial patterns algorithm for feature extraction, and particle swarm optimization for feature selection. The system's performance shows that it has potential to be used for hand rehabilitation of stroke patients. The aim of an article by J. C. Castillo et al. is in the line of robotic therapies in which a robot can perform partially or autonomously a therapy session, endowing a social robot with the ability of assisting therapists in apraxia of speech rehabilitation exercises. The authors integrate computer vision and machine-learning techniques to detect the mouth pose of the user, and on top of that, our social robot performs autonomously the different steps of the therapy using multimodal interaction. Then, M. A. Padilla-Castañeda et al. introduce the development and evaluation of a robotic-assisted rehabilitation system as a new methodology of assisted physiotherapy in orthopedics. The proposal consists of an enhanced end-effector haptic interface mounted in a passive mechanism for allowing patients to perform upper limb exercising and integrates virtual reality games conceived explicitly for assisting the treatment of the forearm after injuries at the wrist or elbow joints. The authors design specific game scenarios enriched by proprioceptive and haptic force feedback in three training modes: passive, active, and assisted exercising. Finally, in a study by E. Beretta et al., a group of 18 children and adolescents with hemiplegia, an impairment due to acquired brain injuries, was enrolled and underwent intensive rehabilitation treatment including flank physical therapy and constrained-induced movement therapy or Armeo®Spring therapy. The effects of the treatments are assessed using clinical functional scales and upper limb kinematic analysis during horizontal and vertical motor tasks. The results of this study may be of help to define the best rehabilitation treatment for each patient, depending on the goal, and may thus support clinical decision.


Incorporating patients’ preferences in the care they receive is an important component of evidence-based practice and patient-centred care.This study assessed stroke patients’ preferences regarding rehabilitation settings.A cross-sectional design was used to examine preferences of stroke patients receiving physiotherapy at three hospitals in Northern Nigeria. Personal factors and preferred rehabilitation setting data were obtained using the Modified Rankin Scale  and a researcher-developed questionnaire. Associations between preferences and personal factors were explored using bivariate statistics.X2 = 6.80; p = 0.03). Similarly, most of the participants (53.3%) who depended on family finances preferred home- or community-based rehabilitation, while most of those who depended on employment income for finances preferred an outpatient rehabilitation setting .Sixty stroke patients whose mean age was 53.6 ± 14.8 years participated in the study. Most of the participants (38.3%) preferred an outpatient setting, 19 (31.7%) preferred rehabilitation in their homes, 14 chose inpatient rehabilitation (23.3%), while 4 (6.7%) preferred the community. Age and source of finance were significantly associated with preferences. The majority (66.7%) of those aged ≥ 65 years expressed a preference for rehabilitation in the home or community (A preference for rehabilitation in outpatient facilities predominated followed by home-based rehabilitation, and preferences varied based on age and source of finance. These variations in preferences have implications for making rehabilitation decisions. Acknowledging patients’ preferences regarding the care they receive is gradually occupying a position of prominence in health care systems. With the advent of shared decision-making, patient-centred care and evidence-based practice, clinical decisions are expected to incorporate patients’ views, values and preferences  and clinical  attributes of the participants. Information on post-stroke duration was obtained from the participants and verified using their medical records while data on the type of stroke, as diagnosed by physicians and neurologists, was solely obtained from the participants’ medical records. Another clinical attribute that was assessed was global disability, using the Modified Rankin Scale (mRS). The mRS is a valid measure of disability on a six-point scale with values ranging from 0 to 5  through face-to-face interviews from May to July 2014.All data  obtained were presented as descriptive statistics.Chi-square statistics were used to examine the associations between participants’ personal factors  and their preferred rehabilitation setting. For the purpose of the analyses, age categories were < 65 years and ≥ 65 years; marital status categories were married and unmarried  while source of finance was categorised as employment income, pension, family and charity. The level of statistical significance was set at α = 0.05.Ethical approval for the study was obtained from the Research and Ethical Committee of the University of Maiduguri Teaching Hospital, Maiduguri, Borno State, Nigeria.Sixty stroke patients, with a male majority (61.7%), participated in the study. The mean (standard deviation [SD]) age was 53.6 (14.8) years. Details of the socio-demographic and clinical characteristics of participants are presented in In terms of preferences for rehabilitation settings, 23 (38.3%) participants preferred an outpatient setting, 19 (31.7%) preferred rehabilitation in their homes, 14 chose inpatient rehabilitation (23.3%), while 4 (6.7%) preferred the community .X2 = 6.80; p = 0.03). Most (53.3%) participants who depended on family finances preferred rehabilitation in the home or community while an outpatient rehabilitation setting was preferred by most of those who depended on employment income for their finances .With the results of the descriptive statistics showing that only four participants preferred the community and given the fact that rehabilitation in the home and community settings both represent domiciliary rehabilitation, the ‘home’ and ‘community’ options were presented as an entity (home or community-based) for the inferential analyses. Among the personal factors explored, only age and source of finance were significantly associated with setting preferences. Incorporating patients’ preferences into the care they receive is a cornerstone of evidence-based practice and represents an important premise of the shared decision-making model of care. An important decision regarding stroke rehabilitation is the setting in which rehabilitation takes place, and this was examined in this study.Most participants expressed their preference for rehabilitation at outpatient facilities, closely followed by those who preferred rehabilitation at home. This finding may be considered as a reflection of the current status of stroke rehabilitation services in Nigeria. As in many developing or African countries, rehabilitation facilities are underdeveloped in the country and the only available settings for stroke rehabilitation are the private residence of the patients and outpatient facilities. Contrary to the findings of this study, however, one study conducted in the United States showed that stroke patients undergoing acute care overwhelmingly (85%) preferred rehabilitation in their homes post-acute care discharge (Gregory et al. Inpatient facility was another rehabilitation setting option in this study and it was preferred by a fifth of the participants. However, it is important to mention here that inpatient rehabilitation centres are not available in Nigeria, although rehabilitation in such centres has been reported to result in far better stroke outcomes compared to rehabilitation in other settings such as the home, outpatient or nursing facilities (Chan et al. Age was a significant personal factor that was found to be associated with preferences, and about 6 out of every 10 stroke patients who were 65 years and above in this study preferred having rehabilitation in their homes or in the community. This observation may indicate that the elderly are more comfortable at home and have psychological and emotional attachments to the family and familiar environments such as their homes. Frailty that often accompanies aging may also contribute to preference. Although the influence of frailty on rehabilitation setting preference was not explored, the study assessed level of disability but found no significant association between preference and disability level. However a previous study showed that older stroke patients were less likely to be discharged home compared to those of younger age (Nguyen et al. Most of the participants who were financially self-sufficient preferred the outpatient setting while those who depended on others for their finances preferred rehabilitation in their private homes or the community. A study that compared stroke rehabilitation in the home versus an outpatient setting in the United States showed that stroke patients with higher incomes had more outpatient visits than those with lower income (Chan et al. Aside from age and source of finance, no other personal factor was found to be significantly associated with preference for rehabilitation setting in this study. However it is important to note that some previous studies did not dwell on the effect of personal factors on rehabilitation setting preference (AL-Oraibi et al. The limitations of the present study include the small size of the sample, which precluded data treatment using more vigorous statistics such as multiple regression analyses to identify independent determinants of preferences. Similarly, the small sample size could have been responsible for the very low number of participants that registered their preference for rehabilitation in the community. This necessitated grouping the home and community settings as an entity for the purpose of the inferential analyses, although it is important to mention that the two settings represent a domiciliary rehabilitation setting. The hospital-based design of the study may also limit the generalisability of the findings and future community-based studies would therefore be beneficial. In the same vein, the fact that the participants were all undergoing outpatient physiotherapy at the time of the study could have biased the results of the study and resulted in some participants’ preference for the familiar. Although the impact of patients’ preference on the outcome of care was outside the scope of this study, it is important to note that patients’ preferences may not always be realistic, feasible or in their best interest and this should be borne in mind when interpreting the findings. Also, given the debilitating effect of stroke on several body functions including mental functions, obtaining information on stroke patients’ preferred rehabilitation setting may not always be possible or feasible.The distribution of the preferences for rehabilitation setting in this study coupled with the variations in preferences based on the stroke patients’ personal factors highlight the variability of individual patient preferences and supports the need for patient-centred stroke rehabilitation procedures that assess patients’ preferences and take them into consideration when decisions about rehabilitation settings are made.Assessment of stroke patients’ preferences for rehabilitation practice should constitute routine practice given the variations in preferred setting among different categories of patients.


Efficient blood supply to the brain is of paramount importance to its normal functioning and improper blood flow can result in potentially devastating neurological consequences. Cerebral blood flow in response to neural activity is intrinsically regulated by a complex interplay between various cell types within the brain in a relationship termed neurovascular coupling. The breakdown of neurovascular coupling is evident across a wide variety of both neurological and psychiatric disorders including Alzheimer’s disease. Atherosclerosis is a chronic syndrome affecting the integrity and function of major blood vessels including those that supply the brain, and it is therefore hypothesised that atherosclerosis impairs cerebral blood flow and neurovascular coupling leading to cerebrovascular dysfunction. This review will discuss the mechanisms of neurovascular coupling in health and disease and how atherosclerosis can potentially cause cerebrovascular dysfunction that may lead to cognitive decline as well as stroke. Understanding the mechanisms of neurovascular coupling in health and disease may enable us to develop potential therapies to prevent the breakdown of neurovascular coupling in the treatment of vascular brain diseases including vascular dementia, Alzheimer’s disease and stroke. The human brain accounts for just 2% of total body weight, yet receives between 15 and 20% of total cardiac output . Neurovarefer to Fig. streptococci, which can cause meningitis IC and glutamatergic activation [2) as a result of basal [Ca2+]IC fluctuations in astrocytic end-feet  concentration of appropriately 50 nM, which is kept constant due to the action of several calcium ion channels and pumps such as the efflux plasma membrane calcium ATP-ase (PMCA) [2+] levels. These indicators can be virally injected into the brain, or genetically engineered. Genetically encoded calcium indicators (GECIs) are bioluminescent protein complexes with integral fluorophores, which when stimulated emit a resonance signal in the form of visible light which can be detected by a camera, allowing quantification of neural activity based on the level of luminescence emitted by the fluorophore. Examples of common GECIs include Yellow Cameleon 3.60 (YC3.60), which contains an enhanced cyan fluorescent protein (ECFP) donor fluorophore, bound to a Venus protein acceptor fluorophore, linked by a calcium-binding domain, calmodulin [At rest, neurons typically have a low basal [Cae (PMCA) . Upon nee (PMCA) . Therefolmodulin . More relmodulin . Specifilmodulin  amongst 2-photon microscopy is a technique in which a single laser emits two infrared (low-energy) photons to a focused region to cause excitation in fluorophores to a higher-energy state . As the Most stimulation paradigms in pre-clinical research involves the mechanical stimulation of either whiskers or fore/hind-paws to evoke a neural and haemodynamic response in the somatosensory cortex to study neurovascular coupling. Optogenetics involves expressing light-responding proteins such as channelrhodopsin 2 (ChR2), a blue-light sensitive protein which upon stimulation causes labelled cells to become active . NeuronsMany neurological conditions including the common neurodegenerative disorders display some level of neurovascular dysfunction; which either initiates or contributes to their pathogenesis, or is caused as a consequence. The neurovascular dysfunction seen in disorders such as Alzheimer’s and stroke can be directly attributed to cardiovascular deficits, however in some cases it may also not be directly associated with any underlying cardiovascular pathology, rather a functional breakdown of the neurovascular unit and signalling pathways.For the first time since medical records began, dementia has become the single-most leading cause of death in the UK surpassing heart disease, strokes and major cancers . With anSwedish K670 N/M671L mutation as well as the Indiana V717F mutation, which both increase β-secretase activity [AD is an age-related neurodegenerative disorder and the most frequent form of dementia worldwide . The priactivity , 48. Theactivity . Figure The vast majority of AD cases are sporadic (~ 90%) and are relatively later-onset, and are therefore not attributable to inherited genetic mutations to amyloid processing genes. It is important to note that even in sporadic AD cases, β-amyloid plaques are still present, and has many similarities to familial AD. Amongst the common risk factors already associated with cardiovascular disease and dementia such as age, hypertension, hypercholesterolaemia, diabetes etc., there are several risk factor gene polymorphisms which greatly enhance the likelihood of developing AD. The most common is the APOEε4 allele which is attributed to up to approximately 50-60% of all AD cases , 52. The3 total capillary length reduction compared to control subjects without the APOEε4 allele [−/− (pericyte deficient) mice have a reduced cerebral oxygenation level compared to controls as well as neurovascular uncoupling leading to neuronal death [There is a growing body of evidence which suggests that neurovascular coupling may be impaired in AD , 53, 54.4 allele . This st4 allele  in whichal death . More real death . The breal death  al death . It is tVascular dementia and other forms of cognitive impairment can be caused directly by having a stroke. Strokes can arise as a direct consequence of arterial pathology with profound changes to the neurovascular unit and the normal physiology of neurovascular coupling. This impaired blood flow to the brain can lead to the death of neural tissue in specific regions where blood flow is disrupted to cause a stroke. Depending on the severity of the stroke, symptoms can be mild ranging from a transient ischaemic attack to a potentially fatal in a stroke where a large are of the brain is affected . Strokes2+-mediated vasodilation, post-stroke, there is a switch in the polarity of neurovascular coupling in which Ca2+ actually causes vasoconstriction pathologically. This then limits CBF and functional hyperaemia to the brain parenchyma resulting in progressive ischaemia after the stroke takes place usually resulting in death of the patient a few days after the stroke [B1 receptors on endothelial cells to reduce overall synthesis of endothelial nitric oxide synthase (eNOS), and hence NO levels. ET-1 also binds to ETA receptors on the tunica media of vessels to upregulate PKC-α, which in turn inhibits the function of K+ channels to contribute to vasospasm, and BBB breakdown causing oedema [Subarachnoid haemorrhages (SAH) as a result of cerebral aneurysm rupture can cause a devastating stroke which usually results in death a few days after the initial rupture, if not fatal during the initial stroke . It has e stroke . A potene stroke . ET-1 big oedema . These sAgeing is itself the biggest risk factor in the development of many diseases including cardiovascular disease, Alzheimer’s and vascular dementia. Normal physiological ageing itself results in progressive neurovascular dysfunction, primarily marked by a reduction of overall CBF , 62. A kA recent study by Duncombe and colleagues explored the pathophysiology of neurovascular coupling associated with ageing . They foOne key pathological change associated with ageing is that of atherosclerosis. Atherosclerosis is a chronic inflammatory syndrome resulting in the progressive thickening and hardening (arteriosclerosis) of major artery walls . This coAtherosclerosis can take decades to take full effect with patients being largely asymptomatic until their mid-50 s, although initiation of disease takes place in early adulthood. The initiation of atherosclerosis is still not fully understood, although general consensus is that atherosclerosis begins with inflammatory changes within the endothelium caused by low-density lipoproteins (LDLs) being deposited , 68, andLDLs have been shown to deposit more readily in atherosclerotic-prone and low blood flow areas of an arterial branch, where they undergo oxidation by reactive oxygen species (ROS), stimulating an immune-response . The artThe solidifying step in atherogenesis is when invading monocytes differentiate into macrophages in the intima after binding to endothelial receptors such as VCAM1 and P-Selectin , Fig. 4.The atheromatous plaque (atheroma) may in itself cause an obstruction of blood flow, or more commonly the atheroma may rupture and form a thrombus with platelets, which can be repeated to form a stable cap disrupting blood flow over time further exacerbating stenosis, or rupturing within the vessel wall to cause either an embolism or haemorrhage. Either of these can contribute to coronary heart disease, myocardial infarction (if in the heart), or a stroke if in the carotid or cerebral arteries.Atherosclerosis affects most large to medium sized systemic arteries including those supplying the cerebral circulation, namely the internal carotid and vertebral arteries . ProgresAtherosclerosis and related cardiovascular diseases and risk factors can cause a wide array of simple and complex vascular lesions in the brain. For example, bilateral carotid occlusion directly affects the blood supply to the brain, and is commonly associated with stroke and vascular dementia . Some ofH1 (CD4+) lymphocytes are also involved in the inflammatory response [H1 cells bind to antigen sites on LDL molecules and subsequently produce inflammatory mediators including tumour necrosis factor (TNF) and interferons e.g. IFNγ [Lipohyalinosis is a brain-specific vessel disease characterised by vascular wall disruption and occurs as a result of systemic atherosclerosis, especially due to extensive inflammation occurring through the accumulation of ROS, in addition to gliosis involving astrocytes, oligodendrocyte precursor cells and microglia which themselves in their reactive state produce ROS . As discresponse , 81. TH1.g. IFNγ , which a.g. IFNγ . This st.g. IFNγ . Recent .g. IFNγ , 84. Cli.g. IFNγ .Vascular dysfunction caused by atherosclerosis and lifestyle/genetic risk factors can directly lead to cerebral vascular damage to cause small vessel disease and a breakdown of the BBB, marked by inflammation and hyper-connectivity . This va−/− and LDLR−/− knockout mice can develop hypercholesterolemia and atherosclerosis and disease severity and progression is exacerbated by being fed a high-fat diet [−/− mice also develop BBB leakage which worsens over time, and may contribute to a breakdown of neurovascular coupling [LDL molecules comprise of an apolipoprotein B-100 protein, in addition to other lipids (including up to 1500 cholesterol molecules), triglycerides and structural protein complexes . LDLs bifat diet , 90. Apocoupling , 92.DY) mouse model  has been used to study atherosclerosis [DY is sufficient to cause long-term atherosclerosis in mice comparable to LDLR−/− strains, without the need to create a transgenic mutant knock-in line [2) by causing their internalisation and degradation within a cell [D374Y mutation, plasma concentrations of cholesterol can exceed 500 mg/dL, where normal levels would be less than 200 mg/dL and pathological being above 250 mg/dL [DY mice, much like the LDLR−/− strains, however normal rodent chow can also result in moderate atherogenesis [−/− mice, and having the added benefit of not maintaining knockout lines, the rAAV-PCSK9DY model is certainly the more economical and more effective model in terms of induction of atherosclerosis as well as a comparable LDL-C profile to that of patients.More recently, an rAAV8 D377Y-mPCSK9  pathology in patients , as wellDY method has many benefits over the conventional knock-out lines of ApoE−/− and LDLR−/−, including the ability to induce atherosclerosis in any mouse model including AD-mouse models. In future studies, tracking haemodynamic changes over time combined with cellular, genetic and in vivo imaging would provide a comprehensive overview and mechanistic insight into how cardiovascular disease affects neurovascular coupling and the brain as a whole in mouse models of atherosclerosis and mixed models of dementia.Neurovascular coupling is critical to the proper functioning of the brain, ensuring that increased neural activity, which itself is incredibly metabolically demanding, is matched with an appropriate change to CBF to both nourish the parenchyma with oxygen and glucose as well as the quick and efficient removal of carbon dioxide and waste products. In order to achieve this, neurons communicate with endothelial cells via the neurogliovascular signalling pathways within the NVU. Pathological changes to the NVU and endothelia impairs the signalling pathways involved in neurovascular coupling, leading to brain pathologies ranging from subtle cognitive deficits to severe AD. Although our knowledge and understanding of the mechanisms behind neurovascular coupling in health, ageing and disease has vastly improved in recent years, much is still to be investigated, especially with respect to the effect of atherosclerosis on cerebral vasculature and neurovascular coupling. Although there are many established models of atherosclerosis, the new rAAV-PCSK9


Chloris barbata, which is a common invasive weed in Thailand, as a feedstock for bioethanol production. Chloris barbata was exposed to autoclave-assisted alkaline pretreatment by using different sodium hydroxide (NaOH) concentrations (1% to 4%) and heat intensities (110 °C to 130 °C) that were dissipated from autoclaving. The optimum condition for pretreatment was determined to be 2% NaOH at 110 °C for 60 min. At this condition, maximum hydrolysis efficiency (90.0%) and glucose recovery (30.7%), as compared to those of raw C. barbata , were observed. Evaluation of glucose production from 1000 g of C. barbata based on material balance analysis revealed an estimated yield of 304 g after pretreatment at the optimum condition when compared to that of raw C. barbata (61 g), an increase of five-fold. Structural analysis by the scanning electron microscopy (SEM) and X-ray diffraction (XRD) revealed the disruption of the intact structure of C. barbata and an increase in the cellulose crystallinity index (CrI), respectively. The results from this study demonstrate the efficiency of using C. barbata as a potential feedstock for bioethanol production.Developing an optimum pretreatment condition to enhance glucose recovery assessed the potential of  Sida acuta, Achyranthes aspera [Eichhornia crasspies, Arundo donax, Lantana camara, Saccharum spontaneum, and Mikania mikrantha [Chloris barbata was identified as one of the most common weed species growing in the area with the potential as feedstock for bioethanol production [Invasive weeds have been identified as a major environmental and socioeconomic problem due to their ability to infest large areas of arable and non-arable lands, as well as water bodies . Howevers aspera , Eichhorikrantha  have beeoduction . This weoduction . Neverthoduction .Phyllostachys edulis [Bioconversion of weed and other lignocellulosic biomass to bioethanol and other high value chemicals involves three main steps, namely; pretreatment, enzymatic hydrolysis, and fermentation . The pres edulis , sugarcas edulis , rice hus edulis , and rics edulis . C. barbata was established by analyzing the effects of different concentrations of NaOH solutions and autoclave temperatures on hydrolysis efficiency and glucose recovery. Scanning electron microscope (SEM) and X-ray diffraction (XRD) were used to analyze morphological changes in the raw and pretreated C. barbata.One of the most important chemical pretreatment factors is the supply of heat to facilitate the destruction of bonds between molecules during chemical action . DiffereC. barbata that was used in this study was sampled from Phitsanulok Province, Thailand. The weed biomass was chopped into pieces and then dried in the open air for one week. The dried samples were then milled in a SM 100, Rtsch, Rheinis-che StraBe 36-D-42781  wood miller and passed through 150–300 µm screen. The sieved biomass was stored in tightly closed plastic bags at an ambient temperature until use. The The National Renewable Energy Laboratory  analytical procedures were used to analyze the chemical composition of the weed biomass before and after pretreatment. The structural carbohydrates, acid insoluble lignin (AIL), and acid soluble lignin (ASL) were determined while following NREL protocol . Acid soChloris barbata (3 g) was added to 24 mL of NaOH in a 125 mL Erlenmeyer flask, mixed well, and tightly sealed with aluminum foil to avoid moisture loss, before heating in the autoclave  for 60 min. A two-stage pretreatment process was carried out by varying one factor at a time. In the first stage, the optimum NaOH concentration(s) was determined by pretreating the biomass with 1% (0.25 mol L−1), 2% (0.5 mol L−1), 3% (0.75 mol L−1), and 4% (0.1 mol L−1) w/v NaOH solutions at a fixed temperature of 120 °C. The optimum pretreatment temperature(s) was then analyzed in the second stage by subjecting the biomass to 2% NaOH solution and then autoclaving at the temperatures of 110 °C, 120 °C, and 130 °C. At the end of the pretreatment process, the slurry was rapidly ice-cooled and then filtered. The solid portion was washed thoroughly with deionized water to attain a neutral pH. The washed autoclave-assisted NaOH pretreated biomass samples were used to further analysis. Autoclave-assisted alkaline pretreatment was carried out while using NaOH as a catalyst. C. barbata was carried out as described in a previous study [C. barbata were hydrolyzed in a 50 mL Erlenmeyer flask with 0.1 g biomass (dry weight) in 10 mL digestion solution. The digestion solution contained 0.05 mol L−1 sodium citrate buffer (pH 4.8), 2% (0.3 mol L−1) w/v sodium azide, and an enzyme mixture of 30 filter paper units (FPU) of cellulase  plus 60 U β-glucosidase  per gram of dry biomass. The reaction was performed at 50 °C for 72 h in a rotary shaker  at 150 rpm (revolution per minute). The hydrolysates were sampled at 12 h, 24 h, 48 h, and 72 h to analyze the concentration of glucose. The enzymatic hydrolysis efficiency and glucose recovery were calculated by Equations (1) and (2), respectively: Enzymatic hydrolysis of us study . The raw is 1.11 . C. barbata was analyzed using scanning electron microscope  at 500× magnification and a beam accelerating voltage of 20 kV. All of the samples were freeze dried and then mounted on aluminum stubs. They were then sputter-coated with gold and observed under the SEM. The representative images were photographed in the study. The effect of autoclave-assisted alkaline pretreatment on the surface morphology of C. barbata. The biomass samples were washed thrice with acetone and then dried at 32 °C. The dried samples were ground to the size of 150 µm mesh screen and scanned at the rate of 0.02° s−1 from 2θ = 10° to 40°. The CrI was calculated following the relationship below (3): I002 and Iam represent the maximum intensity peak at 2θ = 22.6° and the minimum intensity peak at 2θ = 18.6°, respectively. X-ray diffraction (XRD) analysis using PANalytical X’pert Pro, PW 3040/60 Diffractometer  characterized the crystalline structure of cellulose in raw and pretreated All of the analyses were carried out in triplicate. The experimental design was completely randomized in two stages. The first stage  comprised of five experimental conditions of a single factor (NaOH concentration) with four levels  plus the control (untreated biomass). The second stage  was made up of four experimental conditions of one factor (temperature) with three levels , in addition to the untreated biomass control. The data collected were subjected to one-way analysis of variance (ANOVA) using SPSS version 17.0 . Multiply comparison of treatment means was performed with Tukey’s test at a 5% significance level. The average values were presented with their standard deviations. C. barbata (dry weight) is mainly composed of lignin and carbohydrates , xylan (17.23%), and galactan (1.35%), accounted for approximately 55% of the dry biomass. However, mannose and arabinose were not detected. The total lignin fraction, that is AIL (14.78%) and ASL (4.89%), formed about 20% of the total raw biomass, while other minor components, such as ash and ethanol extractives, accounted for nearly 8% and 7%, respectively. The glucan content of C. barbata in this study is in agreement with that of other previously studied invasive weed species, such as; Chromolaena odorata (36.2%), Pennisetum purpureum (32.0%) [A. aspera (45.9%), and S. acuta (46.9%) [Chemical composition analysis revealed that hydrates . The car (32.0%) , A. aspe, xylan 1.23%, and (46.9%) . Besides (46.9%) . C. barbata was evaluated with great emphasis on maximum glucose recovery, in addition to improving enzymatic hydrolysis efficiency. The results (p = 0.0001) decrease in the total lignin content, from 19.67% (raw biomass) to 6.68% (4% NaOH-pretreated biomass), representing approximately 87% lignin removal (p = 0.0001) increased as the NaOH concentration was increased from 1% to 4% (Pennisetum hybrid (P. americanum × P. purpureum) after pretreatment with the higher NaOH concentration (8%) and autoclave temperature (121 °C). Phitsuwan [P. purpureum with lesser NaOH concentration (2%) and a higher autoclave temperature of 121 °C for 60 min. However, considerably higher lignin content (92.2%) was removed after Lv et al. [C. barbata. The efficiency of different NaOH concentrations for the pretreatment of  results  showed a removal . Lignin 1% to 4% , which ihitsuwan  also extv et al.  pretreatp = 0.0001) reduced (p = 0.0001) when the NaOH concentration was increased, reaching a maximum of around 80% (p = 0.0001) increase in the glucan content (p = 0.0001) increased as the NaOH concentration was increased from 1% to 3%, beyond which there was no significant (p = 0.985) increase. The solubilization of lignin and xylan during alkaline pretreatment destroys the complex structure of lignocellulosic biomass and exposes cellulose fibers [p = 0.236) different from that (about 77%) recovered after 2% NaOH pretreatment. However, there was a highly significant (p = 0.0001) increase in glucan solubilization after pretreatment with NaOH concentrations of above 2%  improved relative to raw biomass. Maximum hydrolysis efficiency and glucose recovery were obtained after 72 h (p = 0.6398) different from that (80.63%) of the 3% NaOH-pretreated biomass, which was also not different (p = 0.8595) from that (79.64%) obtained after pretreatment with 2% NaOH concentration. The glucose recovery on the other hand significantly (p = 0.0001) increased, reaching a maximum of 24.96% after pretreatment with 2% NaOH concentration. Further increase in the NaOH concentration above 2% lead to a highly significant (p = 0.0001) decrease in the glucose recovery. It can be observed from this study that high hydrolysis efficiency did not lead to maximum glucose recovery  variation in the chemical composition of C. barbata. Changes in the autoclave temperature also had a highly significant (p = 0.0001) effect on the chemical composition. The lignin and xylan contents of C. barbata were substantially reduced after pretreatment at different temperatures. Lignin reduction at 110 °C was not significantly (p = 0.1635) different from that at 120 °C, but it was significantly (p = 0.0040) different from that at 130 °C. Nevertheless, its reduction at the temperatures of 120 °C and 130 °C were the same  different from that at 120 °C, but was significantly (p = 0.0040) different from that at 130 °C. However, at the temperatures of 120 °C and 130 °C, xylan reduction was the same  increased from 36.88% (raw biomass), reaching a maximum of 59.62% (120 °C). However, further increase in the autoclave temperature up to 130 °C, lead to a highly significant (p = 0.0001) decrease in the glucan content (55.53%) when compared to that of the other operating temperatures (110 °C and 120 °C), but not less that of the raw biomass. This may be due to glucan degradation at this high temperature. About 83%, 77%, and 67% glucan contents were recovered at the pretreatment temperatures of 110 °C, 120 °C, and 130 °C, respectively (p = 0.0001), from 15.15% (raw biomass) up to 90.03% (110 °C), after which there was a highly significant (p = 0.0001) decrease (p = 0.0001) high glucose recovery (30.70%) was obtained after pretreatment at 110 °C in comparison with that at the other temperatures and raw biomass. The optimum autoclave temperature for the pretreatment of r 60 min . Accordiectively . The totectively . Solid ldecrease . SimilarC. barbata in this study was determined to be 2% NaOH concentration at 110 °C for 60 min. Maximum glucose recovery of 30.7% was observed at this condition. The highest hydrolysis efficiency of 90.0% was also obtained at this condition, which is higher than that of other lignocellulosic biomass that was pretreated with NaOH, including rice straw (78.7%) [Phoenix canariensis (87.0%), Opuntia ficus-indic (84.5%) [S. acuta (82.8%) [C. barbata at the optimum pretreatment condition, among other factors, is enough to allow for cellulolytic enzymes to infiltrate the cell wall. Information about the optimal pretreatment condition(s) is very important in the effective conversion of lignocellulosic biomass to bioethanol [Therefore, the optimum pretreatment condition for  (78.7%) , Phoenix (82.8%) . It is aoethanol . C. barbata were analyzed to comprehend the structural alterations during the pretreatment process (C. barbata showed a thick smooth surface (C. barbata was more open exposing the reactive sites of cellulose microfibrils, which enhanced cellulase accessibility. The response of C. barbata to different NaOH concentrations and autoclave temperatures was different. In the case of NaOH concentrations, the linkages between the parallel strips of fibers were broken, causing the fiber bundles to loosen and separate (Morphological changes that were induced by autoclave-assisted alkaline pretreatment of  process . The raw surface a, unlike surface b–d, an iseparate b–e. The separate  caused aseparate b–d. FibeC. barbata during the pretreatment process were monitored while using XRD. Two main diffraction peaks of around 2θ = 15.3° and 22.6°, representing cellulose II and I, respectively, were identified (Cellulose is composed of intra- and inter-molecular hydrogen bonding, which gives it a crystalline structure. Cellulose crystallinity is one of the major causes of biomass recalcitrance . An effeentified . An upsuentified . The CrIentified . It is nentified  contribuC. barbata into bioethanol. Based on 1000 g of C. barbata at the optimum pretreatment condition of 2% NaOH at an autoclave temperature of 110 °C for 60 min, approximately 304 g of glucose can be recovered after enzymatic hydrolysis at the efficiency of 90.03% (C. barbata after enzymatic hydrolysis at the efficiency of 15.15%. Glucose production from 1000 g of C. barbata is estimated to improve by approximately five-fold after pretreatment at the optimum condition in the current study. The material balance for the optimum pretreatment condition in the current study was performed to evaluate the overall process of converting f 90.03% . HoweverC. barbata, exposing the cellulose to enzymatic attack to enhance the hydrolysis efficiency and glucose production. Highest hydrolysis efficiency and glucose recovery were obtained at the optimum pretreatment condition of 2% NaOH at 110 °C for 60 min. At this pretreatment condition, glucose recovery was greatly improved by approximately five-fold when compared to the raw C. barbata. Based on the findings of this study, C. barbata can be confirmed as a potential feedstock for bioethanol production in Thailand.The synergistic effect of mild alkaline solution and autoclaving was able to effectively destroy the lignin-polysaccharide matrix in


The effect of NaOH solution on the formation of nanoparticles has been the subject of ongoing debate in selenium-based material research. In this project, the robust correlation between the mechanistic growth of zinc selenide/graphene oxide (ZnSe/GO) composite and the concentration of NaOH are elucidated. The ZnSe/GO composite was synthesized via microwave-assisted hydrothermal method and the concentrations of NaOH are controlled at 2 M, 3 M, 4 M, 5 M and 6 M. The XRD spectra show that the crystal phases of the samples exhibited a 100% purity of ZnSe when the concentration of sodium hydroxide (NaOH) was set at 4 M. The further increase of NaOH concentration leads to the formation of impurities. This result reflects the essential role of hydroxyl ions in modifying the purity state of ZnSe/GO composite. The optical band gap energy of ZnSe/GO composite also decreased from 2.68 eV to 2.64 eV when the concentration of NaOH increased from 2 M to 4 M. Therefore, it can be concluded that the optimum concentration of NaOH used in synthesizing ZnSe/GO composite is 4 M. This project provides an alternative green method in the formation of a high purity ZnSe/GO composite. Chalcogenide semiconductor nanoparticles have optical properties which are widely used in optoelectronic applications . Zinc seMicrowave technology has been developed dynamically in recent years. It has been used in the synthesis of materials and various applications ,15,16 suThere is limited research on the formation of graphene-based ZnSe materials using the microwave synthesis approach. Therefore, the idea of incorporating graphene in the ZnSe crystal matrix is investigated in this project. By mixing graphene oxide with a ZnSe nanoparticle, it can be observed that the graphene oxide layer can reduce the recombination of electron-electron hole pairs during the photon absorption process. It is believed that the lifetime of the photogenerated carriers in samples also improved along with their potential use in optoelectronic applications ,21. In aHowever, the research related to the effect of NaOH concentration on synthesizing ZnSe materials is yet to be fully explored. Ben Nasr et al. have reported the effect of the concentration of hydroxyl ions on the properties of ZnS thin films . MeanwhiThe chemicals used were sodium hydroxide , hydrazine hydrate (Fisher Scientific), diethanolamine  selenium , and zinc chloride . A graphene oxide sheet was purchased from GO Advanced Solutions Sdn. Bhd.The previously reported method was used in this study . In detaIn this session, the experimental results, such as phase and structural, optical, and morphological results, are presented.2 against energy for all the samples. The data was analysed using the Kubelka–Munk approach. The Kubelka–Munk equation is stated as follows:g is the optical band gap energy. From the graph, the optical band gap energy was obtained by extrapolating the linear part of the plot to the baseline.3)2(H2O)4) (ICSD 98-003-3368) were detected. Moreover, the crystallite size of the sample was also calculated using Scherrer’s equation 2 against energy is presented in The optical band gap energy  particle (dash-border box); and (ii) wrinkle sheet (dot-border box). The particle size of the sample is around (45.8 ± 1.4) nm. The particle is referred to ZnSe while the wrinkled sheet is assigned to graphene oxide sheet. Thus, it can be concluded that ZnSe/GO composites are successfully synthesized using the microwave-assisted hydrothermal method where ZnSe is grown on top of the GO sheet, and confirmed by the Raman characterization.3)2(H2O)4.In conclusion, the optimum NaOH concentration used to synthesize high purity ZnSe/GO composite is 4 M. The XRD result shows that the samples possess the highest percentage of ZnSe. The experiment proved that the hydroxyl ions play an essential role in producing high purity ZnSe/GO composite. The optical band gap energy of ZnSe/GO composite is shifted from (2.68 ± 0.02) eV to (2.64 ± 0.02) eV when the concentration of NaOH increased from 2 M to 4 M. The slight change in the band gap is due to the growth of the crystallite size when a higher concentration of NaOH is used. It is confirmed that the concentration of NaOH at 5 M and 6 M is not suitable to be used as a solution for producing high purity ZnSe/GO nanocomposite. This is due to the formation of ZnO and Zn(HSeO


Hydrogenophaga, Acinetobacter, Dechloromonas, Comamonas, and Rhizobium/Agrobacterium species. In addition, the enriched microbiomes contained genes highly similar to the arsenite oxidase (aioA) gene of chemolithoautotrophic  and heterotrophic arsenite-oxidizing strains. The enriched cultures also contained aioA phylotypes not detected in the previous survey of uncultivated samples from the same wells. Anaerobic enrichments disclosed a wider diversity of arsenite oxidizing aioA phylotypes than did aerobic enrichments. The cultivatable chemolithoautotrophic and heterotrophic arsenite oxidizers are of great interest for future in or ex-situ arsenic bioremediation technologies for the detoxification of drinking water by oxidizing arsenite to arsenate that should then precipitates with iron oxides. The microbial activities required for such a technology seem present, amplifiable, diverse and hence robust.Millions of people worldwide are at risk of arsenic poisoning from their drinking water. In Bangladesh the problem extends to rural drinking water wells, where non-biological solutions are not feasible. In serial enrichment cultures of water from various Bangladesh drinking water wells, we found transfer-persistent arsenite oxidation activity under four conditions . This suggests that biological decontamination may help ameliorate the problem. The enriched microbial communities were phylogenetically at least as diverse as the unenriched communities: they contained a bonanza of 16S rRNA gene sequences. These related to  This is achieved by periodic injection of aerated water into the aquifers surrounding the drinking water wells  per liter of distilled water. The final arsenite concentration was 2.0 mM. The carbon for the autotrophic growth derived from the bicarbonate in the medium (see above) plus the 0.04 % CO2 in the air above it. The modified minimal salt medium for enriching aerobic arsenite-oxidizing microorganisms based on Santini et al.,  consiste3 was increased to 0.5 g/L and NH4Cl (0.3 g/L) instead of (NH4)2SO4 was used, while anaerobic conditions were employed. The final concentration of arsenite was 0.5 mM. In preliminary experiments we had used 0.2, 0.5 and 5 mM, where 0.5 mM appeared to be the best compromise between toxicity and substrate effect. The anaerobic heterotrophic arsenite oxidation (AHAO) conditions were created by supplementation with 3.0 mM acetate. The same medium as used for enriching aerobic arsenite oxidizers was employed, safe a few modifications. The amount of KNO4)2SO4, 0.4 KH2PO4, 0.18 MgSO4·7H2O, 0.875 NaCl and 0.2 MgCl2·6H2O . Am. AmaioA aioA amplicons  from all six positive anaerobic (autotrophic and heterotrophic) arsenite-oxidizing enrichments were cloned into Escherichia coli JM109 using the pGEM-T-vector system . Transformants were checked for correctly sized cloned inserts by a PCR with p-GEM-T-specific primers T7 and Sp6 . For anaerobic arsenite oxidizing enrichments 113 clones were screened, with on average 19 clones per enrichment. Based on differences in restriction profiles, the clones were classified into RFLP types. At least one representative clone per RFLP type and per enrichment was sequenced. The  and Sp6  31]. Pr. PraioA aioA  have been deposited in GenBank under accession numbers KU685236 to KU685320. Sequences were aligned and manually edited with ClustalW using default settings. Primer regions were removed in view of 12 sequence-ill-defined bases at the ends. Phylogenetic analyses were performed with MEGA 4 . A PoissQuantitative analysis of DGGE and RFLP profiles was performed with GelCompar II  . Similar−1dilution-growth iterations (corresponding to enrichments), even though arsenite oxidation potential was not abundant in the samples  microbial growth was observed down to a 10−3 sample dilution, and for eleven out of the twelve groundwater samples tested. However, arsenite oxidation as judged by the permanganate assay, occurred only in half of these enrichments  and electrons  (aioA) genes could be amplified in all these chemolithoautotrophic or heterotrophic, aerobic or anaerobic, arsenite-oxidizing enrichments, whenever arsenite oxidation had been observed  that did not seem to oxidize arsenite ] but aioA‘+’ in column HAO, Aerobic chemolithoautotrophic arsenite-oxidation (CAO) enabling media, inoculated with groundwater samples from any one of twenty-two drinking water wells, oxidized arsenite for all wells except one Mu in B. This wichments B, suggesSelection for a specific type of arsenic metabolism might be expected to lead to impoverishment of the ecosystem with only a few species remaining. We decided to examine this first by using a method that was not specific for arsenite metabolism: 16S rRNA-gene based DGGE analysis of 19 out of the 21 aerobic chemolithoautotrophic arsenite oxidizing (CAO) enrichments  showed phttps://www.bio.vu.nl/~microb/Protocols/DGGE/DGGEhelpV1.pdf, page 18). Performing a phylogenetic analysis on the results, we found that 64% of these were closely related to 16S rRNA gene sequences of Betaproteobacteria, 14% to Alpha- and Gamma-proteobacteria, some 5% to Bacteroidetes, and 1 band to an uncultured bacterium .We sequenced the 36 bands that dominated the DGGE gels a, as sucacterium . Among tHydrogenophaga strains C and NT-6 . This analysis confirmed considerable variation in ty level .aioA sequencing. We focused on these 9 out of the 21 enrichments because most band positions visible for the other 12 samples appeared to be covered by these nine enrichments. This phylogenetic analysis of aioA genes derived from aerobic chemolithoautotrophic enrichments was conducted together with the analysis of aioA genes derived from the aerobic heterotrophic (HAO) and anaerobic arsenite-oxidizing enrichments (AAO and AHAO) (see later sections). Based on an 85% amino acid sequence identity cut-off value, a total of ten distinct phylotypes were distinguished among 350 aioA sequences   enrichments derived from groundwater samples Mn-40.1, K1, Td, Uz, Ts, Mn-40.3, Gp, Sm and NA2 to cloning and chments) . The phybacteria .aioA sequences from aerobic chemolithoautotrophic cultures were most closely related to Alphaproteobacteria and belonged to either phylotype 1, 5 or 7, while the remaining 40% all belonged to the Betaproteobacteria: phylotype 8. For the rDNA—based tree   and chemoorganoheterotrophic [AHAO]) revealed a total of 7 different 3 clones . Alphaprp. DAO10  and Brad8 and 9; b. Phylotdefluvii b. Notabl Mn-40.1 . We had hypothesized a diverse range of active and cultivatable arsenite-oxidizing microorganisms to reside in arsenic-contaminated groundwaters in Bangladesh. We therefore deployed four different types of enrichment conditions (aerobic/anaerobic-chemolithoautotrophic and aerobic/anaerobic-heterotrophic) that may be relevant for arsenite oxidation by microorganisms as well as for the amplification thereof by microbial growth. As we had expected, nearly all wells housed arsenite oxidation activities that could be amplified in enrichment culture, suggesting that virtually all of the investigated wells offered a potential of cultivatable arsenite-oxidizing microorganisms. We found arsenite-oxidation related nucleic acids in addition to the ones already identified in our previous cultivation-independent 16S rRNA and functional gene-based study . These cAlpha-, Beta-, and Gamma-proteobacteria, whereas our previous cultivation-independent study had revealed few Beta- and Gamma-proteobacterial 16S rRNA gene sequences. And, we identified three aioA phylotypes  that we had not detected in our previous cultivation-independent study [It appears we are not looking at a single opportunity for such bioremediation, such as by a single (group of) species. The microbial community obtained differed widely between enrichments from different groundwater wells for the same culturing condition, as well as between different culturing conditions for any single groundwater well, as has also been revealed in other studies ,41. As ent study . Apparennt study  for arse3−, which we considered as an average for Bangladesh groundwaters. In reality the groundwater quality differs between locations . The present study reports considerable differences between the 16S rRNA-based classification and the aioA-based phylogeny. Such differences were also observed when classifying archaeal and bacterial diversity in an arsenic rich hydrothermal system [aioA genes than in terms of rRNA genes, or vice versa system [aioA but were still identical in terms of their rRNA genes, or vice versa. It is of course unclear whether for any particular case, this is actually an issue; sometimes it does not seem to be [The diversity what we observed here is a diversity of genes as well as of species. While we examined the molecular homology-based similarities between the l system . There ml system  quantifia system . Evolutiem to be .aioA genes [aioA trees to differ. And then there are more technical causes such as that not all aioA and rRNA genes for all species are reported in the databases, and the sequence stretch size used for the tree reconstructions was different. A second possible cause, i.e., horizontal gene transfer of oA genes , would aaioA). Yet, 5 out of 11 that did contain the aioA gene, still did not show arsenite oxidation  in enrichment medium, we first tried to grow them on semisolid agar using the same constituents as those of minimal salt enrichment medium containing As(III) and an inorganic carbon source. This failed however. Consequently, we neither isolated nor characterized individual strains in terms of their maximum arsenite oxidizing capacity for energy retrieval or for relief from growth inhibitory conditions. This suggests that the microorganisms we observed, may themselves not be capable of chemolithoautotrophic arsenite oxidation; only the observed co-metabolic communities may be. Phylogenetic analysis of or Japan . Nonetheor Japan ,53. Bose glucose . A simil glucose .Alkalilimnicola erhlichii MLHE-1 paradoxically contains an arxA [arrA than to aioA [arxA might however operate substantially in reverse  which could explain some or all of the arsenite oxidation capability of the strain [Arsenite oxidases (Aio) are involved in both autotrophic and heterotrophic arsenic oxidation under both aerobic and anaerobic conditions . The fac an arxA  that see to aioA ,55. Thise strain . Alternae strain ,39,57,58We conclude, first that in terms of arsenite oxidation there is diversity of all sorts in Bangladesh drinking water wells: (i) diversity in activities  and relevant nucleic acids, (ii) diversity between 16S rRNA gene-based and functional gene based phylotypes, and (iii) diversity within each well, diversity between wells and thereby also of the enriched microbial populations. Second, we conclude that there is considerable metabolic flexibility (mixotrophy) and adaptability at the population level  we identified  well are of interest, as they contain closely related if not identical CAO and HAO isolates  we found here  technology  introducin-situ or ex-situ, much attention should be paid to the anaerobic conditions, which could revamp arsenite from precipitated arsenate [Independent of whether biological arsenic remediation from Bangladesh groundwater through conversion to arsenate would be conducted arsenate . Keeping