The in-network caching capability of CCN ensures higher content availability, lesser network delay, and leads to server load reduction. It was observed that caching the contents on each intermediate node does not use the network resources efficiently. Hence, efficient content caching decisions are crucial to improve the Quality-of-Service (QoS) for the end-user devices and improved network performance. Towards this, a novel content caching scheme is proposed in this paper. The proposed scheme first clusters the network nodes based on the hop count and bandwidth parameters to reduce content redundancy and caching operations. Then, the scheme takes content placement decisions using the cluster information, content popularity, and the hop count parameters, where the caching probability improves as the content traversed toward the requester. Hence, using the proposed heuristics, the popular contents are placed near the edges of the network to achieve a high cache hit ratio. Once the cache becomes full, the scheme implements Least-Frequently-Used (LFU) replacement scheme to substitute the least accessed content in the network routers. Extensive simulations are conducted and the performance of the proposed scheme is investigated under different network parameters that demonstrate the superiority of the proposed strategy  The Internet is initially designed as a “collection of hosts” which is used to access available resources that are distributed in the network. The traditional TCP/IP Internet architecture supports the host-centric content retrieval mechanism, where the contents are accessed using the IP addresses of network nodes. The Internet has become a global infrastructure and with its tremendous growth in applications, the IP-based network traffic is estimated to be 4712 Exabytes per year at the end of 2022  impose iIn this context, the Content-Centric Networking (CCN) is proposed as a clean slate architecture for the future Internet . CCN supThe in-network content caching policy takes decisions related to the selection of suitable locations for the content placement and selection of older contents for replacement operations when the cache becomes full. These caching policies are generally categorized into on-path and off-path caching schemes . In on-pNetwork traffic and redundancy: The conventional on-path caching policy of CCN, called ubiquitous caching [ caching  allows eContent retrieval delay: Most of the existing on-path caching schemes takes autonomous caching decisions. Before forwarding the content to downstream nodes, each on-path router needs to perform certain computations for content caching decisions. This excessive computation for content caching becomes an obstruction in real-time content delivery and also causes excessive consumption of computational resources in the network routers. Therefore, it is essential to reduce the computational delay during caching decisions and the suitable contents need to be placed in appropriate network routers.A clustering-based in-network content caching scheme is proposed for the CCN to improve QoS for end-user devices and comprehensive use of cache space. By clustering the network nodes, the proposed scheme constrains excessive caching operations and content redundancy in the network.The proposed caching scheme considers content popularity and hop-count metrics along with the clusters information for the caching decisions. Using these heuristics, the caching probability increases for the frequently accessed contents near the end-user devices to reduce content access delay.The performance of the proposed caching scheme is examined through extensive simulations on the realistic network topology. Simulations results show the necessity of the proposed clustering-based caching scheme since the conventional scheme does not achieve a considerable hit rate in the network. Moreover, the proposed scheme demonstrates a significant decrease in the content retrieval delay and network traffic from the existing caching strategies.With these motivations, the objective of this paper is to propose an efficient content caching scheme that reduces the content retrieval delay and resource consumptions to offer improved network performance in CCN networks. Towards this, the proposed scheme provides two-folded content caching strategy. First, it partitioned the network nodes into the non-overlapping clusters using the topological information of the network. The clustering is performed to reduce content placement/replacement operations and to decrease computational latency in the network routers. During content retrieval, at most one copy of the incoming content is cached in that cluster from where the request is generated. The intermediate routers that do not belong to requester’s cluster in the path, cannot cache the forwarded contents. Hence, the computational latency is significantly reduced for the network routers. Secondly, to take caching decisions, the proposed scheme considers the content popularity and the hop count information to place popular contents near the end-user devices. When an intra-cluster router cache the incoming content, the remaining routers of that cluster just forward the content towards the requester without further caching operations. Thus, the proposed heuristics also control the excessive content redundancy and lead to comprehensive use of the caching capacities of the network. The major contributions of the paper are as follows:The organization of the remaining paper is as follows. The next section  providesThis section briefly describes the CCN architecture and its operations to provide the foundation for further discussions. As CCN is a data-centric network, the content retrieval mechanism relies on two types of messages: Interest message and Content message . The endOn receiving the Interest message from the end-user device, the network router first searches its CS for the requested content. If a cache hit occurs then the Content message is created by the router and forwarded towards the end-user device using the interface through which the Interest message arrived. If a cache miss occurs, then the router investigates its PIT. If a matching entry is found in PIT then the interface information of the incoming Interest message is aggregated in the PIT and the message is disposed from the network. Otherwise, a record is created in the PIT and the Interest message is forwarded towards the source using FIB.When an intermediate router receives a Content message, it checks its PIT for the matching records. If the entry is found then the router forwards the Content message toward those interfaces that are mentioned in the PIT and cache the Content message in its CS based on the content placement and replacement policies. After content forwarding, the router removes entries for that Content message from the PIT.In-network content caching is an inherent characteristic of CCN architecture that raises several challenges during content placement and replacement operations. To improve the network performance and QoS for the end-user devices, various content caching schemes are proposed by the research community ,26. The For content placement decisions, a random probability-based caching scheme called posed in . The schposed in  that drostrategy  approximstrategy ,27,28,29To increase cache hit probability on those routers that observe high network traffic, various centrality-based caching schemes are also proposed . A betweVarious researchers have also recommended considering the content popularity for caching decisions in the network. Towards this, in the Most-Popular Content Caching (MPC) scheme , each roHowever, most of the above discussed caching schemes ,28,29,34To alleviate the load im-balancing issues and reduction in excessive caching operations, several cluster-based caching schemes are also proposed in the CCN ,36,37,38Once the cache of the network routers becomes full, the older content needs to be evicted to cache the incoming content. Generally, this cache replacement operation is performed using the First-In-First-Out (FIFO), Least-recently Used (LRU), Least-Frequently-used (LFU), and optimal cache replacement strategies ,40. As dThe distinguishing features of the reviewed caching strategies are summarized in Therefore, a novel network clustering scheme is proposed in this paper for efficient use of the caching resources and improved QoS for the end-users. The proposed scheme considers hop-count and link bandwidth information to form tightly coupled clusters. Then, the proposed caching scheme jointly considers the clustering information, content popularity, and the content provider distance for caching decisions. With this, the popular contents are placed near the end-users with fairly multiplexed content redundancy in the path. This makes the proposed scheme suitable for CCN-based applications.V = {E denotes the set of connections that are used for the Interest/Content message forwarding among nodes in the network. ith end-user device and it generates Interest messages in the network. The ith router in the network and these routers perform Interest/Content message forwarding and caching operations. The notation Let To simplify further discussions, the notations used in the model are defined in As shown in For caching decisions, the content popularity and hop count metrics are determined using the following concepts:Content popularity determination using Popularity Table: According to the Zipf distribution, there are always few content requests for the unpopular contents in the network. If the caching scheme does not consider content access patterns during placement decisions, then the unpopular contents may be stored for longer durations in the network routers without being accessed again. This leads to poor use of network resources as cache miss probability increases due to caching of unpopular contents. Moreover, it has also been observed that the few routers with high importance receive more number of Interest messages as compared to other routers in the network. To resolve these issues, our previous work [ous work  has suggHop count monitoring: The hop count is a simple and effective metric to increase caching probability towards the edges of the network [ network ,34. The In this section, the proposed network clustering scheme is discussed in Algorithm 1: Proposed network clustering schemeInput: All the network routers Output: Set of “Sort the routers according to their decreasing order of degree centrality.Designate top “step-3(a), 3(b) and step-4, till there is a change in centroids:Iterate (a)Determine the distance between the routers (b)Assign each router Determine the new centroid Algorithm 1 shows the proposed clustering mechanism to form the clusters. The intra-cluster nodes collaborate with each other to take caching decisions without any additional communication overhead. In the proposed clustering strategy, initially the top “er works ,44 in tha). The distance between a centroid step-3(b)). It improves the efficiency of content forwarding from one node to another node within the clusters using higher bandwidth connections. After each iteration of the cluster formations, the router that has minimum distance  message Structure of Content message:…The name of the requested content is stored in the a)) and insert the requested content name b). Then, d) to (f) and elaborated in Algorithm 2: Interest message forwarding mechanism Then, any intermediate router (a)Update the value of (b)If (c)Algorithm 3: Content message forwarding and caching mechanism.If requested content exists in the (d)Else, if PIT of (e)Else, Search the FIB of (f)Else, discard In this section, the Interest message forwarding and processing mechanism are discussed and summarized in Algorithm 2 (Interest message forwarding mechanism). As shown in Algorithm 3: Content message forwarding and caching mechanism If requested content exists in the (a)Prepare a Content message (b)Replicate the values of (c)Initialize, (d)The content provider (e)Initialize the boolean field (f)Transmit When Update the value in If Else,(a)Compute, (b)Compute, (c)Cache Reset If d) to 1(e). The content provider then forward the message towards its requester This section elaborates Content message forwarding and caching mechanism which is summarized in Algorithm 3: (Content message forwarding and caching mechanism). When requested content is found in the cache of router step-4. Otherwise, if the Interest message is generated from the different cluster than the content provider then, following steps are performed. For caching decisions in a). Then, the b)). The normalized hop count is determined as the ratio of b), the In the proposed caching scheme, at most one copy of the content is cached in those routers b) of Algorithm 3 as follows:As discussed in c), the content would not be cached in Suppose, the value of In this case, the value of On the other side, if the content is cached in As the proposed scheme does not consider the router’s importance  during content placement decisions, the network load is not concentrated on a few network routers. Moreover, the proposed caching scheme does not require cluster heads for Interest/Content message forwarding and caching operations. Thus, the network traffic and computations are distributed among the network routers and the scheme does not suffer from the load balancing and bottleneck issues.This section first discusses the simulation environment and the values of its parameters. After this, the performance of the proposed caching scheme is evaluated in terms of the cache hit ratio, average network hop count, delay, and network traffic metrics. Then, the obtained results are compared with the peer caching schemes such as traditional caching strategy (LCE) , DC-baseThe ndnSIM simulation tool  is used The server er α=0.7 . The Intior work  suggestek”), the cache hit ratio has been computed with Before performance evaluations, the Abilene network topology is clustered into different number of non-overlapping clusters To determine the optimal threshold value A cache hit occurs when the incoming Interest message is satisfied using the cached copy from the network routers. Contrarily, if the requested content is not found in the CS of the router, then the cache miss happens. The network cache hit ratio (%) is the percentage ratio of the number of cache hits and the total number of Interest messages received by all the routers in the network. The increase in the cache hit ratio decreases the content retrieval delay and the load from servers. The cache hit ratio represents the effectiveness of caching scheme to reduce the redundant traffic in the network. The gain in cache hit ratio is computed as the difference between the average cache hit ratio achieved by the proposed scheme and the existing caching schemes.The number of hops traversed by the Interest message for cache hit  is defined as the hop-count to retrieve the requested content. The average hop count is computed as the average number of hops that are traversed to satisfy the Interest messages in the network. It is desired that the value of the average hop count should be smaller for improved QoS for end-user devices. The percentage of hop count reduction is computed using Equation  as mentiThe average network delay is determined as the total time (in microseconds) between preparing the Interest message and receiving the requested content. It also includes the request retransmission delay, if the content is not received within the defined duration. This metric represents the performance of the network from the perspective of end-user devices. The reduction in average network delay signifies improved network performance as the content is retrieved from the nearby routers.The average network traffic is computed as the total amount of data on network connections in per unit time and represented in terms of KB/s. This metric is used to examine the efficiency of the caching schemes and content transmissions in the network. The proposed clustering-based caching scheme does not flood the Interest messages in the network and supports efficient caching decisions using the network clusters, content popularity and distance parameters. Therefore, the network traffic is reduced for identical content transmissions and more diverse contents are accessed from the nearby devices. The percentage reduction in average network traffic is determined using Equation . In thisIt has also been observed that a direct correlation exists between the average traffic and the average network delay metrics. The smaller average network delay implies that the requested contents are found near the end-user devices and thus, a lesser number of hops are traversed to retrieve the content. This leads to decreased network traffic and increases the use of network resources. As the This paper starts with presenting various existing content placement schemes for the CCN environment in the literature. Then, a novel network clustering-based content caching scheme is proposed in which the intra-cluster routers cooperate with each other during content placement decisions. The proposed scheme considers the cluster information, content popularity, and hop count parameters to effectively use the available cache resources. In the proposed strategy, the network routers are clustered based on the joint consideration of hop count and the bandwidth parameters. Using the network clustering mechanism, the excessive cache replacement operations and the computational latency reduces significantly without additional communication overhead. Using proposed caching heuristics, the scheme increases the probability to cache the popular contents close to the end-user devices. Finally, the widespread simulations are performed with realistic network configurations and the performance of the proposed caching scheme is examined on cache hit ratio, average network hop count, network delay, and traffic metrics. The results showed that the proposed scheme outperforms the traditional CCN caching scheme along with peer heuristic-based DC-based, FGPC, PDC, and CPNDD caching strategies.In future works, the performance of the proposed strategy will be analyzed in mobility-based networks and the recent network topologies such as Geant, Tiger2, DTelekom and Internet2 etc. Additionally, more parameters can be integrated with the existing solution for further improvement in network performance.


The World Health Organization recommends increased male involvement to improve maternal and newborn health in low- and middle-income countries, but few studies have measured the impact of male-engagement interventions on targeted men. A trial of universal home visits to pregnant women and their spouses in Nigeria improved maternal and child health outcomes. This analysis examines the impact of the visits on male spouses.t test. Generalised linear mixed modelling accounted for the effect of socio-economic differences on the measured impact.In Toro Local Government Area in Bauchi State, Nigeria, we randomly allocated eight wards into four waves, beginning the intervention at one-year intervals. The intervention comprised two-monthly evidence-based home visits to discuss local risk factors for maternal and child health with all pregnant women and their male spouses. Measured secondary outcomes of the intervention in the men included knowledge about danger signs in pregnancy and childbirth, beliefs about heavy work in pregnancy, discussion with their wives about pregnancy and childbirth, knowledge about causes and intentions about management of childhood diarrhoea, and views about childhood immunisation. The analysis compared outcomes between men in visited wards (intervention group) and pre-intervention wards (control group), using a cluster The analysis included 6931 men in the intervention group and 9434 in the control group. More men in the intervention group knew four or more danger signs in pregnancy (risk difference (RD) = 0.186, 95% confidence interval (CI)  = 0.044 to 0.327), and three danger signs in childbirth , thought pregnant women should reduce heavy work before the third trimester , and had discussed pregnancy and childbirth with their spouse . More knew correct management of childhood diarrhoea with fluids and feeding  and less would give a child medicine to stop diarrhoea . Socio-economic differences did not explain the effect of the intervention on any of the outcomes.Universal home visits improved knowledge of male spouses about maternal and child health, which could contribute to improved maternal and child outcomes.http://www.isrctn.com/ISRCTN82954580ISRCTN, ISRCTN82954580. 11 August 2017. Retrospectively registered.  Maternal and child mortality remain high in many parts of the world ,2. The WA recent stepped wedge cluster randomized controlled trial of home visits to pregnant women and their spouses in Bauchi State, Nigeria  reportedThe conceptual framework underlying the universal home visits to pregnant women and their spouses explicitly recognised male involvement as crucial for household actions to improve maternal and child health. We hypothesized that visiting male spouses would improve their knowledge and attitudes and encourage them to act in support of maternal and child health. The analysis reported here has two objectives. First, we examined the impact of the home visits on male knowledge and attitudes, comparing visited wards (intervention group) with pre-visited wards (control group) in the stepped-wedge design. Second, within the intervention group, we examined changes in male knowledge and beliefs over the course of two or more visits, to look for associations between positive changes and characteristics of the men.Bauchi State in North Eastern Nigeria has a population of about 5 million based on projections from the 2006 census. Over 95% of the population are Muslim and polygamy is common. Some 63% of women and 44% of men in Bauchi have no formal education, compared with 35% and 22% nationally . NationaThe published trial protocol describes the overall trial design and methods in detail . The triAll households in the eight wards participated in the study. The visits were universal by design; an initial mapping in collaboration with community leaders identified all households in each community and all were eligible for visits. Female home visitors visited every household every two months and, on each visit, identified any new pregnancies. No households refused the visits, which were strongly endorsed by community leaders. Home visitors collected baseline socio-demographic characteristics of the households at the pre-intervention visits in wards in waves 2, 3 and 4 and in the first intervention visit in wave 1 wards. For the analysis reported here, the intervention group comprised spouses of pregnant women in the visited wards who were visited by a male home visitor at least once during the pregnancy and who provided data about their knowledge and attitudes in a visit soon after the birth of the child . The control group comprised spouses of women giving birth within the last 12 months or with a child aged 12-18 months at the baseline visit in the pre-intervention wards.The trial protocol describes the intervention in detail . The intThe key outcomes among male spouses were knowledge and attitudes relevant to the targeted maternal and child health outcomes. They included: knowledge of danger signs during pregnancy and childbirth; belief that a pregnant woman should reduce heavy work; discussion with their spouse about pregnancy and childbirth; knowledge of poor hygiene as a cause of childhood diarrhoea; knowledge about giving extra fluid and continued feeding for management of childhood diarrhoea; intention not to give anti-diarrhoea medicines to young children; perception of childhood immunization as worthwhile; and discussion with their spouse about immunization of children. To compare between men in the intervention group and men in the control group, we measured these outcomes on the latest visit prior to the one-year cut-off among visited men and in the baseline visit to households among control (pre-intervention) men.We calculated the trial sample size based on maternal outcomes . Since mWe selected eight out of 18 wards in Toro LGA as eligible for participation in the trial, excluding those where the security situation was too precarious. We divided these eight wards into two sets, geographically apart, and randomly selected four wards from each of these two sets, generating four pairs of wards. An epidemiologist not involved in the fieldwork (NA) randomly assigned the pairs of wards into four waves for implementation, using a computer-generated random sequence.The home visitors could not be unaware of the allocation of the intervention. We used standardised procedures to administer questionnaires and record responses in pre-intervention and intervention visits and the questionnaire used in both contexts was the same. The home visitors had no reason to conduct the process differently in intervention and pre-intervention visits. The households gave their consent to receive visits and to respond to questionnaires, but they were not aware that they were allocated to a treatment or control group.Analysis relied on CIETmap open-source software , which pt test, comparing outcomes between men in intervention and control groups. We report the risk difference (RD), relative risk reduction (RRR) and 95% confidence intervals (CI) of these parameters. To examine whether differences in socio-economic characteristics could explain the intervention effect, we used generalized linear mixed modelling (GLMM) [To assess the impact of home visits on male outcomes we used a cluster g (GLMM) . We inclA supplementary analysis examined changes in male knowledge and beliefs between their first and last visit, categorising these as negative to positive, negative to negative, positive to positive, and positive to negative. Among the subset with negative knowledge or beliefs on the first occasion, we compared those who shifted to positive with those who remained negative, and looked for associations between this “positive change” and baseline socio-economic characteristics of the men in a multivariate analysis , allowinWe followed the principles embodied in the Declaration of Helsinki to conduct the trial. We shared the study protocol and design in non-technical terms with the community, ward and LGA leadership and sought their approval before starting the trial. The home visitors sought informed consent from each household, woman and spouse to be visited and to respond to questionnaires. All responses recorded from participants were confidential and no names or identifying information were linked to the recorded data reaching the server.Data on male knowledge and attitudes were available among 16 365 men. In the intervention group these were 6931 male spouses of pregnant women visited during their pregnancies. In the control group these were 9434 male spouses of women who gave birth within the last 12 months or who had a child aged 12-18 months old. t test. Men in the intervention group had better knowledge about danger signs during pregnancy and childbirth than men in the control group. This was mainly apparent for knowing more than one danger sign, with significantly higher proportions of men in the intervention group knowing three or more danger signs in pregnancy and childbirth. In both the intervention and control groups, nearly all men said that women should reduce heavy work at some point during pregnancy. The proportion who thought women should reduce heavy work before the third trimester of pregnancy was significantly higher in the intervention group. Significantly higher proportions in the intervention group said they had discussed pregnancy and childbirth with their spouses or had discussed these issues with them often.t test. Beliefs about management of childhood diarrhoea were significantly better in the intervention group than in the control group: 30% more thought that children with diarrhoea should be given extra fluids and continued feeding and 21% more said they would not give a child medicine to stop diarrhoea. The perception of childhood immunization being worthwhile was nearly universal in both intervention and control groups, and nearly all in both groups said they had discussed childhood immunization with their family or spouse.Some 11% more men in the intervention group mentioned poor hygiene around the household as a cause of childhood diarrhoea, but this difference was not statistically significant in the cluster Further analysis examined whether baseline socio-economic differences between the intervention and control groups could explain the impact of the intervention on men’s knowledge and attitudes. The results of GLMM analysis for the effect of the intervention using a model including the intervention alone, and a model including the baseline socio-economic characteristics are shown in Table S1 in the Knowledge and beliefs among visited men shifted between first and last visits  the same local evidence about actionable upstream risk factors for maternal health: heavy work during pregnancy, domestic violence during pregnancy, lack of spousal discussion about pregnancy and childbirth, and lack of knowledge of danger signs during pregnancy and childbirth . We prevOur study expands the existing evidence base about the benefits of male involvement in reproductive health. The most commonly reported outcome of studies of male involvement is women’s use of maternal care services, such as antenatal care visits, skilled birth attendance, institutional delivery, and postnatal visits. Male involvement is itself often defined in terms of use of services. A recent systematic review of male involvement, defined as attendance to antenatal care with the partner, reported increased skilled birth attendance, increased institutional delivery and increased postpartum visits among women who attended antenatal care with a partner . Less thStudies often assess the impact of interventions on men indirectly. Studies of community interventions in India and Pakistan assessed the impact on the participating men indirectly, through women’s reports about the behaviour of their partners, such as attendance at antenatal care ,9. Our sIn our study, only 23% of men in the control group knew three or more danger signs in pregnancy and only 5% knew three danger signs in childbirth. This accords with a qualitative study in northern Nigeria which described low levels of knowledge about danger signs among men . Cross-sOur study was a randomized controlled trial that measured outcomes in men directly, increasing our confidence that measured differences were due to the intervention. We demonstrated a significant impact on men in the context of a trial that also found significant impacts on maternal and child health outcomes, and not simply services utilization. Repeated measures of male knowledge and attitudes during the intervention allowed us to examine changes in outcomes over several visits and to explore the association between these changes and individual characteristics.The trial had a small number of clusters (wards) in each group. We cannot exclude unmeasured imbalances between intervention and control clusters. This is less likely in the stepped-wedge design, where the wards in waves 2 and 3 were included in both their pre-intervention and intervention status.All visited households had visits to both pregnant women and their male spouses, so we cannot tell if visiting the male spouses added value to the visits to pregnant women, or even if visiting male spouses could have led to changes in maternal and child outcomes in the absence of visits to pregnant women themselves. Future research could examine which elements of the home visits were most important in improving maternal, child and male outcomes and measure sustainability of the male knowledge changes associated with the visits.Evidence-driven home visits to the male spouses of pregnant women increased their knowledge about maternal, newborn and early child health and associated local risk factors and increased their discussion with their wives about these issues. Together with evidence of impact of the visits on maternal and child health outcomes, this supports the idea that community efforts to improve maternal and child health must involve men.


Multiple myeloma (MM) is the second most commonly diagnosed hematological malignancy. Understanding the basic mechanisms of the metabolism in MM may lead to new therapies that benefit patients. We collected the gene expression profile data of GSE39754 and performed differential analysis. Furthermore, identify the candidate genes that affect the prognosis of the differentially expressed genes (DEGs) related to the metabolism. Enrichment analysis is used to identify the biological effects of candidate genes. Perform coexpression analysis on the verified DEGs. In addition, the candidate genes are used to cluster MM into different subtypes through consistent clustering. Use LASSO regression analysis to identify key genes, and use Cox regression analysis to evaluate the prognostic effects of key genes. Evaluation of immune cell infiltration in MM is by CIBERSORT. We identified 2821 DEGs, of which 348 genes were metabolic-related prognostic genes and were considered candidate genes. Enrichment analysis revealed that the candidate genes are mainly related to the proteasome, purine metabolism, and cysteine and methionine metabolism signaling pathways. According to the consensus clustering method, we identified the two subtypes of group 1 and group 2 that affect the prognosis of MM patients. Using the LASSO model, we have identified 10 key genes. The prognosis of the high-risk group identified by Cox regression analysis is worse than that of the low-risk group. Among them, PKLR has a greater impact on the prognosis of MM, and the prognosis of MM patients is poor when the expression is high. In addition, the level of immune cell infiltration in the high-risk group is higher than that in the low-risk group. In the summary, metabolism-related genes significantly affect the prognosis of MM patients through the metabolic process of MM patients. PKLR may be a prognostic risk factor for MM patients. Multiple myeloma (MM) is a hematological malignant tumor derived from abnormal monoclonal plasma cells. The main clinical symptoms include anemia, infection, bone destruction, and renal insufficiency . Due to The tumor is known for its special function of promoting the unlimited proliferation of tumor cells . Its parMetabolic changes are a common feature of most tumorigenesis. As a nonsolid tumor in the bone marrow, the metabolic process of MM has gradually been clarified with the deepening of research and the development of research methods. Moreover, changes in metabolic pathways also affect the efficacy of different treatment methods in MM. In this process, different targeted treatment methods are also being developed and applied. However, the diagnosis, treatment, and prognosis of MM are still trouble people. Therefore, this study intends to evaluate the impact of metabolism-related genes on the prognosis of MM patients by transcriptome sequencing and predict the risk factors of MM prognosis, in order to provide a theoretical basis for the treatment of MM patients and improve their prognosis and survival rate.| > 1 and P < 0.05. Genes associated with the metabolism were accessed in the MSigDB database.The gene expression profiles of GSE39754 were downloaded from the Gene Expression Omnibus (GEO) database. The GSE39754 included gene expression profiles of CD138 purified myeloma plasma cells from 170 newly diagnosed MM patients and 6 CD138 purified plasma cells from healthy donors. The data were analyzed with the Affymetrix package. The differentially expressed genes (DEGs) were calculated by limma package [The “survival” package of R was used to do the Cox single factor analysis, and then, metabolic-related genes affecting MM patient survival were then screened. Candidate genes were used to build a binomial least absolute shrinkage and selection operator regression (LASSO) model using the glmnet R package . MM sampThe ConsensusClusterPlus package was used to cluster the expression of candidate genes in MM samples. 170 samples of GSE39754 MM were divided into two subtypes (group 1 and group 2).P < 0.05.Gene Ontology (GO) and KEGG pathway analysis of candidate genes were carried out by enrichGO and enrichKEGG functions of clusterProfiler package. Terms in enriched GO and KEGG pathways were identified according to the cutoff criterion of The CIBERSORT was used to quantify the immune infiltration level of MM through the 22 immune cells. The limma package was used to calculate the difference between high and low-risk groups.To identify gene expression abnormalities of multiple myeloma, we identified differentially expressed genes between MM and controls in GSE39754. A total of 2821 differentially expressed genes (DEGs) were identified, including 1317 upregulated and 1504 downregulated expressions . SubsequTo identify the mechanism of action of candidate genes in MM, we performed an enrichment analysis. Results of GO showed that  candidatMM samples were divided into group 1 and group 2 by candidate genes using consensus clustering . Survivaλ) = 78 was selected, a model was developed based on a 78-gene signature in candidate genes (Figures To identify specific prognostic genes among the candidate genes, we performed LASSO regulation analysis. When log( Figures . The top Figures .To evaluate candidate genes, we utilized Cox regression analysis to divide the MM sample into high and low-risk groups by the median of the risk scores . There wThe method of CIBERSORT was applied to detect 22 immune cell infiltration levels in high-risk and low-risk groups of MM . DiffereIn this study, 348 metabolism-related prognostic genes were identified. Further studies showed that the candidate genes were mainly related to the proteasome, purine metabolism, and cysteine and methionine metabolism. In addition, the expression of PKLR was significantly different between the high-risk group and the low-risk group, suggesting that the expression of PKLR plays a regulatory role in the prognosis of MM.In the process of tumor, many kinds of cell processes changed in tumor cells of MM. MM tumor cells multiply rapidly and accelerate the tumor process, which will inevitably lead to changes in some traits in tumor cells. Genes are the individual units that regulate characteristics, and their expression levels also determine the differences in tumor cell characteristics. In addition, in order to meet the needs of cell proliferation in tumor cells, the changes in their metabolic levels must also be regulated by metabolism-related genes. The study of metabolism-related genes can enable us to better understand the occurrence of tumor cells and promote the treatment of tumors, which is of great significance. In this study, 2821 DEGs were identified, of which 576 DEGs are related to the metabolism. Research on these DEGs may have a certain effect on the treatment of MM.Further analysis of 576 DEGs in MM. We found that these DEGs are mainly enriched in the process of regulating the amino acid metabolism. Amino acid metabolism disorder is an important indicator of tumorigenesis . The uptDue to the abnormal chromosomes of tumor cells, the large and rapid proliferation will also lead to excessive protein synthesis . The selGlycolysis is a reaction system for tissue cells in the body to maintain normal physiological functions and provide energy . In the In summary, MM differentially expressed genes are mainly enriched in metabolic signaling pathways. Further research shows that PKLR can be used as a key marker for the prognosis of MM.


While toe fractures are the most common fractures of the foot in the pediatric population, the majority of these injuries do not require aggressive treatment. However, the mechanism of stubbing the great toe with bleeding at the base of the nail, a laceration proximal to the nail fold, or signs of a displaced fracture through the physis of the distal phalanx signal a likely open fracture involving the distal phalanx of the great toe. Unrecognized open fractures can lead to mistreatment and consequently osteomyelitis or growth disturbances. We report a case of a patient who required open reduction and Kirschner wire (K-wire) stabilization of a Salter-Harris I fracture involving the distal phalanx of the great toe after delayed recognition of the initial injury and subsequent failure to obtain closed reduction with a digital block in the clinic setting. We present this case to demonstrate the importance of vigilance in the evaluation of stubbed great toe injuries and propose an algorithm to guide the management of these injuries. Injuries involving a stubbed great toe in the pediatric population are often managed non-operatively with overall successful outcomes and few complications . HoweverThe importance of acute recognition and appropriate management of the open fracture is well established; however, there is scarce literature detailing consensus of management and stabilization of open Salter-Harris fractures of the distal phalanx of the great toe. Due to the intimate relationship of the epiphyseal growth plate and the germinal matrix, there is a risk for the interposition of the germinal matrix within the fracture, which could block reduction. These types of injuries have been closely compared to Seymour fractures of the terminal phalanx of the finger in which the treatment algorithm involves antibiotic administration, open reduction of the fracture, irrigation and debridement of the fracture site, nailbed repair, and possible Kirschner wire (K-wire) fixation across the distal interphalangeal joint (DIPJ) for maintenance of reduction . In avaiA healthy 13-year-old male presented to the orthopedic clinic five days after injuring his left great toe while running on the stairs. Initially, after the injury, bleeding was noted at the base of the nail which, lasted several hours, and he felt something hard “like bone” protruding from the wound. He was seen at an urgent care clinic the day following his injury, where radiographs of the left great toe were obtained, demonstrating a displaced Salter-Harris type I fracture of the distal phalanx of the great toe with dorsal displacement and angulation resulting in widening of the dorsal aspect of the physis  Figure . The naiThe patient was allowed to be weight-bearing as tolerated after surgery in a post-op shoe and instructed to complete the full course of cephalexin previously prescribed for a total course of 10 days. He followed up one week post-operatively, and his wounds appeared to be healing well without signs of infection. He was instructed to follow up at six weeks post-operatively for K-wire removal. He followed up three weeks post-operatively for evaluation after accidental removal of the pin at home. Radiographs at that time demonstrated appropriate healing of the fracture, and physical examination did not demonstrate concern for infection. He remained in a post-op shoe for protection until six weeks post-operatively. Final radiographs at that time demonstrated good healing with anatomic alignment, and he was able to transition into a regular shoe and activities without complication Figure . The patStubbing injuries of the great toe place the distal phalanx into forced hyperflexion and are often associated with physeal injuries of the distal phalanx. The consequence of a missed open physeal fracture of the great toe from a stubbed toe injury has been well described. Pinckney et al.  originalConsistent with our patient, many children sustain this injury while barefoot and report bleeding from the base of the nail after stubbing their toe. Kensinger et al. reported five pediatric cases in which each patient recalled bleeding from around the nail at the time of injury . In threOur patient presented for definitive management sub-acutely after having received initial treatment with oral antibiotics, irrigation with chlorhexidine scrub, and splinting in situ in an urgent care setting. After assessment of the injury in the clinic four days later, a reduction attempt was performed under a digital block, but the fracture was unable to be reduced. It was suspected that the germinal matrix was blocking the reduction, so given the nature of the open fracture, involvement of the physis, and amount of displacement, the decision was made to proceed with open reduction and pinning in the operating room. After addressing the soft tissues, the fracture was able to be successfully reduced and stabilized with a K-wire, which ultimately was accidentally removed at home three weeks post-operatively. At follow-up six weeks post-operatively, the patient was transitioning to regular activities without residual concern for infection and radiographs demonstrated maintenance of the fracture reduction. Ultimately, the patient’s fracture healed appropriately without concern for osteomyelitis or other complications as a consequence of his injury; however, the disorganized management of the patient’s open fracture from initial presentation to surgical treatment demonstrated the need for a clear treatment algorithm for open Salter-Harris fractures of the distal phalanx of the great toe Figure .There are few studies that describe open treatment for these injuries and scant documentation of indications for operative management of open and displaced Salter-Harris fractures of the distal phalanx of the great toe. Conversely, several studies have proposed algorithms for non-operative versus operative management of Seymour fractures, the hand counterpart to the stubbed great toe. Krusche-Mandl et al. analyzed the management of Seymour fractures and long-term follow-up results in a retrospective review of 24 patients treated operatively and conservatively . From thWhile a rare injury, it is important to recognize and appropriately treat patients who present with Salter-Harris fractures involving the distal phalanx of the great toe. We propose a treatment algorithm for the assessment and management of stubbed great toe injuries, which can help guide non-operative and operative treatment decisions Figure . We propAn assessment of the management of our patient’s open distal phalanx Salter-Harris fracture of the great toe highlights the importance of vigilance when evaluating suspicious injuries and provided the opportunity to develop a treatment algorithm to standardize efficient and effective care. We aim to decrease preventable adverse outcomes such as osteomyelitis and growth disturbance by creating a systematic method for approaching these injuries.


The Covid-19 pandemic has highlighted the longstanding under representation of ethnic minorities in clinical research – including migrant populations The NIHR has stated that Covid-19 research needs more ethnic minority participants to ensure the research is representative of the UK population. The inclusion of these groups in Covid-19 vaccine trials is particularly pertinent, with vaccine uptake being lower among ethnic minority groups than White people in the UK despite their increased risk Reasons for the underrepresentation of ethnic minority groups in Covid-19 research are poorly elucidated but are likely due to a combination of personal and structural factors. Socio-political factors may include social deprivation limiting access to health services, and subsequently, participation in – and awareness of – health research. Participant-related factors may include language and cultural barriers, and mistrust towards researchers and research institutions. Fear, mistrust and access barriers are the effects of powerful upstream factors including structural racism, marginalisation and ethnic exclusion. Previous studies have noted that a good researcher-participant relationship is a key facilitator of their participation Factors relating to research design and approach taken by researchers, including the omission of ethnicity from covariate data and non-inclusive recruitment strategies, are also likely to impact negatively on recruitment from these communities. As of 27th March 2021, only 1·43% of US-based studies investigating Covid-19 registered on Clinicaltrials.gov were collecting data about ethnicity, highlighting how this problem continues to go overlooked. Another review examining ethnic minority research participation found that the number of individuals from minority groups actually invited by researchers to participate in research was disproportionately small compared to their representation in the wider population In-depth cross-sectoral qualitative research is urgently needed to better understand the barriers hindering the involvement of people from ethnic minority communities in clinical research and develop robust solutions. Initiatives such as the INCLUDE project, which aims to promote the inclusion of underserved communities in clinical trials, are forming the basis for this much needed work and may provide fresh insight and useful action points The NHS Race & Health Observatory, which has responded to the specific health and wellbeing of ethnic groups in the UK, should now also focus on the equitable inclusion of these groups in Covid-19 clinical research and trials. Ultimately, we should ensure meaningful participation is placed at the heart of public health interventions research, with beneficiaries of the research included in every stage of the research pathway The Authors have nothing to disclose. The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care.


The medial septum (MS), as part of the basal forebrain, supports many physiological functions, from sensorimotor integration to cognition. With often reciprocal connections with a broad set of peers at all major divisions of the brain, the MS orchestrates oscillatory neuronal activities throughout the brain. These oscillations are critical in generating sensory and emotional salience, locomotion, maintaining mood, supporting innate anxiety, and governing learning and memory. Accumulating evidence points out that the physiological oscillations under septal influence are frequently disrupted or altered in pathological conditions. Therefore, the MS may be a potential target for treating neurological and psychiatric disorders with abnormal oscillations (oscillopathies) to restore healthy patterns or erase undesired ones. Recent studies have revealed that the patterned stimulation of the MS alleviates symptoms of epilepsy. We discuss here that stimulus timing is a critical determinant of treatment efficacy on multiple time scales. On-demand stimulation may dramatically reduce side effects by not interfering with normal physiological functions. A precise pattern-matched stimulation through adaptive timing governed by the ongoing oscillations is essential to effectively terminate pathological oscillations. The time-targeted strategy for the MS stimulation may provide an effective way of treating multiple disorders including Alzheimer’s disease, anxiety/fear, schizophrenia, and depression, as well as pain. We first describe the anatomy of the medial septum (MS) in section “Anatomy of the Medial Septum.” We then provide information on how the MS regulates oscillatory activities in the brain in section “Roles of the MS in Physiological Oscillations.” In section “The Medial Septum as a Target for Deep Brain Stimulation for Epilepsy Control and Beyond,” we discuss the possibility of the MS as a target of deep brain stimulation (DBS) for controlling oscillopathies .The septal region is conventionally split into four subregions based on anatomical location: the lateral, medial, posterior and ventral groups. Ample evidence stresses the importance of respecting the distinct nature of the septal region’s subregions. Unfortunately, however, many studies that investigate various septal areas refer to them by using the vague term “septum” and fail to precisely define the actual region within the scope of the study. It is particularly important to separate the medial and lateral septal nuclei because these two regions receive and send different modalities through their afferent and efferent fibers, occasionally to the same brain regions, and these modalities have distinct functional roles in information processing.In this review, we focus on the medial group referred to as the “medial septum (MS),” which consists of the medial septal nucleus and the diagonal band of Broca.The chemoarchitecture of the MS allows us to distinguish at least three major neuronal populations: cholinergic, GABAergic, and glutamatergic neurons .There are approximately 10,000 cholinergic neurons, containing the enzyme choline acetyltransferase (ChAT), in the rodent MS . They arGABAergic neurons in the MS, present at approximately half the number of the MS cholinergic neurons in rodents, express glutamic acid decarboxylase (GAD) . They arN-methyl-D-aspartate (NMDA) receptors  and are either projection or local neurons. Upon activation, MS glutamatergic neurons evoke strong and fast excitation of intermingled cholinergic and GABAergic neurons . In addieceptors . This eneceptors .Note that so far there is no consensus about the exact number and proportion of these neuronal populations of the MS. Particularly the ratio of GABAergic and cholinergic neurons is still in debate. On one hand, studies reported twice as much cholinergic neurons as GABAergic ones . On the In the subsequent subsections, we outline the long-range afferent and efferent connections of the MS neurons. In most cases these pathways consist of fibers operating with multiple neurotransmitters, thus we overview them structure by structure rather than focusing primarily on the types of neurotransmitters . Due to The anatomical and functional interplay between the MS and the hippocampal formation is very important in many cognitive functions, including learning and memory. The MS and the hippocampus (HPC) have reciprocal connections establishing a prominent contribution of the MS to HPC theta oscillations. The MS cholinergic, GABAergic, and glutamatergic neurons all project to the HPC cornu ammonis 1 region (CA1).Projecting axons of all the MS neurons enter the HPC via the fimbria/fornix. The cholinergic projections are nearly 65% of the total MS to HPC projections although the percentage greatly varies depending on the targeted subregion and neuronal type as mentioned above: larger on excitatory neurons than that on inhibitory neurons in CA1 region of the HPC . A lesioA subpopulation of MS GABAergic neurons that expresses PV and hyperpolarization-activated cyclic nucleotide-gated (HCN) channels presumably provides the theta rhythmic drive to the HPC . The sepN-Acetylaspartylglutamate-expressing MS neurons project to the dorsal HPC  . The sepwith ACh . The MS e layers .The MS receives many neuromodulatory afferents including cholinergic ones from the dorsolateral tegmental nucleus, adrenergic ones from the locus coeruleus, serotonergic ones from the raphe nuclei, dopaminergic ones from the ventral tegmental area (VTA), histaminergic ones from the hypothalamus, and GABAergic ones from the HPC and the lateral septum (LS) . The ascThe noradrenergic fibers, originating from the ventrolateral medulla (A1 cell group) and nucleus tractus solitarii (A2 cell group) of the brainstem, project to the MS and modulate septal gonadotropin hormone-releasing hormone (GnRH)-secreting neurons . The loc2A receptors, rather than a direct serotonergic influence of the HPC  of the hypothalamus was identified as a modulator/driver of the HPC theta rhythm generation during some behavioral tasks and during urethane anesthesia as well . NeuronsThe histaminergic neurons of the hypothalamus found in the tuberomammillary nucleus (TMN) innervate the GABAergic and MS cholinergic neurons with particularly dense axon terminals . Their aOther hypothalamic regions, such as the posterior hypothalamus and the nucleus pontis oralis also send afferents to the MS. These afferents are cholinergic and act primarily on the muscarinic receptors of the MS. Their electrical stimulation can evoke theta oscillations through the activation of the MS . DiencepA reciprocal connection between the MS and the nucleus incertus (NI) was proved by retrograde labeling of NI. These projections are passing through the MS  and may Tract-tracing experiments identified further afferents of the MS from the amygdala and the prefrontal cortex .The efferent connections of the lateral and medial parts of the MS are topographically organized. Regarding the hippocampal formation, the lateral part of the MS preferentially projects to the ventral parts of the subiculum, the HPC, the MEC and the LEC. On the other hand, the medial parts of the MS mainly project to the dorsal and ventral HPC, and the dorsolateral EC . The latInvestigation of the cholinergic system and projections from the pontomesencephalic tegmentum to the thalamus and basal ganglia revealed information about the septal efferent connections . Woolf eAlthough research interest regarding the efferent connections of the MS was biased toward the cholinergic system and its role in attention, the MS GABAergic and glutamatergic projections should not be neglected. A significant portion of the non-cholinergic fibers project to the thalamus, the hypothalamus and the brainstem. The cholinergic fibers targeting cortical areas are frequently coupled with GABAergic fibers, but cholinergic axons outnumber the GABAergic fibers in these bundles .A receptors, combined with a delayed inhibition through GABAB receptors. These septal fibers are under massive control in the medial habenula by endocannabinoid signaling, which is hypothesized to be important in anxiety and depression . The MS is considered to be a key structure in generating theta oscillations . They emnetworks . Howevernetworks . For exanetworks  and coolnetworks .in vivo injection of a H current blocker into the MS did indeed reduce discharge frequency of the PV/HCN MC GABAergic neurons and power of theta oscillations in the HPC  are episodes caused by highly synchronous excitation in the HPC, each of which consists of a single high-amplitude wave followed by a fast 110–250 Hz oscillatory event at the pyramidal cell layer . They ocIt is known that the majority of the MS neurons are inhibited during SPW–Rs, when the HPC neurons fire in a high probability . On the As we described in the previous sections, the MS governs physiological oscillatory brain activities, which are closely related to normal functions of the brain. In particular neurological and psychiatric disorders where normal oscillations are disrupted, the normal functions of the brain are also disrupted . If the In this section, we briefly review pathological changes of the MS in epilepsy and other oscillopathies, and possible MS-mediated intervention strategies for these oscillopathies. Note that the roles of MS in diseases discussed here are mainly based on results of experiments using animal models. Their validity for human disorders is uncertain and thus the proposed therapeutic strategies are hypothetical.Epilepsy is a neurological disorder characterized by an enduring predisposition to generate epileptic seizures . EpileptIn the healthy septo-hippocampal axis, the rhythms in the MS (LFP and unit firings) are very coherent and strongly coupled to the HPC, mainly in the theta frequency range (5–12 Hz). This coherent coupling is disrupted in epileptic conditions of animals. The amplitude of the theta oscillation in the septo-hippocampal axis is significantly reduced in animal models of TLE . This diEpileptic brains have at least two distinct stable oscillatory states: interictal (resting) and ictal (hypersynchronous) states . These sDuring the interictal state , theta rhythm stimulation of the MS can be suggested to reduce seizure susceptibility . The oriIt is noteworthy to mention that the MS stimulation might be employed to prevent development of epileptogenesis after for example traumatic brain injury . This is® System) in patients, although its stimulation target is not the MS   . The pret the MS . Howevereffects) .Once seizures have already developed  , the resWhen ictal episodes have finished, convulsions cease and the LFP traces become flat  . NormallThe DBS electrode in the MS could be used both for theta rhythm stimulation during the preictal  state and for seizure rhythm stimulation during the ictal state . The samThe MS stimulation has been shown to be effective in rodent models of TLE with and without obvious damages of HPC , which correspond to human epilepsy with and without sclerosis . In addiThe closed-loop on-demand brain stimulation technology has several advantages compared with conventional open-loop DBS: it can be more effective ; it can Alzheimer’s disease (AD) is a chronic neurodegenerative disease with well-defined neurological characteristics: amyloid beta plaques, neurofibrillary tangles, and neuronal loss . AD accoOscillatory disturbances in the brain have been characterized in AD patients . Disruptions of theta oscillations, gamma oscillations and theta–gamma cross-frequency phase–amplitude coupling are commonly observed in the HPC  and the Thus, it is possible to raise a hypothesis that cognitive disfunctions of AD are alleviated by restoring theta and gamma oscillations in the septo-hippocampal axis using DBS.To date, many preclinical studies have provided evidence that supports this hypothesis. For example, electrical stimulation of the MS (MS-DBS) improved the performance of MS cholinergic neuron-lesioned rats in the Morris water maze task , 2017. CFor human applications, the nucleus basalis of Meynert (NBM) and the fornix have already been investigated as DBS target in AD patients with promising outcomes, technical feasibility, and good tolerance . DBS of Although it is only speculative whether the MS is involved, gamma frequency sensory stimulation has been shown to effectively prevent AD pathology and to improve cognitive functions in animal models of AD . This fiChronic and exaggerated anxiety and fear are symptoms of some psychiatric disorders, including generalized anxiety disorder and post-traumatic stress disorders . It has As noted in section “Roles of the MS in Physiological Oscillations,” many animal studies have provided evidence that the MS is related to anxiety/fear generation and its regulation. For example, electrolytic lesion of the MS, which decreased AChE activity in the HPC, reduced anxiety of rats during successive alleys tests (innate anxiety) and reduced freezing in contextual conditioned fear (learned fear) . PharmacDBS has been investigated to alleviate various neurological and psychiatric disorders, including anxiety disorders . HoweverSchizophrenia is a severe psychiatric disorder characterized by positive symptoms  and negative symptoms  . Schizop2 receptors (neuroleptics) alleviate symptoms of schizophrenia patients, especially positive symptoms  induces schizophrenia-like psychosis in humans . The admsymptoms .There is accumulating evidence to suggest that the MS is involved in the schizophrenia-like phenotypes in animals. For example, sensorimotor gating deficit is evaluated as prepulse inhibition (PPI) and auditory sensory gating in rodent models, which are closely related to theta and gamma band oscillations in the septo-hippocampal axis . PsychoaIn the DA hypothesis of schizophrenia, the positive symptoms of schizophrenia are thought to be caused by hyperactivity of midbrain dopaminergic neurons, which is positively modulated by pyramidal neurons in the ventral HPC via NAc and the ventral pallidum (VP) . Brain iFor clinical application, Heath, a psychiatrist of Tulane University, performed initial studies of brain stimulation as a therapy of schizophrenia in the 1950s . His stuMajor depressive disorder (MDD) is a common and persistent mental illness with extreme sadness and low mood disproportionate to any possible causes . MDD lowRecent studies have suggested there are oscillatory disturbances in the limbic brain areas of MDD patients and rodent models of depression . The oscRecent advances of biological studies have shown that the oscillations in the septo-hippocampal axis are affected by depression and involved in its symptoms. For example, olfactory bulbectomy, a model of depression, decreased the number of cholinergic neurons in the MS . SystemiThe feeling caused by MS electrical stimulation has been reported in earlier studies of depression, epilepsy, schizophrenia, and refractory pain patients . Their rPain is an important function that alerts individuals to, for example, a tissue injury with nociception and unpleasant feelings. Pain normally disappears when the tissue injury is cured. However, if pain persists and becomes chronic, the chronic pain  significantly decreases the quality of life of patients. The tremendous pain of, for example, cancer patients with continuous tissue invasion should be properly controlled as well. Existing therapy, including analgesic drugs , cannot control every type of pain, including chronic and continuous pain. Therefore, DBS has been investigated for those treatment-resistant types of pain . The sepThe MS is a part of the pain system in the brain. The MS receives afferents from the nociceptive system/pathway  and an electrophysiological study showed that more than 50% neurons in the MS are activated by peripheral nociceptive stimulation . AnotherAccumulating evidence from rodent studies has implicated the MS in both processing and regulation of pain . For encA receptor agonist) or AMPA/NMDA antagonist reduced experimental neuropathic pain of mice (A receptors) suppressed formalin-induced licking and flinching  in anesthetized rats and cats . Those aIn humans, Heath and Mickle found that septal stimulation induced an immediate relief of chronic pain in patients . In 1967YT and AB developed the idea. YT, AN, LB, and QL prepared the figures. YT, AN, and LB wrote the original draft. MO, KM, and AB discussed and commented on the manuscript. All authors contributed to the article and approved the submitted version.AB and AJN are the owner of Amplipex Llc. and a shareholder of Neunos Ltd., Szeged, Hungary, manufacturers of signal-multiplexed neuronal amplifiers and neurostimulator devices. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Reoviridae virus family and are classified into G and P genotypes based on the outer capsid proteins VP7 and VP4, respectively. To date, 36 G types and 51 P types have been characterised from humans and varied animal species , which was identified from an unvaccinated 12 month old male who had been hospitalised for gastroenteritis in Zambia . MultiplLong-term surveillance is critical in understanding the trends in rotavirus genotype diversity and distribution, as overinterpretation of short-term fluctuations in genotype prevalence can be misleading with without a greater context of the natural, cyclic patterns in genotype replacement over time. This is especially critical when comparing trends in genotype distribution pre- and post-vaccine introduction. Genotype surveillance data in Australia and elsewhere has revealed changes in diversity, as well as temporal and geographic fluctuations over time following vaccine introduction ,26. FurtYandle et al. described changes in the burden of rotavirus disease and genotype distribution in Ireland following vaccine introduction . RotavirRotarix was introduced into the Fiji National Immunisation Program in 2012 and has reduced the burden of rotavirus disease and hospitalisations in children under 5 years of age. Ongoing rotavirus surveillance has been conducted in Fiji to investigate changes in genotype diversity, and Thomas et al. described patterns of rotavirus genotype diversity from 2005 to 2018 . Prior tLong-term surveillance pre- and post-vaccine introduction was also conducted at five sentinel sites in India between 2012 to 2020 by Varghese and colleagues . The RotJoão et al. also reported the prevalence of rotavirus genotypes, pre- (2012–2015) and post-vaccine (2016–2019) introduction in Mozambique . In the The constant alternations in rotavirus genotype diversity and prevalence within countries and globally highlights the importance of on-going epidemiological and molecular surveillance programs. The increasing detection of unusual zoonotic and reassortant strains emphasises the necessity of whole genome sequencing and detailed phylogenetic analysis. Concerns remain that widespread vaccine use may shape the diversity of rotavirus strains and that vaccine-escape variants may emerge in some settings. Understanding the long-term patterns of rotavirus genotype distribution and evolution is critical in order to assess any changes observed following vaccine introduction given the tendency for natural temporal and geographic fluctuations in the absence of vaccines.The continued surveillance and characterisation of rotavirus genotypes circulating in the vaccine era globally will provide important insights into epidemiology and strain diversity, ensuring the success of current and future vaccination programs. We express our sincere thanks to the authors and reviewers for their contribution to this very important topic. We also express our sincere gratitude to the Bill and Melinda Gates Foundation for providing funds to support the publication fees for the articles in this special edition.


Micro-organisms colonized the world before the multi-cellular organisms evolved. With the advent of microscopy, their existence became evident to the mankind and also the vast processes they regulate, that are in direct interest of the human beings. One such process that intrigued the researchers is the ability to grow in presence of toxic metals. The process seemed to be simple with the metal ions being sequestrated into the inclusion bodies or cell surfaces enabling the conversion into nontoxic nanostructures. However, the discovery of genome sequencing techniques highlighted the genetic makeup of these microbes as a quintessential aspect of these phenomena. The findings of metal resistance genes (MRG) in these microbes showed a rather complex regulation of these processes. Since most of these MRGs are plasmid encoded they can be transferred horizontally. With the discovery of nanoparticles and their many applications from polymer chemistry to drug delivery, the demand for innovative techniques of nanoparticle synthesis increased dramatically. It is now established that microbial synthesis of nanoparticles provides numerous advantages over the existing chemical methods. However, it is the explicit use of biotechnology, molecular biology, metabolic engineering, synthetic biology, and genetic engineering tools that revolutionized the world of microbial nanotechnology. Detailed study of the micro and even nanolevel assembly of microbial life also intrigued biologists and engineers to generate molecular motors that mimic bacterial flagellar motor. In this review, we highlight the importance and tremendous hidden potential of bio-engineering tools in exploiting the area of microbial nanoparticle synthesis. We also highlight the application oriented specific modulations that can be done in the stages involved in the synthesis of these nanoparticles. Finally, the role of these nanoparticles in the natural ecosystem is also addressed. Nanotechnology is the branch of science that supports the designing and manipulation of organic and inorganic matter to the nanoscale levels  or below the cell wall surface , and palladium (Pd) NPs of different size. Amorphous Se NPs (10–80 nm) are formed by Nostoc linckia by selenite reduction involving thiol groups of cyanobacterial proteins (3O4) particles using yeast cells. Saccharomyces cerevisae have been found to produce ~20 nm antimony trioxide (Sb2O3) and titanium dioxide (TiO2) NPs. Other examples include zinc oxide (ZnO) NPs by Aspergillus fumigatus and Candida albicans  and lithium (Li)] and extracellular locations  in crystalline as well as amorphous state at room temperature  was used to examine crystallites of 100–300 nm covering the periplasmic space of the bacteria  at pH 6. However, Au nanowires were produced at increasing concentrations of AuCl4 at the constant pH. It was observed that when pH changed to 4, both triangular and spherical shaped NPs were formed at diluted concentrations of salt , derived from specialized intracellular vesicles and water-soluble acidic polysaccharides, called coccoliths. Emiliania huxleyi, is a dominant coccolithophore used as a source of mineralized material for nanotechnology applications such as nano-fluidics  NPs have been found to reduce the content of chlorophyll in Chlorella spp. and Scenedesmus spp. (2) NPs affect the photosynthesis process in Scenedesmus spp., by reducing the content of chlorophyll a and b without affecting the carotenoid content (Production of reactive oxygen species (ROS) by NPs is another mechanism of toxicity as ROS oxidizes numerous biomolecules such as proteins, lipids, nucleic acids, and carbohydrates leading to loss of function . It alsomus spp. . It has  content .S. cerevisiae are shown to be useful in the photo catalytic degradation of azo dye used in textile industries  and 4-nitrophenol  was successfully used as a reduction system for the synthesis and biosorption of intracellular Ag and Se nanoparticles  to Ag NPs under illumination with fluorescent tubes at ambient temperature  using another isolate H. salifodinae BK18. The mechanism of NP synthesis is similar to the one involved in the synthesis of Ag NPs using BK3 isolate  homeostasis in hydrothermal ecosystems as greigite is the major catalysts for CO2 reduction (Sulfolobus tokadaii for the reduction of Pd (II) to Pd (0) NPs in a redox reaction (2O3) NPs has been achieved using Halobiforma sp. N1 with better properties suited for localized hyperthermia therapy used in cancer treatment , not observed in ferritin templates from other microbial systems  NPs rather than a mixture of different oxidation states of Au metal with unusually strong paramagnetic properties  has been an area of extensive research. Using the S-layer from operties . The fer systems . The samtivities . This properties .The ability of biological molecules such as proteins, viruses, and DNA to self-assemble in a solution is a promising approach for predesigned engineering nanostructures. These engineered nanostructures self-assemble to form functional nanoconstructs and are used in biomedicine field for the targeted drug delivery, vaccine preparation, gene therapy, bioimaging, tissue engineering, and specific cell targeting .Viruses are nanosized entities consisting of a capsid, a protein shell enclosing a viral genome . They reSalmonella typhimuriam bacteriophage P22 are being focused more as they are safe to be used in humans  and self-assemble to form stable polysheaths with potential applications in different fields of nanosciences , it was observed that these biofilms consists of a combination of P. aeruginosa, Inquilinus limosus, and Dolosigranulum pigrum. The combination of these species within the biofilms varied under stresses of oxygen concentration and presence of antibiotics (Cytophaga/Flexibacter/Bacteroides (CFB group), Planctomycetales, Spirochaetales, Clostridia, and green non-sulfur bacteria. The bacteria belonging to family Desulfobacteriaceae predominated the older and mature biofilms that are ZnS rich, whereas fresh and ZnS poor biofilms have a more diverse proportion of individual communities  niche in which single or multiple bacterial species can co-exist within the framework formed by the extra cellular polysaccharides (EPS) secreted by its habitants. EPS can consist of xanthan, dextran, succinoglycan, hyaluronic acid, alginate, and several other polymeric substances, depending on the type of microbes involved in biofilm formation. Xanthan gum is produced by acterium . Recent f carbon .P. aeruginosa were found between 2 and 600 times more resistant to the heavy metals such as copper (Cu), lead (Pb), and zinc (Zn) than free swimming cells  operon that is positively regulated by rcsA gene. Escherichia coli overexpressing rcsA cloned in pSB1A2 overproducing colonic acid based EPS was found to exhibit better resistance to the activity of Ag NPs. Addition of exogenous xanthan based EPS to bacteria that lacked rcsA also confers the similar advantage by increasing the aggregation of NPs and also reducing the surface area of bacterial cells exposed to NPs  to Pd (0) NPs was found to be affected by the nature of the electron donor and was significantly enhanced by the addition of external reducing agents  and 0.004% by weight of Se (Sulfur reducing bacteria (SRB) has been used for many years for bioremediation of metal polluted soils and water bodies. Majority of studies involving SRB have been performed using ter spp. . These bter spp. . The natter spp. . Synthesht of Se . Thus, bLeuconostoc mesenteroids T3 have better sensitivity and selectivity toward detection of cysteine in aqueous solutions  NPs and their orderly arrangement, and positioning inside the magnetosomes. The smaller operons, namely mamGFDC, mms6, and mamXY, play accessory roles in this process of biomineralization. Using a phage vector, a set of 29 genes from these four operons was stitched together in an expression cassette and introduced into photosynthetic bacterium Rhodospirillum rubrum. This study opened a new era of tailor made optimized synthesis of magnetic nanostructures in recombinant host of interest using synthetic biology approaches. Another study shows the importance of redox control mechanism along with carbon metabolism and iron supply in inducing synthetic bio magnetization in model organism S. cerevisiae. Screening of knockout mutants for inducing magnetization identified TCO89, a component of target of rapamycin complex 1 (TORC1) along with several genes for carbon metabolism in yeast as essential in inducing the bio magnetization  solution. This self-assembled biofilm is multilayered with a pore diameter of 150–200 nm and a depth of 15–20 nm. It consists of alternating units of M13 phage along with polydiallyldimethylammonium chloride (PDDA) formed by pulling in and out method. The role of PDDA is to reduce the surface roughness of M13 phage allowing, thus facilitating the smooth assembly of biofilm. Using genetic engineering tools, negative charge of the bacteriophage is increased by inducing the expression of glumatic acid. Using this biofilm, NaA layers . This meA layers .in vivo usage of this therapy due to the poor control over the replication and spread of phages post clearance of bacterial infection and also the chances of facilitating horizontal gene exchange via the process of transduction  are luminiscent NPs that has revolutionized the field of nanotechnology because of its unparalleled importance in biomedical fields and diagnostics. They are made up of semiconducting material that has unique electronic and optical characteristics and can also transfer electrons. A typical QD consists of a core made of semiconducting material, a shell made of metallic layer such as ZnS to enhance the optical properties and a cap to increase the solvent accessibility. Several types of QDs have been synthesized by now are made up of hetero nano-structures such as CdS, PbSe, CdTe, ZnS, and PbS. These QDs range in size from 1 to 10 nm and exhibit quantum effect owing to the quantum confinement.S. pombe and Candida glabrata. These species accumulated CdS crystallites in the presence of Cd salts. A short peptide was used to control the nucleation and the growth of these nanocrystals. The nanocrystals synthesized by this green approach were uniform in size (monodisperse) compared to those synthesized chemically  also cannot restore the reduced ability of yeast cells to synthesize CdSe nanocrystals. However, addition of exogenous glutathione restored this ability to a great extent  using the toxic solvents . By usint extent . This st. Therefore, in depth understanding of the factors that govern the size and shape of NPs is essential to achieve consistent homogeneity. Other parameters which need more attention are the drug loading and release capacity of these NPs.Beyond this review highlighting the advancements in the field of microbial nanotechnology, further research is essential for the widespread applications of nanomedicine while preserving the ecosystem. The concept of controlled drug release at target sites, bio distribution of these drugs and their intended effects at the tissue/cellular level, as well as theoretical mathematical prediction models yet need to be improved. Many studies in nanomedicine field focus on biomedical and formulation studies that appear in the early stages of biomedical applications. Therefore, it is essential to implement these approaches using de novo protein engineering and in silico techniques have developed rapidly and can play an important role in supramolecular nanomaterial engineering for specific applications. Several challenges remain, including the difficulty of synthesizing non-aggregating NPs that have coherent and desirable properties, a fundamental lack of understanding of how the physical properties of NPs affect their bio-distribution and orientation, and how these properties affect their interactions with the biological system at different levels from cell to the tissue and organ. Other state-of-the-art techniques, such as computational design, material genomes, and artificial intelligence, can be integrated to discover more effective and translational NPs based on bioengineering strategies.Nowadays, SARS, Ebola, and Covid-19, and repair mechanisms with complete external control mechanisms. This is not yet a reality and is still a future investigation that perhaps humanity can achieve in the very near future.With better control over nanomaterials synthesis, researchers can make the nanomaterial world more amazing. However, current synthesis technology remains bottleneck that prevents in-depth exploration of the properties and applications of nanomaterials. Therefore, there is a lot of scope for improvement. There has been great enthusiasm for the simple approach toward the development of nano-robots (and nano devices) acting in tissue diagnosis, drug delivery, combating deadly virus like However, along with its benefits, the potential risk of nanomedicine toward both humans and the environment also needs long-term studies. Since nanomedicine has revolutionized the field of drug discovery and its administration in biological systems, the need for the regulation over its usage in healthcare and environmental systems is also increasing. Therefore, the appropriate acute or chronic toxicity effects of new nanomaterials on humans and the environment should be analyzed appropriately. Finally, the disposal of such nanomaterials into environment should be dealt with strict guidelines and regulations.RD, TH, PT, and SK wrote the manuscript. SK and TH conceived the idea. SK compiled the manuscript. KS made the illustrations. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Widespread vaccination provides a means for countries to lift strict COVID-19 restrictions previously imposed to contain the spread of the disease. However, to date, Africa has secured enough COVID-19 vaccine doses for less than 5% of its population. With widespread vaccination not on the horizon for Africa, there is a strong emphasis on non-pharmaceutical interventions which include movement restrictions (lockdowns). This general COVID-19 pandemic response of imposing lockdowns, however, neglects to factor in non-fatal consequences leading to disruption socio-economic wellbeing of the society at large. The economy in most African countries can no longer sustain lockdown restrictions. Some studies have indicated that a hard lockdown statistical value of the extra lives saved would be dwarfed by its long-term cost. At the same time not responding to the threat of the pandemic will cost lives and disrupts the social fabric. This paper proffers ways to mitigate the both and advocate for better policymaking that addresses specific challenges in defined communities thus yield higher population welfare. Public health and economic impact of COVID-19 lockdowns: despite the low number of reported cases in Africa [n Africa , the effn Africa . In Rwann Africa . During n Africa .The closure of borders and businesses has disrupted economies in African states with a regional average of about 5% in public revenue losses and total merchandise exports contracting by about 17% . FurtherRoll out of COVID-19 vaccination programs: the roll-out of COVID-19 vaccine programs has brought a glimpse of hope, globally, as populations are expecting a shift in the trajectory of the pandemic and ease of lockdown restrictions as more people attain herd immunity. However, this glimmer of hope seems to be elusive to many in low-income countries particularly in Africa as they are far from realising the potential positive effect of a COVID-19 vaccine due to limited access and supply. As the vaccination campaigns roll out in other parts of the globe, Africa faces a mammoth task of ensuring the availability and accessibility of a safe, inclusive and sustainable distribution process that reaches frontline health workers, at-risk groups, and eventually all individuals to attain herd immunity [immunity . It was immunity . This eximmunity .Africa, with the huge informal economy, what are the potential strategies to ease lockdown restrictions? The impact of COVID-19 lockdown restrictions on the public health programs and economy underscores the need for urgent innovative strategies to ease restrictions while avoiding a resurgence of the disease. Widespread vaccination, a pathway exploited by richer countries as a means to ease restrictions through the attainment of herd immunity, is not on the horizon for Africa. In this perspective, we offer recommendations on how the continent can strategically ease the lockdowns that have slumped economic growth and disrupted other essential public health programs. a) Smart Lockdowns: the adoption of lockdowns as a standard response to the COVID-19 pandemic has caused socio-economic turmoil on the continent. Scientists have proposed intermittent lockdowns commonly referred to as “smart lockdown” to lessen the impact of general restrictive measures [measures . The smameasures . The pubb) Digital contact tracing and surveillance: the success of smart lockdown hinges on the implementation of robust surveillance mechanisms aided with the adoption of telehealth. Effective surveillance on the continent has already been achieved in Nigeria and South Africa making use of mobile network positioning data and application. In Nigeria, partnerships with mobile operators to develop contact tracing and surveillance application of known contacts through mobile positioning data significantly improved decision making, capacity and scope of contact tracing, and surveillance. We noted that collaborations between the government, mobile network operators, and technology companies were utilised for successful mobile positioning data tracing interventions for COVID-19 patients. Mobile applications such as the Vula platform in South Africa were also noted as effective in providing psychological support to health care workers attending to patients [patients . While hpatients . Grantedpatients .c) Adoption of alternative education platforms: education is a human right and the COVID-19 pandemic is threatening this right. Schools where applicable must be encouraged to adopt the use of digital technologies for education and promote home-schooling for the delivery of lessons. International support for digital education is forthcoming on different platforms including United Nations Educational, Scientific and Cultural Organization WhatsApp platform and Dzidzo Paden|Imfundwe´ndlini App for remote teaching [teaching . Also, steaching ,18. Wherd) COVID-19 vaccine passports: the vaccine rollout at the end of 2020 to fight the pandemic has provided a glimmer of hope to eliminate the virus. However, Africa and most low to middle-income countries are significantly constrained with access to and uptake of the vaccines [vaccines , but in vaccines . Emerginvaccines . Some covaccines . TherefoMany governments responded to the COVID-19 pandemic by imposing lockdown and restrictive measures to curb the spread of the disease. Consequently, there was remarkable disruption of ongoing public health programmes, social and economic activities. In light of the negative impact that lockdowns have had on the African continent, we recommend a shift in policy from the imposition of hard lockdowns to smart and targeted lockdowns on COVID-19 hotspots and allow normalcy in less-affected regions. There is an urgent need for the scale-up digital-based models of learning and service delivery to enable continuity of learning.