Frequency-modulated continuous wave radar techniques typically have inadequate angular resolutions due to the limited aperture sizes of antenna arrays in spite of employing multiple-input multiple-output (MIMO) techniques. Therefore, despite the existence of multiple objects, angularly close objects with similar distances and relative velocities are recognized as one single object. Autonomous driving requires the accurate recognition of road conditions. This requirement is one of the critical issues to be solved to distinguish significantly close objects. This paper proposes a technique referred to as an antenna element space pseudo-peak suppressing (APPS) method to resolve angularly close targets. The proposed APPS method aims to identify closely spaced objects on roads. These angularly close targets cause a single peak in a spatial spectrum obtained by a beamformer-based angle estimation. The APPS considers this single peak as pseudo. The APPS radar cancels this pseudo peak from the spatial spectrum. Then, the obtained residual received signal is analyzed. With these procedures, the APPS identifies the number of targets. The APPS also estimates the target angles. The proposed APPS is experimentally validated using a typical single-chip MIMO-based radar evaluation board with three transmit (TX) and four receive (RX) antennas. The experimental results confirm that the proposed APPS successfully resolves angularly close pseudo targets with an angle difference of approximately  Compared with LiDAR (light detection and ranging), cameras and optical sensors, the cost of millimeter-wave radar is low, since it only consists of an integrated circuit (IC) and printed antennas. Moreover, compared to traditional camera imaging systems, millimeter-wave radars are more effective in bad weather conditions (fog and rain) and nonline of sight (NLOS) target detection (curves on roads)4. Frequency-modulated continuous wave (FMCW) and fast chirp modulation (FCM) radars can be used to inherently measure ranges and relative velocities between a radar and targets6. In addition, these radars can be used to measure target angles if equipped with arrayed antennas and multiple-input multiple-output (MIMO) radar functions7. The abovementioned angle estimation ability is required to resolve multiple targets when these targets have almost the same ranges and relative velocities. In particular, the detection and characterization of multiple targets on a road will play significantly important roles in ADAS and autonomous driving applications.Because of increasing demand and interest in advanced driver assistance systems (ADASs) and autonomous driving systems, millimeter-wave radars are attracting much attention9. In these 4D radar systems, angularly close targets need to be individually resolved because the radars need to identify all the possible objects and obstacles on roads. To recognize targets, various kinds of higher resolution millimeter-wave radars have been studied. A beam switching method was introduced in10, a radar imaging and neural network were used in11 to characterize road surfaces, and wide aperture and cascading radar units were employed in13 and15, respectively. Leak mitigation has been considered in16. Human recognition has been investigated in17 and18. A calibration method and an indoor radar imaging system to suppress the influence of surrounding clutter have been shown in19 and20. However, all of these studies require large-scale hardware and complicated signal processing. For example,10 need additional circuitries for transmit antenna beam switching.14 and15 are much larger than a 3TX–4RX MIMO radar with a single radar IC.17 and18 use high computational complexity artificial intelligence (AI) algorithms.21 also uses a high computational complexity AI algorithm: convolutional neural network (CNN) for multi-target classification. Moreover, some classical radar signal processing techniques such as virtual antennas22, and multiple signal classification (MUSIC) algorithms23, are also adopted to the FMCW radar. However, beamformer-based angle estimations are typically used in practical automotive millimeter radars5, where these angle estimation methods are sometimes referred to as Angle Fast Fourier Transform (FFT). Maximum likelihood estimation is also a classic radar signal processing technique, but it has high computational complexity and cannot resolve extremely close targets24. Therefore, radar with low-complexity signal processing and small form factors is urgently needed.To equip various kinds of vehicles with millimeter-wave radars, including cars, tricycle, scooters, and bicycles, their production costs need to be low, and the form factors should be small. Furthermore, four-dimensional (4D) radar measuring ranges, relative velocities, azimuthal and elevation angles have been actively developed worldwideX–4RX MIMO radar with a single radar IC25. The proposed method is based on an antenna element space interference canceling (AIC) technique that was proposed in26 for automotive millimeter wave FMCW radars. The original proposed AIC technique has been previously introduced into multiple target radar imaging for suppressing the influence of side-lobes from large targets. We extend this method and refer to it as antenna element space pseudo peak suppression (APPS) in this paper. RELAX and CLEAN algorithms29 first introduced the signal canceling concept to array antenna processing. On the other hand, the proposed APPS for automotive millimeter-wave FMCW radars focuses on realizing the detection of significantly angularly close targets and higher angular resolution performance. These requirements are recent urgent problems to be solved in automotive FMCW radars. If two targets are significantly angularly close, an ordinary beamformer method observes a single peak in a spatial spectrum because of the inadequate aperture of the antenna array. The APPS processing considers this single peak as pseudo and cancels this pseudo peak in the antenna element space of the antenna array. The beamforming-based angle estimation is applied for the resulting input signal vector again to analyze the residual received signal component. With this procedure, the APPS radar system identifies the number of targets by the residual signal after canceling the pseudo peak. Furthermore, the APPS also estimates the angles of the targets. The proposed APPS is experimentally validated using a typical single-chip radar evaluation board, where a multiple-input multiple-output radar architecture is equipped with three transmit (TX) and four receive (RX) antennas. The experimental results confirm that the proposed APPS can successfully resolve ground-placed targets that have an angle difference of only To this end, this paper proposes a higher angular resolution technique by introducing an antenna element space pseudo peak suppressing method, which has extremely low signal processing cost and just use 3TAPPS is developed to resolve angularly extremely close targets, as shown in Fig. As previously stated, because FMCW radars can distinguish between multiple targets in terms of range and relative velocity, most targets can be resolved based on these inherent features of FMCW radars. Hence, the number of targets that need to be resolved by angle estimation only occurs when multiple targets have the same distance and velocity, and, thus, is very rare in reality. Therefore, the number of targets that require angle estimation is assumed to be at most two in this paper. An ordinary beamforming method cannot be used to distinguish targets when they are angularly close to each other. In this case, a pseudo peak appears in the spatial spectrum despite the presence of two targets. To overcome this problem, APPS is introduced to FMCW radar systems.26. AIC radar processing enables the provision of a replica received signal vector for the maximum peak angle of a spatial spectrum, which is generated by beamforming-based angle estimation.APPS processing is based on the antenna element space interference canceling radarN is the number of receive antennas, and Figure 26APPS is applied to each peak. A replica signal is generated for the maximum peak angle based on AIC radar processing, and then this replica is subtracted from the received signal vector. A coefficient for an incident plane wave for the pseudo peak, Figures The proposed angle estimation mechanism can be explained as follows: If the number of targets is one, the pseudo peak angle corresponds to the estimated angle. However, for two targets, the angles cannot be directly estimated. Therefore, we first obtain the relationship between the average residual levels for the spatial spectra and angle differences by computer simulation, as shown in Fig. Moreover, an antenna array with a smaller number of antenna elements widens a null width in a spatial spectrum, reducing the residual incident wave level. The obtained residual incident wave level must be higher than background noise to identify the presence of incident waves in APPS processing. Therefore, the number of antenna elements impacts APPS processing. Figure Figure Estimate a pseudo peak angle Create a replica signal vector for Apply the beamformer method for the received signal vector after subtracting the replica and obtain a spatial spectrum again,Assess the residual levels for the incident waves,Repeat (1) to (4) for multiple chirps or chirp frames to obtain an average residual signal level for the incident waves,Compare the obtained average residual signal level with a threshold determined based on computer simulations or experimental realizations, and estimate the number of incident waves: one or two,Let the incident angle be Based on these characteristics, if a residual component is observed in a spatial spectrum after replica subtraction, the presence of two incident waves is identified. In addition, because an average residual incident wave level is proportional to the difference in incident angle, this characteristic makes it possible to estimate the incident angle. The APPS scheme after obtaining a first spatial spectrum can be summarized as follows: A 79 GHz band MIMO radar-based FMCW/FCM millimeter-wave radar evaluation board, T14 produced by S-TAKAYA, was used in our experiments. This radar is equipped with three TX and four RX RF circuits and antennas, enabling a universal linear array antenna with an antenna element spacing that avoids the generation of grating lobes. The radar offered a 3 TX-4 RX MIMO function. This radar works as a receive antenna array with 12 elements. An inset in Fig. Figure Figures In addition, several experiments for detecting human and bicycle on an asphalt roadway were conducted. Figure Figure Figures For the single incident wave case shown in Fig. For the two incident waves shown in Fig. We also verify the performance of long-distance. Figures Moreover, Fig. For more practically validating the APPS processing, experiments for detecting plastic water bottles on an asphalt roadway were conducted. The plastic water bottles were placed on the asphalt roadway as on-road obstacles in Fig. The angle estimation performances were also experimentally evaluated, where the experimental environment is the same as that shown in Fig.Figure This paper proposes a method to estimate the number of targets in a field of view, i.e., one target or two extremely close targets, along with their respective angles using a typical single-chip millimeter-wave radar IC. Two extremely closely located targets with an angle difference of approximately


Bicyclusbutterflies within a phylogenetic framework, we inferred the group’s origin at theOligo-Miocene boundary from ancestors in the Congolian rainforests of central Africa.Abrupt climatic fluctuations during the Miocene (ca. 19–17 Ma) likelyfragmented ancestral populations, resulting in at least eight early-divergent lineages.Only one of these lineages appears to have diversified during the drastic climate andbiome changes of the early Miocene, radiating into the largest group of extant species.The other seven lineages diversified in forest ecosystems during the late Miocene andPleistocene when climatic conditions were more favorable—warmer and wetter. Our resultssuggest changing Neogene climate, uplift of eastern African orogens, and bioticinteractions have had different effects on the various subclades ofBicyclus, producing one of the most spectacular butterfly radiations inAfrica. Compared to other regions, the drivers of diversification in Africa are poorlyunderstood. We studied a radiation of insects with over 100 species occurring in a widerange of habitats across the Afrotropics to investigate the fundamental evolutionaryprocesses and geological events that generate and maintain patterns of species richness onthe continent. By investigating the evolutionary history of  Understanding the processes that promote diversification and their role in shaping currentspecies distributions is a central theme in evolutionary ecology . The manRecent advances in mechanistic modeling of macroevolution allow the relative effects ofintrinsic biotic interactions and extrinsic abiotic factors on diversification to be evaluatedby modeling diversification rates as a function of time, diversity, environmental changes, andcharacter states using time-calibrated phylogenies  butsee Bicyclus Kirby 1871, an African-endemic butterfly genus, is an ideal taxonfor evaluating the contributions of biotic and abiotic factors in the generation andmaintenance of Afrotropical biodiversity. With 102 currently recognized species   , and it Poaceae) . DiversiPoaceae) .ca. 14Ma . The included samples covered all16 currently recognized species-groups and 11 other closely related satyrinae taxa as outgroups, selected on the basis ofevolutionary relationships recovered in two earlier higher-level phylogenetic studies were included in this study .Supplementary AppendixS1 available on Dryad).Most sequences used in this study were obtained from Phylogenetic relationships were inferred under a maximum-likelihood (ML) framework withIQ-TREE 2.0.6 . The conWe assessed nodal support of recovered relationships using 1000 ultrafast bootstrapreplicates, UFBoot , and theWe estimated divergence times in a Bayesian framework using BEAST 1.10.4 . Similarce prior . To examce prior . The modBicyclus or closely related taxa, so werelied on secondary calibration points from the large-scale fossil-based dating frameworkof (Supplementary AppendixS2 available on Dryad).There are no known fossils of eworkof  inferredBEAST dating analyses were run for 50 million Markov chain Monte Carlo (MCMC) generationswith model parameters and tree sampled every 1000 generations, yielding a total of 50 000samples per analysis. Convergence and performance of the MCMC were assessed using Tracer1.7.1 based on the ESS threshold values of >200 for each parameter . FollowiBicyclus are frequently misidentified in books, databases, and museumcollections. Therefore, we only used data for which voucher specimens or photos wereavailable to verify the validity of individual records. When using literature records, weonly relied on sources for which we personally knew the identification skills of theauthor(s). We screened the collections of nine museums , and the personal collections of Oskar Brattström,Thomas Desloges, Szabolcs Sáfián, Robert Tropek, Robin van Velzen, and Robert Warren.Online sources that provided clear photos of specimens were also used—primarilyinaturalist.org and several Facebook groups. By supplementing these location data withinformation from two reliable published sources  model implemen sources , we extr sources .Supplementary AppendixS3a available on Dryad). In the second constrained model (M2), we estimatedthe scalar multiplier as Supplementary AppendixS3b available on Dryad). The maximum number of areas per ancestral state wasset to seven. Finally, to quantify the relative role of dispersal and vicariance ondiversification, we implemented 1000 stochastic biogeographic mappings using the best-fitDEC model. From these mappings we obtained the overall probabilities of the anagenetic andcladogenetic events conditioned on the geographic distributions, the phylogeny, and thebest-fit DEC model.We ran BioGeoBEARS analyses with three dispersal rate scalar matrices. The first analysis(M0) assumed that dispersal between any two geographic regions was equally probable andused a single dispersal rate scalar value throughout time. The other two dispersal ratescalar matrices presumed that dispersal events between adjacent areas were not scaled orpenalized  recapitulated the major taxonomic groups by applying a nonparametric method to ourtime-calibrated phylogeny . We compTo understand the relative contribution of intrinsic biotic interactions and extrinsichistorical factors, we characterized diversification dynamics of each clade by fitting aseries of macroevolutionary models to the data. To take into account uncertainty in treetopology and divergence time estimates, we fit all of the macroevolutionary models exceptthe density-dependent diversification models first to the Bayesian consensus MCC tree, andthen to 200 trees randomly sampled from the posterior distribution of the BEAST analysis(after a 50% burnin-in with outgroups removed). A sampling fraction of 0.92 was used forall analyses to take into account missing or unsampled taxa. We compared the fit of eachmodel to the data using the AIC for each analysis. The AIC scores also allow computationof ng model .Bicyclus and its componentclades.We assessed variation in species diversification over time by performing a time-dependentdiversification analyses using RPANDA . We eval<inline-formula><tex-math notation="LaTeX" id="ImEquation24"              ><![CDATA[$\quad$]]></tex-math></inline-formula>   evolution  to four diversification rateshifts during the evolutionary history of the group. For these analyses, we set the “grid”to 0.1 million years for a fine-scale estimation of rate shifts, the “end” as the crownage of the group and “start” as the present . We also relaxed the assumption of equal rates in the Unconstrained sixrates model in which all transitions could be different from one another. Inthe Constrained single rate model, we assumed that forest and openhabitats are the end points of a gradient and species cannot change from one habitat tothe other. Instead, lineages must transition through an intermediate state . In this model, all events happen at thesame rate. The Constrained four rates model is similar to the previousmodel because change in habitat preference requires transition through an intermediatehabitat state, but the transition rates are all different  tointermediate and then open habitat species will all be the same but differ from transitionrates in the opposite direction.Because habitat preferences are not static over time, we defined five models thatdescribe transitions across habitats. Under the Unconstrained single rate,Unconstrained six rates, Constrained single rate,Constrained four rates, Constrained specialist, andConstrained openness) and ran ML optimizations using three differentstarting points for each analysis to prevent finding only local optima. Because the numberof free parameters varied across models, we compared their AICc scores. With theparameters that maximize the likelihood of the best supported model, we obtained theprobability of habitat preference state at each internal node. This procedure is similarto ancestral state reconstruction, but a more conceptually accurate interpretation wouldbe that the probability of habitat preference at a given node reflects the probability ofthat node producing descendant lineages with their observed (or inferred) habitatpreferences given the model and its parameters. To further validate these SecSSE models,we used the speciation rates and transition matrix parameters of the best performing CTDand ETD models to simulate 30 data sets of phylogenetic trees for extant species withcorresponding habitat preferences. Simulated time (crown age) was adjusted such thatsimulated trees were similar in size to the empirical one. We then fitted SecSSE CTD andETD models to the simulated data sets in a manner similar to the main analysis.Afterwards, we compared the loglikelihood values of both models to find the number ofsimulations where the analysis chooses the right model. In other words, we counted thenumber of ETD-simulated data sets in which SecSSE found ETD as the best performing model.Similarly, we counted the number of cases where ETD was (erroneously) selected as the mostlikely model when in fact CTD was the generating model.We set up 36 models that combine trait dependence , speciation , and habitat evolution  diversified under a differentdiversity-dependent scenario by decoupling its dynamics from Clade I in key innovation(KI) models  using thncorrect . The proFinally, to assess the relative contribution of time-specific global events includinghistoric changes in climate, biotic interactions among conspecifics, and time-dependentprocesses on diversification, we compared the AIC scores of the best-fit models of eachmacroevolutionary analysis. We then computed the Akaike weights across the differentmacroevolutionary best-fit models to determine the relative roles of each process in thediversification of the group. The SecSSE models were excluded from this comparison becausedifferences in the conditioning and normalization factors used in the likelihoodcomputation made them not directly comparable with the other approaches . All othSupplementaryAppendix S4 available on Dryad). Dating analyses using different clocks andtree models estimated similar ages with broadly overlapping credibility intervals . The diversification ofBicyclus started about 5 Ma after the split of the MRCA of the groupfrom Hallelesis in the mid-Miocene . Theinitial events of cladogenesis within the crown group are inferred to have occurredrapidly and contemporaneously among eight major lineages between 19 and 17 Ma. Theevolutionary relationships among these eight early-divergent lineages are largelyunresolved  with relatively short branches. Consequently, we present results of the latteranalysis (M2) in Bicyclus and its sister taxonHallelesis. The initial burst of diversification that produced theeight early-divergent lineages are all inferred to have occurred in this region in theMiocene. Range expansion is the main driver of Bicyclus’ geographicdistribution on the continent . We infer range expansion from the origin tothe Shaba region in the mid-Miocene, which resulted in Clade II. Some ancestors of CladeII further dispersed towards mountains in the Zambezian region of Eastern Africa,resulting in an East African endemic montane clade (saussurei species-group).The remaining ancestors appear to have diversified mainly in the Shaba region until a lateMiocene/Pliocene dispersal event back west to the Congolian region and then to rest of thecontinent  areresults of more recent Pleistocene range expansion events of some ancestors of Clade II.Within Clade I, we infer multiple independent range expansion towards Western Africa fromthe geographic origin starting in the Early Pliocene.The constrained BioGeoBEARS ancestral range analysis (M2) with estimated dispersal scalarontinent . BicycluThe spectral density profile of the modified graph Laplacian identified two clusters,which correspond to the two major clades identified above: Clades I and II. When weiteratively applied the eigengap heuristic to the phylogeny specifying two clusters, wefound that 74 Supplementary AppendicesS7–S9 available on Dryad).Similar results of non-constant temporal diversification were found when Clades I and IIwere analyzed separately. Null models of constant speciation and extinction through timewere outperformed by most time- and paleo-environment-dependent models (Supplementary AppendicesS10–S12 available on Dryad).The best fitting episodic birth-death model for the entire phylogeny—as indicated by theLRT and AIC—is the one with two progressive decreases in diversification rates andrelative extinction (turnover) at 1.5 Ma and 15.7 Ma (Supplementary AppendixS7 available on Dryad). When analyzed separately, Clade I was also best fitby a model with two decreases in diversification, one in mid-Miocene (17 Ma) and the otherin Pleistocene (2 Ma). This is similar to the phylogeny of the entire genus, but with a13-fold lower diversification rate in the mid-Miocene (compared toSupplementary AppendixS8 available on Dryad). Clade II was, however, best fit by a model with aconstant diversification rate of 0.089 since its origin, followed by a 3-fold downshift inrate in the late Pliocene (2.6 Ma) .The relative extinction rate in Clade I was 0.503 after the origin of the clade, 0.758after 17.0 Ma and 0.163 after 2 Ma), while in Clade II it was 0.089 after the origin with0.143 turnover after 2.6 Ma. When compared to the entire Among the paleoenvironment-dependent likelihood models where diversification rates arefunctions of a time-dependent environmental variable, we found that the best fitdiversification model (present . Across present a.Supplementary Appendices S11 andS12 available on Dryad).We also found that the Clade I and II consensus phylogenies, albeit with aComparisons of the habitat transition rate models revealed that the most complex model(ARD) provides a significantly better fit to our character matrix than a simple one-ratemodel , our results suggest that habitats had an importantinfluence on <inline-formula><tex-math notation="LaTeX" id="ImEquation90"              ><![CDATA[$\quad$]]></tex-math></inline-formula> diversification. The speciation rateincreased or decreased every time there was a shift in habitat preference. The speciationrate associated with forest habitats is 0.136, savannahs or open habitats have aspeciation rate of 0.080, and intermediate habitats have a speciation rate of 0.211.Extinction rate was estimated at 1.16eBecause the Examined-Trait-Dependent (ETD) models are strongly supported (Constrained with four rates model best explains the evolution ofhabitat preferences (SupplementaryAppendix S15 available on Dryad). This model outperforms theUnconstrained version, which suggests that Bicyclusspecies are unlikely to switch from forest directly to savanna (Supplementary AppendixS16 available on Dryad). Hence, our method will detect ETD almost always whenit is present, but will occasionally detect it when it is not.The  savanna . They fiis 0.026. Our resSupplementary AppendixS17a available on Dryad). This was, however, not substantially different fromthe second and third best-fit models, KI2 and KI3 where the Clade II not only differs fromthe ancestral clade in the carrying capacity but also in the extinction rate or theintrinsic speciation rate. This suggests that not only did the decoupled Clade IIexperience a range of new niches, it was also able to colonize these niches more rapidlythan would be expected from a reduction in the influence of diversity-dependence alone.The assumed key innovation is estimated to have occurred about 19.2 Mya, regardless of thescenario of the KI Model (but note that this decoupling time is restricted to lie betweenthe stem and crown age of Clade II) (Supplementary Appendix S17a available on Dryad).Among the key innovation models, most support is lent to the KI1 model where the subclade(Clade II) differs from the main clade (Clade I) in the carrying capacity K (The indirect comparison of the key innovation models (KI) with the clade-wide shifts indiversification rates (SR) via the CR versions of the KI and SR models , suggests,however that a clade-wide shift in the clade-level carrying capacity around 9.59 Ma is amore likely explanation of the phylogenetic patterns . However, when the standard constant-rate diversificationmodels were applied separately to Clade I and Clade II, the best-fit model was adiversity-dependent diversification process in which speciation rate decreased asdiversity increased. This model received 55% and 69% support for Clade I and IIrespectively .When comparing the best-fit models for the radiation across our differentmacroevolutionary analyses , an episncreases .Bicyclus, Africa’s most species-richsatyrine butterfly genus, originated in the Congolian rainforest block in Central Africaaround the late Oligocene (ca. 25 Ma) when it diverged from its sistertaxon Hallelesis. Initial diversification of extantBicyclus, however, began ca. 5 million years laterwhen eight major extant lineages diverged in rapid succession. The antiquity and rapidityof these divergences in the Miocene hampered attempts to untangle the precise topology ofthis portion of the tree with confidence. Ancestral range and habitat reconstructionanalyses suggest that these early rapid divergences all occurred within ancient CentralAfrica forests  the seven early-divergent lineages comprising Clade Idiversified recently in the late Miocene to early Pliocene (ca. 8–2.5Ma); and 2) Clade II diversified steadily when the lineage split from the rest ofBicyclus after the mid-Miocene (ca. 17.5 ma), reachingits current size of >50 extant species. The hypothesis that Clade I and Clade IIdiversified under two distinctly different modes is further supported by ancestral habitatstate and geographic range reconstructions. Except for the milyas-groupcomprising B. milyas and B. pavonis, all extant taxa inClade I are strict forest-interior adapted species and most live in Western andWest-Central Africa forests , reveals a significantly high (0.76) backgroundextinction within Clade I beginning at the peak of the MMCO,18–15 Ma. This peThe late Miocene to early Pliocene (9–3 Mya) is thought to have been characterized byintermittent moist climates in Africa, resulting in alternating expansion and contractionof forested habitats . We hypoThe second mode of diversification describes the rapid radiation of a single parentlineage ,which coincides with the onset of uplift in the western branch of the East African RiftSystem (ca. 10 Ma (ca. 16 Ma) and became widespread in the late Miocene(ca. 8 Ma)  . The fina. 8 Ma) . We hypoa. 8 Ma) . Similara. 8 Ma) , birds (a. 8 Ma)  and, outa. 8 Ma) , the Hima. 8 Ma) b, and tha. 8 Ma) .Radiation within the remaining taxa of the Clade II, although rapid, allowed confidentinference of tree topology. The onset of diversification within this clade is estimated at15 Ma, around the end of the MMCO. During the MMCO, eastern Africa became drier, causingforest habitats to become grassland  ostensibB. anynana  and inMelanitis leda  , suggesting they are notgeneralist herbivores of Poaceae as larvae. They may not feed on grasses at all, whichmight explain the dearth of host–plant records for this group of butterflies despite theirabundance and popularity among collectors: field biologists may be looking on the wrongplants to find eggs and larvae. The vast majority of Satyrinae feed only on monocots, andthe only host plant records for species in Clade I, include Zingiberaceae and Marantaceaefor the hewitsonii-species group. Bicyclus iccius, alsoa member of the hewitsonii-species group has been recorded on Poaceae(The widely held viewpoint that Poaceae)  maynot  Poaceae, but LarBicyclus are documented from species in this clade, giving the generalimpression that Bicyclus are grass feeders. However, this clade is justone of eight early-divergent Bicyclus lineages emerging in the earlyMiocene.As expected of taxa with specialized host–plant associations, allopatric differentiationis presumed to be the dominant mechanism of divergence since colonization of novelenvironments requires the presence of suitable hostplants. Recent taxonomic scrutiny tendsto find that isolated populations in Clade I are novel species . This suBicyclus documenting a series of rapid early divergences beginning 5Ma after divergence from its sister genus, Hallelesis. The firstbifurcation within Bicyclus resulted in two clades with contrasting habitatrequirements that responded differently to environmental change. Most of the100B. anynana, has been a model for studying development andevolution for almost four decades. The phylogeny and macroevolutionary hypotheses providedhere now serve as a scaffold to extend the rich arsenal of ecological and evolutionaryinformation gained from this species to the remainder of the taxa in the radiation.Allopatric divergence has also clearly been important, especially along the backbone of thetree, most probably in response to mesic forest fragmentation during the peak of themid-Miocene global warming. Lastly, although we do not assume to have captured all thefactors and processes that culminated in radiation of Bicyclus on theAfrican continent, our results provide strong arguments for the influence of biotic andabiotic factors on the diversification of Bicyclus and represent a stepforward in understanding the mechanisms that led to the rich extant biodiversity ofAfrica.We present a robust, time-calibrated phylogenetic hypothesis for the African butterflygenus adiation . The mos


In clinical practice, bee venom injection into acupuncture points is used to relieve various types of musculoskeletal pain. In the current study, we describe a patient who had persistent severe muscle pain caused by mistakenly injected high-dose bee venom.A 63-year-old woman mistakenly received an injection of high-dose  bee venom . Immediately after the injection, extreme burning pain developed at the injection site, which persisted for 1 month with a mean pain score of 9 on the numeric rating scale. T1-weighted gadolinium-enhanced axial lumbar magnetic resonance imaging revealed increased intensity in the medial part of the left psoas muscle around the L4-5 intervertebral disc level.This finding indicated the presence of inflammation in the left psoas muscle, which was thought to be associated with pain.A buprenorphine transdermal patch (releasing 5 mcg/hours) was applied to alleviate the pain.Six months after the bee venom injection, the pain completely resolved.Bee venom has a strong toxic effect; therefore, only a minimal dose of diluted bee venom should be administered for musculoskeletal pain control. Special caution is required during bee venom injection to avoid excessive doses of bee venom. Additionally, it can be applied to reduce various types of musculoskeletal pain, mainly in East Asian countries.,3Injection of diluted bee venom is 1 of the most commonly used traditional complementary and alternative therapeutic methods, which has been believed to be effective in the treatment of several diseases, such as rheumatic arthritis, tendinitis, herpes zoster, bursitis, gout, and burns. Lower back pain frequently limits daily activities and lowers the quality of life. Additionally, it is a leading cause of absence from work worldwide. Several therapeutic tools, such as oral medications, physical therapies, and injection procedures, are used to control lower back pain.,7 Bee venom injection into acupuncture points has also been used for relieving lower back pain.,9Lower back pain is 1 of the most common complaints of patients who visit hospitals or clinics. An individual has a 60 to 85% lifetime chance of experiencing lower back pain. An epidemiological investigation reported that approximately 3% of adults had a systemic reaction to the bee sting. Second, even though venom has been reported to have anti-nociceptive and anti-inflammatory effects, it can cause pain and inflammation in the injected areas as it is harmful to the human body. Many studies have demonstrated the strong toxicological effects of bee venom and its major polypeptide (melittin) on pain-producing and pain-conducting systems. Due to allergic reactions and the strong toxicological effects of bee venom, only a minimal dose of highly diluted bee venom is injected into acupuncture points.However, bee venom injections can cause a few complications. First, in sensitized patients, it may cause allergic reactions, including systemic or anaphylactic reactions, which can be life-threatening.Here, we describe a patient who had persistent severe muscle pain caused by mistakenly injected high-dose bee venom during the treatment of lower back pain.A 63-year-old woman presented to the pain clinic of a university hospital with severe left lower back pain. Owing to severe pain, the patient was unable to stand or walk. The patient had no specific medical history or psychological disorder. Her left lower back pain started 1 month prior to the mistakenly injected high-dose bee venom in the local clinic. The physician at the local clinic conducted a trigger point injection (TPI) into the left psoas muscle. When the physician injected the TPI solution, the patient complained of extreme burning pain at the injection site. The physician realized that he had injected 2 mL bee venom  diluted in saline at a 1:2000 ratio instead of the TPI solution of normal saline and 2% lidocaine.Physical examination revealed no motor weakness or sensory deficits in the lower extremities. Knee and ankle jerks were normal bilaterally. The patient’s left lower back pain aggravated during passive or active flexion or extension of her left hip joint. Her pain was burning and sharp in nature, with a pain score of 9 on the numeric rating scale (NRS). An electrophysiological study (nerve conduction study and electromyography) performed 1 month after the bee venom injection showed normal findings. T1-weighted gadolinium-enhanced axial lumbar magnetic resonance imaging (MRI) revealed increased intensity in the medial part of the left psoas muscle around the L4-5 intervertebral disc level  with a dose of 20 times more than the standard dose into the left psoas muscle. The pain caused by the bee venom injection persisted for several months. It induces pain associated with bee stings through direct and indirect actions on primary nociceptor cells. Its direct action is the activation of the thermal nociceptor transient receptor potential vanilloid 1 through the phospholipase A2 cascade pathway, causing sensitization of the primary nociceptor.,14 Additionally, melittin indirectly induces the release of pain-inducing substances such as adenosine triphosphate, 5-hydroxytryptamine, H+, histamine, and bradykinin.,14 Furthermore, mast cell-degranulating peptide is another important peptide in bee venom that induces histamine release from mast cells and plays a central role in the inflammatory process. Hyaluronidase is an enzyme found in bee venom that induces the fast spread of the toxin. The above-mentioned inflammation- and pain-producing materials in the venom seem to induce inflammation or tissue damage in the patient’s psoas muscle and cause severe pain.The main and most toxic compound in bee venom is melittin, a major polypeptide consisting of 50 to 60% of the whole bee venom. Therefore, it is widely used to control inflammation and alleviate musculoskeletal pain in Asia, Eastern Europe, and South America. Several clinical trials have been conducted to evaluate the effectiveness of bee venom for managing lower back pain and have demonstrated its positive therapeutic effect.,9,18 As bee venom has a strong toxic effect, only a minimal dose of diluted bee venom is allowed in clinical practice. Bee venom is colorless and can be confused with other injection materials such as normal saline or dexamethasone. When it is injected with a dose beyond the allowed range, severe pain, as in our case, or a life-threatening systemic reaction can occur. Therefore, special caution is required during pain control treatment using bee venom.Bee venom has promising anti-inflammatory and immunomodulatory characteristics.In conclusion, we described a case of severe pain around the injection site after mistakenly injecting 2 mL of bee venom (diluted at a 1:2000 ratio) for the lower back pain treatment. To the best of our knowledge, this is the first report of a side effect following bee venom injection at a dose exceeding the permitted dose range.Conceptualization: Min Cheol Chang.Data curation: Min Cheol Chang.Formal analysis: Min Cheol Chang.Investigation: Min Cheol Chang.Methodology: Min Cheol Chang.Resources: Min Cheol Chang.Supervision: Min Cheol Chang.Validation: Min Cheol Chang.Visualization: Min Cheol Chang.Writing – original draft: Min Cheol Chang.Writing – review & editing: Min Cheol Chang.


Editorial on the Research TopicCardio-circulatory support of neonatal transitionvia the ductus arteriosus into the systemic circulation. Following the elimination of the low-resistance placenta by clamping the umbilical cord and the decrease of pulmonary vascular resistance with the first breaths after birth, major hemodynamic changes occur. The consecutive fall in pulmonary vascular resistance and increase in systemic vascular resistance further induces the reversal of ductal shunting from right-to-left to left-to-right, with functional closure of the ductus arteriosus within 24–48 h after birth in healthy term infants  after birth (intervention group) or according to the standard procedure at 58 s (IQR: 35–86 s) after birth (control group). The authors concluded that deferring cord clamping ≥1 min following a physiological-based approach offered no additional benefits regarding cerebral oxygenation and perfusion compared to the routinely used time-based cord clamping approach in a group of healthy neonates after uncomplicated vaginal delivery. Further studies focusing on PBCC in different populations such as neonates requiring initial respiratory support or with adverse conditions such as prematurity, asphyxia, or sepsis are warranted.Clamping of the umbilical cord eliminates placental circulation, and switches gas exchange to the lungs. Although the procedure of umbilical cord clamping is quick and simple, the timing may have a substantial impact during the immediate neonatal transition period . Recent ncreases . Therefoncreases . Whereasncreases . SchwabeBerisha et al. performed a feasibility study using impedance cardiography for NICOM in asphyxiated piglets to evaluate its potential to improve monitoring and treatment of sick neonates. Half of the registrations in asphyxiated piglets were of adequate quality, albeit signal quality was highly variable between individuals. The presented data was interpreted to be physiologically plausible and support further research evaluating this method for NICOM in clinical studies. The signal quality was superior to recently published delivery room studies in human neonates using electrical velocimetry for NICOM in which about ¾ of the registrations were excluded due to poor signal quality , additional cardio-circulatory monitoring may offer further insights into cardiovascular transition . Ideally quality , 13. Howansition , 10, 11.Ho et al. published a literature review about the pathophysiology of patent ductus arteriosus, and some in silico models used for blood flow and arterial wall modeling. They stress that a multiscale paradigm needs to be adopted when developing future in silico models for patent ductus arteriosus to optimize treatment strategies.Once neonates in need of cardio-circulatory support are identified, establishing vascular access, administering emergency drugs and fluid therapy , and performing chest compressions in cases of severe bradycardia are vital interventions during the immediate neonatal transition –16. The During immediate neonatal transition, optimally timed umbilical cord clamping, improved cardio-circulatory monitoring and properly applied cardio-circulatory support have the potential to reduce mortality and morbidities, while suboptimal care could cause long-term sequelae. The research topic may contribute to consolidating the current body of information with the hopes of enhancing and stimulating further studies on all spectra of “Cardio-Circulatory Support of Neonatal Transition”.


Antiphospholipid syndrome (APS) is a rare coagulopathic disorder diagnosed with a combination of clinical/imaging findings with specific antibody titer elevations over a period of 12 weeks. The following case report will discuss the unusual and challenging hospital course of a patient with extensive autosomal dominant polycystic kidney disease (ADPKD) being treated for a multi-drug resistant urinary tract infection (UTI). The patient later developed multiple deep vein thrombosis (DVT) and was found to have antiphospholipid syndrome. Warfarin, the anticoagulant of choice for antiphospholipid syndrome, has a higher likelihood of intracerebral hemorrhage than direct oral anticoagulants. This is particularly challenging since patients with autosomal dominant polycystic kidney disease have a higher propensity to develop intracranial aneurysms (ICA). Antiphospholipid syndrome (APS) is a rare disorder in which inflammatory antibodies produced by the body interact with the lining of endothelial cells, causing widespread venous or arterial thrombosis. The exact etiology of APS remains unclear; however, genetic predisposition appears to be a contributor, as well as environmental exposures that may lead to the increased production of specific antibodies. The diagnosis of APS involves an unexplained thrombotic event accompanied by an elevation in either lupus anticoagulant, anticardiolipin, or anti-beta-2-glycoprotein antibodies. The titer elevations must be measured at least 12 weeks apart to exclude transient increases. Although there is no direct correlation between autosomal dominant polycystic kidney disease (ADPKD) and APS, ADPKD has been known to increase the likelihood of urinary tract infections (UTI) [We describe the case of a 51-year-old man with a past medical history significant for ADPKD with extensive liver involvement. The patient was originally admitted for treatment of a complicated urinary tract infection (UTI) with Proteus sp. organisms isolated on urine culture. Despite the completion of a treatment course consisting of intravenous (IV) cefepime, the patient remained febrile. A computed tomography (CT) of the abdomen and pelvis showed innumerable cysts in both the liver and kidneys, which were previously known to the patient. The liver cysts varied from 2 mm to 6 cm and extended throughout the entirety of the liver  in the popliteal vein. Further duplex ultrasound studies revealed extensive DVTs in the right femoral and right subclavian veins Figure .The sudden onset of DVTs, despite being on subcutaneous prophylactic anticoagulation, prompted a hypercoagulable workup, which was remarkable for an elevation in beta-2-glycoprotein immunoglobulin M (IgM) and anticardiolipin antibodies, suggesting a preliminary diagnosis of APS. The patient was treated with an unfractionated heparin drip of 25,000 units with a target-activated partial thromboplastin time (aPTT) of 60-80 seconds. The patient’s fever resolved, and he was eventually discharged on apixaban, which was the preferred direct oral anti-coagulant (DOAC) for DVTs in our institution. On follow-up 12 weeks later, the patient's serology panel was repeated, which showed persistence in the elevation of beta-2-glycoprotein IgM and anticardiolipin antibody titers, completing the diagnosis of APS. The patient's anticoagulation has now been transitioned to warfarin, with a target INR of 2.0-3.0, the standard of therapy for patients diagnosed with APS [APS is an autoimmune disorder characterized by the persistence of antiphospholipid antibodies (APA) in the presence of arterial thrombosis, venous thrombosis, small vessel thrombosis, and/or pregnancy loss . AntiphoThe pathophysiology of APS is not entirely clear. It is generally accepted that these antibodies interact with phospholipids, phospholipid-complexes, and phospholipid-binding proteins on endothelial cells. Antibodies against beta-2-glycoprotein-1 seem to be the main pathogenic player, activating endothelial cells, neutrophils, and platelets, which transform the regularly anticoagulant endothelial surface into a procoagulant phenotype . The oriThe patient, with a known history of autosomal dominant polycystic kidney disease, was initially treated for a complicated UTI with Proteus mirabilis isolated from the urine culture. UTIs are quite common for ADPKD patients, with one study estimating that 30-50% of ADPKD patients eventually develop a UTI . PatientThe choice of anticoagulation for our patient is worth discussing. Warfarin is the anticoagulant of choice in APS, and a recent study has demonstrated a threefold reduction in recurrent thrombosis in patients treated with warfarin versus DOACs . HoweverThe patient’s unusual hospital course and challenging investigative workup led to a great teaching opportunity. Our ADPKD patient was treated for a complicated UTI and was found to have multiple DVTs, uncovering a diagnosis of APS. The choice of anticoagulation was challenging, as warfarin is preferred for APS, yet it has a higher risk for ICA. This is particularly concerning for ADPKD patients, who have a propensity to develop an ICA. This current gap in the literature deserves to be explored in future studies.


Chronic diseases like cardiovascular disease or type two diabetes mellitus are related to work exit types such as unemployment and work disability. It is unknown whether metabolic syndrome as a risk factor preceding these chronic diseases is related to work exit type. Metabolic syndrome is present when people have at least three out of the following five risk factors: hypertension, abdominal obesity, raised triglycerides, raised blood glucose, and reduced HDL-cholesterol. We examined the association of metabolic syndrome with work exit type while controlling for socio-demographic and occupational factors.The sample included 55,817 Dutch workers aged 40-65 years from the Lifelines Cohort Study and Biobank. We used data from five measurement waves with a mean follow-up time of 4.1 years. Metabolic syndrome was measured at baseline and based on physical examinations, blood markers, and medication use. Follow-up work exit types were self-reported and included unemployment, work disability, early and regular retirement. Competing risk regression analysis was used.PreliminaryMetabolic syndrome increased the risk of work disability  and unemployment , and was not associated with early and regular retirement.Metabolic syndrome increases the risk of work disability and unemployment. More awareness about and prevention of metabolic syndrome is needed among general practitioners and occupational physicians. Early detection of metabolic syndrome as a risk factor preceding chronic diseases might prevent premature work exit in middle- and late-career.• Metabolic syndrome increases the risk of work disability and unemployment among middle-aged and older workers.• More metabolic syndrome awareness and prevention might help to extend healthy working years.


They evaluated the mean effective analgesic concentration (MEAC) when treating ARCR patients with 10 mL of ropivacaine. They also assessed sufentanil consumption, the onset time of sensory block and motor block, and some other parameters.Liqiong Yu and the team summarized the studies on traditional Tibetan medicine and published a review article entitled “Traditional Tibetan medicine: therapeutic potential in rheumatoid arthritis.” In the manuscript, they analysed the common pathways regulating the aberrant pathophysiology in rheumatoid arthritis. They have also made a comparative analysis between the 27 species that were documented as traditional Tibetan medicines and had the potential to manage rheumatoid arthritis cases. The data gathered from various Tibetan medicine monographs and online Chinese and international databases.Yuan Kang and the team published their research article entitled “Anti-oxidative and anti-inflammatory activities of the ethanol extract of edible flower from Chimonanthus praecox.” Chimonanthi Praecocis Flos is commonly known as the wintersweet flower, and it is an edible flower. The ethanolic extract obtained from these flower buds was subjected to HPLC for component analysis. To validate the anti-oxidative and anti-inflammatory activities, they have performed varied in vitro and in vivo studies, especially that related to the measurement of intracellular reactive oxygen species (ROS), intracellular NADPH oxidase, mtROS, supernatant pro-inflammatory mediators, iNOS, NLRP3 inflammasome activation, luciferase reporter gene, and mouse endotoxemia model.Reshmi Akter and the team published their research article entitled “Pomegranate juice fermented by tannin acyl hydrolase and Lactobacillus vespulae DCY75 enhance estrogen receptor expression and anti-inflammatory effect.” With the aid of tannin acyl hydrolase and Lactobacillus vespulae, they transformed hydrolyzable tannins present in pomegranate juice into ellagic acid. Fermented pomegranate juice was thus enriched in ellagic acid. They conducted various in vitro experiments to validate the upregulation of estrogen receptor expression as well as anti-inflammatory potential.Keun-Tae Park and the team published their research article entitled “The involvement of the noradrenergic system in the antinociceptive effect of cucurbitacin D on mice with paclitaxel-induced neuropathic pain.” Paclitaxel, a widely known anticancer drug, is also well known for its inducing effect as peripheral neuropathy. They studied the alleviating effects of cucurbitacins B and D on paclitaxel-induced neuropathic pain in mice. But cucurbitacins B expressed higher cytotoxicity for the non-cancerous cell line, which resulted in removing this molecule from further experiments. Without affecting the anticancer potential of paclitaxel, cucurbitacin D expressed potential in reducing neuropathic pain, and authors have validated it with a series of experiments and explored the mechanism of action of cucurbitacin D in peripheral neuropathy.This Research Topic thus covered one clinical trial, one review, and three original research articles. This Research Topic offers a comprehensive overview of the potential of natural products in managing various forms of neuropathic pain and neuroinflammation. Further translational studies are necessary to ensure the clinical applicability of these natural products. Such studies may shed light on the safety, efficacy, and optimal dosage of these natural products, as well as possible drug-drug interactions. This will help advance the development of alternative or complementary therapies to manage pain.


Curcuma longa, have been investigated in several studies. The aim of this study is to explore the therapeutic potential of curcumin in attenuating post-operative adhesion band (PSAB) formation in both peritoneal and peritendinous surgeries in animal models.Adhesion band formation is a common cause of morbidity for patients undergoing surgeries. Anti-inflammatory and anti-fibrotic properties of curcumin, a pharmacologically active component of Bio-mechanical, histological and quantitative evaluation of inflammation, and total fibrosis scores were graded and measured in the presence and absence of phytosomal curcumin.Results showed that phytosomal curcumin significantly decreased severity, length, density and tolerance of mobility of peritendinous adhesions as well as incidence and severity of abdominal fibrotic bands post-surgery. Curcumin may decrease inflammation by attenuating recruitment of inflammatory cells and regulating oxidant/anti-oxidant balance in post-operative tissue samples. Moreover, markedly lower fibrosis scores were obtained in the adhesive tissues of phytosomal curcumin-treated groups which correlated with a significant decrease in quantity, quality and grading of fibers, and collagen deposition in animal models.These results suggest that protective effects of phytosomal curcumin against PSAB formation is partially mediated by decreasing inflammation and fibrosis at site of surgery. Further studies are needed to investigate the therapeutic potential of this molecule in preventing PSAB. Post-operative adhesion bands (PSAB) are fibrotic tissues generated by impaired fibrinolysis and cellular exudates following injury to operated areas such as abdominal cavity , flexor Similarly, peritendinous adhesions are a serious complication of flexor tendon injury associated with a high personal and economic burden for patients . TendonsCurcuma longa L. rhizome) and has been used for centuries in traditional Chinese and Indian medicine [Currently, various complementary and alternative medicine methods have been used for human diseases , 15. In medicine . Curcumimedicine . Howevermedicine . To enhamedicine –23.Aberrant regulation of the inflammatory response and fibrosis are major factors in adhesion band formation , 16. AntPhytosomal curcumin was obtained from Sami Labs Ltd. . All reagents for malondialdehyde (MDA), total thiol, catalase (CAT), and superoxide dismutase (SOD) were purchased from Sigma-Aldrich Chemical Co. .Male Wistar rats (weighing 200–250 g) were obtained from the laboratory animals center of the medical school at Mashhad University of Medical Sciences. Animals were housed according to the protocol approved by Institutional Animal Care Guidelines. All animals had free access to drinking water and were fed standard rat chow. They were kept at a normal temperature of 22–25 °C and a standard 12-hr light/dark cycle.General anesthesia was induced with an intraperitoneal injection of ketamine/xylazine. Post-operational peritendinous adhesion model was induced according to a protocol established by Tang et al . Brieflyn = 6) as described below: (A) sham group with surgical incision but no adhesion, (B) positive control group with total surgical transection and adhesion receiving normal saline daily, (C) phytosomal curcumin group which is the same as group B except that rats were treated with 25 mg/kg/day curcumin orally [Animals in each model were randomly divided into 3 groups  or Masson’s trichrome staining. H&E staining was performed to analyze general tissue structure and the inflammatory cells infiltration whereas trichrome staining was utilized to explore the collagen deposition, reflecting the severity of fibrosis. Inflammatory cell infiltration was quantified using Moran et al.  scoring Assessment of the antioxidant effect of phytosomal curcumin was performed by measuring MDA and total thiol concentrations as well as SOD and catalase enzyme activities in tissue samples as described , 9.Achilles tendon tissue mechanical properties were analyzed as described previously , 40. In P-value of < 0.05 was considered significant. All statistical assessments were performed using the SPSS software .Results were expressed as mean ± standard error of the mean (SEM). Statistical analysis was performed using one-way ANOVA and Shapiro-Wilk normality test. A Anesthetic induction and surgical procedures were successful with all rats surviving to the end of the study. Results showed that curcumin significantly decreased adhesion band formation in both the tendon Fig. A and abdUsing Tang macroscopic grading scores of peritendinous adhesion bands , curcumiNext, Nair  and LeacInflammation is one of the key factors in the pathogenesis of post-surgical adhesion band formation. To determine the protective effect of phytosomal curcumin on adhesion band-associated inflammation, either tendon or abdominal adhesion tissues were stained with H&E to examine the morphological and histological changes in adhesion rat models. It has been suggested that 25 mg/Kg phytosomal curcumin has potential therapeutic effects , 23. ResEvaluation of oxidative stress markers was used to further investigate the anti-inflammatory activity of curcumin in post-surgical adhesion band models. Compared to the positive control group, curcumin treatment significantly reduced the level of MDA, an oxidative marker of fatty acid peroxidation, in both peritendinous Fig. A and abdNext, total thiol Fig. B, and F Tissues were stained with Masson’s trichrome to determine the effect of curcumin on fibrosis and collagen deposition at the surgery site. Results showed that compared to the positive control rats, curcumin suppressed fibrosis and collagen content in peritendinous adhesion tissues Fig. A. MoreovTo ascertain the effects of phytosomal curcumin on structural and mechanical properties of tendon adhesion tissues, the load-elongation and the stress-strain curves were plotted. Cross-sectional area (CSA) was also measured to evaluate possible modifications in mechanical features of the Achilles tendon in response to phytosomal curcumin. Results showed that in comparison to the positive control group, phytosomal curcumin improved the structural properties of damaged tissues, including maximum load (N) in tendons Table . HoweverSince the load-elongation curve depends on size, volume, and shape of tissue samples, the quantities were normalized by using the stress-strain curve analyzing the ultimate stress (MPa), ultimate strain (%), and tangent modulus (MPa) indexes. As expected, in the sham group, the level of ultimate stress and ultimate strain is higher and lower than positive control group, respectively. Moreover, phytosomal curcumin improved ultimate stress and strain when compared to the positive control group. Consistently, compared to the positive control group, treatment with phytosomal curcumin increased tangent modulus which is an index indicating the ability of specimens to resist deformation Table .In this study the protective effects of oral phytosomal curcumin in decreasing adhesion formation post tendon and abdominal surgeries were investigated in animal models. Results suggested that phytosomal curcumin significantly decreased post-operational adhesion band formation in both rat models. Moreover, phytosomal curcumin reduced adhesion-related inflammatory responses by decreasing infiltration of inflammatory cells and regulating the oxidant/anti-oxidant balance at surgery sites. Furthermore, phytosomal curcumin potently exhibited anti-fibrotic activities by attenuating fibrotic bundle thickness and collagen deposition. These findings support the therapeutic potential of phytosomal curcumin in decreasing post-surgical adhesion band formation.Adhesion band formation post tendon and abdominal injuries are common surgery-associated complications in patients worldwide –46. InflAlthough surgically-induced adhesions and inflammatory responses occur early during the adhesions formations, fibrosis appears as a late event with a major impact on tissues dysfunction , 16. In These results suggest that protective response of phytosomal curcumin against PSAB formation is partially mediated by decreasing inflammation and fibrosis at site of surgery. Further studies are needed to investigate therapeutic potential of this molecule in preventing PSAB.”. The exact protective mechanisms of phytosomal curcumin in adhesion models have yet to be understood. Further animal and clinical studies are needed to clarify these underlying mechanisms and validate these results in patients.Results of this study suggest that the protective response of phytosomal curcumin against PSAB formation is partially mediated by decreasing inflammation and fibrosis at surgery sites. Supplementary animal and clinical studies are required to elucidate these underlying mechanisms and confirm these results in patients.


While there are a growing number of human pluripotent stem cell repositories, genetic diversity remains limited in most collections and studies. Here, we discuss the importance of incorporating diverse ancestries in these models to improve equity and accelerate biological discovery. Despite these advances, a well-documented issue in existing genomic data is the inadequate representation of samples with non-European ancestries2. For instance, in genome wide association studies (GWAS), commonly used to discover connections between disease risk and genomic variants, most participants are of European ancestry (95.76%)3. Participants of Asian ancestry collectively represent 2.9%, while participants of African ancestry and participants of Hispanic/Latin American ancestries make up less than 1%  registry, 93% are derived from donors of European ancestry8. Similarly, samples from donors of reported European ancestry on average contribute to more than 67% of iPSC lines across multiple larger repositories compared to cell lines derived from donors of reported African ancestry, Asian ancestry, or Hispanic or Latino ethnicities , including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), can be used to generate such systems as they can be obtained from individuals of diverse genetic backgrounds, differentiated into disease-relevant cell types, and engineered to harbor one or more genetic variants of interest. Recognizing the utility of such cellular models, several large-scale stem cell repositories have emerged worldwide to ensure standardized collection, reprogramming methods, storage, and distribution of thousands of hPSC lines Fig. . While hies Fig. . By compion Fig. . While t9. These advances, however, also underscore the importance of expanding the genetic diversity of stem cell models to ensure equitable, timely, and broadly applicable scientific insights and medical benefits. Genetically diverse cellular models can be important drivers of scientific and therapeutic discovery for several reasons. First, including cell lines from diverse genetic backgrounds in functional studies can facilitate the identification of genetic variants underlying specific traits and disorders. Populations of African ancestry for instance have more genetic variation compared with the rest of the world. African genomes have shorter haploblocks and exhibit less linkage disequilibrium (LD) which allows causal alleles for a particular trait or disorder to be mapped at higher spatial resolution11. Indeed, despite a relatively small sample size, the H3Africa study5 discussed above successfully identified several novel disease-associated variants, a higher number than what is typically discovered in similar studies based on participants of European ancestry. Second, certain disease-associated risk variants may be more or less prevalent in individuals from specific ancestries. For example, a genetic variant that doubles the risk of respiratory failure from COVID-19 occurs in 60% of individuals of South Asian ancestry compared to 15% of individuals of European ancestry12. Studying the functional impacts of such variants would therefore benefit from greater access to cell lines generated from donors of South Asian ancestry. As efforts to increase the diversity in genomic studies come to fruition, such examples will only increase. Third, the growing number of iPSC-based clinical trials13 highlights their potential in cell replacement therapy. Since autologous transplantation is neither time nor cost effective, efforts have focused instead on the matching of HLA haplotypes to reduce immune rejection. In this approach, iPSC lines are derived from donors homozygous for a few HLA haplotypes common in a given population. For example, a selection of 10 iPSC lines with the most common HLA types in the United Kingdom (UK) would provide a beneficial match for two-thirds of the UK population14. As the most common HLA types vary across ancestries, it is critical to ensure that similar efforts include HLA haplotypes representative of additional global populations through either cell collection efforts or gene editing15, such as the efforts of the CiRA Foundation in Japan discussed above. Finally, non-genetic modifiers, such as co-infection, microbiomes, diet, and socio-cultural factors may also contribute to phenotypic differences across populations. Some of these effects can be tested in hPSC-based models to disentangle genetic versus non-genetic contributors.In recent years, the use of hPSC-based models has rapidly expanded beyond basic research and has begun to revolutionize the pharmaceutical industry, with applications in drug discovery and toxicity7. Another study of 110 iPSC lines identified rare deleterious non-synonymous single nucleotide variants (nsSNVs) that were associated with outlier cellular phenotypes, which deviated significantly from modal phenotypic values16. Of note, the iPSC lines utilized in both studies were predominantly from donors of European ancestry. Increasing the number of cell lines from donors of additional ancestries are thus needed to enable similarly scaled analyses across populations. In that regard, one study using 72 dermal fibroblast-iPSC pairs from donors of African American ancestry and donors of European ancestry identified ancestry-dependent transcriptional profiles that were associated with reprogramming efficiency17. Additionally, a study of iPSC-derived hepatocytes from 28 individuals revealed that donor genetic background was sufficient to drive differences in hepatocyte differentiation propensity across cell lines18, highlighting the potential for inter-individual genetic variation to impact cell type-relevant preclinical toxicity measurements. Together, these reports suggest that some genetic differences across individuals from diverse ancestries have the potential to manifest as measurable differences in cellular phenotypes in iPSC-based in vitro models.While the extent to which different ancestral backgrounds might impact phenotypes in iPSC-based in vitro models is not well understood, studies suggest that inter-individual genetic changes, including common genetic variation, can result in quantifiable differences in cellular phenotypes. One study of iPSC lines derived from 301 donors found that genetic differences between individual donors affected most cellular traits in iPSCs, including differentiation capacity and cellular morphology19. For example, as discussed above, treatment response to Warfarin shows greater variation in individuals of North Asian ancestry as compared to individuals of European ancestry6. This presents a challenge for scientific and pharmaceutical industries, as experimental validation and patient recruitment for clinical trials are predominantly performed in the United States and Europe13. While establishing clinical trials across the globe would be costly and time-consuming, iPSCs can be readily generated using samples from ancestrally and geographically diverse donors, which could pave the way for the investigation of pharmacogenetic effects on iPSC-derived cell types to optimize drug treatments. Given the importance of HLA haplotypes discussed above, one study screened HLA haplotypes from 1000 participants in Taiwan, identifying 13 homozygous HLA haplotypes, and subsequently generating iPSC lines from all of them. Cardiomyocytes and neurons derived from the 13 lines were then used in toxicity screens, illustrating the utility of a population-based hPSC approach to assess cytotoxicity20. Second, introducing protective or pathogenic variants by gene editing on multiple genetic backgrounds is also critical. Given the potential for modifier alleles to influence phenotypes, evaluating the effects of variants across multiple human genetic backgrounds, rather than a single isogenic background, may generate novel insights. Third, studying the effect of natural genetic variation on cellular phenotypes such as expression quantitative trait loci is important. The availability of large-scale hPSC repositories . Self-reported race or ethnicity reflects identity categories that can change over time, while genetically inferred ancestry  reflects aspects of underlying biology which remain static for a given individual. Both types of data provide relevant information, but reliance upon self-reported race or ethnicity alone presents several specific limitations. The 2020 United States Census provides a timely example of how shifting social, political and cultural factors can influence self-reporting29. Here, the allelic variant does not segregate within the population terms used in any hPSC banks. While different studies will require varying levels of granularity in the populations under investigation, current estimates put the number of subcontinental ancestries at a minimum of 21, with 97.3% of individuals harboring ancestral heterogeneity30. In other words, while many hPSC collections were launched prior to or simultaneous with large-scale genomic initiatives , laboratories within Brazil have derived iPSC lines and performed ancestry analyses to establish cellular resources that better reflect the Brazilian population, linked with clinical phenotyping data32. Alternatively, as groups like TOPMed33 and 1000 Genomes34 are expanding their reference populations, and cell banks like the California Institute for Regenerative Medicine are utilizing SNP microarrays to assay genomic integrity, these data could be combined to ascertain more refined ancestry predictions as opposed to relying solely on self-reported race or ethnicity. These more diverse reference panels will also further enable analyses of experimental models from underrepresented populations. At a minimum, standardized and more accurate descriptions of race, ethnicity, and ancestry should be used in future hPSC collections. Here, frameworks developed for reporting data in genomic studies could be leveraged to provide greater harmonization across disciplines 35.In a best-case scenario, for participants who have consented to iPSC derivation, material for iPSC reprogramming and banking would be collected alongside material for genomic and/or phenotypic studies, thus providing a direct link between these data and available cellular resources Fig. . NotablyMoving forward, increasing genetic diversity in hPSC collections and experimental paradigms has the potential to both accelerate basic biological discovery and facilitate more equitable distribution of scientific benefit. Beyond the discrete points discussed above, this will require a deliberate shift in the mentality investigators use to structure experiments and address the pressing scientific questions in their respective fields.Further information on research design is available in the Reporting Summary


Rectal tonsils are an abnormal reactive proliferation of lymphoid tissue in the rectum. Typical lymphoid tissue of the colon and rectum can proliferate with an increased number of germinal centers in response to exposure to an antigen in the GI tract. This response, in rare cases, escalates to the proliferation of a lymphoid mass known as a rectal tonsil. Here, we present a case of a 46-year-old woman with iron deficiency anemia who underwent colonoscopy with incidental finding of a rectal tonsil. This report discusses initial clinical workup, colonoscopy findings, pathological analysis, subsequent testing, and surgical removal of the identified mass and this rare finding. Treponema pallidum antibody-positive until now [Rectal tonsils present when there is abnormally increased growth of reactive lymphoid tissue in the rectum resulting in the formation of a mass. The etiology of this occurrence is unknown but has been found to occur in patients with other underlying disorders like diverticulosis -3. The cntil now ,4,5.A 46-year-old African American woman with history significant for iron deficiency anemia was referred for colonoscopy to evaluate for a potential source of bleeding. Prior to scope insertion, rectal exam identified a 1cm submucosal mass just inside the anal verge on the left side proximal to the anorectal ring , though rapid plasma reagin (RPR) testing was negative. Upon further questioning, the patient denied any signs or symptoms of a previous sexually transmitted infection (STI). Three weeks after colonoscopy, the colorectal surgeon removed the submucosal mass via transanal excision and sent the specimen (Sample 2) to pathology. The patient was also treated with a course of bicillin 2.4 million units administered intramuscularly at three separate visits at roughly one-week intervals. This was done due to her reactive treponemal tests, and her denying having had previous treatment.The patient was referred to a colorectal surgeon for complete excision of the mass. Histopathology of the initial sample (Sample 1) collected at colonoscopy showed an atypical lymphoid population, described below. Serological testing was positive for HistopathologySample 1 consisted of two portions of tan-brown soft tissue obtained during colonoscopy. It was significant for atypical lymphoid proliferation demonstrating extensive submucosal proliferation of predominantly mature small lymphocytes that infiltrated and caused expansion of the lamina propria. No lymphoepithelial lesions were seen. Immunohistochemistry showed expanded B-cell follicles with associated follicular dendritic cell networks, expanded marginal zones, and no aberrant immunophenotypic expression. B-cell gene rearrangement studies revealed no evidence of clonality.Sample 2 obtained during transanal excision of the rectal mass provided a greater volume of the lymphoid population for evaluation, which showed organized and expanded lymphoid follicles with loose fibrous banding. Flow cytometry showed polyclonal B-cells and unremarkable T-cells. Immunohistochemistry demonstrated findings in keeping with reactive lymphoid tissue, including well-demarcated BCL6+/BCL2- germinal centers associated with CD23+ follicular dendritic cell networks, surrounded by mildly expanded but polarized marginal zones (Figure Chlamydia trachomatis, and tuberculosis [The report done by Hong et al. demonstrated that rectal tonsils were found in patients of all ages, races, and sexes and that the morphology and presentation can vary widely . The finrculosis ,4,5. A prculosis . Rectal rculosis . FurtherIn conclusion, the diagnosis of rectal tonsils should be considered in the differential diagnosis when a submucosal mass is discovered in the colon or rectum. The additional testing needed to rule out malignancy should include wide margin excision as well as immunohistology and flow cytometry. This interesting case of a rectal tonsil being diagnosed in a patient with positive syphilis serology with no prior treatment is a unique addition to the literature.


Evidence-based colorectal cancer screening (CRCS) interventions have not been broadly adopted in rural primary care settings. Co-production of implementation strategies through a bundled approach may be promising in closing this gap by helping rural healthcare practitioners select and implement the best fitting CRCS interventions to the local context. This paper describes the process and outcomes of co-development and delivery of the bundled implementation strategy to improve adoption and implementation of CRCS interventions with two rural clinics.n = 7) were asked to evaluate the bundled approach based on overall reactions and perceptions of innovation characteristics using 5-point Likert scale. After completing the bundled approach, we collected implementation outcomes and limited intervention effectiveness of the CRCS evidence-based interventions (EBIs) developed through the process.We used a bundle of implementation strategies with a core focus on academic-clinical partnership development (strategy 1) and Plan-Do-Study-Act cycles (strategy 2) to identify clinical partner interests/preferences on delivery methods and content needed to facilitate intervention identification and implementation that improves CRCS. We also developed an implementation blueprint for each clinic (strategy 3) through an online blueprinting process based on adapted “Putting Public Health Evidence in Action” (PPHEA) training curriculum. Clinic physicians and staff  to tailor selected CRCS EBIs. As a result of the bundled strategies, each clinic selected and adopted specific EBI(s) with the varying degrees of implementation and CRCS outcomes.The bundle of implementation strategies used were feasible and acceptable in rural primary care practices to facilitate the use of EBIs to improve CRCS.The online version contains supplementary material available at 10.1186/s43058-022-00375-2. Contributions to the literatureFew studies report on the co-production of implementation strategies for evidence-based colorectal cancer screening (CRCS) in rural primary care clinics.A bundle of implementation strategies that include (1) academic-practice partnership, (2) development of implementation blueprint through distance-learning training modules, and (3) plan-do-study-act cycle for continuous refinement resulted in selection and adoption of evidence-based approaches for the two rural clinics with varying levels of implementation and modest improvements in CRCS outcomes.This bundle of strategies has potential to improve adoption, implementation, and sustainability of CRCS EBIs in rural primary care settings.Colorectal cancer (CRC) is the fourth most common cancer in the USA . In 2022p < 0.001) [Rural communities experience much lower CRCS rates than urban counterparts. In 2013, two studies found rural-urban disparities among CRCS. Studies reported that rural patients were less likely to receive screening recommendation by physicians, and less likely to have completed screening or the stool-based test compared to their urban counterparts –11. For < 0.001) . This ru< 0.001) .A plethora of evidence-based intervention (EBI) strategies and programs to promote CRCS are available. The Community Guide recommends more than 15 intervention strategies to increase community demand , community access , or increase provider delivery  –16. The Despite these strong evidence and a wide range of selections, adopting and implementing CRCS EBIs is still challenging for rural health practitioners . Some ofTo address these challenges in rural practices, the study introduced the co-production of implementation strategies to facilitate CRCS EBI uptake in the rural health systems. Co-production  is “a collaboration between researchers and end users  from the onset, in question framing, research design and delivery, and influencing strategy, with implementation and broader dissemination strategies part of its design from gestations.” . Co-prodParticipatory approaches also tend to bundle implementation strategies to help facilitate the translation of EBI to practice. Using Expert Recommendations for Implementing Change (ERIC) taxonomy, we bundled three implementation strategies: (1) developing an academic-clinical partnership, (2) creating a formal implementation blueprint through an online blueprinting process, and (3) using plan-do-study-act (PDSA) cycles to identify clinical partner interests and preferences on delivery methods, content refinement, and system change processes that improve CRCS EBI adoption implementation and sustainability . In thisThe study was conducted with two rural primary care clinics as a part of an accountable care organization (ACO) located in a rural county with a population of 34,914 (2019 estimates by US Census) with a Rural-Urban Commuting Area (RUCA) Codes of “4 = micropolitan area core with primary flow within an urban cluster of 10,000 to 49,999” or “5 = micropolitan high commuting with primary flow 30% or more to a large urban cluster” [Among the six clinics, the two clinics showed interests in participating in the study. Clinic A provides essential primary care services to the community through six providers  and additional 17 staff members. Clinic B is the largest primary care practice in the ACO network and provides services through 11 providers  with the support of 22 non-medical staff. Both clinics have participated in the co-production approach from July 2019 to March 2022  aim/purpose of the implementation, (2) scope of the change, (3) timeframe and milestones, and (4) appropriate performance/process measures . Once den = 8) to assess clinic representatives’ opinions regarding the most suitable methods to receive information on EBI characteristics and blueprints for implementation, systems-change strategies to facilitate implementation, and tools to identify intervention strategies that are both efficient and effective (Plan). Each focus group session took about an hour and conducted in person at each clinic’s conference room. Based on information from the planning session, we delivered a prototype distance-learning module of PPHEA training to clinic participants (Do). Upon completing each session (n = 8 sessions), participants provided feedback using surveys to assess perceptions of the bundled approach and potential adaptations for the next session (Study). Based on the feedback, modifications were made for the next modules (Act). After delivering all eight sessions, academic facilitators continued to hold monthly meetings with clinic participants to facilitate the implementation of the EBIs selected from the training . Clinic B provided number and proportions of CRCS eligible patients who completed the screening during the flu vaccination season (August to February of 2020 and 2021).We used descriptive statistics, including Means, Standard Deviations, Frequencies, and Percentages, to analyze quantitative data. Qualitative data  were analyzed by inductive and deductive development and organization of thematic codes. The research team (JK and AA) developed a coding structure, which includes key conceptual domains and participant perspectives. Minor modifications were made iteratively until the model was saturated. Facilitation notes were carefully reviewed and summarized. Data were analyzed using SAS version 9.3 and NVivo qualitative analysis software (QSR NVivo 11).Through the initial focus group, we identified clinic participants’ interests and preferences on the content tailored to CRCS and delivery methods using synchronous  and asynchronous (pre-recorded lecture video) distance learning technologies. Based on these initial preferences, we converted the original eight PPHEA sessions to pre-recorded, online video sessions followed by online discussion forums or live-streaming conference videos/calls facilitated by the academic team. Following the PPHEA training facilitator’s guide, we included all core contents in each training session and customized contexts/supplemental materials  specific to CRCS EBIs. This resulted in the integration of 6 EBI strategies recommended by the CommunityGuide  and three packaged programs introduced by Research Tested Intervention Programs  as well as the recent evidence of mailed stool-based approaches and multi-component strategies –21. Origgroup learning” approach (viewed online lectures together at a reserved conference room followed by live streaming discussion). In contrast, clinic B used a “hybrid learning” approach . After implementing the first module (defining EBI), clinic A provided constructive feedback regarding the video lecture presentation quality and content . Clinic A also requested to skip the session on community assessment and move directly to the module session that included the CRCS EBI examples. After receiving clinic’s feedback, we improved the video presentation quality and skipped the module 2 (community assessment). As a result, clinic A completed seven sessions, while clinic B completed all eight sessions as planned  and a nurse clinical manager from the ACO administrative team. Clinic B’s team included a physician, a clinical data coordinator, a nurse care coordinator, a referral/schedule coordinator, and a care manager. The two clinical teams also showed different learning styles. Clinic A used a “Despite some negative feedback from clinic A for the first session, participants in both clinics reported overall high mean scores  on the five items of reactions: overall satisfaction with the session, knowledge enhancement on CRCS interventions, relevance to job, worth the time invested, and credibility of information  followed by Trialability (Diff = 1.34). In clinic B, the largest improvement was seen in their perception of Compatibility of the bundled approach (Diff = 0.72) . As shown in the Table The selection and implementation of CRCS EBIs in rural primary care clinics are critical given the geographic disparities in cancer screening and outcomes. Equally important is understanding how these clinics perceive co-production of implementation strategies intended to facilitate the uptake of CRCS EBIs. In this study, we began our approach by building an academic-practice partnership in the process of problem prioritization (CRCS), strategy selection, adaptation, and implementation. We developed implementation blueprints by providing training and education using the adapted PPHEA modules specifically designed for, and delivered to rural primary care practitioners. We also used a rapid improvement cycle (PDSA) to make iterative changes to the implementation strategy based on clinic staff feedback. Our approach was perceived positively to clinic participants and resulted in an adoption of EBIs in each clinic with varying levels of implementation. Our project provides preliminary information about the potential of the bundled implementation strategy as feasible and acceptable in rural primary care practices.While this is preliminary and pilot from two rural clinics, our findings align with research that suggests the likely success of implementation strategies that bundle activities such as academic-practice partnerships, implementation blueprint, and quality improvement strategies with regular feedback with iterative adaptations , 42. AdaIt is worth noting that our co-production approach was well received by our rural primary care practitioners given that most participants reporting positive satisfaction and relative advantage of the bundled implementation strategy approach. Specifically, participants from both clinics reported significant improvements in relative advantage and compatibility, which indicates a relative benefit of our bundled approach compared to other implementation strategies and a better fit with the clinics’ needs and capacity. While previous studies focus on the volume of system-level implementation strategies  , 42, ourInterestingly, the two clinic teams showed varying levels of implementation. Besides the struggle that the clinic A faced due to the loss of a lead physician in the midst of pandemic, clinic A did not engage any support staff or “doers” in the module training process, which could be the reason for delayed and partial implementation. This goes back to the literature highlighting the importance of involving interdisciplinary staff  in the implementation team composition depending on the scope and complexity of the project . Daly etrange of examples are provided to fit differential resources across rural clinics. Interestingly, our rural clinic partners prefer EBIs that include colonoscopy as a major test option and stool-based tests as alternatives, rather than selecting stool-based test as a single main intervention . We found the similar preferences in another rural-focused study reporting that rural clinics are more likely to prefer the use of colonoscopy alone or prefer to use both colonoscopy and stool tests [While only a pilot, the process supported both clinics in identifying and initiating implementation of evidence-based approaches to increase CRC screening. Both clinics selected EBIs that had been included as examples, underscoring the need to do preliminary work to ensure that a ol tests , 45. ThiOf course, as a pilot project, there are several limitations. These include a small sample of clinics and staff from each clinic, and both clinics also came from the same region, which limits generalizability. We did not include patient/community representatives in our co-production process, which may lack their perspectives on the best fitting CRCS EBIs for the community’s needs. Finally, our quantitative data is limited to descriptive considerations rather than inferential statistics due to the small sample size. Nevertheless, our qualitative data indicated that this participatory approach fits well with the clinical resources, time, and interest. Future work will include testing the newly developed modules in a broader range of rural clinics to determine the utility of this co-production of the bundled implementation strategies in supporting EBI selection, adoption, implementation, and sustainability to promote CRCS.Little is known about the co-production of implementation strategies for evidence-based CRCS in rural primary care clinics. We used a bundle of strategies  to help rural clinics identify the best fitting EBI to their practice context. We developed eight distance-learning modules to build an online blueprint for CRCS EBI selection and implementation combined with monthly live-streaming conferences to allow for CRCS tailoring. After completing all the modules, participants in two rural clinics reported positive reactions toward the bundled approach. Both clinics reported improvements in how they perceived the characteristics of the bundled approach to tailor selected CRCS EBIs. Through the process, both clinics developed and adopted the EBIs with varying degrees of implementation and modest increase of CRCS outcomes. Our preliminary data showed that our bundle of implementation strategies were feasible and acceptable in rural primary care practices with modifications for the local context to facilitate the use of evidence-based approaches to improve CRCS.Additional file 1.. Project MilestoneAdditional file 2.. Survey InstrumentsAdditional file 3.. Original and Modified PPHEA Modules with video linksAdditional file 4.. TIDIeR checklist