In this paper, we present very recent results regarding the latency characterization of a novel bidirectional visible light communication (VLC) system for vehicular applications, which could be relevant in intelligent transportation system (ITS) safety applications, such as the assisted and automated braking of cars and motorbikes in critical situations. The VLC system has been implemented using real motorbike head- and tail-lights with distances up to 27 m in a realistic outdoor scenario. We performed a detailed statistical analysis of the observed error distribution in the communication process, assessing the most probable statistical values of expected latency depending on the observed packet error rate (PER). A minimum attainable observed round-trip latency of 2.5 ms was measured. Using our dataset, we have also estimated the probability to receive correctly a message with a specific average latency for a target PER, and we compare it to the ultra-reliable low-latency (URLL) 5G communications service. In addition, a mobility model is implemented to compare the VLC and radio frequency (RF) technologies  to support an automated braking systems for vehicles in urban platooning. In recent years, the integration of assisted driving and advanced communication technologies in modern vehicles has significantly increased, aiming at reducing fatalities and injury rates caused by accidents in urban scenarios . IntelliBefore 5G-mobile communication systems, the Vehicle-to-Everything (V2X) communications were standardized by IEEE using wireless access for vehicular environments (WAVE) and the set of protocols of dedicated short range communication (DSRC). In particular, the radio frequency (RF) based physical layer in DSCR is defined in IEEE 802.11p, while the cellular-based competing technology for vehicular application (C-V2X) has been defined within Long-Term Evolution (LTE) technology as the LTE-V2X. Currently, the Third Generation Partnership Project (3GPP) is working on 5G-V2X standard, but in literature there are only some tests on the 5G network for vehicular applications [Orthogonal frequency division multiplexing (OFDM) is often proposed to provide higher data rate in VLC systems. For example, in , a novelVehicle to vehicle communication using VLC is a topic recently quite investigated by the scientific community. In , an analLatency performance of RF-based technology in vehicular applications is usually affected by the density of vehicles . In thisHowever, whilst most of the initial works in the field of VLC for ITS applications focused on the implementation of unidirectional links ,25,26,27In future 6G networks, the two technologies, RF and VLC, could coexist. An hybrid RF-VLC solution could be used to reduce the high density of RF users and to increase the range limitation of VLC technology . While cThis paper presents an experimental implementation of a bidirectional V2V communication based on VLC technology for the low latency exchange of messages. Experimental measurements have been carried out in a real urban scenario. Characterization of the transmission latency observed using a recently developed VLC system implementing a bidirectional link realized via modulation of light intensity emitted by motorbike head- and tail-lights  is reporFirst characterization of latency in a bidirectional VLC-based communication system for ITS application;Experimental measurements campaign have been carried out in a real urban scenario with real head- and tail- motorbike LED-based lamps;The results demonstrate that our low-cost system guarantees good latency performance, compared with the URLL 5G communication threshold ;The simulation results using the experiments-based mobility model can highlight the benefit of VLC systems for future vehicular applications, as the communication interface currently planned by 6G.The main contributions of this paper can be summarized as follows:The rest of the paper is organized as follows. The bidirectional VLC link is established performing an On-Off Keying (OOK) 0–200% modulation of the light intensity of a commercial headlamp  and of a tail light  of a motorbike for distances up to 25 m with baud rates up to 57 kBaud . Both laFlat configuration), which is the most realistic situation occurring in a real road scenario. A receiving system (RX) is placed besides each lamp, thus each lamp has one transmitter and one receiver. We named TX1 and RX2 the transmitter and receiver, respectively, of the first motorbike, while we named TX2 and RX1 the transmitter and receiver of the second motorbike, respectively. For a detailed analysis of the luminous pattern emitted in terms of the measured experimental SNR, considering the combination between the radiance map of the light sources and the FoV of the detectors, see [The experiment has been carried out in a realistic outdoor configuration in the central hour of the day under sunlight exposure with a luminous flux intensity of around 100 klux. Referring to ors, see . Figure l system . The diml system . The RX In case all packets of the message have been correctly received by the second motorbike (RX1), a message is generated and sent through the rear light back to the first motorcycle. Here, an analogous RX system, placed onboard to the headlights of the first motorbike, performs an active decoding of the received data stream, retrieving the number of correctly received packets . ThiInter-Packet Delay (IPD) slightly greater then the Packet Time (PT) along the x-axis direction towards the second motorbike (TX2/RX1). If a perfect reception occurs, the TX2 replies back to the first motorbike and, again in the case of perfect reception, we find the minimum latency Statistically Averaged Latency—SAL) corresponding to a certain PER in the communication. Indeed, assuming to have a data stream with a certain PT, IPD and PER = 0.5, several situations could arise depending on the size of error packets clusters and their occurrence. For example, the case where good packets and bad packets are evenly alternating is very different from the case where an initial perfect reception of packets is followed by a continuous stream of error packets, forming a very large error cluster. Even intuitively, these cases could tremendously differ in terms of expected latency.The latency could be defined as the time interval elapsed between the first bit of the transmitted packet and the last bit of the received packet after a round trip. As pointed out by smission b and in smission c. In botl aspect ,31, as iA thorough post-analysis of data is carried out to retrieve the errors’ distribution observed in the packet delivery process considering the bidirectional communication. In particular, we analyzed the occurrences and sizes of clusters of errors for different PER values in order to estimate the statistical value of the latency occurring in the complete round-trip communication. By observing the positions of the packets over time containing at least one error, we can measure the latencies to receive each packet correctly. The latencies obtained in such way can be used to derive the probability to reach a target average latency. In other words, we aimed to derive the probability that a specific latency occurs, i.e., that the message is correctly received by that time. The procedure is similar to the one reported in  for an IFlat configuration with a data rate of 57 kBaud. Since the number of transmitted packets is The communication performance of our bidirectional VLC system is evaluated through PER measurements in a As discussed in Following a procedure which has been recently employed in infrastructure-to-vehicle (I2V) applications , the numservices  is only It is important to remark that a significant aspect to be addressed in view of extensive deployment of bidirectional VLC links in ITS is represented by possible effects of unbalance in the quality between the forward and backward channels on the round-trip SAL values. We will provide further theoretical and experimental characterization of such aspects in future works.In this section, a mobility model for the vehicles is provided and used to compare VLC and RF connection capabilities to enable fast braking automated systems. A platooning scenario is assumed to compare the braking capabilities of VLC- and RF-connected vehicles.The mobility model is important to compare latency performance of RF communication standards and the proposed VLC system and to evaluate the limits of VLC in typical vehicular applications. In particular, we focused on the platooning case in an urban scenario, where the high density of vehicles makes RF communication not reliable enough to assure high-safety performances , mostly We considered two vehicles running over a road in a platooning situation. We aim to calculate the safety distance  which allows the vehicles to avoid collision. The latency of the communication link between vehicles is critical to start braking on-time and avoid collisions. Platooning in this case means that each vehicle is obligated to follow the previous vehicle, witth no possibility to change line. This is a frequent situation, in particular in urban areas. We aim to find which is the safety distance that two consecutive vehicles have to maintain in order to avoid collisions in case the first one suddenly starts a critical braking. When many vehicles are involved, it is important that each vehicle is connected to the previous and the following ones since this is what impacts the collision.The implemented model simulates the dynamic trajectory of two vehicles during platooning. The model is used to define the minimum safety distance between the vehicles for different velocities of the vehicles. Different braking methods between the vehicles are compared: human reaction and automated/assisted braking systems based on communication technologies. The communication technologies that we considered are: RF standards such as 802.11p and C-V2X (LTE-based and 5G-based) and the proposed VLC system. The speed of the vehicles is assumed to be the same and both vehicles are assumed to be equipped with an automated braking system, activated by an alert message coming from the other vehicle.Phase 1.The first vehicle starts to brake and its velocity decreases following the equation of uniformly decelerated motion. In the meanwhile, the second vehicle continues to move following the equation of linear motion with constant speed. This step lasts until the automated braking system engages.Phase 2.After the latency time of the communication system, the automated braking system engages and the second vehicle starts to brake following the equation of uniformly decelerated motion. The first vehicle continues to brake as in the previous point.To obtain the dynamic model of the two vehicles’ motion in the platooning scenario, we split the motion in two phases:a used in uniformly decelerated motion is linearly proportional to gravitational acceleration gThe deceleration value In o 1.37 s , while w50 ms for IEEE802.11p standard;40 ms in C-V2X based on LTE network;10 ms in C-V2X based on 5G network.For the VLC communication system, we considered SAL To define the latency of the RF technologies for vehicular communications, the experimental results in  was used 9 m see a,c. As iThe results of x-axis in y-axis in The same mobility model is used to evaluate the performance of the proposed VLC system for two vehicles with different initial velocity. In this case, the two vehicles are assumed not to be connected yet. As it can be derived from In this work, a first characterization of latency times in a bidirectional VLC link in a realistic outdoor scenario for V2V applications is presented. In particular, our system includes real motorbike LED head- and tail-lights as VLC sources, and experimental measurements have been conducted in an outdoor urban scenario. We characterized the communication performance of the system in terms of PER up to 27 m for a baud rate of 57 kBaud. Our results demonstrate an error-free bidirectional communication for distances up to 9 m, and a PER The analysis reported in this paper shows that our bidirectional V2V system is capable of establishing a VLC link with a minimum attainable bidirectional latency as low as 2.5 ms at 57 kBaud, yet granting latencies of the order of 5 ms for distances up to 10 m between vehicles. We highlight that this value is compatible with 5G URLL pillar for ITS, including road safety. We also present a mobility model aimed at comparing the braking distance attainable if two moving vehicles are equipped with VLC, 5G-URLL, LTE-V2X, and IEEEE802.11p communication technologies, respectively. Our model, in combination with our data, shows that VLC technology attains a reduction of the braking safety distance in a platooning scenario by nearly one order of magnitude as compared to what, e.g, IEEE802.11p RF-based systems, could allow. Our results pave the way towards the integration of VLC technology as a reliable communication platform in future ITS services, opening for future works aiming to provide more insight into further relevant aspects, such as tests in real driving conditions, effects of atmospheric conditions on performances of bidirectional VLC systems, or effects of relative unbalance between the forward and backward channel quality on the bidirectional round-trip latency.


The therapeutic effects of cannabinoid compounds have been the center of many investigations. This study provides a synthesis on all Cochrane systematic reviews (SRs) that assessed the use of cannabinoids as a therapeutic approach. Review of SRs, conducted in the Discipline of Evidence-Based Medicine, Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP). A broad search was conducted in the Cochrane Database of Systematic Reviews to retrieve any Cochrane SRs that assessed the efficacy and safety of cannabinoids as a therapeutic approach. The results and key characteristics of all reviews included were summarized and discussed. Eight SRs were included. They assessed the use of cannabinoids for the following types of conditions: neurological (two SRs), psychiatric (two SRs), rheumatological (one SR), infectious (one SR) and oncological (two SRs). There was moderate-quality evidence showing that the use of cannabinoids reduced nausea and vomiting among adults, compared with placebo. Additionally, there was moderate-quality evidence showing that there was no difference between cannabinoids and prochlorperazine regarding the number of participants who reported vomiting, in this same population. This review identified eight Cochrane systematic reviews that provided evidence of unknown to moderate quality regarding the use of cannabinoids as a therapeutic intervention. Further studies are still imperative for solid conclusions to be reached regarding practical recommendations. More than 500 natural compounds  have been identified in the cannabis plant. The recreational and therapeutic effects of cannabinoid compounds (there are nearly 100 of these compounds) have been the center of many investigations. The most common constituent of cannabis is delta-9 tetrahydrocannabinol (THC), the substance that is considered to be the primary psychoactive agent in cannabis.However, it has been hypothesized that not only THC but also a huge number of other cannabinoids  such as cannabidiol, cannabinol, nabilone, dronabinol and levonantradol have therapeutic effects. The route of administration may play an important role in the effect that cannabis has, and this needs to be considered in designing health interventions. These possible routes involve smoking, vaporization, oral ingestion, passive exposure, intravenous injection and administration of rectal suppositories.In 2018, a committee designated by the National Academies of Sciences, Engineering and Medicine (NASEM) of the United States described the following as health-related endpoints from medical use of cannabis: therapeutic effects; mental health effects; cannabis abuse; problems relating to cannabis use; cardiometabolic risks; incidence of cancer; and death.A quick search for cannabis-related trials in the ClinicalTrials database  in July 2018 showed that 432 studies are currently registered as trials in this database alone. Most of these are investigating the use of cannabis as an intervention for a variety of conditions, such as anxiety, pain, nausea and vomiting, depression and attention-deficit hyperactivity disorder.Despite the high amounts that have been invested in research on this topic, the relevance of cannabinoids as a therapeutic approach is still a matter of debate. Because these compounds may form a reasonable alternative for treating numerous conditions, it is imperative to assess the efficacy and safety of cannabinoids through well-designed and well-conducted randomized controlled trials.To present the evidence from Cochrane systematic reviews that assessed the therapeutic use of cannabinoids for any disease or condition.This was a review of Cochrane systematic reviews.This review was conducted within the Discipline of Evidence-Based Medicine of Escola Paulista de Medicina (EPM), Federal University of São Paulo .We included the latest version of full Cochrane systematic reviews (SR). We did not consider protocols or any SRs that had the status “withdrawn” in the Cochrane Database of Systematic Reviews (CDSR).We considered participants with any clinical condition, regardless of age or sex.We considered any intervention derived from cannabis and its synthetic analogues. The cannabinoid compounds considered in these reviews included cannabidiol, cannabinol, nabilone, dronabinol, levonantradol and delta‐9‐tetrahydrocannabinol (THC), in any regimens or doses, when used for therapeutic purposes. We considered any pharmacological or non‐pharmacological intervention as comparators.We considered any clinical, social, laboratory or economic outcomes, as assessed and reported in the systematic reviews included.We conducted a broad systematic search in the Cochrane Database of Systematic Reviews (via Wiley) on July 10, 2018. The search strategy is shown in full in Two researchers (RLP and COCL) independently read all the abstracts that were retrieved, to check their eligibility in relation to the inclusion criteria. Any disagreements during the selection phase were resolved by a third author (RR).We produced a synthesis of the key results and characteristics of all the reviews included, using a narrative approach .For each SR included, we identified the respective population, intervention, comparator and outcomes (PICO); methods for searching for and selecting studies; methods for and results from critical assessment; methods for pooling results ; quality of the body of evidence for each outcome; and applicability. In situations in which multiple interventions were addressed by a single SR, we considered only those that were relevant for the present study.,,,,,,,The initial search retrieved 139 systematic reviews (SRs). After the screening process, 8 SRs were included and brought together in the form of a synthesis of the data.,,,The SRs addressed the following types of conditions: neurological (n = 2),There is some evidence that the cannabinoid system may play a role during the regulation of neurodegenerative processes, including in relation to excessive glutamate production, oxidative stress and neuroinflammation. Neurodegeneration is a feature common to various types of dementia. These findings have led to interest in whether cannabinoids might be useful for treating dementia.http://cochranelibrary-wiley.com/doi/10.1002/14651858.CD007204.pub2/full.The objective of this reviewThis reviewhttp://cochranelibrary-wiley.com/doi/10.1002/14651858.CD009270.pub3/full.Overall, the reports were very poor and precluded any conclusion relating to practice. For further details, refer to the original abstract, available from: Cannabis compounds have been used to reduce pain and other somatic and psychological symptoms. This reviewPain reduction: greater with nabilone than with placebo; no difference between nabilone and amitriptyline.Quality of life: better with nabilone than with placebo; no difference between nabilone and amitriptyline.Fatigue and depression: no difference between nabilone and placebo.Sleep pattern: greater improvement with nabilone than with amitriptyline.Mood: no difference between nabilone and amitriptyline.Withdrawal due to adverse events: higher in the nabilone groups (4/52 participants) than in the control groups (1/20 in placebo and 0/32 in amitriptyline group).Adverse events: the most frequent adverse events were dizziness, nausea, dry mouth and drowsiness (six participants in the nabilone groups). Neither study reported any serious adverse events.http://cochranelibrary-wiley.com/doi/10.1002/14651858.CD011694.pub2/full.It was concluded that there was no convincing unbiased high-quality evidence that might suggest that nabilone was useful for treating fibromyalgia. Moreover, its tolerability was low in this population. For further details, refer to the original abstract, available from: There have been claims that cannabis improves the appetites of people with AIDS, results in weight gain and lifts mood, thus improving the quality of life. This reviewhttp://cochranelibrary-wiley.com/doi/10.1002/14651858.CD005175.pub3/full.It was concluded that even though dronabinol has been registered by at least some medicine regulatory authorities for treatment of AIDS-associated anorexia, and even though some jurisdictions make allowances for “medical” use of marijuana by patients with HIV/AIDS, evidence to show that cannabis and cannabinoids would be effective and safe for this purpose is lacking. For further details, refer to the original abstract, available from: This reviewCannabinoid versus placebo:Complete absence of vomiting: more frequent with cannabinoid ;Complete absence of nausea and vomiting: more frequent with cannabinoid ;2 = 0%; very low-quality evidence);Withdrawal due to adverse event: more frequent with cannabinoid .Cannabinoid versus prochlorperazine2 = 63%; low-quality evidence);Participants reporting no nausea: no difference between groups ;Participants reporting no vomiting: no difference between groups ;Complete absence of nausea and vomiting: no difference between groups ;Withdrawal due to adverse event: more frequent with cannabinoid ;2 = 12%), dysphoria , euphoria , “feeling high”  and sedation  were more frequent in the cannabinoid group;Adverse events: dizziness .Personal preference: people reported a preference for cannabinoids rather than prochlorperazine  compared an antiemetic drug alone with cannabinoid added to the antiemetic drug and did not show any differences between the groups.http://cochranelibrary-wiley.com/doi/10.1002/14651858.CD009464.pub2/full.It was concluded that cannabis-based interventions might be useful for adults with refractory chemotherapy-induced nausea and vomiting. However, the methodological limitations of the RCTs reduced the confidence regarding these findings. Future research considering the current chemotherapy regimens and newer antiemetic drugs is likely to modify these conclusions. For further details, refer to the original abstract, available from: This reviewTetrahydrocannabinol versus prochlorperazine/metoclopramide: two RCTs showed conflicting results and the heterogeneity of the studies included meant that no data could be pooled.Nabilone versus domperidone: cannabinoid showed benefit regarding reduction of nausea .http://cochranelibrary-wiley.com/doi/10.1002/14651858.CD007786.pub3/full.It was concluded that cannabinoids might be effective but that they produced frequent side effects. The current evidence relating to the use of cannabinoids for this purpose is too scarce for any sound conclusion to be reached regarding the implications for clinical practice. For further details, refer to the original abstract, available from: This reviewBrief Psychiatric Rating Scale-E (BPRS): no difference between the groups at 7 days , 14 days , 21 days  or 28 days .Average overall score : no difference between the groups at 14 days  or 28 days .Average negative symptom score : no difference between the groups at 14 days  or 28 days .Average positive symptom score : no difference between the groups at 14 days  or 28 days .Adverse events: poorly reported and no analyses were performed.http://cochranelibrary-wiley.com/doi/10.1002/14651858.CD004837.pub3/full.It was concluded that the evidence so far is insufficient to show that cannabidiol has any antipsychotic effect. For further details, refer to the original abstract, available from: Gilles de la Tourette syndrome is a developmental neuropsychiatric condition characterized by chronic motor and phonic tics. Currently, the drugs used for Tourette’s syndrome either lack efficacy or are associated with intolerable adverse events.This reviewhttp://cochranelibrary-wiley.com/doi/10.1002/14651858.CD006565.pub2/full.It was concluded that so far there is not enough evidence to support the use of cannabinoids for treating tics and obsessive-compulsive behavior among people with Tourette’s syndrome. For further details, refer to the original abstract, available from: This review included eight systematic reviews (SRs) that assessed the use of cannabinoids for neurological, psychiatric, rheumatological, infectious and oncological conditions. Only two of the SRs included assessed the overall quality of the evidence through using the GRADE approach. The only moderate-quality evidence found was related to the use of cannabinoids to treat chemotherapy-related nauseas and vomiting among adults, showing that the use of cannabidiol reduces nausea and vomiting among adults, in comparison with placebo. Additionally, there was moderate-quality evidence showing that there was no difference between cannabinoid and prochlorperazine regarding the number of participants who reported vomiting. All other evidence ranged in quality from low to very low. These findings were similar to those of a previous overview of SRs that addressed only the effects of cannabinoids for nausea and vomiting related to chemotherapy.The benefits and harm of any therapeutic intervention, including use of cannabinoids, need to be properly addressed through randomized controlled trials (RCTs). Thus, the scope of this review did not extend to presenting results from primary observational or animal experimentation studies. The results from such studies are more susceptible to bias and should always be taken to be exploratory. These studies may nevertheless be useful for guiding well-designed RCTs.Regarding the implications for practice and research, the results presented in According to these Cochrane SRs, use of cannabinoids to treat medical conditions is not supported by high-quality evidence. The scarcity of data precludes any solid conclusions regarding the efficacy and, especially, the safety of cannabinoids as therapeutic interventions. Further updating of the presented Cochrane systematic reviews also needs to carefully assess the quality of evidence, in order to better support healthcare decisions.This review identified eight Cochrane systematic reviews (SRs) that provided evidence of unknown to moderate quality regarding the use of cannabinoids as a therapeutic intervention. These SRs found moderate-quality evidence regarding (a) benefits provided by cannabinoids (compared with placebo) for reducing nausea and vomiting that related to chemotherapy among adults and (b) lack of difference between cannabinoids and prochlorperazine regarding the number of participants in this subgroup who reported vomiting.


The current paper examines the level of use of evidence and factors affecting the use of evidence by frontline maternal, newborn and child health (MNCH) and reproductive and child health (RCH) staff in practice decisions in selected health facilities in Ghana.Data on use of evidence and its correlates was collected from 509 frontline healthcare staff drawn from 44 health facilities in three regions in Ghana. Means were used to examine the level of use of evidence, whiles cross-tabulations and Partial Least Squares-based regression were used to examine factors associated with the use of evidence in practice decisions by frontline MNCH/RCH staff.The findings suggest a high level of use of evidence by frontline MNCH/RCH staff in practice decisions (score of 3.98 out of 5), albeit that evidence use is skewed towards the use of practice guidelines and policies. For the antecedents of evidence use, attitude had the highest score (3.99), followed by knowledge (3.8), access to evidence (3.77) and organizational structure (3.57), using a threshold of 5. The regression results indicate that attitudes and knowledge of frontline MNCH/RCH staff, organizational structure (strongest association), years of experience, being a male and working in a mission health facility are significantly positive correlated with evidence use, whiles working in a private health facility or in the post-natal clinic is negatively correlated with the use of evidence.We argue that any effort to improve the use of evidence by frontline MNCH/RCH staff in practice decisions should focus on improving attitudes and knowledge of staff as well as challenges related to the structure of the organisation. Given however that the score for attitude was relatively high, emphases to improve evidence use should be on access to evidence and organizational structure in particular, which had the lowest score even though it has the strongest association with the use of evidence.The online version contains supplementary material available at 10.1186/s12961-022-00881-8. The health of mothers, children and infants have been prioritized by both international and national organizations. Per Millennium Development Goal (MDGs) 4 and 5, Under-5 and maternal mortality were expected to be reduced by two-thirds and three-quarters respectively by 2015 [Notwithstanding the above, reproductive and child health (RCH) outcomes in Ghana have improved substantially over the last two decades. Nationally, the maternal mortality rate (MMR) reduced by 59.2% from 760/100,000 live births in 1990 to 310/100,000 live births in 2017 . AdditioIn terms of infant and child health, similar gains have been recorded over the last two and half decades. There has been 61% increase in basic vaccination coverage between 1988 47%) and 2014 (77%) 7% and 20.Notwithstanding these improvements, Ghana did not meet MDG 4 and 5. Besides, quality of care in many Ghanaian health facilities, especially in maternal, newborn and child health (MNCH) and RCH remains a challenge . As it iNotwithstanding potential benefits associated with the use of these instruments, evidence in the literature suggest that they are hardly used by frontline health workers, thereby limiting their potential for realizing associated benefits –9 such aExamines the level of use of evidence by frontline MNCH/RCH staff in making practice decisions in selected health facilities in GhanaExamine factors that influence the use of evidence in practice decision-making by frontline MNCH/RCH staff in selected health facilities in Ghana.The current paper therefore examines the level of use of evidence  by frontline MNCH/RCH staff in making practice decisions. Specifically, the paper.Although it is not standard for frontline health staff to synthesize evidence from academic research findings for purposes of decision making, it is common for higher end health facilities to synthesize research findings to inform team decisions or also use the findings of facility conducted operations research to inform decisions. Thus, evidence in the context of this study is operationalized to include evidence products such as health policies, guidelines, protocols as well as use of evidence from academic and facility-based operations research.The study is based on a cross-sectional design with health facilities carefully selected to include those in rural and urban areas as well as areas that do not have demographic surveillance sites. Demographic surveillance sites were excluded from the study sample to ensure that on-going interventions in those sites do not confound/interfere with the findings of the current study.The study targeted frontline health workers  in the Greater Accra, Ashanti and Eastern regions of Ghana, who are involved in the provision of MNCH/RCH services. The choice of the 3 regions is on the basis that they have the highest number and types (i.e. by ownership and hierarchy) of health facilities. Having the highest number of health facilities was important for having an adequate sample size for the study, whiles diversity in ownership and hierarchy was essential for selecting health facilities that are representative of the population of health facilities in Ghana.Health facilities for the study were conveniently selected from the three regions based on accessibility and willingness to participate in the study. The selection was however done in a way to account for diversity in the types (ownership and hierarchy) of health facilities in a region. The health facilities selected include Ashanti: 8 Community Health and Planning Services (CHPS) Compounds, 6 Health Centers, 3 Maternity Homes/Clinics, 4 Private Hospitals, 2 Mission Hospitals, 2 District Hospitals, 1 Poly Clinic and 1 Regional Hospital. Although a tertiary hospital was originally part of the Ashanti sample, conditions for securing access was almost impossible to achieve and so the tertiary facility was dropped. In Greater Accra, there were 4 CHPS Compounds, 5 Health Centers, 2 Private Hospitals, 1 Mission Hospital, 1 Regional Hospital, 1 Tertiary Hospital and 1 Qusai-Government Hospital. In the Eastern region, 1 Mission Hospital, 1 Regional Hospital and 1 Poly Clinic were selected. The relatively smaller number of health facilities from the Eastern region is based on health facility willingness to participate. Overall, there were 12 CHPS compounds, 11 Health Centers, 3 Maternity Homes/Clinics, 6 Private Hospitals, 3, Mission Hospitals, 2 District Hospitals, 2 Poly Clinics, 3 Regional Hospitals, 1 Quasi Government Hospital and 1 Tertiary Hospital.After the selection of health facilities, the total number of MNCH/RCH staff in each health facility selected was ascertained and a representative sample calculated using 95% confidence level and 5% margin of error. Proportional representation was then used to distribute the calculated sample across the different cadre of MHNCH/RCH staff  in each health facility. However, the selection of staff from each cadre to respond to the survey was done based on availability and willingness to participate due to the shift system operated by frontline MNCH/RCH staff, in addition to the fact that their heavy workload meant it was impossible to interview some of them. Overall, a total of 509 staff from all the health facilities selected completed a questionnaire each administered by enumerators.The instrument contained questions on socio-demographic characteristics of respondents , use of evidence, and key factors that influence the use of evidence by frontline MNCH/RCH staff in making practice decisions. Specific questions used to capture the use of evidence and key factors influencing the use of evidence were based on instruments used by prior authors –22. The Ethical clearance  was sought from the Ethical Review Board of the Ghana Health Service in addition to administrative approval from relevant regional and district Directors of Health Services as well as heads of participating facilities. Prior to the main data collection, the instrument was amended to reflect the results of a pre-testing of the instrument at the MNCH/RCH unit of the University of Ghana Hospital. The instrument was administered by trained data collectors to respondents using a Computer Assisted Personal Interviewing (CAPI) technology. The CAPI was used to ensure that data collected is transmitted directly and stored in a central data repository. This limited the possibility of errors normally associated with manual capture of data.Descriptive statistics based on frequencies and means were used to examine the extent of use of evidence. In addition, an ANOVA test was used to examine differences in the use of evidence between different groups . A Partial Least Squares approach via structural equation modeling was used, with the help of version 3 of SmartPLS software to examine factors that influence the use of evidence by frontline MNCH/RCH staff in their practice decisions. In estimating the structural equation, measurement models that account for reliability of the constructs were assessed, starting with convergent validity, which examines the internal consistency of the constructs. This was assessed using the Cronbach Alpha scores, the composite reliability scores and the AVE scores. An internally consistent construct must have a Cronbach Alpha and a composite reliability score to be at least 0.7. Also, indicators for each construct must explain at least 50% of the total variation (AVE score of at least 0.5). Table Secondly, discriminant validity assessments were done to examine the ability of an indicator to uniquely describe its own construct, and not another construct. The Fornel–Larcker criterion assesses the inter-construct correlations, against the square root of the AVE scores of each of the construct. Discriminant validity is established if the square root of the AVE score for a construct is greater than its correlation with other items. Table Henseler et al.  providesThe different health facilities were re-categorized  for ease of analysis as in Table In this section, we present results on the extent of use of evidence by frontline MNCH/RCH staff as well as factors  that are correlated with the use of evidence in MNCH/RCH practice decisions. As indicated in the methodology section, each factor/construct was measured using several questions. However, only those indicators with factor loadings of 0.5 and above through a confirmatory factor analysis were retained. Table Table F = 8.95, p = 0.00) and therefore key in the estimated model. Overall, the attitude construct had the highest average score, followed by the construct on use of evidence. Organisational structure recorded the least average score, which was followed by the construct on access.An examination of the overall mean of each of the constructs suggest that the constructs are significantly different from each other  captured in Table In this section, the determinants of use of evidence in practice decisions by frontline MNCH/RCH staff was examined. Using the structural model , a stepwise structural regression model was used to examine the association between attitude, access, knowledge and organizational structure on the use of evidence by frontline MNCH/RCH staff in their practice decisions as per Table R2 and the coefficients of the key independent variables . The results of the additional controls shows that the use of evidence is significantly higher among workers in mission health facilities, but lower in private facilities compared to government facilities. Additionally, the use of evidence is significantly higher among males compared to females and lower among staff who work in a post-natal clinic compared to those in ANC, child welfare clinic and the delivery unit. Also, the length of time in terms of years worked in the health facility is also significantly positively correlated with the use of evidence by frontline MNCH/RCH staff in their practice decisions.In Table The study sought to examine the extent of use of evidence by frontline MNCH/RCH staff for practice decisions, as well as the factors that influence the use of such evidence. From the findings, the use of evidence for practice decisions had a score of 3.89 out a total of 5, with the use of both manual and online practice guidelines, protocols and policies as key sources of evidence. In terms of the factors that affect the use of evidence by frontline MNCH/RCH staff, attitudes of staff towards the use of evidence had the highest score (3.99), with organizational structure and processes to facilitate the use of evidence having the lowest score (3.57). The regression results further suggest that frontline MNCH/RCH staff’s attitude towards evidence-based practice, access to relevant information, knowledge of evidence-based practices in MNCH/RCH service provision and organizational structure and processes have a significant positive effect on the use of evidence by frontline MNCH/RCH staff. However, when socio-demographic characteristics of respondents  were introduced into the model, access to information remained positive but insignificant. Additionally, the results showed that an MNCH/RCH staff from a mission health facility, being a male and having worked for a longer time was significantly positively correlated with the use evidence for practice decisions.The overall score of 3.89 out of a total of 5 for use of evidence by MNCH/RCH staff for practice decisions suggest that there is a high level of use of evidence by MNCH/RCH staff, although it can be improved. As expected, evidence seems to be dominated mostly by the use of practice guidelines, protocols and policies. There may be the need to improve the skills of staff to also search for synthesized findings of academic and operations research and use that to inform practice decisions. Within the literature, the use of research evidence (academic or operations research) does not seem to be as popular as practice guidelines, protocols and policies, given the complexities associated with the use of research-based evidence. Issues that have been raised include timeliness and relevance of research evidence to specific practice challenges, relationship between producers and users of evidence and the ability of users to adapt research-based evidence to their context . This maThe results of the study resonate with findings from other jurisdictions . PositiThe findings also suggest that practice-relevant research evidence is available and accessible to MNCH/RCH staff to some degree. Such availability and accessibility create opportunities for the implementation of strategies that can help to improve MNCH/RCH staff’s understanding of what constitute practice evidence, and consequently ensure that research findings take into consideration the reality of practice. This can be key in improving access to relevant information for evidence-based practice decisions. The difficulty in having access to relevant information for decision-making has also been identified in the literature as a barrier to the use of evidence , 32–34. The findings also suggest that knowledge of evidence-based practices in MNCH/RCH is key to using evidence in practice decisions. However, apart from the confidence of staff in sharing and disseminating information with or to their colleagues, all the other indicators on knowledge of evidence-based practices by the MNCH/RCH staff interviewed had relatively lower scores and will therefore need some improvements. Effort to improve knowledge will ensure that staff have the requisite skill and experience needed to utilize available evidence to inform practice decisions. The existing literature suggest that healthcare workers have shortfalls in skills and experience especially in areas of research literacy and research utilization , 32, 37.The results under organizational structure suggest that only 2  out of the 14 indicators retained in the organizational structure constructs had higher scores. Although the scores of the other 12 indicators were not extremely low, the key message is that the other 12 indicators need some improvement. This will be essential in ensuring that the use of evidence to inform practice decisions among MNCH/RCH staff sees a significant positive change. For example, issues of research leadership, strong professional leadership that facilitate research, putting in place systems that encourages external learning and benchmarking and communication of key research priorities and strategies are essential in this direction. Prior studies have argued that structural rigidities in organizations constitute key constraints to the use of evidence in decision-making . These rThe study sought to examine the level of use of evidence, as well as factors that influence the use of evidence by frontline MNCH/RCH staff in their practice decisions. The results of the study suggest that the level of use of evidence  by MNCH/RCH staff to inform practice decisions is high (3.98 out of 5). In addition, attitude, knowledge, organisational structure, health facility ownership, gender, unit/area of work and years of work experience of individual employees were also found to be correlated with the use of evidence by MNCH/RCH staff for practice decisions.Notwithstanding the results, it is unlikely that policy makers will seek to change the ownership of health facilities, gender and unit/area of work of individuals in the MNCH/RCH unit of health facilities in other to sustain and improve the use of evidence to inform decisions by frontline MNCH/RCH staff. It will therefore be important that existing and new capacity building programmes that seeks to sustain and improve the use of evidence by frontline health staff emphasise attitudes, knowledge and organisational structure related constraints. It will also be important to ensure that factors such as years of work experience and area/unit where an individual MNCH/RCH staff works is taken into consideration when selecting staff for capacity improvement in relation to the use of evidence. Given that the result of the study shows that organisational structure and access to relevant evidence are the weakest link among the three key determinants, it will be important that management focuses its attention on improving structural challenges within the health facility in general and the MNCH/RCH unit in particular. Indeed, the issues captured under organisational structure /reproductive & child health (RCH) services


The current strategy of searching for an effective treatment for COVID-19 relies mainly on repurposing existing therapies developed to target other diseases. Conflicting results have emerged in regard to the efficacy of several tested compounds but later results were negative. The number of conducted and ongoing trials and the urgent need for a treatment pose the risk that false-positive results will be incorrectly interpreted as evidence for treatments’ efficacy and a ground for drug approval. Our purpose is twofold. First, we show that the number of drug-repurposing trials can explain the false-positive results. Second, we assess the evidence for treatments’ efficacy from the perspective of evidential pluralism and argue that considering mechanistic evidence is particularly needed in cases when the evidence from clinical trials is conflicting or of low quality. Our analysis is an application of the program of Evidence Based Medicine Plus (EBM+) to the drug repurposing trials for COVID. Our study shows that if decision-makers applied EBM+, authorizing the use of ineffective treatments would be less likely. We analyze the example of trials assessing the efficacy of hydroxychloroquine as a treatment for COVID-19 and mechanistic evidence in favor of and against its therapeutic power to draw a lesson for decision-makers and drug agencies on how excessive hypothesis testing can lead to spurious findings and how studying negative mechanistic evidence can be helpful in discriminating genuine from spurious results. SARS-CoV-2 belongs to a family of human coronaviruses that cause the common cold and more severe conditions such as breathing difficulty and acute respiratory distress syndrome (ARDS)  have been conducted, ongoing, or in preparation   as treatments for COVID-19 to argue that false-positive results should be expected to emerge from the vast number of repurposing trials and argue that using mechanistic evidence from laboratory and animal studies can help in resolving inconsistencies in the results of clinical trials and telling genuine from spurious results. The purpose of our case study is twofold. First, we offer an explanation of why clinical trials assessing the efficacy of repurposing candidates reported inconsistent results, which can also deepen our understanding of conflicting evidence delivered by normal science. Second, we support applying the approach of Evidence Based Medicine Plus (EBM+) to asses clinical evidence and argue that it is particularly useful when the results of clinical trials are conflicting or of low quality, but decisions nevertheless need to be made.The approach of EBM+ can be considered as an extension to the standard view on assessing medical evidence known as evidence-based medicine (EBM) . It denotes a situation when conducting more than one statistical inference from the same or dependent datasets. It is well known that, in such cases, p-values are underestimated, and type-I errors (rejecting the null hypothesis of no effect when it is true) happen more often than could be expected Dunn, . In SectIt is well known that false-positive results can also emerge when several research teams address the same research question  are taken as evidence for treatment’s efficacy. The statistical significance threshold The threshold of statistical significance Tannock, :\documenSuppose this pessimistic scenario is accurate and none of the candidates for repurposing tested in more than two thousand active RCTs is effective. In that case, the number of false positives can be expected to be as high as 100  when The formula shows that for as little as 283 trials (compared to the number of studies in the field of drug repurposing for COVID-19), the chances of Still, it is possible that some treatments will turn out to be genuinely effective and some RCTs will report true positive  equals zero  as treatments for COVID-19. Our take on the positive results of HCQ repurposing trials does not exclude other reasons that may have played a role since biases resulting from poor research design or scientific misconduct could also shape the results of some studies. Due to the number of clinical trials, the use of HCQ in clinical practice (with the Food and Drug Administration’s emergency use authorization being the prime example), and heated debates about the treatment’s purported efficacy, HCQ is a paradigmatic example of the problem we approach in the paper.The hype for hydroxychloroquine had been started by a few positive results of observational studies and was fueled by the urgent need for any therapy in the face of the pandemic. The simplified view on the chronological order of published results is that first positive results of small and biased studies [e.g., the randomized controlled trial from China  or used doses that cause side effects and increase mortality in treatments arms. Million et al.  explaineEven though vested interests and inferior design of some studies seems a plausible explanation of the conflicting results at first glance, detailed analysis of the designs shows that methodological problems populate studies reporting both positive and negative results. For example, it is true that antiviral agents that target replication mechanism are only effective at earlier stages of the disease and first studies tested the efficacy of HCQ in hospitalized patients, but more recent reports showed that HCQ does not treat early COVID-19 too. For example, Skipper and Boulware  conducteThis negative result can be contrasted with the Henry Ford Study  scientific misconduct, the view that spurious findings have only been reported by methodologically unsound studies is not warranted. As we argued in Sect. Despite these characteristics of the field of studies repurposing HCQ for COVID-19, most studies do not control for base rate fallacy and those exceptional cases that control for the number of statistical comparisons and lower the significance thresholds accordingly, take into account only the number of trial arms and not the base rate of the whole field. The question of whether an alpha correction for a field’s false discovery rate should be introduced at the level of individual studies remains debatable. Rubin  argued rSystematic review with meta-analysis allows for obtaining an estimate of average treatment effect more accurate than the results of individual studies  can be useful and is more believable than mechanistic evidence for treatment efficacy. Our stance results from the belief that high-quality mechanistic evidence in favor of a drug’s efficacy can support weak difference-making evidence and they, jointly, may warrant authorizing a treatment. This approach can speed up the assessment process as waiting for the results of high-quality RCTs or meta-analyses of interventional studies is not needed. But negative mechanistic evidence can also outweigh the results of clinical trials in cases when difference-making evidence is of low quality due to, for example, base-rate fallacy. However, negative mechanistic evidence is, all else being equal, more believable because there are many mechanisms operating in vivo and interacting with each other and therefore showing that a mechanism by which a drug acts is screened off or not working in the target is more plausible than results describing a treatment’s mechanism, especially if the cell lines used in in vitro research are not the same as those targeted by the treatment under consideration. We have used the example of HCQ to argue that difference-making evidence is of low quality because of the high likelihood of false-positive results. Here, we claim that supporting decisions regarding the authorization of HCQ as a treatment for COVID-19 with an in-depth assessment of the quality of mechanistic evidence would save FDA from authorizing the use of an ineffective treatment. However, we believe that our approach is applicable to other treatments.Initial in vitro studies of HCQ efficacy had been conducted on the African green monkey kidney Vero cells  are more helpful in producing difference-making evidence. In contrast, others  are more suitable for inferring mechanisms  and the evidence stemming from clinical trials were assessed appropriately then less faith would be put in HCQ.Our case study of the attempt at repurposing HCQ as a treatment for COVID-19 has shown that false-positive results have emerged due to the number of clinical trials because HCQ is unable to stop SARS-CoV-2 from replicating in human lungs. One could also ask why false-positive results have emerged from the perspective of the sociology of knowledge or economics of the pharmacological industry. While further research is still needed to gather conclusive evidence, our analysis sheds light on some plausible explanations. Considering that HCQ is a cheap drug that is not protected by intellectual property law, vested interests of the industry are an unlikely factor driving starting so many trials. Therefore, researchers’ presupposition regarding HCQ efficacy seems a more likely factor influencing decisions to start clinical trials. Although we do not argue that clinical trials should not be conducted without strong mechanistic evidence for treatment’s efficacy , we need to observe that excessive testing of the efficacy of one drug can possibly be used by the pharmaceutical industry instead of other questionable or fraudulent practices to spuriously show a compound’s efficacy and profit from selling stockpiled treatments. For example, remdesivir was first tested as a single therapy and later jointly with various generic drugs, what raised the chance for false-positive results to emerge. It received FDA’s emergency use authorization despite conflicting evidence, the price of over 2000 euro per therapy (Rezagholizadeh et al., Furthermore, our analysis supports the use of the program of EBM+ to assess the efficacy of repurposing candidates. If false-positive results are expected because of the number of clinical trials testing the same compounds, negative mechanistic evidence for drugs’ efficacy can influence the inferences drawn from positive results and prevent authorizing treatments based on spurious outcomes. We believe that the program may prove fruitful, especially in public health emergencies such as the SARS-CoV-2 pandemic, due to the pressing need for undertaking decisions with limited evidence. Furthermore, our case study has shown that negative mechanistic evidence is more trustworthy than in vitro results demonstrating a treatment efficacy.Our analysis was focused on the process of drug repurposing for COVID-19. But discovering pharmaceutical treatments is not the only strategy against the COVID-19 pandemic. Developing vaccines and novel drugs are the other possibilities. The former falls beyond the scope of the paper, but the latter should be discussed briefly. Should our worries be limited solely to the case of drug repurposing or maybe as well to novel drugs explicitly developed to treat patients suffering from COVID-19? The two procedures seem to be quite different. When a new drug is being developed, several steps are required (for full comparison, see Ashburn & Thor, However, we do not argue that we should refrain from clinical studies when there is insufficient mechanistic evidence in favor of certain drugs given the extraordinary circumstances. First of all, we should distinguish between the state of regular science and science doing in the state of global crisis. The current situation raises many concerns and issues, both scientific and social (Chellappoo, At the same time, we should be aware that going through shortcuts might be problematic in many ways. It might generate false hopes that might influence the way the public behaves. For instance, some people have mistakenly taken other drugs that sounded like hydroxychloroquine after hearing about it to prevent covid-19 (Samuels & Kelly,


Drosophila melanogaster Nora virus (DmNV) is a novel picorna-like virus first characterized in 2006. Since then, Nora virus has been detected in several non-Drosophila species, including insects in the Orders Hymenoptera, Lepidoptera, Coleoptera, and Orthoptera. The objective of this study was to determine if DmNV could infect individuals of other species of invertebrates besides D. melanogaster. The presence of DmNV in native invertebrates and commercially available stocks was determined. Laboratory-reared D. yakuba, D. mercatorum, Gryllodes sigillatus, Tenebrio molitor, Galleria mellonella, and Musca domestica were intentionally infected with DmNV. In addition, native invertebrates were collected and D. melanogaster stocks were purchased and screened for DmNV presence using reverse transcription-polymerase chain reaction (RT-PCR) before being intentionally infected for study. All Drosophila species and other invertebrates, except M. domestica, that were intentionally infected with DmNV ended up scoring positive for the virus via RT-PCR. DmNV infection was also detected in three native invertebrates  and all commercially available stocks tested. These findings suggest that DmNV readily infects individuals of other species of invertebrates, while also appearing to be an endemic virus in both wild and laboratory D. melanogaster populations. The detection of DmNV in commercially available stocks presents a cautionary message for scientists using these stocks in studies of virology and immunology. Drosophila melanogaster Nora virus (DmNV) is a positive-sense single-stranded RNA virus that is spread via the fecal-oral route. DmNV has genetic similarity to viruses of the Picornaviridae and Iflaviridae families, specifically regarding the helicase and RNA polymerase regions, respectively. The DmNV genome has 4 open reading frames (ORFs), with ORF2 encoding a helicase-protease-replicase cassette, characteristic of picornaviruses [ORF1 encodes an RNAi inhibitor [ORF4 encodes a polyprotein that, after proteolytic cleavage gives rise to 3 mature capsid proteins. Finally, ORF3 appears to encode a capsid-stabilizing protein [Picornavirales [aviruses . Meanwhi protein . The stravirales .Picornaviridae family [Nasonia vitripennis virus 3 in Nasonia parasitoid wasps [Haematobia irritans [Spodoptera exigua Nora virus [Agrotis ipsilon Nora virus [Apis mellifera Nora virus [Helicoverpa armigera Nora virus. These viruses are all genetically similar to the DmNV and appear to cluster together in a previously unknown family of insect viruses distinct from Picornaviridae [D. melanogaster is possible.While DmNV is picorna-like, it does not appear to fall neatly within the e family . In factid wasps , an unnairritans , the Spora virus , Agrotisra virus , Apis mera virus  and Heliaviridae . Given tD. melanogaster research, as unrecognized DmNV infection in laboratory stocks could, in theory, confound experimental results. For this reason, stocks ordered from a stock center were tested to investigate the prevalence of DmNV in laboratory-reared D. melanogaster.Finally, the only overt pathogenic effects of DmNV are minimal decreases in survivability and a locomotor deficiency that is apparent through geotaxis assays with large sample sizes . HoweverGryllodes sigillatus (crickets), Tenebrio molitor , Galleria mellonella (wax moths), and Musca domestica (house flies) in the laboratory setting via an established infection protocol. Further, to investigate the potential for DmNV cross-species infection in the wild, insects were collected and tested for the presence of DmNV infection via RNA extraction and reverse transcription-polymerase chain reaction (RT-PCR) using primers specific for DmNV ORF1. Finally, the presence of DmNV infection in laboratory stocks of D. melanogaster from a stock center was performed to raise awareness of the prevalence of DmNV in D. melanogaster populations within the research laboratory.The purpose of this study was to investigate the ability of DmNV to infect individuals of other species of Drosophila, and other insects, including RelE23  D. melanogaster were maintained at 25 °C on standard cornmeal, molasses and torula yeast medium with diurnal light. Flies were either infected fecal-orally to establish Nora virus-infected (NV+) stocks or maintained uninfected for further analysis. Once adequately established, stocks were expanded for fly collection by transferring flies into new bottles. The Witi RelE23 stocks were checked weekly via RT-PCR for NV infection via Witi 2·H2O, 182 mM KCl, 46 mM NaCl, 10 mM Tris base, pH adjusted to 7.2, and was sterilized by autoclaving), placed on fresh food, and reared under standard conditions. Once the adults had developed, they were subjected to testing for NV infection via Stock bottles were routinely checked to verify uninfected status. If a stock became NV+, embryos were collected on apple juice agar plates and dechorionated in 2.7% hypochlorite for 2–5 min. The dechorionated embryos were washed with 1X Drosophila Ringer’s solution . After 96 h, the NV+ D. melanogaster males were removed and replaced with 5 Nora virus negative (NV−) males and 5 NV− females of each species: D. yakuba, D. mercatorum, and D. melanogaster. Flies were collected after 5 days and stored at −80 °C. Three biological replicates with ten flies each were tested in duplicate for NV infection via ablished ,16. For ® Instant Drosophila Medium . The males were allowed to defecate on the food for 96 h to ensure adequate transfer of Nora virus to the surface of the food. After 96 h, 10 M. domestica pupa  and 10 G. mellonella larva  were added to individual vials and allowed to eclose at room temperature. The mature M. domestica and G. mellonella remained on the infected food for 7 days to allow for a productive infection. Collected M. domestica and G. mellonella were stored at −80 °C. Three biological replicates with ten M. domestica or G. mellonella each were tested in duplicate for NV infection via Ten NV+ D. melanogaster males were placed into 10 vials with Formula 4-24D. melanogaster were placed into vials containing a 1:1 combination of Formula 4-24® Instant Drosophila Medium  and Instant Dry Cricket medium . The males were allowed to defecate on the food for 96 h to ensure adequate transfer of Nora virus to the surface of the food. After 96 h, 5 nymph G. sigillatus  were added to pre-prepared bug containers with sterile sand and the infected food was placed inside. The G. sigillatus remained on the infected food for 7 days. Collected G. sigillatus were stored at −80 °C. Three biological replicates with ten G. sigillatus each were tested in duplicate for NV infection via Ten NV+ T. molitor  were placed into each container. Every third day, the T. molitor were fed 30 NV+ D. melanogaster for 14 days. After 14 days, the T. molitor were collected and stored at −80 °C. Three biological replicates with ten T. molitor each were tested in duplicate for NV infection via Ten small plastic insect containers were prepared with soil and potato pieces. Five Native insects were randomly collected from counties Hamilton, Butler, Buffalo, Merrick, Hall, and Polk in Nebraska. Invertebrates were placed in sealable baggies and placed on ice. Once collected, insects were stored at −80 °C. Native insects were individually tested in duplicate for NV infection via D. melanogaster. Canton Special wild-type , Oregon-R wild-type , vestigial , and apterous  D. melanogaster stocks were purchased. Upon receipt of the stocks, they were quarantined in a separate room isolated from the other stocks maintained for research purposes that were known to be infected with Nora virus. Ten emergents from each stock vial were collected within 24 h of eclosion and tested in duplicate for Nora virus infection per Drosophila stocks were purchased from the Bloomington Drosophila Stock Center (BDSC) to investigate the prevalence of DmNV in commercially available/laboratory-reared ® per manufacturer’s instructions . Each sample was quantitated using a NanoDropTM ONE spectrophotometer (ThermoFisher Scientific) to assess RNA purity (260/280 ≈ 2.0) and concentration. Samples were analyzed for the presence of Nora virus using Nora ORF1 55–844  primers and qScript XLT 1-Step RT-PCR  according to manufacturer’s instructions. The positive controls were an RNA extraction that previously tested positive for Nora virus. Reactions using 250 ng of total RNA were set-up under the following conditions for Nora virus: 50 °C for 30 min, 94 °C for 2 min,  for 30 cycles, 68 °C for 5 min, and hold at 4 °C. Samples were analyzed on a 1.0% agarose gel in a TAE buffer solution at 50 V for 3 h. A positive reaction yielded a product at approximately 790 bp for DmNV [https://blast.ncbi.nlm.nih.gov/Blast.cgi; accessed on 1 June 2022) to determine sequence identity. All sequenced products were identified as DmNV glycoprotein 1 .Total RNA extraction was performed using TRIzolfor DmNV ,14,15. DORF1. A 790 bp product for ORF1 verified DmNV infection  was selected because it is a non-Drosophilidae Dipteran species. G. sigillatus (house cricket), G. mellonella (wax moth), and T. molitor  were selected because they are commercially available, easy to rear, and easily adaptable for the infection protocol. M. domestica were not able to be infected with DmNV  and/or are currently being used in the laboratory . Using RT-PCR, DmNV infection was found in all stocks purchased , G. sigillatus (Order: Orthoptera–cricket), and T. molitor (Order: Coleoptera–beetle) . The viral connections between some of these species and D. melanogaster have been shown in past studies with different viruses. For example, G. sigillatus is susceptible to Cricket Paralysis virus (CrPV) [D. melanogaster, and G. mellonella [D. melanogaster, G. mellonella [T. molitor [G. sigillatus can be infected by a related iridovirus, Cricket iridovirus (CrIV) [H. irritans  [S. exigua (Order: Lepidoptera) [A. ipsilon (Order: Lepidoptera) [A. mellifera (Order: Hymenoptera) [H. armigera (Order: Lepidoptera) [S. virginica from Order Lepidoptera and O. disjunctus from the Order Coleoptera, as well a non-Insecta class species, Diplopoda (Class: Diplopoda). This indicates that invertebrates in the Orders Lepidoptera and Coleoptera, may share similar virus receptors to Drosophila, specifically when considering DmNV. This also demonstrates the breadth of organisms that can be infected by DmNV, both in the laboratory and in nature. These data corroborate the hypothesis that DmNV may represent the first member of a widespread family of viruses [M. domestica was not able to be infected in the laboratory setting . Therefore, not being able to infect individuals of this species with DmNV is not surprising.Not only was DmNV able to infect individuals of other Drosophila species, it was also able to infect laboratory-reared insects. DmNV was able to infect –beetle) , but nots (CrPV) , which illonella . In addillonella , and T.  molitor , whereass (CrIV) . Additiouscidae) , S. exigdoptera) , A. ipsidoptera) , A. mellnoptera)  and H. adoptera) . Further viruses . Interes setting , nor was setting . Upon anORF1 of these stocks indicates that DmNV is present within all 4 stocks purchased  identified as candidate markers of DmNV infection [D. melanogaster [D. melanogaster, especially those examining gene regulation or innate immunity. The data presented suggests that DmNV has the ability to cross-infect not only individuals of other Drosophila species but other organisms, and unknown contamination of stocks from either stock centers or other laboratories. It is important to quarantine stocks upon receipt and subsequently test for infection before carrying out experiments. Fortunately, since DmNV is horizontally transferred, infected stocks can be easily cured by dechorionation of embryos and transferring these to fresh food.As noted earlier, the only observed phenotype with DmNV infection is a geotaxis defect detectable when large sample sizes are analyzed via a geotaxis assay . What isnfection , similarnogaster ,26. Thisnogaster ). All ofD. melanogaster. Lastly, this investigation offers a cautionary tale for those who perform research in the realm of virology and immunology using stocks from a stock center or donated from other laboratories, as DmNV infection appears to be endemic to many laboratory-reared stocks.In conclusion, this study demonstrates that the host range of DmNV is perhaps larger than previously expected. Namely, DmNV has the ability to infect not only Drosophila species but other invertebrates as well. Further examination of DmNV or DmNV-like viruses in these native invertebrate species needs to be performed. In addition, the receptor that gives DmNV access to host cells needs to be elucidated. This would give a better understanding as to how DmNV is able to infect other organisms besides