To evaluate measurement discrepancies by race between pulse oximetry and arterial oxygen saturation  among inpatients not in intensive care.Multicenter, retrospective cohort study using electronic medical records from general care medical and surgical inpatients.Veteran Health Administration, a national and racially diverse integrated health system in the United States, from 2013 to 2019.Adult inpatients in general care , in Veteran Health Administration medical centers.2) of <88% despite a pulse oximetry (SpO2) reading of ≥92%), and whether rates of occult hypoxemia varied by race and ethnic origin.Occult hypoxemia ) patients; non-Hispanic black patients and Hispanic or Latino patients accounted for 6498 (21.6%) and 1623 (5.4%) pairs in the sample, respectively. Among SpO2 values greater or equal to 92%, unadjusted probabilities of occult hypoxemia were 15.6%  in white patients, 19.6% (18.6% to 20.6%) in black patients , and 16.2% (14.4% to 18.1%) in Hispanic or Latino patients . This result was consistent in SpO2-SaO2 pairs restricted to occur within 5 minutes and 2 minutes. In white patients, an initial SpO2-SaO2 pair with little difference in saturation was associated with a 2.7%  probability of SaO2 <88% on a later paired SpO2-SaO2 reading showing an SpO2 of 92%, but black patients had a higher probability (12.9% (−3.3% to 29.0%)).A total of 30 039 pairs of SpO2) and pulse oximetry (SpO2) were obtained, black patients had higher odds than white patients of having occult hypoxemia noted on arterial blood gas but not detected by pulse oximetry. This difference could limit access to supplemental oxygen and other more intensive support and treatments for black patients.In general care inpatient settings across the Veterans Health Administration where paired readings of arterial blood gas (SaO Among critically ill patients, patients with occult hypoxemia detected by arterial blood gas but missed by pulse oximetry have recently been shown to have worse clinical outcomes by detailed analyses—including higher mortality and greater incidence of organ failure—as might be expected given the central role of oxygen delivery in healthy cellular functioning.Pulse oximetry is a ubiquitous technology with applications in both ambulatory and inpatient settings. Despite its widespread use, variation in device accuracy by patient race in critically ill patients has been reported as early as 1990.2 <85%.2 readings of ≥92%, acting on existing data could lead to unnecessary invasive measurement of arterial blood gases or unnecessary expenditures on new pulse oximeters. Finally, while SpO2-SaO2 discrepancies are more common in black patients than in white patients, the stability of such discrepancies within an individual patient is unclear. Thus, if a clinician has documented such a discrepancy (or its absence) and the same patient later shows signs or symptoms compatible with arterial hypoxemia, the next step is unclear: whether to repeat an arterial blood gas measurement, or to assume the same direction and magnitude of the SpO2-SaO2 discrepancy as recently documented.Policy and regulatory interest over the technology has increased,2-SaO2 measurements collected during routine care from inpatients in the Veterans Health Administration from 2013 to 2019.We sought to correct these gaps in this study of medical and surgical inpatients in general care in the Veterans Health Administration, a large and diverse health system serving veterans across the US. We hypothesized that black patients in general care  would have an increased frequency of occult hypoxemia compared with white inpatients in general care. We also hypothesized that a blood gas measurement showing the absence of occult hypoxemia would be associated with a low probability of occult hypoxemia on subsequent arterial blood gases, and that this probability would not vary by race. As in past work, we analyzed spontaneously recorded SpO2 and SaO2 data from the Corporate Data Warehouse for all hospital stays in acute medical and surgical care in the Veterans Affairs Patient Database (VAPD). 2 values and time of pulse oximetry measurement on general care floors are generally entered into the electronic health record by the clinician measuring them; SaO2 values of arterial blood gas and timing of the blood draw come directly from laboratory reporting systems.The nationwide Veterans Health Administration provides comprehensive inpatient and outpatient medical care to US veterans in over 100 hospitals. During the study period, Veterans Health Administration hospitals used one electronic health record system (Computerized Patient Record System), which archived data to a central repository (Corporate Data Warehouse). During the study (2013-19), we extracted SpO2 <88%) with a pulse oximetry reading of SpO2 ≥92% recorded at the same time. This range of SpO2 was selected because clinicians would probably not increase oxygen levels in response to SpO2 ≥92%.2 was included as a continuous variable. All pairs of SpO2 and SaO2 values that were measured within 10 minutes for patients in the VAPD were identified. We excluded SaO2 measurements that were labeled as temperature corrected or calculated, and information on whether the SaO2 reading was measured by co-oximetry was not available. To remove critically ill patients and isolate a sample consisting of patients in general care for these analyses, we excluded pairs of readings that were measured on days when patients were in intensive care or transferred in or out of intensive care. For the same reason, we also excluded pairs of SpO2-SaO2 readings from the rare days when more than two blood gases were drawn. We removed SaO2 and SpO2 values lower than 70% to reduce the possibility of mislabeling a venous blood gas as an arterial blood gas or data entry error, because such values seemed unlikely to be accurate in patients who were not critically ill.We defined occult hypoxemia as a low saturation of arterial oxygen ; therefore, these data were complete for all analyses. For race and ethnic origin, adjusted models that predicted occult hypoxemia included non-Hispanic black (referred to in this article as “black”), non-Hispanic white (“white”), and Hispanic or Latino (“Hispanic”), consistent with previous research on racial and ethnic differences of occult hypoxemia being most reproducible between those groups.2-SaO2 reading on that day; χ2 and independent t tests were conducted to compare patient characteristics by race and ethnic origin. We fit a multivariable logistic regression model to predict the odds of occult hypoxemia (SaO2 <88% for patients with SpO2 ≥92%). Models were adjusted for patient level characteristics that included black race (v white race), age, sex, patient comorbidities (appendix 1), supplemental oxygen (as a continuous variable), and diagnoses on admission .2 readings to account for non-linearity in the difference between SpO2 and SaO2 values, and we included an interaction term for black race and SpO2 readings to test the hypothesis that the relation between race and occult hypoxemia is different across values of SpO2. Standard errors robust to clusters were used to account for clustering at the patient level.Analyses included all patient days with one or two pairs of SpOv white patients, and separately for Hispanic patients v white patients).Because of the complex non-linearities and interactions in the model, we did not report the specific coefficients for race. Instead, we reported predictive margins and average marginal effects of occult hypoxemia for each category of race included in the model  with SpO2 values ≥92%. The number of predicted instances of occult hypoxemia, and not unique hospital admissions or unique patients, is rounded to the nearest thousand to emphasize to readers that this number is an estimate.We estimated the potential population level burden of occult hypoxemia among general care inpatients in VA hospitals across the system under the simple assumption that the probability of occult hypoxemia was the same in SpO2 value minus the SaO2 value), precision (standard deviation of the differences), limits of agreement (bias ±1.96×standard deviation), and accuracy (root mean square error (ARMS) is the square root of the sum of squared bias and squared precision) by race and ethnic origin for the entire dataset according to previously described methods.2 and SpO2). A root mean square error of ≤2-3% is required by the US Food and Drug Administration (FDA) for initial approval pulse oximetry devices.We calculated bias  and Stata/MP version 16.1 . The analytic code is included in appendix 2, and is available through GitHub. This study followed the STROBE statement  guidelinesNo members of the public were formally involved in the conception of this study, but our interest in the phenomenon of pulse oximetry measurement bias stems from caring for patients from different racial and ethnic backgrounds during the covid-19 pandemic.2-SaO2 readings recorded within 10 minutes of each other during the study period. From these, 33 556 SpO2-SaO2 pairs were excluded, representing patients who received intensive care at any point on that calendar day. We also excluded 204 pairs from patient days with more than two pairs of readings in one day. The cohort included 30 039 SpO2-SaO2 pairs ). SaO2 readings were slightly more common on days when an SpO2 reading was recorded for white patients than for black patients ), but if an SaO2 reading was available on an SpO2 monitored day, no substantial difference by race was seen in whether that reading was close enough in time to be in a pair with a recorded SpO2 reading .Characteristics of included pairs of SpO2-SaO2 pairs by race and ethnic origin at the SpO2 range of 89-100% are summarized in appendix 3. Among all values of SpO2 ≥92%, unadjusted probabilities of occult hypoxemia  were 15.6%  for 2823 pairs in white patients, and 19.6% (18.6% to 20.6%) for 1144 pairs in black patients , and pulse oximeters had better precision in white patients than in black patients , when the paired SaO2 values were used as the gold standard across the entire range of SpO2 values in the data (appendix 5).White patients faced less measurement bias in pulse oximetry than black patients  for 24 009 pairs, absolute adjusted increased probability of occult hypoxemia 4.0% ), for pairs no more than 5 minutes apart (2.6 minutes (1.0-4.0) for 12 603 pairs, 3.7% ), or for pairs no more than 2 minutes apart (1.0 minute (0.2-1.5) for 5305 pairs, 4.6% ; This estimate of racial differences in occult hypoxemia was not sensitive to time between SpO2 readings of 92-100% for black and white inpatients in the study. If occult hypoxemia occurred at the same rate as occult hypoxemia in the SpO2-SaO2 pairs analyzed in the study, then an estimated 573 000 additional instances of occult hypoxemia would have occurred in black patients during the study period and would have been detected if pulse oximeters performed as well in black patients as in white patients.In total, we observed 54 048 788 SpO2-SaO2 measurements recorded on the same day. We aimed to determine whether the difference within SpO2-SaO2 readings from the first pair was associated with the odds of occult hypoxemia in the second pair. In 2-SaO2 differences from the first reading of the day into equal groups by tertiles. These groups ranged from having the lowest SpO2-SaO2 difference (SaO2 reading is 0.1 percentage points lower than or any amount higher than SpO2 reading) to having the largest SpO2-SaO2 difference (SaO2 reading is at least 2.5 percentage points lower than SpO2 reading).We recorded a total of 3016 patient days  that had two pairs of SpO2 of 92% in the second pair of readings were 2.7%  if the first pair was in the group with the smallest SpO2-SaO2 difference, 2.4% (−0.1% to 4.8%) if the first pair had an SpO2-SaO2 difference in the intermediate group, but 32.0% (15.2% to 48.8%) if the first pair in the group with the largest SpO2-SaO2 difference. Similar patterns in white patients were seen with SpO2 readings of 98% in the second pair. Probabilities of occult hypoxemia of 2.4% (0.4% to 4.5%) and 2.1% (0.3% to 3.9%) were recorded in the groups with the smallest and intermediate SpO2-SaO2 differences, respectively, but 33.8% (26.1% to 41.50%) if the SpO2-SaO2 difference was in the highest group  probability of hypoxemia. The probability of occult hypoxemia in black patients with the largest SpO2-SaO2 differences on the first pair was as high as in white patients if the SpO2 value in the second pair was 92% (39.6% (11.5% to 67.7%), and even when the SpO2 value was 98% (28.4% (18.5% to 38.2%); The probabilities of occult hypoxemia on the second SpO2-SaO2 readings varied depending on magnitude of the difference in the first pair, SpO2 value in the second pair, and patient race. For example, for white patients who did not show occult hypoxemia in the first pair of SpO2-SaO2 readings of a given day, occult hypoxemia was present in only 2.5% of the second pair of SpO2-SaO2 measurements. Among black patients with two pairs of SpO2-SaO2 readings in a given day and with no occult hypoxemia shown in the first pair, occult hypoxemia was present in 6.5% of the second pair of SpO2-SaO2 measurements. While the two SpO2-SaO2 pairs had, on average, the same difference between SaO2 and SpO2 (mean 0.05 for white patients and 0.15 for black patients), the standard deviation in differences between the pairs was 4.1 for white patients and 4.9 for black patients, indicating that the possibility of wide discrepancy was more common in black patients.To put it another way, the odds of occult hypoxemia on the second pair of SpO2-SaO2 measurements from patients identified as Hispanic (or Latino). Among all pairs with SpO2 readings of ≥92%, the unadjusted probability of occult hypoxemia (SaO2 <88%) was 16.2%  in these patients , and pulse oximeters had similar precision in Hispanic patients (6.6 v 6.7), when the paired SaO2 was used as the gold standard across the entire range of SpO2 in the data (appendix 5). Because of the small sample size, we did not further analyze this subgroup of patients.We also analyzed the results of 1623 pairs of SpO2 readings of 92-100%. Including all readings did not meaningfully change the results (appendix 7). Controlling for clustering within hospitals as a random effect did not meaningfully change the results (appendix 8). Analyses testing for a difference in the observed race differences in measurement bias in surgical and non-surgical patients yielded inconsistent results that were sensitive to subtle parameterization choices in the small (<10% of sample) population of patients in surgical general care with SpO2-SaO2 pairs, but did not change the estimated differences meaningfully in the large majority of patients who were not in surgical care (appendix 9).Focusing on differences between black and white patients, our primary analyses in 2-SaO2 readings, white patients could have some reassurance that a later normal SpO2 reading was unlikely to be associated with a SaO2 reading of <88%; however, less reassurance was available for black patients. The overall prevalence of occult hypoxemia could be considerable and racially biased.This study of inpatients in general care in the Veterans Health Administration indicates a significant difference in the ability of pulse oximetry to detect clinically relevant hypoxemia in patients of different races. Black patients were found to have more occult hypoxemia than white patients. Pulse oximetry readings had greater bias and worse precision among black inpatients in general care than among white inpatients in general care. This greater bias and worse precision meant that on receiving a recent and well correlated pair of SpO2 measurement bias between black and white patients, where the odds of occult hypoxemia were higher in black preterm infants,Our findings accord with the most recent data on racial bias in pulse oximetry focused on more ill patients, often in intensive care.Our results add to the literature by showing that the concerns about worse bias and noise  in black patients being monitored with pulse oximetry are also relevant to patients in general care who often receive no or little supplemental oxygen. They also suggest the possibility that this differential pulse oximeter performance affects Hispanic patients, although those results need additional confirmation. Two independent groups have recently shown that arterial hypoxemia undetected by pulse oximetry is associated with worse clinical outcomes.2 and SaO2 readings. These data indicate that excess occult hypoxemia is present in black patients relative to white patients, regardless of whether the SpO2-SaO2 pairs are measured within 2, 5, or 10 minutes apart, suggesting that time between measurements is not a driver of these results—consistent with other recently published work.Our study looks at concerns about possible differential bias being introduced because of the potential interval between SpO2-SaO2 readings measured on the same day, a well aligned SpO2-SaO2 pair for white patients was associated with low levels of occult hypoxemia on subsequent pairs; such concordance might be reassuring in many clinical scenarios. This concordance was less true for black patients, and these differences should be considered in deciding whether to obtain an arterial blood gas reading in appropriate clinical situations until non-racially biased pulse oximeters are in use.Errors in pulse oximeters could be due to a combination of systematic error or bias, which is reproducible across measurements, as well as random error or noise.2 reading of ≥92% is truly associated with an SaO2 reading of <88%. However, for this non-random arterial blood sampling to explain the apparent differences between racial groups, it would be necessary that clinicians are better at making the bedside diagnosis of occult hypoxemia in black patients than in white patients—and, at the same time, that they order confirmatory tests less often but more accurately in black patients. We are unaware of data that support such an alternative mechanism.This study had several limitations. Arterial blood gases are measured less frequently than pulse oximetry. If clinicians are more likely to obtain an arterial blood gas when they have concerns about the accuracy of pulse oximetry, then the probabilities of occult hypoxemia noted in our study would overestimate the frequency with which an SpOSkin color is not consistently recorded as part of the medical record, so we used race as a surrogate, which might not fully reflect the skin tone diversity within each patient group or other differences that might contribute to pulse oximetry bias. Data on Hispanic patients suggest a similar problem, but the smaller sample size resulted in more imprecise estimates; these data should be considered as hypothesis generating unless replicated. Likewise, the potential instability of estimates of racial differentials in surgical patients warrants additional exploration, although should be taken in the light of other published findings.An additional limitation was that nearly all patients had a military service history. No oximeter brand information was available for this study. However, most commercially available oximeters use similar technology.Black patients under the inpatient care of the US Veterans Administration had excess episodes of undetected hypoxemia in 2013-19 compared with white patients. Other scientists have reported increased morbidity and mortality associated with occult hypoxemia.2) is a method used to non-invasively measure arterial oxygen saturation (SaO2); the accepted most accurate method is by invasive arterial blood gasPulse oximetry (SpO2 <88% despite an SpO2 ≥92%) experience worse organ dysfunction and mortality rates in hospital than those without occult hypoxemiaPatients in intensive care with occult hypoxemia  care in hospital, occult hypoxemia was more common for black patients than for white patients; differences in occult hypoxemia by race from real world data did not change according to differences between the recording of the SpO2-SaO2 readings in a day were unlikely to have it on a second pair of SpO2-SaO2 readings on the same day; black patients had less consistency across different pairs of SpO2-SaO2 measurements on the same day White patients not showing occult hypoxemia on a first pair of SpO2 reading does not show occult hypoxemia in a black patient, an elevated index of suspicion might be warranted in black patients with compatible signs and symptoms until unbiased pulse oximeters are routinely availableThese results suggest that if a recent SaO


RVFV causes severe and recurrent outbreaks in Africa and the Arabian Peninsula with a significant risk for emergence into new locations. Although there are a variety of RVFV veterinary vaccines for use in endemic areas, there is currently no licensed vaccine for human use; therefore, there is a need to develop and assess new vaccines. Herein, we report a live-attenuated recombinant vaccine candidate for RVFV, based on the previously described genomic reconfiguration of the conditionally licensed MP12 vaccine. There are two general strategies used to develop live-attenuated RVFV vaccines, one being serial passage of wild-type RVFV strains to select attenuated mutants such as Smithburn, Clone 13, and MP12 vaccine strains. The second strategy has utilized reverse genetics to attenuate RVFV strains by introducing deletions or insertions within the viral genome. The novel candidate vaccine characterized in this report contains a two-segmented genome that lacks the medium viral segment (M) and two virulence genes . The vaccine candidate, named r2segMP12, was evaluated for the production of neutralizing antibodies to RVFV in outbred CD-1 mice. The immune response induced by the r2segMP12 vaccine candidate was directly compared to the immune response induced by the rMP12 parental strain vaccine. Our study demonstrated that a single immunization with the r2segMP12 vaccine candidate at 10 Phenuiviridae, Phlebovirus) is a clinically important mosquito-borne pathogen causing disease in both humans and ruminants. Although most humans have no clinical sign, others develop flu-like symptoms with headaches, fever, or myalgia  gene  for a neutralizing antibody titer at or above the threshold antibody level for protection.The neutralizing antibody response produced by the different titers of r2segMP12 was compared between the use of a single dose versus a single dose followed by a booster dose. Neutralizing antibodies produced following the r2segMP12 vaccine and the rMP12 parental strain vaccine were also examined. Taken together, these data demonstrated that the double deletion of NSs and NSm genes does not reduce the immunogenicity of the MP12 vaccine strain.The r2segMP12 recombinant vaccine candidate for RVFV was produced in a previously published study as summarized in 3, 104, or 105 plaque-forming unit [PFU]) of infectious viruses in a single immunization or two immunizations  were subcutaneously immunized with one of the following: r2segMP12 vaccine candidate, rMP12 parental strain vaccine, or sterile L-15 media. Animals were randomly assigned into 10 groups of 5, representing 8 experimental regimens, using increasing doses (10izations .n = 5) was included to determine the correlation of neutralizing antibodies produced by different dosages of the r2segMP12 strain. Three groups of mice (n = 5) received a second immunization of the r2segMP12 strain to evaluate the effect of a booster immunization using the dose of the vaccine that produced the highest neutralizing antibody titer after the primary vaccine (105 PFU). In addition to the experimental groups that received the r2segMP12 vaccine candidate, four additional groups (n = 5) were designated as control groups, with two positive control groups receiving the rMP12 vaccine at 105 PFU and two negative control groups receiving an equal volume of sterile culture L-15 media.The regimen of a single immunization administered at an increasing dosage per group . Before the initial immunization, all mice were determined to be healthy and seronegative to RVFV through the analysis of collected serum using PRNT (data not shown). Animals were maintained for 6 weeks after the initial immunization. Mice were immobilized using an isoflurane vaporizer before blood collection. Whole blood samples of 0.1 mL were collected from the lateral saphenous vein from immunized animals at 20 days after initial immunization using a 22 g needle. Serum samples were obtained through centrifugation of coagulated blood at 2,000 PRNT were used to determine which vaccination regimen produced the highest titer of neutralizing antibodies following the protocol previously described . Seroconversion was defined using the cutoff of 1:10 PRNT50 titer, a seropositive threshold commonly used for assessing the neutralizing antibody responses elicited by arbovirus vaccines . Finally, PRNT50 titers of animals receiving only a single dosage of the r2segMP12 vaccine candidate at 105 PFU were compared to animals receiving the rMP12 vaccine at 105 PFU at 42 dpi with a Mann–Whitney test. All tests were performed using the GraphPad Prism (version 8.1.2) program .Using a Kruskal–Wallis test followed by a Dunnett's test 50 > 10). All, but one mouse immunized with the r2segMP12 vaccine candidate seroconverted after a single immunization , demonstrating a dose–response relationship in the vaccine immunogenicity. Importantly, the comparison of immunogenicity with the rMP12 vaccine strain suggests the superior immunogenicity of the r2segMP12 strain. Mice immunized with r2segMP12 at a titer of 105 PFU had a significantly higher neutralizing antibody response compared to mice that received the rMP12 vaccine at the same titer (p = 0.0079).Therefore, the r2segMP12 vaccine candidate was capable of eliciting neutralizing antibody responses in CD-1 mice at dosages between 1050 titers were measured in mice that received varying initial titers of the r2segMP12, followed by a booster at 21 dpi. Animals in groups 6, 7, and 8 . In addition, serum neutralizing titers in four out of five mice immunized with the rMP12 vaccine wane below PRNT50 titer of 10, demonstrating the need for a booster immunization. These data suggest that the r2segMP12 strain is superior to the rMP12 vaccine in eliciting neutralizing antibody responses in mice.Given the observation that a single dose of the r2segMP12 vaccine candidate at 10Collectively, these data suggest that the r2segMP12 strain is immunogenic and can elicit neutralizing antibody responses in CD-1 mice that received one single immunization. In addition, the lack of the NSs and NSm genes ensures the safety, but does not compromise the immunogenicity of the r2segMP12 vaccine strain.Due to the impact of RVFV on both human and livestock health, efforts to prevent and control RVFV have been continuous, however, the limitations of each vaccine, multiple doses required, and expenses to maintain these regimens have made it difficult  (McElroy et al, We conclude that the superior immunogenicity of the r2segMP12 strain warrants its advancement in the process of vaccine development, including challenge protection studies conducted in mice and then sheep, which are the amplifying hosts for RVFV. Future experiments will focus on the characterization of the immune response induced by r2segMP12 and its ability to protect against a lethal RVFV challenge.


Keap1 mutations regulate Nrf2 activity and lead to chemoresistance in cancers. Yet the underlying molecular mechanisms of chemoresistance are poorly explored. By focusing and genotyping head and neck squamous cell carcinoma (HNSCC) that had available pathologic and clinical data, we provide evidence that Keap1 displays frequent alterations (17%) in HNSCC. Functional loss of Keap1 results in significant activation of Nrf2 and promotes cancer cell growth, proliferation, and elevated cancer stem cell (CSCs) self-renewal efficiency and resistance to oxidative stress. Furthermore, decreased Keap1 activity in these cells increased nuclear accumulation of Nrf2 and activation of the Notch pathway, causing enhanced transcriptional alterations of antioxidants, xenobiotic metabolism enzymes, and resistance to chemotherapeutic treatment. Limiting the Nrf2 activity by either Keap1 complementation or by Nrf2 silencing increased the sensitivity to chemotherapy in Keap1-mutated cells and repressed the CSC self-renewal activity. Our findings suggest that Keap1 mutations define a distinct disease phenotype and the Keap1-Nrf2 pathway is one of the leading molecular mechanisms for clinical chemotherapeutic resistance. Targeting this pathway may provide a potential and attractive personalized treatment strategy for overcoming chemotherapeutic resistance conferred by Keap1 mutations. Head and neck squamous cell carcinoma (HNSCC) is the utmost global health concern and affects >890,000 patients, and over 450,000 HNSCC-related deaths occur every year , 2. To dKeap1, and Nrf2- pathway mutations were found in patients with HNSCC.The Cancer Genome Atlas (TCGA) project has profiled an extensive landscape of somatic genomic alterations in HNSCC . This reKeap1) strongly induces NF-E2-related factor 2 (Nrf2), and acquires malignancy in several types of cancer ). The patient had rapid disease progression and underwent biopsy and genotyping of lung metastasis that revealed Keap1 mutations  tumors out of 24 HNSCC patient tumors sequenced, which is much higher than the TCGA data set. The possible putative reason for the higher incidence of Keap1 mutations in our case is unknown however, we speculate that demographic and genetic makeup may play roles in the higher incidence of Keap1 mutations in HNSCC patients. In this study, we found that all Keap1 mutated tumors exhibited nuclear accumulation of Nrf2 as assessed by immunohistochemical analysis. Keap1 mutations result in the accumulation and activation of Nrf2 and may partly confer the resistance to chemotherapeutic treatment in HNSCC patients and reduce drug-induced ROS production. Importantly, all these mutations involved functionally relevant domains of Keap1 protein, including BTB (c.403 C > T and c.1129 G > A), IVR domain (c.1111 G > A), and KR3 region (c.1766A > G) of Keap1, which are responsible for the ubiquitination and binding of Nrf2 [Nrf2 pathway has been proposed to be the leading cause of chemoresistance in several cancers [Nrf2 pathway activation in HNSCC, we sequenced the Keap1 and Nrf2 genes and identified mutations in Nrf2 immunopositive tumors. Interestingly, the Nrf2 mutations involved the Neh2 domain where Keap1 binds. Importantly, we did not detect any mutations in the matched normal tissues, confirming that the mutation is somatic in origin. Furthermore, the overall frequency of mutations  for the Keap1 gene in HNSCC tissues suggests that Keap1 mutations are likely a frequent genetic alteration in HNSCC at least in our case which is much higher than that of data reported in TCGA. In addition, we assessed whether the Nrf2 activation pathway is engaged in mediating chemotherapeutic resistance in HNSCC. We used human HNSCC primary tumor cells and HNSCC cell lines to evaluate the functional relationship between Nrf2 activation and chemotherapeutic resistance. Knockdown of Keap1 by siRNA in HNSCC cells demonstrated enhanced Nrf2 pathway activity, which led to enhanced transcriptional activity thereby rendering HNSCC cells resistant to chemotherapy. We have achieved a comparable result in HNSCC tumor and normal tissue, where the loss of functional Keap1 gene and subsequently increased staining intensity of Nrf2 corroborate the above findings. In concordance with the above findings, as expected, GST, NQO1, and SOD1 enzyme levels and GSH levels were highly significantly elevated in the tumor tissues compared with matched normal individuals. This suggests that upregulation of these Nrf2-dependent genes likely contributed to the resistance to chemotherapy treatment and cell survival. This result is in agreement with previously published results, where high antioxidant capacity increases cell survival and proliferation and protects against oxidants, radiation therapy, and chemotherapies [Loss of ux pumps . In the  of Nrf2 , 47. The cancers , 49. To herapies .Keap1 and Nrf2 overexpression induces many stress resistance genes and can restore cancer cell proliferation. It has previously shown that constitutive activation of Nrf2 contributes to tumorigenesis, ROS detoxification, and modulation of redox state and also contributes to resistance to many anticancer drugs [Keap1 has been identified as a possible mechanism of chemoradiation resistance in many cancers [Keap1 mutations showed more resistance to etoposide and carboplatin than to Keap1 wild-type cells [Nrf2 due to Keap1 loss may confer resistance to cisplatin, a widely used chemotherapy regimen for head and neck cancer, by regulating ROS and cancer stem cell pathways. In concordance with these previous results, our results demonstrated that downregulation of the Nrf2 expression in Keap1 mutated cancer cells or introduction of Keap1 cDNA in Keap1 mutant cells significantly enhances the sensitivity to cisplatin. In analyzing the clinical cases, all four Keap1 mutated HSNCC patients had a history of recurrence within a year all cases were treated with 2 and 3 lines of chemotherapy and exhibited poor clinical response, suggesting therapeutic resistance due to Keap1 mutations as well as the potential existence of CSC populations in these tumors.Loss of er drugs , 50. Los cancers . In the pe cells . Thus, oKeap1 mutations during HNSCC oncogenesis due to the deletion of Keap1 led to the increased self-renewal activity of cancer cells and subsequent therapeutic resistance. Importantly the impact of Keap1 loss leads to increased self-renewal activity and therapeutic resistance compared to Keap1 wild-type cells or Keap1 reintroduced clones which define the greater clinical relevance. Previous reports have demonstrated the molecular mechanisms of cisplatin resistance in cancer cells and cisplatin treatment increased mitochondrial ROS generation [Keap1 or silenced Nrf2 observed increased mitochondrial ROS generation and limited the cell growth in cells treated with cisplatin. This suggests that constitutive Nrf2 activation due to Keap1 loss influences oxidative stress and reduces the cellular damage induced by the increase of ROS, therefore, triggering the resistance of chemotherapy, particularly for cisplatin. Han and colleagues [Nrf2 and Notch signaling in OCSC  cells. Activation of Nrf2 in Keap1 deleted cells resulted in hyperproliferation of squamous epithelial cells and activation of Notch signaling [Nrf2 activation in Keap1 mutated cells, suggesting that Notch also plays a role in mediating the effects of Nrf2 activation in HNSCC cells.Our observation from this study suggests a pivotal role for neration  and trigneration , 54. In lleagues  recentlyignaling . CongrueKeap1, activation of Nrf2 and the Notch signaling pathway promotes cellular metabolic reprogramming that sustains cellular proliferation. This reprogramming feature leads to the clonal expansion of mutant cells thus acquiring self-renewal capacity and triggering resistance pathways and outpacing the Keap1 WT cells and potentially acquiring additional genetic changes and subsequently leading to therapeutic failures. All these features explain the therapeutic failure and adverse and poor survival outcomes conferred by Keap1 mutations in rapidly progressing HNSCC cells. In a recent study, it was reported the Keap1-Nrf2 double mutations in lung cancer leads to poor outcome and severe therapeutic resistance in radiation therapy [Nrf2 mutation and none of the patients had concurrent Keap1-Nrf2 mutations. This may be due to the small number of patients assessed in this study and we cannot rule out that Keap1-Nrf2 dual mutations may have somatic or epigenetic consequences and may play a devastating role in therapeutic failures and poor outcomes, disease recurrence in Keap1-Nrf2-mutant tumors. Thus, future studies should include large numbers of patients’ samples to examine the effects of Keap1 mutation as well as to explore the role of Keap1-Nrf2-dual mutation in chemoresistance.Reviewing all our results, it appears that once HNSCC cells acquire a mutation in  therapy . In our Keap1–Nrf2 alterations are known to play roles in chemotherapeutic resistance particularly cisplatin resistance in HNSCC, surprisingly, the mutation status is not widely used to make a treatment decision in head and cancer. It would be interesting to investigate whether loss of Keap1 and overexpression of Nrf2 status in tumor samples are clinically relevant mechanisms of chemotherapeutic resistance in HNSCC and develop alternative therapy to counter Nrf2 activation. This may improve personalized therapy in a subset of HNSCC patients who are prone to therapeutic resistance.In conclusion, although Supplementary Figure LegendsSupplementary Figure S1Supplementary Figure S2Supplementary Figure S3Supplementary Table S1Supplementary Table S2Supplementary Table S3Supplementary Table S4Supplementary Table S5Supplementary Table S6Supplementary Table S7Reporting summer/Reproducibility checklistOriginal Data File


Bacillus vallismortis have been reported to be efficient plant-growth-promoting bacteria as well as inducers of systemic resistance. Here, we report the draft genome sequence of Bacillus vallismortis strain BL01, isolated from the roots of Artemisia lerchiana Web.Some strains of  Bacillus vallismortis can produce metabolites with strong growth inhibition activity against phytopathogenic fungi . Plant roots were disinfected with tap water for 30 s followed by 70% ethanol for 5 min and then 15% H2O2 for 10 min and sterile water for 2 min 5 times and subsequently crushed with a mortar and pestle under sterile conditions. Aliquots of 100 mL of the resulting plant juices were plated onto a 1/20 dilution of tryptic soy agar  plates. The sterility check consisted of aliquots of water from the last rinsing that were plated onto 1/20 TSA according to methods reported previously (de novo assembled using SPAdes v3.14.1 (https://github.com/lh3/seqtk). The contigs were mapped to the Bacillus vallismortis strain Bac111 (GenBank accession number CP033052.1) genome by CONTIGuator v2.7   . A totalPRJNA809498. The raw reads were deposited in the Sequence Read Archive under accession number SRR18107534. The GenBank nucleotide sequence accession number is CP092751.All data are available in the National Center for Biotechnology Information database under BioProject accession number


Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer with high heterogeneity, rapid progression, and paucity of treatment options. The most effective chemotherapeutic drug used to treat TNBC is doxorubicin (Doxo) which is an anthracycline antibiotic. However, Doxo treatment alters cytosolic calcium dynamics leading to drug-resistance condition. The aim of this study is to capture the alterations in the activity of various calcium channels and pumps during Doxo treatment and their consequences on cytosolic calcium dynamics that ultimately result in drug resistance.In the present study, a mathematical model is proposed to capture the complex dynamical landscape of intracellular calcium during Doxo treatment. This study provides an insight into Doxo remodeling of calcium dynamics and associated drug-resistance effect. The model was first analyzed analytically and then explored through numerical simulation using techniques like global sensitivity analysis, parameter recalibration, etc.2+ channels and pumps might provide efficient chemotherapy in TNBC. It was also observed that the indispensability of calcium influx rate is paramount in the Doxo drug resistance. Finally, three drugs were identified from existing literature that could be used as a combination therapy along with Doxo.The model is used to predict the potential combination therapy for Doxo that can overcome Doxo associated drug resistance. The results show targeting the dysregulated CaThe investigation highlights the importance of integrating the calcium signaling of various calcium regulating compounds for their effective anti-tumor effects deliverance along with chemotherapeutic agents. The results from this study might provide a new direction to the experimental biologists to explore different combination therapies with Doxo to enhance its anti-tumor effect. Breast cancer is the leading cause of cancer-related deaths diagnosed among women –3. Tripl3R) channels ER reduction. These channels are known as SOCE. This process is intervened by STIM1 of the ER membrane and is sensitive to ER Ca2+ levels. STIM1 proteins bind to ORAI1, the proteins embedded in the plasma membrane that forms the pore of SOCE and triggers their opening. The inward calcium flux through the SOCE channel is defined by the term β[Pso] [2+ influx through TRPC1 channel is dependent on the difference between the Ca2+ concentration of extracellular space (ces) and cytosol ([Ca2+]c). Therefore, the Ca2+ influx through TRPC1 channel is modeled as γ (esc – [Ca2+]c). We assumed the Ca2+ concentration of extracellular to be a constant as taken by Mirzakhalili et al. [ξ and δ respectively [We developed a three-dimensional model using ordinary differential equations (ODEs) to capture the complex dynamical landscape of intracellular calcium during Doxo treatment. The first equation of the model describes the rate of change of cytosolic calcium level. The inward flux of Cat rate α . SOCE ism β[Pso] . As mentm β[Pso] , the Ca2i et al. . The plaectively .2+ flux into cytosol from ER through IP3R2 channel  describes the fraction of STIM1 proteins binding to ORAI1 proteins to activate the SOCE channel for calcium influx into the cytosol from extracellular space. This fraction adjusts depending upon the changes in ER calcium levels because the diffusion of STIM1 within the ER membrane is a slow process ER, and is represented by Hill function (with Hill coefficient 4), considering affinity ks for [Ca2+]ER [The steady-state function [Ca2+]ER . Based ot > 0 : 0 < [Ca2+]c (t) <, Pso] (t) < 1}, with Result 1. (i) The solution of the system  exists aResult 2. The system  has inte21 > 0 and A1A2 > A3 holds simultaneously , γ ([Ca2+] influx rate through TRPC1 channel), δ(leaking rate), ξ([Ca2+] export through NCX), p1 ([Ca2+] influx rate through IP3R2), q1 , p2 (flux rate through SERCA1 channel) were estimated using experimental data points by Bong et al. [2+]ER and [PSO] are also obtained and shown in We explored different literatures and obtained few parameters while the rest of the parameters namely, g et al. . Here weg et al. , see FigE* was satisfied, and the interior equilibrium point E* =  is locally asymptotically stable as the eigenvalues of the characteristic equation is . This follows that CRD shows stable dynamics through damped oscillations. SOCE is an important plasma membrane channel of calcium entry in the cytosol that opens in response to ER calcium depletion. Entry through this channel is mediated by STIM1 proteins that disseminate from the ER membrane and binds to ORAI1 proteins and activates SOCE. IP3R2 channel plays an important role in the exit of calcium from ER into cytosol. The involvement of IP3R2 channel in mediating SOCE has been established earlier thereby influencing cytosolic calcium dynamics [β channel’s influx rate) and p1 ([Ca2+] influx rate through IP3R2) and its role in establishing CRD. We varied these two parameters 2-fold up down from their base values and generated the 2D parameter space shown in β and setting p1 to its baseline value, the model dynamics changes from stable focus to stable node dynamics.We also observed that for this baseline parameter set  the exisdynamics , 42. To β , γ ([Ca2+] influx rate through TRPC1 channel), p1 ([Ca2+] influx rate through IP3R2), p2 (flux rate through SERCA1 channel), p3 (flux rate through PMCA2 channel), and they were simultaneously and step-wise increased by 25% (Doxo 1), 50% (Doxo 2), and 75% (Doxo 3). The calcium profile generated through elevated parameter ranges was given in Doxo works in a concentration-dependent manner . An alteIn this section, we explored the effect of parameter variations on the system’s dynamics. We first varied single parameters through robustness analysis and observed their effect, and then we performed a global sensitivity analysis (GSA) to capture the effect of multi-parameter variation.ξ, p1, q3  method, which is based on sampling, for GSA. In our study, we used the LHS method as it compactly differentiates the input parameters . PRCC meβ), maximum Ca2+ influx through TRPC1 channel (γ), Ca2+ export through NCX (ξ) and calcium concentration in extracellular space (ces) were significantly correlated with cytosolic calcium (see 2+ influx through TRPC1 channel (γ), Ca2+ export through NCX (ξ) and calcium flux rate (p1) and half saturation constant (q1) of IP3R2 channel, calcium flux rate (p2) and half saturation constant (q2) of SERCA1 channel were significantly correlated with [Ca2+]ER  associated with these calcium channels and pumps in our model , γ , p1 , p2 , p3 . We picked one parameter from the above and reduced its value by 20%. After that, all other parameters were varied 100% up and down from their respective values (i.e. in the Doxo 3 parameter set) as done in the single parameter variation. Out of the five parameters related to Doxo associated proteins, the reduction of β  by 20% turned out to be the only parameter that can restore CRD in combination with other model parameters. Except for β , reduction in other parameters related to Doxo associated proteins was not sufficient to restore CRD in combination with any other model parameter. The results of double parameter variation in 75% increased case are given in We performed a single parameter variation where each parameter was perturbed 100% up-down from their respective values and we filtered out parameters that retained the Doxo profile to the control profile (CRD). This exercise was repeated for different Doxo treatments where associated parameters were increased by 25% (Doxo 1), 50% (Doxo 2), 75% (Doxo 3). Single parametric variation was able to restore the CRD only in the case of 25% and 50% increase. The results of single parameter variation for the first two cases of Doxo treatments are given in We filtered out those restoration approaches which robustly produced CRD once obtained in our parametric variation range with 90% CI of readout values  used in the recalibration exercise done in the previous section . We downloaded a list of drugs with their corresponding drug targets from Drug Bank database  influx), the proliferation of invasive tumor cells was reduced [Doxo is a broad-spectrum chemotherapeutic drug used in the clinic . Targetinowadays . The in gularity . In a re binding , 66. The binding . SKF9636 reduced . Moreove reduced . This unThe present study focused on the mechanistic viewpoint of reduced sensitivity of TNBC towards Doxo. The investigation highlights the importance of integrating the calcium signaling of various calcium regulating compounds for their effective anti-tumor effects deliverance along with chemotherapeutic agents. Here, we proposed few potential strategies which can be used in combination with Doxo. We further used a drug dataset to identify three drugs from existing literature that could be used individually along with Doxo as a combination therapy. Before ending this article, we would like to mention that our work lacks experimental validation, but the proposed approach delineates the complex nature of Doxo drug-resistance and can help in developing effective therapeutic strategies for TNBC.


In 2021, 20-valent pneumococcal conjugate vaccine (PCV) (PCV20)  and 15-valent PCV (PCV15) (Merck Sharp & Dohme Corp.) were licensed by the Food and Drug Administration for adults aged ≥18 years, based on studies that compared antibody responses to PCV20 and PCV15 with those to 13-valent PCV (PCV13) . Antibody responses to two additional serotypes included in PCV15 were compared to corresponding responses after PCV13 vaccination, and antibody responses to seven additional serotypes included in PCV20 were compared with those to the 23-valent pneumococcal polysaccharide vaccine (PPSV23) (Merck Sharp & Dohme Corp.). On October 20, 2021, the Advisory Committee on Immunization Practices (ACIP) recommended use of either PCV20 alone or PCV15 in series with PPSV23 for all adults aged ≥65 years, and for adults aged 19–64 years with certain underlying medical conditions or other risk factorsPPSV23 has been recommended for use in the United States since the 1980s for adults aged ≥65 years and for younger adults with underlying conditions that increase their risk for pneumococcal disease . In 2012Recent systematic reviews continue to support the effectiveness of PCV13 against invasive pneumococcal disease (IPD)During February–October 2021, ACIP reviewed the epidemiology of pneumococcal disease and considerations for use of PCV15 and PCV20 in adults. The ACIP Pneumococcal Vaccines Work Group (Work Group) evaluated the quality of evidence for PCV15 and PCV20 immunogenicity and safety using the GRADE approach.Pneumococcal disease incidence in adults. During 2018–2019, the incidence of all IPD in adults aged ≥65 years was 24 per 100,000 population  . The most frequently reported adverse reactions were injection site pain, fatigue, and myalgia. The rates of serious adverse events (SAEs) within 6 months of vaccination were 2.5% among PCV15 recipients and 2.4% among PCV13 recipients. No SAEs or deaths were considered to be related to the study vaccines . When PCV15 is used, it should be followed by a dose of PPSV23 . When PCV15 is used, it should be followed by a dose of PPSV23.Dosing schedule. When PCV15 is used, the recommended interval between administration of PCV15 and PPSV23 is ≥1 year. A minimum interval of 8 weeks can be considered for adults with an immunocompromising condition, cochlear implant, or cerebrospinal fluid leak to minimize the risk for IPD caused by serotypes unique to PPSV23 in these vulnerable groups (Adults with previous PPSV23 only. Adults who have only received PPSV23 may receive a PCV (either PCV20 or PCV15) ≥1 year after their last PPSV23 dose. When PCV15 is used in those with history of PPSV23 receipt, it need not be followed by another dose of PPSV23.Adults with previous PCV13. The incremental public health benefits of providing PCV15 or PCV20 to adults who have received PCV13 only or both PCV13 and PPSV23 have not been evaluated. These adults should complete the previously recommended PPSV23Coadministration with other vaccines. PCV15, PCV20, or PPSV23 can be coadministered with QIV in an adult immunization program, as concomitant administration  has been demonstrated to be immunogenic and safe. However, slightly lower pneumococcal serotype-specific OPA GMTs or geometric mean concentrations were reported when pneumococcal vaccines were coadministered with QIV compared with when pneumococcal vaccines were given alone  (PCV13) and the 23-valent pneumococcal polysaccharide vaccine (PPSV23) are recommended for U.S. adults. Recommendations vary by age and risk groups.On October 20, 2021, the Advisory Committee on Immunization Practices recommended 15-valent PCV (PCV15) or 20-valent PCV (PCV20) for PCV–naïve adults who are either aged ≥65 years or aged 19–64 years with certain underlying conditions. When PCV15 is used, it should be followed by a dose of PPSV23, typically ≥1 year later.Pneumococcal vaccination recommendations were simplified across age and risk group. Eligible adults may receive either PCV15 in series with PPSV23 or PCV20 alone.


The 12hr panels for EPC cells in PLOS ONE editors have no concerns about the MCF-10A panels in There appear to be similarities between the 12hr, 24hr and 48hr Hoechst panels for MCF-10A cells in Raw data underlying all the results reported in the article are available from the corresponding author, except for FCS files underlying the flow cytometry experiments presented in Figs 4, 5, and S1 FileImages were captured using BD Pathway Bio-imager 435 and each file is a composite of four separate images captured as a 2x2 montage.(ZIP)Click here for additional data file.S2 File(DOCX)Click here for additional data file.


The in ovo injection of vitamin D3 sources has been shown to enhance immunity as well as histomorphological variables in unchallenged conditions. One of the main diseases affecting poultry production is coccidiosis. Because of this, the current study was designed to determine the effects vitamin D3 (D3) and 25-hydroxyvitamin D3 (25OHD3) alone or together on the inflammatory reaction and small intestine morphology of broilers that were challenged with coccidiosis. In this study, it is shown that the in ovo administration of 2.4 μg of 25OHD3 alone increased villus length to crypt depth ratio (VCR) with the D3 + 25OHD3 treatment being intermediate two weeks post-challenge (28 day of age). Furthermore, chickens that received of 25OHD3 alone experienced lower plasma nitric oxide concentration as a systemic inflammatory indicator in comparison to all other treatments. It is concluded that the in ovo injection of 2.4 μg of 25OHD3 at 18 days of incubation can enhance intestinal histomorphology as well as inflammatory reaction of broilers when infected with coccidiosis.Vitamin D3 (D3) and 25-hydroxyvitamin D3 (25OHD3) administration on their inflammatory response and small intestine morphology were evaluated. At 18 d of incubation (doi), a 50 μL volume of the following 5 in ovo injection treatments was administrated: non-injected (1) and diluent injected (2) controls, or diluent injection containing 2.4 μg D3 (3) or 2.4 μg 25OHD3 (4), or their combination (5). Four male broilers were randomly allocated to each of eight isolated replicate wire-floored battery cages at hatch, and birds were challenged at 14 d of age (doa) with a 20x live coccidial vaccine dosage. One bird from each treatment–replicate (40 birds in each of 8 replicates per treatment) was bled at 14 and 28 doa in order to collect blood for the determination of plasma IL-1β and nitric oxide (NO) concentrations. The duodenum, jejunum, and ilium from those same birds were excised for measurement of villus length, crypt depth, villus length to crypt depth ratio (VCR), and villus surface area. In ovo injection of 2.4 μg of 25OHD3 resulted in a reduction in plasma NO levels as compared to all other treatments at 28 doa. Additionally, duodenal VCR increased in response to the in ovo injection of 25OHD3 when compared to the diluent, D3 alone, and the D3 + 25OHD3 combination treatments at two weeks post-challenge (28 doa). Therefore, it can be concluded that 2.4 μg of 25OHD3, when administrated in ovo at 18 doi, may be used to decrease the inflammatory reaction as well as to enhance the small intestine morphology of broilers during a coccidiosis challenge. In broilers challenged with coccidiosis, effects of in ovo vitamin D Coccidiosis is a parasitic infection that still is considered one of the main diseases affecting the performance of poultry reared under intensive production systems. Györke et al.  estimate3 (D3) and its metabolite, 25-hydroxyvitamin D3 (25OHD3), have been shown to affect small intestine morphology and the humoral and inflammatory responses of broiler chickens , which is the active form of vitamin D3. Increased levels of 1,25(OH)2 D3 alone, with or without associated changes in VDR, resulted in an increase in breast meat yield [3-injected treatment groups in a companion study, the expression of 1α-hydroxylase and immune suppressor genes increased 14 d post-challenge in broilers that were in ovo-injected with of 25OHD3 (unpublished data). Therefore, improvements in broiler small intestine morphology during a coccidiosis challenge in response to 25OHD3 alone might have been due to increased intestinal expression of 1α-hydroxylase and an enhanced immunity. In comparison to Der diets . Additiodiets [3 . The effrs (VDR) . The enzat yield  and imprat yield  and immuat yield  of broilat yield . Coccidiat yield . Additio3 reduced NO production during the coccidiosis challenge. Nitric oxide is an important indicator of on inflammatory response and is produced by the oxidation of L-arginine by the action of NO synthase. Inducible NO synthase is considered more important in immunity and inflammation [Eimeria-infected chickens produced higher serum NO concentrations compared with non-infected controls [3 reduced pro-inflammatory responses and increased anti-inflammatory response genes during a coccidiosis infection in comparison to vitamin D3 at the same level of activity [3 modulates the effects of immunoregulatory T-cells during a coccidiosis infection. Dietary 25OHD3 has been shown to increase the number of CD4 + CD25 + cells in mucosal regions, such as cecal tonsils, of Eimeria-infected chickens [3 has the potential to reduce bird losses due to a compromised immunity caused by a coccidia challenge.In the current study, the in ovo injection of 25OHDammation . The expammation . In addicontrols . Dietaryactivity . In addichickens . These r3 sources. Our findings suggest that coccidiosis has detrimental effects on the small intestine morphology and immunity of broilers. However, the in ovo injection of 2.4 μg of 25OHD3 reduced NO production and small intestine CD and increased the small intestine VL and VCR of challenged broilers. However, the other in-ovo-injected treatments tested in this study did not exhibit beneficial effects on the small intestine morphology and inflammatory response of the challenged birds. These results therefore indicate that in ovo injection of 25OHD3 may reduce negative effects on the inflammation and small intestine morphology of broilers caused by coccidiosis. These benefits may have been due to the expression of genes that contributed to the improved intestinal development and subsequent health of the broilers. Further research is required to determine the regulatory effects of vitamin D3 sources on immunity and intestinal development during a coccidiosis infection. In conclusion, the aim of this study was to determine the small intestine morphology and inflammatory response of broilers challenged with coccidiosis when birds were injected in ovo with two vitamin D


Dugesia japonica. Importantly, we found that knockdown of DjMeis1 inhibits the differentiation of neoblasts into eye progenitor cells and results in an eyeless phenotype with normal central nervous system. Furthermore, we observed that DjMeis1 is required for the activation of Wnt signaling pathway by promoting the Djwnt1 expression during posterior regeneration. The silencing of DjMeis1 suppresses the expression of Djwnt1 and results in the inability to reconstruct posterior poles. In general, our findings indicated that DjMeis1 acts as a trigger for the activation of eye and tail regeneration by regulating the differentiation of eye progenitor cells and the formation of posterior poles, respectively.As a member of TALE family, Meis1 has been proven to regulate cell proliferation and differentiation during cell fate commitment; however, the mechanism is still not fully understood. The planarian, which has an abundance of stem cells (neoblasts) responsible for regenerating any organ after injury, is an ideal model for studying the mechanisms of tissue identity determination. Here, we characterized a planarian homolog of Meis1 from the planarian  Meis1 results in severe limb agenesis [Meis1 and associated co-factors can promote the skin tumorigenesis. Knockdown of Meis1 induces a significant decrease in benign and malignant tumors in mice [Meis1 protein is a member of the three amino acid loop extension (TALE) transcription factors family. Previous studies have demonstrated that Meis1 plays essential roles in the embryonic development across metazoans. For example, Meis1 participates in the embryonic cortical development by promoting neuronal proliferation and migration [agenesis . In addiagenesis , eyes, aagenesis ,5. With agenesis ,7. Upreg in mice . Meanwhi in mice ,10. In gDjwnt1 and β-catenin, act as promoters for the construction of posterior poles [Djwnt1 can cause planarians that are unable to regenerate tails or regenerate heads in the place of tails. β-catenin acts as a downstream gene of Djwnt1. Silencing of β-catenin can result in the reversal of posterior poles and cause planarians to regenerate posterior heads from posterior blastema [notum serves as an inhibitor of Djwnt1 during anterior poles regeneration [Notum RNAi can upregulate Wnt signaling and cause animals to regenerate tails in the place of heads [Djpbx and Djislet have been proven to be positive regulators of Djwnt1 during posterior poles regeneration. Knockdown of Djpbx and Djislet can cause planarians that are unable to regenerate their tails [Planarians can regenerate a complete individual from any fragment of their body ,12,13. Tor poles ,25. Knocblastema . Furtherneration . Notum Rof heads . Recentlir tails ,30. In gMeis1 in planarian Dugesia japonica. We observe that DjMeis1 RNAi planarians cannot form mature eye cells or reconstruct the posterior poles. DjMeis1 RNAi decreases the number of eye progenitor cells and suppresses the expression of Djwnt1. Herein, we propose that DjMeis1 serves as a positive factor in the regulation of the differentiation of eye progenitor cells and the establishment of posterior poles during planarian regeneration.In our work, we identify a homolog of Dugesia japonica based on our previous transcriptome [DjMeis2 and DjMeis3 have been previously reported as SmedMeis and SmedMeis-like in planarian Schmidtea mediterranea  and PC2 ) probes to detect the reconstruction of brain nerves [DjMeis1 RNAi animals was normally reconstructed compared to the control animals at 7 dpa  to examine the regeneration of eye progenitor cells in DjMeis1 RNAi animals [+Djovo cells in DjMeis1 RNAi animals was significantly reduced  to determine whether the posterior wound in DjMeis1 RNAi animals could be healed [DjMeis1 RNAi animals showed a normal expression pattern of LaminB compared to the control groups at 7 dpa  reconstruction. We observed that DjMeis1 RNAi animals failed to regenerate VNC posteriorly, which was consistent with the result that DjMeis1 RNAi animals healed the posterior wound at the amputated site without activating the regeneration of tails  of the head fragments and observed that DjMeis1 RNAi animals regenerated incomplete pharynx or failed to regenerate pharynx  at 7 dpa and found that sFRP1+ cells existed both in anterior and posterior poles in double RNAi animals  in hypoxia [DjMeis1 RNAi planarians displayed normal mitotic activities compared to the control groups  used for the RNAi experiments were synthesized by in vitro transcription as previously described [β-catenin and DjMeis1 maintained a concentration of 2 μg/μL dsRNA. The water treated by DEPC was injected to the control group animals. Head, trunk, and tail fragments were amputated from the animals from the anterior and posterior sites of the pharynx at 24 h after the last injection.All the dsRNA  for 10 min at 37 °C and fixed in 4% paraformaldehyde for 20 min at room temperature. Then, the animals were hybridized with DIG-labeled probes at 56 °C for 16-17 h and washed in 2× SSC  and 0.2× SSC three times for 20 min, respectively. Antibody incubation  and colorimetric (NBT/BCIP) were used for in situ detection. Different concentrations of SSC were prepared by dissolving 20× SSC in deionized water.Whole-mount ISH (WISH) was performed as previously described . In eachWhole-mount immunostaining was performed as previously described . In eachEye resection was performed as previously described . In eachBrdU was performed as previously described . In each2O2 in deionized water) was used to bleach the animals under bright light for 2 h. After washing in 1× SSC and PBST for 5 min and incubating in Proteinase K (20 mg/mL in PBST) for 10 min at 37 °C, the animals were fixed in 4% paraformaldehyde for 20 min and washed in PBST for 10 min. Then, the animals were hybridized with DIG-labeled PC2 probe at 56 °C for 16–17 h and washed in 2× SSC and 0.2× SSC three times for 20 min, respectively. PBST with 5% goat serum and 5% Western Blocking Reagent  was used to block the animals for 5 h. Next, the animals were incubated in 1:500 anti-Digoxigenin-POD  overnight. Finally, the animals were treated with tyramide conjugated to Alexa568  and were observed by NIS element software . Different concentrations of SSC were prepared by dissolving 20× SSC in deionized water.Fluorescence in situ hybridization was performed as previously described . In eachp < 0.05.Data were shown as means ± SD, and statistical analyses were performed by students. One-way analysis of variance (ANOVA) was used to analyze the data of two groups. A statistically significant difference was defined as


Moreover, the Δatg1Δpmk1double mutant resembles the single Δpmk1 mutant, suggesting that Atg1 functions in the Pmk1 pathway. In addition, the growth defect induced by overexpression of Pck2, an upstream activator of Pmk1 MAPK was alleviated by the deletion of+atg1. Finally, the deletion of autophagy-related genes recapitulates Pmk1 MAPK signaling inhibition. Our data suggest a novel role for autophagy in MAPK signaling regulation.Apart from the highly conserved role in the cellular degradation process, autophagy also appears to play a key role in cellular proliferation. Here, we describe the genetic interaction of autophagy-related genes and Pmk1 MAPK signaling in fission yeast. Although starvation potently induces autophagy, the basal level of autophagy is maintained even in normal growth conditions, thereby controlling development, cellular metabolism, and proliferation in all eucaryote cells . However, how autophagy organizes cellular signaling networks to exert these functions is not fully understood.Autophagy is an evolutionarily highly conserved mechanism to maintain cellular homeostasis via waste clearance using the lysosomal machinery  phenotype, which recapitulates Pmk1 MAPK signaling inhibition (Methods). The rationale of thevicmutant screening is based on the antagonistic relationship between Pmk1 MAPK and calcineurin, a highly conserved serine/threonine protein phosphatase, in the Cl−homeostasis in fission yeast. Knockout of theS. pombecalcineurin geneppb1+ or inhibition of calcineurin activity by the immunosuppressant FK506, a specific inhibitor of calcineurin, results in hypersensitivity to Cl−. This phenotype associated withppb1deletion was reversed by the inhibition or gene knockout of the components of the Pmk1 MAPK signaling pathways, including Pmk1 MAPK, Pek1 MAPKK, Mkh1 MAPKKK, and Pck2 Protein kinase C . Our previous genetic screening to isolatevic mutants identified upstream activating regulators of MAPK signaling such as geranylgeranyl transferase  and farnesyl transferase .Here, we identified2, whereas knockout of thepmk1+gene makes cells grow much faster in the same condition . Δatg1 cells grew faster than the WT cells in the presence of FK506 and 0.09 M MgCl2, although the growth of Δatg1 cells was slower than that of Δpmk1. The WT, Δpmk1 and Δatg1 cells exhibited essentially the same pattern of sensitivity and resistance to FK506 and Cl−on EMM, YPD and YES plates, although the sensitivity of the WT cells and Δatg1 cells was more enhanced on YES than the other plates. Genetic interaction between Δatg1and Δpmk1was further examined by constructing Δatg1Δpmk1double mutant cells. The degree of thevic phenotype in Δatg1Δpmk1double mutants and Δpmk1cells was almost equivalent. These results are consistent with Atg1 working upstream of the Pmk1 pathway. The difference in growth of these strains is almost indiscernible in the absence of FK506 and MgCl2.The growth of the wild-type (WT) cells was significantly inhibited in the presence of the calcineurin inhibitor FK506 and 0.09 M MgClet al.2006). This phenotypic evaluation also led to the identification of an SH3 adaptor protein Skb5 as a negative regulator of Pck2/Pmk1 signaling . As shown in Figure 1B,atg1deletion significantly suppressed the toxicity induced by Pck2 overexpression, although the impact ofatg1deletion on the suppression of the toxicity of Pck2 overexpression was smaller than that achieved bypmk1deletion. Thus,atg1 deletion is likely to ameliorate Pck2-mediated Pmk1 MAPK hyperactivation.To further explore the functional relationship between Atg1 and Pmk1 MAPK signaling, we utilized the cell growth inhibition associated with Pck2 overexpression. Pck2 overexpression in the WT cells leads to Pmk1 MAPK hyperactivation and cytotoxicity, which can be suppressed by the inhibition or knockout of the components of the Pmk1 MAPK pathway  . The degree of thevic phenotype of theseatgmutants was similar to that of Δatg1 cells. These results suggest that the autophagy system as a whole may be involved in the Pmk1 MAPK signaling regulation.Next, we confirm if the loss-of-function mutants of other autophagy-related genes also display theet al.2006), as well as the role of autophagosome as a scaffold to facilitate spatial coordination of RAF/MEK/ERK phosphorylation . Our epistasis analysis, showing the nonadditivevicphenotype of Δatg1Δpmk1double mutant and the resemblance of the Δatg1Δpmk1double mutant to the Δpmk1single mutant, suggests that Atg1 acts upstream of the Pmk1 signaling pathway. Furthermore, suppressioin of the toxicity induced by Pck2 overexpression byatg1deletion suggests that Atg1 acts downstream of Pck2. MAPK signaling cascades consist of a core module of three tiers of protein kinases MAPK, MAPKK, and MAP3K, and often an additional upstream MAP4K. It would be intriguing if Atg1 serves as an additional layer of kinase mediating MAPK signaling activation. Future studies will elucidate the mechanism and the functional significance of the genetic interaction between autophagy and MAPK signaling revealed by our yeast genetic screen. Given the highly conserved nature of autophagy and MAPK signaling in the fate of cell death and proliferation, this study will provide valuable information to understand human diseases associated with aberrant regulation of MAPK signaling and autophagy.In summary, our genetic screen revealed a functional interaction between autophagy-related genes and Pmk1 MAPK signaling in fission yeast. Several studies report functional crosstalk between MAPK signaling and autophagy, including the role of MAPK ERK in the maturation of autophagosomes . FK506 was provided by Fujisawa Pharmaceutical Co. . Standard genetic and recombinant-DNA methods  were used except where noted.vicmutant screeninget al. was analyzed on the YPD plates with or without FK506 and 0.09 M MgCl2by spotting growth assay.The growth of 89 viable kinase knockout strains generated by BimboSpotting growth assay660/ml, from which five 10-fold serial dilutions were prepared and then spotted onto the indicated plates. Plates were incubated at 27˚C for 3 to 7 days.Yeast cells were cultured in 20 ml of liquid media at 27˚C till mid-log phase and diluted to 0.6 OD


The transformation of Cs4PbBr6 nanocrystals into CsPbBr3 takes place in two steps, the first step being the surface modification of the Cs4PbBr6 nanocrystals with Zn2+ ions and the second step being extraction of CsBr by the Zn2+ ions resulting in the formation of composite Cs4PbBr6/CsPbBr3 nanocrystals. The transformed composite nanocrystals were found to have a PL QY exceeding 90% and the shape of the nanocrystals also changed from hexagonal to cubic shaped. Owing to the highly ionic nature of the nanocrystals, complete anion exchange could be also realized using ZnI2 salt. In the case of the iodide post-treated samples, nanorods were obtained which exhibited bright red photoluminescence. Photodetectors based on the ZnI2 treated Cs4PbBr6 NCs were fabricated, and the photodetectors exhibited a high on/off ratio with a fast response time. The excellent optoelectronic properties make this treatment versatile for a wide range of functional optoelectronic devices like light emitting diodes and photovoltaic devices.Herein we demonstrate a facile approach for the synthesis of all inorganic cesium lead halide perovskite nanocrystal composites CsPbX 3  with high quantum yield by post-synthetic modulation of zero dimensional Cs4PbBr6 nanocrystals with ZnX2 salts.Herein we demonstrate a facile approach for the synthesis of all inorganic cesium lead halide perovskite nanocrystal composites CsPbX If all the corners of the lead hexahalide octahedra are shared, then they are electronically coupled in all three dimensions and referred to as 3D perovskites. If all the corners of the lead hexahalide octahedra are not shared, they are referred to as 0D perovskites.25Lead halide perovskite NCs  of the NCs.26–33 Different groups have recently synthesized 0D colloidal Cs4PbBr6 NCs and it has been found that these nanocrystals don't exhibit any photoluminescence.25 However, the non-luminescent 0D Cs4PbBr6 NCs can be converted to 3D CsPbBr3 NCs using additives like water,34 PbBr2 35 ligands,36 and Prussian blue.37 The non-luminescent nature of Cs4PbBr6 at room temperature is attributed to the origin of structural and point defects.38 Nikl et al. observed the true emission of Cs4PbBr6 at low temperatures (4 K) and the true emission is centered at 375 nm. In the case of thin films at room temperature, Cs4PbBr6 is mixed with traces of the CsPbBr3 phase, which in turn results in the emission at 545 nm. However in the case of nanocrystals, no emission has been observed so far for the pure Cs4PbBr6 nanocrystals.It is extensively recognized that the excellent optical properties in lead halide based perovskite materials come from the high defect tolerance.3 nanocrystals by improving the surface capping. Herein we demonstrate ZnX2 mediated post-synthetic transformation of Cs4PbBr6 nanocrystals (NCs) into CsPbX3 nanostructures through an intermediate composite Cs4PbX6/CsPbX3. We find that this procedure results in a highly reproducible increase of the PL QY of up to 90% for the ZnX2 treatment. The transformation takes place through the surface passivation of Cs4PbBr6 nanocrystals with ZnX2 followed by anion exchange (in the case of ZnI2 and ZnCl2) resulting in the formation of an intermediate Cs4PbX6@CsPbX3 passivated by ZnX2 followed by the extraction of CsBr from the composite nanostructures. Interestingly, as soon the ZnX2 is added to the nanocrystals, excitonic absorption resulting from the Cs4PbBr6 particles passivated with ZnX2 is observed. This is the first time that the true emission of the Cs4PbX6 crystal structure in the nanocrystalline phase was observed at room temperature.Recently it has been shown that addition of appropriate metal halide salts significantly enhances the photoluminescence intensity of CsPbBr4PbBr6 nanocrystals were synthesized according to the hot injection strategy as described in the Experimental section. The Cs4PbBr6 nanocrystal dispersion in hexane is completely transparent and as shown in the TEM images in 4PbBr6 nanocrystals consist of hexagons of around 30 nm in size. The powder diffraction pattern indicated that the nanocrystals crystallized in the rhombohedral Cs4PbBr6 phase in the space group R3c.The Cs4PbBr6 nanocrystals (NCs) were post treated with a ZnX2/hexane solution and used for further characterization. Upon addition of ZnX2  solution, clear color change was observed  in comparison to that in the ZnBr2 treatment was observed. Along with the red shifted absorption, a sharp peak at 550 nm can be seen.The absorption spectra of the ZnX4PbBr6 NCs was taken in a cuvette, different concentrations of ZnBr2 were added to the sample and the absorption spectrum of the ZnBr2 treated NCs was measured. 2/hexane solution added to the cuvette containing NCs. A sharp absorption peak at 425 nm along with a long tail extending up to 510 nm was observed. The absorption intensity of the tail increased gradually and a sharp band edge at 510 nm, which is a characteristic peak of CsPbBr3, was observed. The peak at 425 nm is blue shifted to 420 nm and is masked (though visible) by the strong absorption of CsPbBr3 in this region. Fig. S13 nanocrystals synthesized and the ZnBr2 treated Cs4PbBr6 NCs. A similar concentration dependent study was carried out in the case of ZnI2 treated Cs4PbBr6 NCs. 2 treated NCs with different amounts of ZnI2 added. A sharp absorption at 540 nm is evident even with high amounts of added ZnI2. ZnBr2 treated NC samples were found to be highly stable even after 2 months of maintaining under 70% RH conditions. Fig. S22 treated NC sample and the as-prepared CsPbBr3 NCs after storing under 70% RH conditions.To confirm if the peak observed at 420 nm arises from the sample, an aliquot of the Cs2 treated NCs. 2 treated NCs. No strong PL emission was observed, which is consistent with the previously observed decay of excitonic PL above temperatures of 100 K. Very weak PL emission centered at 515 nm was observed which can be attributed to the CsPbBr3 phase embedded in the PL-inactive Cs4PbBr6 crystalline matrix. With the ZnBr2 post-treated NC solution, strong PL was observed at 518 nm. From the literature, it is known that the excitonic absorption peak of pure CsPbBr3 NCs is located at around 510 nm. So, it is reasonable to speculate that the strong excitonic absorption peak and strong PL peak at 518 nm in 3 NCs, indicating that a transformation might have taken place from Cs4PbBr6 to CsPbBr3.To corroborate the observed absorption behavior, photoluminescence (PL) studies were also carried out on the parent NCs and ZnX2 concentration dependent emission studies. With the addition of ZnBr2, two emission peaks were observed, one at 430 nm and the other at 510 nm, similar to the behavior observed in the absorption spectrum. The emission at 430 nm could be attributed to the 425 nm peak observed in the absorption spectrum and the peak at 510 nm could be attributed to the band edge emission corresponding to the absorption edge shown in We also carried out ZnBr3/Cs4PbX6 composites were formed after the post-treatment with ZnX2. In the case of ZnBr2, the excitonic CsPbBr3 absorption was observed at 510 nm and the absorption peak observed at a lower wavelength could be attributed to the Cs4PbBr6 phase. It has been previously reported that defect states created due to the surface halide vacancies could be responsible for quenching the true emission in Cs4PbBr6 and were only observed at low temperatures (4 K). When the Zn2+ salts were added to the Cs4PbBr6 NCs, the surface halide defect states could be passivated and the true emission from the surface passivated/modified Cs4PbBr6 nanocrystals could be observed at 430 nm. The true emission of Cs4PbBr6 is observed for the first time at room temperature when Zn2+ salts are used to passivate the surface defects; however, the Zn2+ salts also triggered the transformation of 0D Cs4PbBr6 NCs into 3D CsPbBr3 NCs. So, the intensity of the true emission of Cs4PbBr6 NCs gradually decreased as the concentration of the zinc salt is increased. When the concentration of the zinc salt was kept constant at 50 μL of ZnBr2, the intensity gradually decreased indicating that the whole process takes place through a two-step mechanism. Similar studies carried out using other salts like Cu2+ and Mn2+ didn't result in the enhancement of the PL QY. In the case of Mn2+ a slight increase was observed but Cu2+ didn't show any improvement in the PL QY. Fig. S32 and MnBr2 treated Cs4PbBr6 NCs.Together with the results obtained from the XRD studies and UV-vis and PL spectra, we speculate that CsPbX4PbBr6 NCs show hexagonal shaped particles of around 30–35 nm in size similar to the particles reported previously. 4PbBr6 NCs and the ZnBr2 treated samples. When the samples were treated with 50 μL of ZnBr2, initially hexagonal shaped particles were converted mostly to cubic shaped particles as shown in 3 NCs  plane corresponds to the presence of Cs4PbBr6, indicating the formation of a composite of CsPbBr3/Cs4PbBr6. A plausible two step formation mechanism can be inferred from the observed TEM images. As the Cs4PbBr6 nanocrystals were treated with the zinc halide salts, in the first step Zn2+ ions could passivate the surface of the nanocrystals and then result in the extraction of excess Cs+ ions, and an increase in the dimensionality from 0D to 3D is observed. In the second step we believe that an inside out diffusion of Cs+ ions takes place resulting in the formation of CsPbBr3/Cs4PbBr6 composites.TEM images of the as-synthesized Css Fig. S4. The HR-s Fig. S4 shows a 4PbBr6 NCs were treated with 40 μL of ZnI2 solution, the shape of the particles changed from hexagonal to rod shaped. As seen in the TEM images shown in 2 solution to 100 μL the length of the rods increased to nearly 250 nm and the width increased to 20–30 nm.When the Cs7/2) and 143 eV (4f5/2) with a spin–orbit splitting of nearly 5 eV, which corresponds to Pb in the 2+ oxidation state. Quantitative XPS analysis indicates that Cs : Pb : Br = 1.5 : 1 : 4.2 for untreated Cs4PbBr6 NCs, which is Pb rich in composition. However, for ZnBr2-Cs4PbBr6 NCs, the composition becomes anion-rich (Cs : Pb : Br = 0.6 : 1 : 5.5), which was also repeatedly observed  is much higher compared to that in the bromine case  measurements have been used to assess the surface modification of the NCs and to understand the mechanism of PL enhancement. After measurements, all core level spectra were calibrated using the C 1s peak at 285.0 eV. The high-resolution XPS spectrum of untreated NCs for Pb 4f yields 2 peaks as shown in Fig. S5.d Fig. S6. A smalle Fig. S7.2 treated Cs4PbBr6 NCs, we have explored their potential application in optoelectronic devices. To this end, we have fabricated photodetectors by dropcasting the ZnI2 treated NCs in hexane between two fluorine doped tin oxide (FTO) electrodes. 2 treated nanocrystals, the currents obtained were lower compared to those using the ZnI2 treated samples. Fig. S82 treated NCs in the same configuration. This indicates that the performance of the photodetectors could be improved if the length of the nanorods could be made longer than the width of the channel used and also by removing the surface capping ligands by appropriate treatments.Owing to the one-dimensional nature of the nanorods in the case of ZnI4PbBr6 into highly luminescent CsPbBr3/Cs4PbBr6 composites using ZnBr2 treatment. The transformation with ZnBr2 treatment proceeds through a two-step process, where in the first step, zinc doped/modified Cs4PbBr6 NCs are formed which then results in the conversion to CsPbBr3/Cs4PbBr6 nanocomposites. The composite of CsPbBr3/Cs4PbBr6 NCs is found to be very highly luminescent with a PL QY of 90%. In the case of ZnI2 treatment, the post synthetic treatment also resulted in morphological transformation from the hexagonal shaped nanocrystals to nanorods. The length of the rods could be varied depending on the concentration of the nanocrystals and the zinc iodide salt added. In both the iodide and bromide case an intermediate is formed upon treatment with the zinc salt. The intermediate was observed in the time/concentration dependent UV-vis and PL studies, and from the X-ray diffraction analysis, we presume that it is the 0D structure modified/doped by the zinc cations. Photodetectors based on the ZnI2 treated Cs4PbBr6 NCs were fabricated, which exhibit a high on/off ratio with a fast response time. The excellent optoelectronic properties make this treatment versatile for a wide range of functional optoelectronic devices like light emitting diodes and photovoltaic devices.In conclusion, we have developed a simple and convenient post synthetic strategy for the transformation of 0D Cs2CO3, 99.9%), lead(ii) bromide (98%), zinc(ii) iodide (98%), zinc(ii) chloride (99%), oleylamine , oleic acid (90%), and 1-octadecene (90%) were purchased from Sigma Aldrich. Zinc(ii) bromide (98%) was purchased from Alfa Aesar.Cesium carbonate , oleic acid (1 mL), and octadecene (16 mL) were added to a three-neck flat bottom flask and kept under vacuum for 30 min at 120 °C. After 30 minutes the temperature was increased to 150 °C and the system was kept under a N2 atmosphere until a clear solution of cesium oleate was obtained.Cesium carbonate , oleylamine (1 mL), oleic acid (1 mL) and lead bromide  were loaded into a three-neck round bottom flask and degassed for 30 min at 120 °C. The flask was then filled with N2 and oleylamine (1.5 mL) and oleic acid (1.5 mL) were added. After complete solubilisation of PbBr2, the temperature was increased to 165 °C and finally the as-prepared Cs-oleate (1.2 mL) was injected. The reaction was immediately quenched in an ice-water bath.Lead bromide (207 mg) and octadecene (15 mL) were loaded into a three-neck round bottom flask and degassed for 1 hour at 120 °C. The flask was then filled with NThe crude solution of nanoparticles was first centrifuged at 6000 rpm for 10 min. After that, the supernatant was discarded and the precipitate was redispersed in hexane solution. Then again, the large particles present in the solution were removed by centrifugation at 6000 rpm for 10 min. The supernatant was discarded and a colloidal solution of nanoparticles was obtained in hexane.−1) were treated with the pre-made zinc halide solution. Zinc halide solutions were prepared by dissolving zinc halides  (0.05 M) in the mixture of hexane (2 mL) and oleylamine (40 μL) at room temperature. Then the required zinc halide solution was added to 1 mL of the Cs4PbBr6 NCs in hexane solution (8 mg mL−1) under constant stirring. Excess zinc halide salts are removed by centrifugation and the NCs can be stored in organic solvents like hexane, toluene, etc. for a few months.The NCs dispersed in hexane (∼8 mg mLλ = 1.541). PL spectra were collected by using a Horiba FluoroMax spectrometer with 400 nm excitation wavelength, while UV-vis absorption measurement was performed using a UV-vis-NIR spectrometer (PerkinElmer LAMBDA 950). The room-temperature PLQY was calculated according to the following equation38High resolution transmission electron microscope images were obtained using a Tecnai G2 F30 300 kV transmission electron microscope. X-ray diffraction spectra were obtained by using a Rigaku Japan SmartLab X-ray diffractometer  were chosen to be the standards for green and red spectral regions and the standard solutions should be freshly prepared. Attention was paid to keeping the optical densities of the sample and standard below 0.1 at the excitation wavelength.In this equation, QYLaser patterned FTO coated glass with an etched area was used as the photodetector. Before deposition, the laser patterned FTO was cleaned thoroughly by using a soap solution, DI water, acetone, and ethanol for 10 minutes, respectively, by ultra-sonication. Perovskite nanocrystals in hexane were drop cast on the etched area of the FTO coated glass by masking the unetched area with scotch tape. The drop cast substrate was then subjected to vacuum drying. Then the photoresponse was measured under a 617 nm LED light.There are no conflicts to declare.NA-001-C9NA00244H-s001