With evolving patient characteristics and patterns of ICU utilization, the impact of dysnatremias on patient outcomes and healthcare costs in the present era have not been well studied. Patients ≥ 18 years admitted to the ICUs in public hospitals in Hong Kong between January 2010 and June 2022 and had at least one serum sodium measurement obtained within 24 h prior to or following ICU admission were stratified into normonatremic (135-145 mmol/L), hyponatremic (< 135 mmol/L) and hypernatremic (> 145 mmol/L) groups. A total of 162,026 patients were included—9098 (5.6%), 40,533 (25.0%) and 112,395 (69.4%) patients were hypernatremic, hyponatremic and normonatremic at the time of ICU admission, respectively. The odds of patients with hypernatremia and hyponatremia dying in the ICU were 27% and 14% higher , and 52% and 21% higher for dying in the hospital , hospital LOS, and higher healthcare costs than the normonatremic group. Dysnatremias at ICU admission were associated with increased ICU and in-hospital mortality and overall healthcare burden. The presence of serum sodium (Na) abnormalities prior to or following ICU admission is clinically important, as both hypernatremia and hyponatremia have been denoted as independent predictors for increased ICU mortality3. The management of dysnatremias, especially severe ones, are challenging as overzealous correction may lead to serious neurological sequelae such as osmotic demyelination or cerebral edema4.Dysnatremias are common electrolyte disorders in the Intensive Care Unit (ICU) setting. The prevalence of dysnatremias has been reported to be up to 24.6% upon ICU admission, depending on the type of dysnatremias, severity of dysnatremias, and patient population5. The overall number of ICU admissions have also increased by 25% in the last 10 years6. Based on these observations, we postulate that the clinical epidemiology and impact of dysnatremias have changed considerably in critically ill patients admitted to the ICU. Indeed, a shift in the incidence from hyponatremia to increasing hypernatremia in the ICU has been reported in one study7. Furthermore, existing literature often focuses on ICU-acquired dysnatremias or certain ICU sub-cohorts with limited patients numbers, rendering the results less generalizable8. For example, important consequences of dysnatremias such as adverse neurological sequelae are largely studied in neurosurgical patients but not in other ICU patients with non-neurological conditions4.Although the association between dysnatremias and clinical outcomes have been studied in different clinical scenarios, the impact of dysnatremias on patient outlook after ICU admission in the recent era have not been well characterized. Over the past two decades, there have been temporal changes in the pattern of ICU utilization, owing to the ageing population and evolving patient characteristics, advances in the treatment for medical and surgical conditions, e.g. novel anti-cancer therapies, advanced life support systems and surgical techniques, as well as the emergence of new disease entities, e.g. COVID-19 diseaseIn view of these knowledge gaps, we used data from a territory-wide electronic health system over a 12-year period to evaluate the impact of dysnatremias on mortality, neurological complications, and healthcare costs in adult patients admitted to the ICU. The hypothesis was that dysnatremias at ICU admission are associated with increased ICU mortality, worse neurological outcomes, and increased healthcare costs in mixed surgical and medical ICU patients.This retrospective cohort study used data from a territory-wide electronic health record system in Hong Kong from 1st January 2010 to 30th June 2022. The study conforms with the principles outlined in the Declaration of Helsinki and was approved by the Institutional Review Board of the University of Hong Kong/Hospital Authority Hong Kong West Cluster (HKU/HA HKW IRB) (IRB Reference Number: UW 22-650) with a waiver of signed informed consent.Patients who were ≥ 18 years at the time of ICU admission and had at least one serum Na measurement obtained within 24 h prior to or following ICU admission were included. Patients with missing Acute Physiology and Chronic Health Evaluation IV (APACHE IV) predicted risk of death or missing creatinine measurement within 24 h prior to or following ICU admission were excluded from this study.Patients were stratified according to their closest serum Na measurement obtained within 24 h prior to or following ICU admission into the normonatremic (serum Na 135–145 mmol/L), hyponatremic (serum Na < 135 mmol/L), and hypernatremic (serum Na > 145 mmol/L) groups, respectively.9.The co-primary outcomes were ICU and in-hospital mortality. Secondary outcomes included adverse neurological outcomes, hospital discharge destination, ICU length of stay (LOS), hospital LOS, and healthcare costs. An adverse neurological outcome was defined by a clinical surrogate of requiring one or more computed tomography or magnetic resonance imaging scans of the brain performed between 24 h after ICU admission and hospital discharge. Hospital discharge destination was categorized into “death”, “home”, “transfers” which included patients who were transferred to convalescent beds, or “others” which included patients who did not have documented hospital discharge destination information. Healthcare costs were estimated using the bed costs with respective to the number of bed days from ICU admission to hospital discharge. Bed costs were based on 4 types of bed status provided in public hospitals in Hong Kong—intensive care bed , acute general or convalescent bed , and psychiatric bed 10. Patients’ demographics, diagnoses, hospitalization, prescriptions, laboratory results and outcomes were retrieved. All data were de-identified to ensure patients’ privacy and confidentiality. Comorbidities were identified on the basis of routinely recorded codes from the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) , a central de-identified data repository comprising electronic health records from all public hospitals in Hong Kong. Previous data validation for use in cohort studies has shown high coding accuracyCategorical variables were described as frequencies with percentages (%), and continuous variables as median with interquartile range (IQR). Categorical variables were compared with Fisher-exact test or Pearson’s chi-squared test, as appropriate. ANOVA test was used to compare the differences among the exposure groups. Demographics and clinical variables that were significantly different between the exposure groups with a P-value < 0.05 and other confounders selected a priori were included in multivariable logistic regression. The variables including APACHE-IV predicted risk of death, age, sex, creatinine level obtained within 24 h prior to or following ICU admission, cardiovascular diseases, respiratory diseases, kidney diseases, liver diseases, cerebrovascular diseases, diabetes mellitus (DM), dementia, immunosuppressive therapy, presence of COVID-19 infection, emergent ICU admission, and surgical ICU admission were included in the final multivariable logistic regression model. All variables included in the regression model were tested for collinearity.The association between exposure groups and the outcomes of length of stay and medical costs were examined using multivariable adjusted negative binomial regression and generalized linear model, respectively. A local polynomial smooth plot was used to model the changes in plasma Na level over the first 7 days of ICU admission by fitting the moving weighted average of serum Na levels for all 3 groups (details are included in Supplementary Method). All analyses were performed with two-tailed tests and a P-value < 0.05 was considered statistically significant. All analyses were done using STATA MP, version 16.1.Subgroup analyses were performed to assess the effect of serum Na on ICU mortality in the surgical and medical cohorts and patients admitted to ICU for neurological and non-neurological conditions. Detailed definitions of subgroups are included in Supplementary Sect. 8. Second, the effect of hyperglycaemia on serum Na was accounted for by calculating corrected serum Na levels using the following formula: Corrected [Na] = Serum [Na] + 0.4 x (Paired blood glucose – 5.5 mmol/L)11. Paired blood glucose level was defined as the blood glucose measurements taken within 3 h prior to or following the corresponding serum Na measurement. Patients were reclassified into three groups according to the corrected Na, and ICU mortality was compared between groups.Sensitivity analyses included, first, redefining the cut-off value for dysnatremias – normonatremia (serum Na level 130–150 mmol/L), hyponatremia (serum Na level < 130 mmol/L), and hypernatremia (Na level > 150 mmol/L)Subjecting to observed heterogeneity between subgroups of patients with and without DM, interaction analysis was performed by including the interaction term “dysnatremia x diabetes” to the main regression model and re-examining the association with ICU mortality.This study was approved by the Institutional Review Board of the University of Hong Kong/Hospital Authority Hong Kong West Cluster (HKU/HA HKW IRB) (IRB Reference Number: UW 22-650) with a waiver of signed informed consent. This study complies with the Declaration of Helsinki 1975 and its later amendments.A total of 163,493 patients were admitted to the ICUs in public hospitals in Hong Kong between 1st January 2010 and 30th June 2022. After excluding patients with missing data and those younger than 18 years, a total of 162,026 patients were included in the final analysis. The median age was 64 (52–74) years, and 99,804 (61.6%) patients were male. The median APACHE IV predicted risk of death was 0.12 (0.04–0.33). There were 70,080 (43.3%) and 91,946 (56.8%) surgical and medical ICU patients, respectively. A total of 78,453 (48.4%) patients had cardiovascular diseases, 18,534 (11.4%) had kidney diseases, 21,329 (13.2%) had cerebrovascular diseases, and 34,278 (21.2%) had DM. The study flow and baseline characteristics are shown in Fig. At ICU admission, a total of 9098 (5.6%), 40,533 (25.0%), and 112,395 (69.4%) patients were hypernatremic, hyponatremic, and normonatremic, respectively. Hypernatremic and hyponatremic patients were older  compared with normonatremic patients , and had higher APACHE IV-predicted risk of death . Patients with hypernatremia and hyponatremia had higher rates of emergency ICU admission , COVID-19 infection , and sepsis  compared with patients with normonatremia.Unadjusted and adjusted outcomes are shown in Table Adverse neurological outcomes were observed in 23,562 (14.5%) patients and was more frequent in patients with severe hypernatremia (serum Na > 155 mmol/L) and hyponatremia (serum Na < 125 mmol/L) . Multivariable regression showed that the risks of adverse neurological outcomes were higher in patients with hypernatremia and hyponatremia than those with normonatremia upon ICU admission .The median ICU LOS and hospital LOS were 2.0 (1.0–4.6) and 16.0 (8.0–35.0) days, respectively. Patients with hypernatremia and hyponatremia at ICU admission had significantly longer median ICU LOS than those with normonatremia . In adjusted analysis, patients with hypernatremia or hyponatremia had increased risks for longer ICU LOS . There were similar associations of patients with hypernatremia and hyponatremia with longer median hospital LOS, when compared with patients with normonatremia. Furthermore, patients with hypernatremia and hyponatremia at ICU admission were more likely to be transferred for convalescence care than discharged home compared with patients with normonatremia . The median healthcare cost (in thousand United States (U.S.) dollars) in the cohort was $18.6 (11.3–36.6). Patients with hypernatremia and hyponatremia were associated with higher healthcare costs compared with patients with normonatremia . Detailed results on clinical outcomes are presented in Table The changes in plasma Na level from Day 0 to Day 7 following ICU admission stratified by exposure groups and ICU mortality are presented in Fig. The results of subgroup analyses are shown in Table The results of sensitivity analyses are shown in Table Other sensitivity analyses such as examining the use of ICD diagnostic codes to define adverse neurological outcomes are included in the Supplementary Sect. The results of exploratory analyses are shown in Table Dysnatremias are common and important electrolyte abnormalities in critically ill patients. Our data suggested that dysnatremias are prevalent among critically ill patients at the time of ICU admission. Importantly, the presence of hypernatremia or hyponatremia early in the course of critical illness were both associated with various adverse clinical outcomes including increased ICU and in-hospital mortality, neurological complications, longer ICU and hospital stay, worse discharge outcomes, and consequentially higher healthcare costs.16. In this large multicenter cohort of mixed surgical and medical ICU patients, dysnatremias occurred in 30.6% of patients upon ICU admission, with rates of hypernatremia and hyponatremia at 5.6% and 25.0%, respectively. The discrepancy in the frequency of dysnatremias reported in different studies may be explained by the differences in case mix and cut-off values for normal Na level. The incidences for dysnatremias were higher in studies that included primarily medical or mixed medical-surgical cohorts, suggesting that the burden of pre-existing medical comorbidities may predispose to dysnatremias in critically ill patients. Indeed, patients with dysnatremias in our study were older and had higher APACHE IV scores and rates of sepsis compared to those with normonatremia. Notably, there is a temporal shift to growing proportion of hypernatremia over time7, possibly a result of changes in critical care practices such as fluid management18. The rise in proportion of patients with hypernatremia in the ICU has paralleled a growth of ICU admissions in many developed countries6.The incidences of hypernatremia and hyponatremia reported in previous series were variable, ranging from 0.5–9.0% and 13.7–34.4%, respectively19. Moreover, the risk of death escalated with the magnitude of the deviation from normal values20. One should also note that even if these patients survived their ICU stay, the overall in-hospital mortality remained alarmingly high . Furthermore, hypernatremia appeared to have a greater impact on clinical outcomes compared with hyponatremia. In our cohort, patients with hypernatremia at ICU admission had a 27% increased odds of ICU mortality and 52% increased odds of hospital mortality compared to patients with normonatremia. Indeed, hypernatremia, often an indication of volume depletion, is associated with hypotension, hypoperfusion, and subsequent increased mortality in patients with fulminant sepsis or after major operations18. Our finding that hypernatremia conveys an increased risk of mortality in surgical and neurologic patients echoes previous findings in liver transplantation and neurosurgical cohorts22. On the contrary, hyponatremia often occurs in ICU patients who are fluid overloaded and occasionally in those with syndrome of inappropriate antidiuretic hormone due to severe chest infection, such as recently reported in COVID-19 disease5.Mounting evidence have demonstrated the impact of dysnatremias on ICU mortality, even after adjustment for case mix and severity of illness24. There is also evidence from animal studies that osmotic stress impairs immune function by affecting activation and differentiation of B lymphocytes, which are key effector cells for fighting against infections25. Further complicating the situation are the competing priorities for treatment in patients who were admitted to ICU for other conditions but had hypernatremia at presentation, as fluid therapy may contradict with the management strategy of concurrent illnesses. The strong association between dysnatremias at the time of ICU admission and mortality highlights the importance of proper fluid and electrolyte management when patients are managed at the emergency department, general medical/surgical wards, or during the peri-/intra-operative periods. In this study, we observed that hypernatremia was associated with increased ICU mortality in patients without diabetes but not in diabetic subjects. The finding was intriguing but it is possible that diabetic patients have many other medical co-morbidities and impaired immune responses that attenuated the effect of dysnatremias on mortality.The pathophysiology of dysnatremias in increasing patient mortality remains to be thoroughly examined. Putative mechanisms for dysnatremias  to increase mortality include its effects on the neurological, cardiovascular and immunological systems. On a cellular level, Na plays an important role in maintaining tonicity in body compartments and providing optimal enzymatic conditions for essential metabolic processes. Hypernatremia causes serum hyperosmolality, which impairs normal metabolism and transmission of nerve signals. Hypernatremia also decreases cardiomyocyte contractility, thereby compromising cardiac output28. In this study, despite steady correction of the Na level over 75 h in most patients with dysnatremias, up to 15% of patients had evidence of adverse neurological events. Hypernatremia appeared to confer higher risk of subsequent neurological complications compared to hyponatremia. Disturbance in osmoregulation, abnormal thirst sensation and aberrant anti-diuretic hormone response all exacerbate the effects of hypernatremia. The use of mannitol in patients with raised intracranial pressure may also precipitate a hypertonic milieu for neuronal cells29. While previous studies mostly reported the effect of hypernatremia in ICU patients admitted for neurological causes, we showed that hypernatremia at the time of ICU admission was predictive of adverse neurological sequelae in patients with non-neurological conditions. Such finding calls for closer neurological monitoring in ICU patients admitted for non-neurological causes who had abnormal Na levels at the time of admission.Neurological sequelae are important complications of dysnatremias, and are attributable to the Na disturbance per se or overzealous correction. It is well recognized that the tempo of Na correction is important as overly rapid normalization can lead to seizures and potentially life-threatening complications such as cerebral oedema or central pontine demyelinationApart from affecting mortality and neurological outcomes, the impact of dysnatremias also translated to longer duration of stay in ICU and hospital, as well as higher healthcare costs. The burden on healthcare resources are related to the metabolic and neurological sequelae of dysnatremias, which often require prolonged duration on life support and more convoluted process of recovery. Indeed, the downstream effects of adverse neurological outcomes were exemplified by a 50% increase in the requirement for rehabilitation before discharge in patients with hypernatremia at the time of ICU admission.One limitation was the retrospective nature of this study, which may lead to potential biases in other aspects of patient management and problems of residual confounding. For instance, data on fluid management and relevant intervention after ICU admission were not systematically recorded. The measurement of secondary outcomes such as adverse neurological outcomes were not prospectively recorded and may be subjected to errors in coding. It was also not possible to quantitatively gauge the severity of neurological impairment. Lastly, healthcare costs were estimated by the average cost of different types of hospital beds, and outliers in cost of management such as requirement for advanced life support and expensive medications may lead to over/under-estimation of cost data. Notwithstanding, our study included territory-wide data of almost all patients admitted to ICUs across a ten-year study period. The large study population  with mixed medical/surgical cohorts and few exclusion criteria allowed us to conduct relevant stratified analyses. Our findings are therefore representative of real-world data and are highly generalizable to other ICU cohorts.Dysnatremias are common in critically ill patients at the time of ICU admission, and are robust predictors for mortality, neurological complications and increased healthcare costs.Supplementary Information.


Fetal cerebral aneurysm other than aneurysm of vein of Galen aneurysmal malformation (VGAM) is extremely rare. This report describes prenatal features of aneurysm of the posterior cerebral artery (APCA) with rapid progression and its natural intrauterine course of the disease, which has never been reported.This is the first report of prenatal features of APCA, detected at 34–36 weeks of gestation, simulating choroid plexus cyst or arachnoid cyst. The diagnosis was based on color flow ultrasound with tracing along the course of cerebral arteries. Also, rendered 3D color flow ultrasound was helpful in demonstrating course of the vessels feeding the aneurysm and supporting the diagnosis. The aneurysm showed nature of rapidly progressive changes, leading to leakage resulting in intracerebral and intraventricular hemorrhage as well as high output state associated with anemia. Prenatal diagnosis and management are very challenging. This case ended up with planned delivery at 37 weeks, giving birth to a surviving male newborn, weighing 2600 g. The neonatal CT brain scans and CTA confirmed the prenatal findings. The prognosis was relatively poor because of extensive intracerebral hemorrhage with severe hydrocephalus and brain midline shift. The couple opted for neonatal palliative care without neurosurgical correction.This study demonstrate that the most important tool for prenatal diagnosis is color Doppler ultrasound, which will demonstrate turbulent blood flow. Three-dimension color Doppler ultrasound is helpful in supporting the diagnosis. The case presented here suggests that the disease has a natural course of rapid progression and massive brain destruction or high output congestive heart failure can be expected.The online version contains supplementary material available at 10.1186/s12884-023-06056-9. Fetal cerebral aneurysm is a rare but very challenging. Nearly all cases of prenatal diagnosis of cerebral aneurysm ever reported were vein of Galen aneurysmal malformation (VGAM), with the estimated incidence of 1 in 10,000 to 1 in 25,000 births . Though +5 weeks at our hospital, the size of lateral ventricles, third ventricle and fourth ventricle were relatively the same. The round -shape anechoic cyst at the left parieto-occipital lobe was increased in size, 32 × 27 × 25 mm, with expanding hyperechoic area surrounding the cyst, representing pericystic hemorrhage. The left lateral ventricle was filled with hyperechoic irregular mass (hemorrhage) in most part. Color and spectral Doppler ultrasound showed turbulent flow in the cyst. The originating point of arterial jet arose from the left posterior cerebral arteries at left lateral border of temporal-parietal area of the hemisphere  were also documented. Peak systolic velocity of the middle cerebral artery was 103.5 cm/s (greater than 1.5 MoM). After multidisciplinary consensus  and patient counseling about poor prognosis in case of no intervention, planned delivery for postnatal management was performed by induction of labor at 37 weeks of gestation. However, cesarean section was performed due to failure of induction, giving birth to a male newborns, weighing 2,600 gm with Apgar scores of 9, 10 and 10 at 1, 5 and 10 min, respectively. Grossly, the newborn was not obviously abnormal except for pale skin. The occipitofrontal circumference was 37 cm. The fetal movement was normal. Neonatal CT brain scans on day 1 , third ventricle and fourth ventricle, and a round shape, anechoic cyst (13 × 10 mm), at the left parieto-occipital lobe adjacent to the left lateral ventricle  Because of no anomaly seen at mid-pregnancy screening and first detection as a small cyst at 34 weeks, it suggests that APCA likely occur in the third trimester, as commonly seen in VGAM . 2) APCAThe differential diagnosis of a fetal intracranial vascular mass includes hemangioma, Kaposiform hemangioendothelioma, angioblastoma, angiosarcoma, congenital hemangiopericytoma, and arteriovenous malformation / aneurysm . DopplerNatural course of progression of the aneurysm in utero has never been studied. Nevertheless, our evidence supports the progressive nature as described in postnatal life. In adults, most cerebral aneurysms develop rapidly in hours, days or weeks, to achieve a size, of which elasticity in the aneurysmal wall can no longer expand, leading to either rupture or undergoing stabilization and hardening –11. ThisIn summary, fetal aneurysm of the APCA is extremely rare, possibly occurring in the third trimester of pregnancy as commonly seen in aneurysm of the vein of Galen. This disorder must be differentiated from other cystic brain anomalies. The most important tool for prenatal diagnosis is color Doppler ultrasound, which will demonstrate turbulent blood flow. Three-dimension color Doppler ultrasound is helpful in supporting the diagnosis. The case presented here suggests that the disease has a natural course of rapid progression and massive brain destruction or high output congestive heart failure can be expected. However, more several case reports/series are warranted to establish the true natural intrauterine course and prognosis.Additional file 1.


Bioconsolidation treatment using bacterial carbonatogenesis has been proposed as an environmentally friendly strategy for the efficient preservation of damaged stones, particularly suitable for carbonate stones. The study presented here deals with the evaluation of the performance of this treatment, applied to damaged carbonate stones in two historical buildings in Spain. The methodology applied in this research serves as a reference for future similar studies. Results showed significant improvement in the petrophysical and mechanical properties of the damaged stone following the treatment through the production of calcite and vaterite by the abundant carbonatogenic bacteria inhabiting the stone. These bacteria were able to effectively consolidate weathered areas if an adequate nutritional solution was employed, thereby augmenting the stone’s resistance, as evidenced by the Drilling Resistance Measurement System (DRMS). FESEM images showed calcified bacteria and calcified exopolymeric substances (EPS) consolidating stone minerals without blocking their pores. In addition to consolidation, this biotreatment improves the stone’s behavior against water absorption and increases the contact angle of water droplets without significant modifications in the pore size or diminishing vapor permeability. No color changes are observed. Overall, these results show that the application of the nutritional solution (M-3P) for in situ consolidation of different types of porous carbonate building stones is a highly effective conservation method, with no modification of the chemical composition of the treated materials. For example, The bioconsolidation treatment employed here had been successfully applied in some historical buildings in Granada, like the Royal Chapel, San Jeronimo Monastery, and the Royal Hospital , all of The aims of the present study are directed at assessing, on the one hand, the effectiveness of the bioconsolidation treatment on calcarenites with distinct compositions like those employed in the construction of SM and IB. On the other hand, to broaden the scope of existing research on bioconsolidation by exploring its application to other varieties of calcarenite and/or other ornamental materials ,38,39. IFollowing the recommendations of the Spanish Institute of Cultural Heritage (IPCE) , block rThe stone blocks from both buildings were carefully cut into pieces according to the dimensions specified in the different standards ,43,44,45Petrographic characterization of the treated and untreated samples was performed using:Polarizing petrographic microscope (MOP): Zeiss Axioskop transmitted light optical microscope has been used, with a Zeiss Stemi SV 6 magnifying glass and a Photometrics Cool SNAP-CF camera. Thin sections of untreated stone have been observed. This study was carried out at the University of Alicante.Scanning electron microscope (FESEM): Samples have been observed in two different microscopes, at the University of Granada with a Zeiss Model SMT AURIGA Field Emission Scanning Electron Microscope and at the University of Alicante with a ZEISS model Merlin VP Compact. The samples studied were small and prismatic, less than 0.7 cm × 0.7 cm × 0.7 cm. Untreated and treated stone samples have been observed.X-ray diffraction (XRD): The Bruker D8-Advance equipment with the Göebel mirror has been used. Two pulverized grams of each type of untreated stone have been analyzed with the following conditions: Cu Kα radiation, 40 kV, 40 mA, 0.05 2Theta degrees steps, 3 s per step, and 4–60 scan angle. This analysis was carried out at the University of Alicante.Mercury intrusion porosity (MIP): The equipment used was a POREMASTER-60 GT, QUANTACHROME INSTRUMENTS. The samples studied were small and prismatic in shape, less than 0.7 cm × 0.7 cm × 2 cm. This study was carried out at the University of Alicante.The different standard tests (described below) were performed both before and after treatment application, with dried samples taken to constant weight. Non-destructive tests were conducted on the same samples before and after the bioconsolidation treatment to assess their effectiveness.3COO)2·4H2O , 0.2% K2CO3·1/2H2O , and 10 mM phosphate buffer in distilled water (pH 8). Both during treatment and up to two/three days after its completion, the treated blocks remained covered with aluminum foil  to avoid the direct effect of sunlight and to minimize the evaporation of the M-3P solution. The treatments were conducted with strict temperature control to maintain a range of 20 to 30 °C on the treated areas. All samples were kept for at least 30 days before performing any treatment evaluation. The sterile nutritional solution M-3P  was applThe most important stone property that should be improved after successful stone consolidation is the cohesion of disaggregated particles. This enhanced cohesion leads to a significant increase in the stone’s resistance to external forces and environmental deterioration. Many authors consider that the most appropriate method for the evaluation of grain cohesion, and thus of the consolidation performance, is the application of the Drilling Resistance Measurement System  ,48,49,50Both untreated and treated stone cubes measuring 4 cm × 4 cm × 4 cm in size were used for the evaluation of the drilling resistance of the stones. DRMS is designed by SINT Technology  to perform a precise ‘‘drilling resistance’’ test through the continuous measurement of the force necessary to drill a hole in the material under specific operating conditions. The test conditions were as follows: (i) a 4.8 mm diameter flat-edged diamond-tip drill bit ; (ii) a This test was performed following the recommendations of RILEM 25 PEM test II.8 standards . The tesThe stone samples were subjected to this test as recommended in UNE-EN 15802 . ControlFollowing the recommendations of RILEM 25 PEM test II2 , this teE = (ΔL*2 + Δa*2 + Δb*2)½, where ΔL*, Δa*, and Δb* are the difference between untreated and treated stone of L* (lightness: 0 being black and 100 being diffuse white), a* , and b* . This study was carried out at the University of Alicante.This test was carried out according to the UNE-EN 15886 standard  with theData obtained from visualizing the samples using a petrographic polarizing microscope  and a scGiven these specific characteristics and vulnerabilities of both types of calcarenite, bioconsolidation treatments are essential for the conservation of buildings constructed with such stones. Furthermore, these stones present a high susceptibility to deterioration caused by salts, atmospheric agents, and capillary absorption of moisture ,7,8,9,30SEM photomicrographs of treated stone samples showed newly formed cement of calcium carbonate in both historical buildings made with SM and IB stones . In both2s0.5 and 30 g/m2s0.5, respectively.Among the petrophysical characterization study, the most relevant data belongs to porosity and capillary absorption. Results obtained from the Mercury Intrusion Porosity (MIP) test  indicate3 cement consolidated the stone without pore blocking, as was previously shown by SEM analyses. The same effects had been previously demonstrated after the application of this bioconsolidation treatment on other historical buildings [Comparing the MIP data of treated and untreated samples, a reduction of hardly 1% porosity was observed in SM samples, while no detectable reduction was observed in the case of IB samples, maintaining the average stone total porosity. MIP curves moved slightly toward lower diameters but remained in similar ranges so that the behavior against capillary phenomena or salt crystallization, or freeze–thaw cycles were not observed to be affected . These ruildings ,54. The uildings ,56, beinDrilling resistance curves of SM samples and IB samples are presented in These results further corroborate that the strengthening achieved is crucial for the successful consolidation of the stone. Here, once more, the effectiveness of the bioconsolidation treatment applied to calcarenite samples was confirmed, also being aligned with the previous studies ,37,38,39−4 (g/m2), and that of the treated sample was δ = 6.36 × 10−4 (g/m2s). Concerning IB samples, after 30 days, the untreated sample allowed the transit of 5.76 g of water vapor, while the treated one allowed 5.53 g, so the permeability was reduced by about 4 %. The coefficient of water vapor conductivity of the untreated sample was δ = 6.30 × 10−4 (g/m2s), and that of the treated sample was δ = 6.05 × 10−4 (g/m2s).The results of water vapor permeability are shown in These data indicated and confirmed the good behavior of SM and IB stones after the bioconsolidation treatment tested here. The water vapor permeability in the studied stones was reduced much below the limits recommended by experts, who fixed it at 50% . These r3 cement that incorporated bacterially derived organics. This may also be due to modification of the topography of the stone surface [This test helps describe how a liquid droplet interacts with a solid surface and refers to the angle formed between the tangent line of the liquid droplet’s surface where it meets the solid surface and the stone surface itself. This angle is measured within the liquid droplet at the point of contact. The results obtained by the static contact angle measurements are shown in  surface , with thThe results of this test are presented in E values after treatment were 1.08 and 1.90 in SM and IB, respectively. These values are below 2, which is considered an acceptable threshold value for any stone treatment, and they are regarded as extremely low according to the ISO 12647 standard [standard ,60,61, astandard . These fstandard ,37, all The consistent and unaltered color results across different types of calcarenite stones in various historical buildings underscore the effectiveness and reliability of the bioconsolidation treatment as a safe and aesthetically sensitive conservation approach for preserving such important cultural heritage.The study presented here encompasses a comprehensive exploration of various aspects related to the preservation and conservation of historical calcarenite stone structures of different compositions from those previously studied. This study engages in a comprehensive exploration of a promising conservation solution: bioconsolidation treatment. The ongoing results highlight the significance of the innovative approach, its application, and the implications for the broader field of heritage preservation.Historical buildings, often constructed with calcarenite stones, stand as a testament to human history and culture. However, they are confronted with the relentless impact of time, weathering agents, pollution, and salt crystallization, particularly in humid environments. As a result, the gradual increase in material porosity leads to a cascade of issues, including decohesion, mineral constituent disintegration, and, ultimately, structural decay. Preserving these types of historical architectural structures commonly presents a multifaceted challenge.Therefore, central to this study is the concept of bioconsolidation treatment, a novel approach that harnesses the potential of indigenous carbonatogenic bacteria within the stone. The patented nutritional solution, M-3P, acts as a catalyst, prompting these bacteria to precipitate calcium carbonate crystals, thereby fortifying the stone’s structural integrity. This strategy proves effective in consolidating weathered areas, as substantiated by the Drilling Resistance Measurement System (DRMS) results. The efficacy of this bioconsolidation treatment was also evidenced through various analyses, including petrographic and petrophysical characterization, water vapor permeability, and color analysis. Results indicated enhanced stone cohesion, reduced water permeability, and improved hydrophobicity without compromising vapor permeability or altering the stones’ color. These outcomes were consistent with previous studies conducted on similar historical structures, further validating the method’s reliability and potential for broader application.E values. This work highlights the significance of long-term effectiveness, particularly for clay-containing calcarenite substrates. By comparing and contrasting this innovative approach with traditional consolidants, the advancement in conservation strategies ensures the longevity and integrity of our cultural heritage. For instance, comparative analysis with conventional consolidants, such as nanolimes and ethyl silicate, offers insights into the advantages and limitations of each method. While traditional consolidants have demonstrated efficacy, they also raise concerns regarding color changes and potential side effects on the stone, resulting in the formation of cracks and fissures within the stones altering thus the effectiveness of this treatment. In contrast, bioconsolidation treatment distinguishes itself with its non-destructive nature and minimal impact on stone aesthetics, thereby mitigating apprehensions associated with visual alterations.Additionally, through FESEM analysis, the presence of calcified bacterial cells and the detection of bacterial calcium carbonate covered by calcified EPS were evidenced. The method’s effectiveness in consolidating calcitic substrates is further corroborated, and concerns about potential color changes are alleviated, as demonstrated by minimal ΔFurthermore, the consistent success of the treatment across various historical buildings underscores its broader potential. By comparing outcomes to prior studies, including the San Jeronimo Monastery, Royal Hospital, and Royal Chapel, this study establishes the method’s reliability across diverse stone compositions. The implications are far-reaching, potentially revolutionizing the conservation landscape by offering a sustainable and effective alternative for preserving calcarenite stones in an array of architectural and historical contexts.Finally, this innovative approach not only addresses the tangible issues of deterioration but also offers a glimpse into the future of heritage preservation by expanding to the consolidation of different types of historical calcarenite stone substrates. The crossroads of science, conservation, and cultural heritage converge, presenting a formidable opportunity to safeguard our shared history for generations to come.The comprehensive study and analysis of the results, along with visual inspections of the samples before and after the bioconsolidation treatment, confirm the effectiveness of the bacterial carbonatogenesis achieved by the application of the nutritional solution M-3P in strengthening the tested stone. In the case of SM samples, an increase in the drilling resistance was reached in the first millimeters of the stone surface. For the IB samples, the resistance notably improved up to 14 mm depth, doubling its surface resistance, as indicated by the DRMS results. This difference in consolidation depth is attributed to the faster penetration of the M-3P nutritional solution in the case of IB samples, possibly due to the combined effect of the spraying method used for treatment application and the low porosity of this particular sample. Moreover, in both cases, the static contact angle increased after the treatment exceeded double its original values. This indicates that water drops have greater difficulty adhering to the consolidated surface, making the stone more hydrophobic. The increased absorption time of water drops further demonstrates the treatment’s effectiveness in improving the stone’s performance against disturbances generated by humidity.Regarding water vapor permeability, the bioconsolidation treatment to produce the new cement was successful in consolidating the stone without blocking its pores. As a result, there was no significant difference in water vapor permeability between the treated and untreated samples. This is a positive outcome as many conventional consolidation treatments often reduce water vapor permeability , which i


Management of the Mediterranean fruit fly (medfly) with the application of the sterile insect technique depends on the production and release of high quality sterile males that can compete with the feral males to induce sterility in the wild population. Larval rearing should be cost-effective and practical. The right balance between the cost of the protein source (the most expensive ingredient) in larval diets and the quality of the adults produced is a key requirement for effective and economically viable rearing. Two types of diet, a liquid and a gel one, were evaluated across different brewer’s yeast concentrations (protein source) based on survival and developmental duration of larvae and survival of adults. Overall, regardless of the type of diet, brewer’s yeast reduction to 50% of its initial quantity did not affect the quality of the adults produced. Although the performance of immature medflies from the gel diet was inferior compared with the liquid diet, an interesting range of developmental duration phenotypes was observed, opening interesting questions for future experimental work.Ceratitis capitata (Wiedemann) (Diptera: Teprhitidae). There are several efforts to reduce the cost of rearing and optimize the quality of the produced sterile males that are destined for release in sterile insect release programs. Survival, growth, longevity and reproductive capacity of sterile males are strongly connected with the most expensive ingredient, the brewer’s yeast (protein), in the larval diet. The current study focused on settling the optimal content of brewer’s yeast in a liquid diet and a gel diet. Egg hatch rates, developmental duration of immatures, pupation rate, pupae and adult survival were recorded as indicators of quantity and quality of the produced adults. Egg hatch was higher and larval developmental duration longer in the gel diet. In contrast to the liquid diet, an increase in brewer’s yeast concentration was correlated with increased pupation rate and pupae survival in the gel diet. Reducing brewer’s yeast up to 50% of its initial quantity had no significant effect on the survival of the emerging adults regardless of the diet type. Our findings may contribute to the production of low-cost and effective diets for use in mass-rearing facilities of medflies.Several artificial larval diets have been developed, evaluated and used for mass-rearing of the Mediterranean fruit fly (medfly),  Nutrition is a fundamental determinant of insects survival, profoundly impacting their development, growth, behavior and ecology ,2. In frCeratitis capitata (Wiedemann) (Diptera: Tephritidae) is as a highly polyphagous pest that can infest more than 300 different hosts and is considered one of the most threatening pests of the fruit producing industry worldwide [The Mediterranean fruit fly (medfly), orldwide ,16. Becaorldwide ,18,19. Forldwide .SIT is an environmentally friendly control method involving mass-rearing, sex separation and sterilization of males . SteriliOver the last two decades, liquid, meridic and oligidic larval diets have been formulated and tested for medfly rearing ,26,29,30Bactrocera tryoni [Anastrepha ludens as a model system. Amino acids, vitamins and sucrose are the key nutritional components for medfly larval development [A. ludens adults. Developing more efficient and cost-effective diets for mass-rearing is extremely challenging and requires continuous developments and further examination.Gel diets are becoming a preferred alternative to liquid ones not only because they provide similar advantages to liquid ones, but also, they are characterized by physical consistency . Gel diea tryoni . Then, Pa tryoni  evaluateelopment . Yeast ielopment ,46. Althelopment . As a reelopment ,47. For elopment  evaluateOur study evaluated different brewer’s yeast concentrations on a standard liquid diet and a gel diet, aiming on settling the optimum concentration among those tested. The diets were evaluated based on survival and developmental duration of medfly’s immatures and survival of the produced adults. Our results may contribute to the production of low-cost and effective diets for use in mass-rearing facilities of medfly.The experiments were carried out in the Laboratory of Entomology and Agricultural Zoology at the University of Thessaly, Greece. We used eggs from a laboratory adapted strain, named “Benakeio”, which has been maintained under laboratory conditions for more than 30 years .Flies were kept in wooden-framed, wire-screened cages (30 × 30 × 30 cm) under constant laboratory conditions, 25 ± 1 °C, 65 ± 5% RH and a photoperiod of L14:D10 with photophase starting at 7:00 and ending at 21:00. Flies were provided with ad libitum access to water and standard adult diet  . FemalesDomes were placed in rearing cages for 24 h to collect eggs for experimental needs. Collected eggs were placed on Petri dishes containing either liquid diet or gel diet . Petri dThe standard liquid diet consists of 250 mL water, 50 g brewer’s yeast, 50 g sugar, 25 g soy flour, 1 g of salt mixture , 4 g citric acid, 4 g ascorbic acid and 0.75 g sodium propionate [To prepare the liquid formulation, all ingredients A were miUsing a stereomicroscope, 100 eggs were randomly selected and placed on the respective larval diet regime. Five replicates were run for each treatment (brewer’s yeast content) and each diet (100 eggs/replicate: 500 eggs/treatment). In total we used 7000 eggs for the evaluation of both liquid and gel diets. Egg hatch was recorded under a stereomicroscope 3-day post oviposition.Egg -to-pupa developmental duration was calculated (in days) as the time from the day eggs were collected and deposited on larval diet until the day pupation observed. Larval survival was estimated by recording pupation daily. Pupae sieved out from the sand were transferred to plastic Petri dishes and placed in transparent Plexi-glass cages (20 × 20 × 20 cm) that contained adult food and water ad libitum. Pupal survival was estimated by the number of emerging adults. Dead adults were counted and removed each day until the fifth day post emergence .Binary logistic regression and the chi-squared test were used to compare egg hatching, pupation rate, pupa and adult survival rates between the two types of diet and across brewer’s yeast concentrations. Effect sizes are presented as odds ratios (ORs) with 95% confidence intervals (CI), which are the odds of an event  in a group of interest to the odds of the same event occurring in the group used as a reference. ORs greater than 1 indicate greater odds for the group of interest, while ORs less than 1 indicate greater odds for the reference group. Furthermore, Cox proportional hazards regression models were used to infer whether the proportion of brewer’s yeast, the type of diet and their interaction were significant predictors of immature developmental duration. Kaplan–Meier estimates were used to depict pupation progress on different brewer’s yeast concentrations between the two types of diet. The analysis was carried out using R v4.2.2 . The pacp < 0.001). Gel diet had a significantly higher predicted probability (0.95) of egg hatching than liquid diet (0.90) (Predicted probability of hatching as modeled through binary logistic regression was higher than 90% in both diets (liquid and gel). The brewers’ yeast concentration was a non-significant factor on egg hatch; thus, it was removed from the final model. Egg hatching odds increased by a factor of 2.01 for gel diet compared to liquid diet (OR (95% CI) = 2.01 , t (0.90) .2χ = 1440.188, df = 1, p < 0.001) and the brewer’s yeast concentration  were significant predictors of larval developmental duration. The interaction of the aforementioned factors had a significant effect on larval developmental time . Overall, larval developmental duration was longer on the gel diet than on the liquid diet . All larvae completed development and pupation within 10 days from the egg collection day in the liquid diet regardless of brewer’s yeast concentration    . In the p < 0.001) and diet type (OR (95% CI) = 0.0571 , p < 0.001) were significant predictors of pupation (p < 0.001). For the gel diets, the predicted probability of pupating increased with an increase in brewers’ yeast concentration. However, for the liquid diets the predicted probability decreased as the concentration of brewer’s yeast increased. The odds to pupate for each additional unit percent of brewer’s yeast for gel diets increase by a factor of 4.67%  = 0.499 , pupation . In the p < 0.001) was a significant predictor of pupal survival, unlike the diet type (OR (95% CI) = 0.87 , p = 0.2) (p < 0.001). The interaction between the two predictors was significant (OR (95% CI) = 4.92 , p < 0.001). For the gel diets, the predicted probability of pupating increased with an increase in brewers’ yeast concentration. However, for the liquid diets the predicted probability decreased as the concentration of brewer’s yeast increased. The odds to pupate for each additional unit percent of brewer’s yeast for gel diets increase by a factor of 1.6% (Brewers’ yeast concentration (OR (95% CI) = 0.48 , p = 0.2) . The oddp < 0.001). Specifically, the odds for adult survival when reared from the gel diet were 93.17% of those reared from the liquid diet. The survival of adults was also affected by the concentration of brewer’s yeast in each of the diets. The odds of five-day adult survival increased by 5% (p < 0.001). The survival of adults was affected significantly by the type of diet (OR (95% CI) = 0.0683 , Our results reveal that both liquid and gel diets could be used to successfully rear medfly from egg to adult. Reducing brewer’s yeast to 50% of its initial amount on both diet types had no negative effect on the survival and fitness of the adults. Further decreases in brewer’s yeast to levels below 50% in the gel diet resulted in growth retardation. In the gel diet, no immatures survived at the lowest concentrations of 1 and 2% of the initial amount of yeast. However, in the liquid diet, immatures survived in the lowest concentrations, and no negative effects of yeast reduction on developmental duration were found. It seems there is a strong association between brewer’s yeast availability and the diet type. This result may be based on the fact that the use of agar in the gel diet distributed the amount of brewer’s yeast evenly throughout the diet , in contPrevious studies suggested that both liquid ,41,57,58Egg hatching rate was higher on the gel diet than the liquid one. Agar has the ability to modify the amount of water in the gel, providing greater water availability and higher humidity levels for eggs to hatch, while liquid diet tends to dry out faster . Water, The protein amount in larval diet regulates body size, developmental duration, survival and mating success of emerging adults ,60. TherC. capitata. An important requirement for the SIT is to produce qualitative insects at a lower cost. Brewer’s yeast is the most expensive ingredient on most applied diets. Lowering brewer’s yeast concentration up to 50% of the initial brewer’s yeast quantity did not affect the survival of immatures and adults on both types of diets. An increase in brewer’s yeast concentration was correlated with increased pupation and adult emergence rates in the gel type of diets. However, concerning liquid diets, some difficulties such as lower moisture retention should be overcome to be applied in a larger rearing scale. Brewer’s yeast availability is higher on liquid diets than on gel diets because of agar’s properties. Those properties make agar-based diets suitable to be applied in food-stress experimental procedures.Both liquid and gel diets, tested in the current study, could sustain the rearing of


However, their low colloidal stability under storage and operation reduces the potential use of FAPbBr3 perovskite nanocrystals (PeNCs) in various applications. In this study, we prepared the poly(L–lactic acid) (PLLA) nanofibrous membrane embedded with FAPbBr3 perovskite nanocrystals by electrospinning the perovskite and PLLA precursor solution. This is a simple and low-cost technique for the direct confinement of nano-sized functional materials in the continuous polymer nanofibres. PLLA as a polymer matrix provided a high surface framework to fully encapsulate the perovskite NCs. In addition, we found that FAPbBr3 PeNCs crystallize spontaneously inside the PLLA nanofibre. The resultant PLLA-FAPbBr3 nanofibrous membranes were stable and remained in the water for about 45 days without any evident decomposition. The results of this research support the idea of new possibilities for the production of air-stable FAPbBr3 PeNCs by forming a composite with PLLA polymer. The authors believe this study is a new milestone in the development of highly stable metal halide perovskite-based nanofibres, which allow for potential use in lasers, waveguides, and flexible energy harvesters.Formamidinium lead bromide (FAPbBr Most reMAPbBr3) . These NMAPbBr3) , core/shnce (PL) 30–535 nmMAPbBr3) , and encMAPbBr3) ,8. Amongproduced . In anotproduced .Polymer nanofibres (PNFs) are one-dimensional long filaments with diameters in the range of tens to hundreds of nanometers. They offer unique properties of high porosity, bending elasticity, and high encapsulation capacity. Among several techniques to produce PNFs, electrospinning is the most cost-effective and efficient way to process a wide range of polymers . Further3NH3PbBr3 as NCs and PLLA as the matrix. The fabricated perovskite–polymer composite films showed uniform morphology with outstanding emission characteristics of PeNCs and high transmittance of PLLA matrices [Polylactic acid (PLA) is the most commonly used bioplastic. Lactic acid, the precursor of PLA, can be derived from various raw materials including corn, starch, and sugarcane, which is then polymerized to PLA. Among different kinds of PLA, poly (L–lactic acid) (PLLA) has been an object of many studies due to its biocompatibility, biodegradability, and eco-friendly nature . It has matrices . In anotmatrices . The res3 NCs. We mentioned in situ room temperature fabrication of FAPbBr3 NCs–PLLA nanofibres with FAPbBr3 NCs as an emitter and PLLA as the matrix. FAPbBr3 NCs–PLLA nanofibres can be easily produced by the electrospinning technique, which is low-cost, versatile, and allows for the control of NFs composition. The as-produced composite film shows a homogeneous morphology, combining the excellent properties of both materials. The outstanding properties of PLLA help to make highly stable and efficient electrospun nanofibrous films with excellent stability in ambient environments, retaining >50% of PL emission intensity. Overall, this study established a straightforward and low-cost method to prepare the composite film of FAPbBr3 NCs–PLLA for strong stability in air and water.The present research explores, for the first time, the use of PLLA nanofibres to encapsulate FAPbBr6) was supplied by PURAC biochem, Holland. Chemicals including lead bromide , formamidinium bromide , oleic acid , oleylamine , dimethylformamide , and dichloromethane  were purchased from Sigma-Aldrich.Poly (L–lactic acid) (PLLA)  and FABr (0.0124 g) were dissolved in 0.5 mL of DMF. Then, 200 µL oleic acid and 15 µL oleylamine were mixed and poured into the perovskite solution. Next, this perovskite precursor solution was added dropwise into a vigorously stirred PLLA solution at room temperature (25 °C) for 24 h. Then, the mixture was filled into a plastic syringe using a metal needle (19G) and was loaded into a syringe pump. The distance between the syringe tip and the collector was fixed at 30 cm and a high voltage of about 23 kV was applied to charge the precursor solution. The electrospinning solution was released towards the moving roller (200 rpm) to collect NFs. The collected NFs were allowed to dry overnight.The FAPbBrications . PLLA  and FABr (0.1 mmol) were dissolved in 0.5 mL of DMF to prepare the precursor solution. Then, 250 µL of oleic acid and 15 µL of oleylamine were mixed separately in a vial and added to the solution. After that, 130 µL of this freshly prepared precursor solution was dropped into the 10 mL of chloroform under constant stirring and we observed the immediate formation of a green solution. Purification of PeNCs was completed by introducing a small amount of acetonitrile followed by centrifugation at 1200 rpm for 10 min. The collected PeNCs precipitate was further dispersed in 4 mL hexane for characterization.We followed the ligand-assisted reprecipitation (LARP) method for the synthesis of FAPbBr changes . PbBr2  and SEM (FEI Inspect F), respectively. The chemical composition of samples was determined by EDS (attached to FEI Inspect F) at a voltage of 10 kV. The confocal microscope was used to record the fluorescence images with a 488 nm laser as the excitation light source. X-ray diffraction analysis of as-prepared NFs and FAPbBr3 NCs was performed by PANalytical diffractometer with an angular range of 5 < 2θ < 70°.The morphology of FAPbBr−1. The composition of as-prepared samples was confirmed by X-ray photoelectron spectroscopy (Thermo Scientific Nexsa XPS system) with an excitation source of AL Kα X-rays.The UV–Vis absorption and PL spectra were recorded using Perkin Elmer Lambda 35 UV–Vis spectrophotometer and Perkin Elmer LS55 spectrofluorometer, respectively. Fourier transform infrared (FTIR) spectra were recorded using a Burker model FTIR with a range of 4000–400 cm3 NCs due to its outstanding resistance against water and ambient conditions. As schematically shown in 3 NCs–PLLA nanofibres. A precursor solution of PbBr2, FABr, and PLLA in DMF/DCM was used for electrospinning.PLLA was used as a matrix for FAPbBbr3 NCs took place within the PLLA polymer.A liquid is electrified to generate a jet that was directed toward the grounded collector, and the solvent evaporated simultaneously during the electrospinning. The high electric potential between two electrodes able to overcome the surface tension inside the electrospinning solution allows the facile synthesis of continuous fibres. When the concentration of the electrospinning solution reached the critical saturation level, crystallization of the FAPbBbr3 NCs–PLLA nanofibres reveals the smooth nanofibre structure with the FAPbBr3 NCs embedded homogeneously in the PLLA polymer matrix, as shown in 3 NCs. Many recent studies have shown that well-distributed NCs inside the polymer matrices do not affect the diameter of the nanofibres [The average diameter of as-synthesized nanofibres is 1.45 µm, and not influenced by the incorporation of FAPbBrnofibres .3 NCs–PLLA nanofibres show that the fabricated PeNCs are well dispersed inside the PLLA matrix, and have a different shape, from cubic to spherical, depending on the amount of precursor solution concentration. Respective confocal fluorescence images exhibit strong PL emission  was used for the analysis of FAPbBr3 NCs are homogeneously distributed in the PLLA matrix network through the in situ ambient fabrication technique.Therefore, the above-mentioned discussions demonstrate that the FAPbBr3 NCs are synthesized with FABr/PbBr2 using the LARP technique and purified according to the previously mentioned report by Chen et al., with rational changes [3 NCs via UV absorption and photoluminescence (PL) measurements. In 3 NCs dispersion in hexane. From the normalized UV–Vis spectra, it demonstrated a strong light absorption edge at 512 nm.FAPbBr changes . In this3 NCs. The PeNCs crystallize into a cubic shape, monodispersed, with an average size of 10–12 nm. The PeNCs follow the typical perovskite structure ABX3, as shown in 3 NCs was further analysed by X-ray diffraction (XRD) patterns.The crystallinity of FAPbBr3 NCs, and FAPbBr3 NCs–PLLA nanofibres are displayed in 3 NCs , (210), (220), and (300) planes of FAPbBr3 NCs. Nevertheless, the diffraction peak intensity is very low due to the low concentration of FAPbBr3 NCs in the PLLA matrix, as well as the coverage effect of the PLLA matrix [3 NCs have been successfully fabricated inside the PLLA matrix.The XRD patterns of PLLA nanofibres, FAPbBrin 3 NCs  consistsA matrix . These f3 NCs, and FAPbBr3 NCs–PLLA nanofibres are demonstrated. The spectrum shows bands at 2944 and 3000 cm−1 for PLLA, 2850 and 2924 cm−1 for PeNCs, and 2850 cm−1 for FAPbBr3 NCs–PLLA nanofibres. These bands belong to the C–H stretch from CH3. The presence of this stretching confirms the encapsulation of FAPbBr3 NCs into the polymer matrix. Compared to PeNCs bands at 3270 cm−1 for N–H stretching, there is negligible vibration for FAPbBr3 NCs–PLLA. This is due to the lower amount of PeNCs and coverage effects of the PLLA polymer matrix as confirmed by the X-ray diffraction patterns of these samples. In addition, the bands at 1745 cm−1 for PLLA and FAPbBr3 NCs–PLLA correspond to the C=O extension of the ester. For FAPbBr3 NCs, the C=O stretch can be ascribed to the presence of oleic acid at the surface of perovskite NCs.Fourier transform infrared spectroscopy (FTIR) has been widely used to study the chemical changes and conduct the quantitative analysis of polymers as well as serving to investigate the functional groups of the attached molecules. In 3 NCs inside the PLLA matrix. As can be seen in 3 NCs–PLLA nanofibres. However, there is no obvious peak of elements of N (≈401 eV) present on the composite film due to the lower concentration of PeNCs as confirmed by the XRD analysis. Regarding the Pb 4f and Br 3d spectra, the intrinsic peaks show the presence of encapsulated NCs inside the PLLA matrix (3 NCs fabricated by other techniques.X-ray photoelectron spectroscopy (XPS) is extensively employed to measure the binding energy and composition of constituents. As shown by the XPS spectra, it further confirms the presence of Pb and Br in the PLLA matrix, suggesting the formation of FAPbBrA matrix b,c. In aPeNCs can be embedded in a variety of polymers to fabricate nanocomposites that show an array of advantageous properties. The polymer matrix provides stability and flexibility while the PeNCs maintain their size, morphology, and composition-dependent characteristics. Polymers offer unique attributes, such as the ability to interact with the perovskites, low diffusion rates of moisture and oxygen, passivation of NCs surface defects, and decreased agglomeration of PeNCs in the solid state. The use of polymers provides strong chemical interaction with PeNCs, hence reducing the chances of moisture-induced degradation. In addition, the morphology and shape of polymer-encapsulated NCs do not change and PL intensity decreases considerably slower .3 NCs–PLLA nanofibrous films in water and air. The PL peak position and intensity were changed slightly after 10 days of storage in air (3 NCs–PLLA nanofibrous films after storage in air and water. The change in PL intensity value as a function of storage time in air and water was summarized in 3 NCs–PLLA nanofibrous films, and their water immersion is displayed. A comparison of the most recent research studies available on the FAPbBr3 PeNCs embedded into the polymers using the electrospinning method for water and air stability is summarized in This section of the study was concerned with the stability of FAPbBre in air a, which e in air b, the PL3 NCs–PLLA electrospun composite films were prepared in ambient or room temperature conditions and tested for air and water stability. We established that this technique is simple, low cost, and robust enough to fabricate FAPbBr3 NCs that are homogenously distributed into the PLLA polymer nanofibres. These PeNCs are effectively protected and exhibited similar stabilities against air and water stress as those mentioned in the previous studies. Moreover, more than 70% of the PL intensity is still maintained after 45 days of storage. The findings reported here shed new light on the use of biopolymers for optoelectronic applications including lasers, waveguides, and flexible energy harvesters. In addition, the flexible nature of nanofibres films may open the door for the production of flexible and bendable optoelectronics. However, PeNCs-based composite films with tunable emissions in the visible spectrum remain to be explored. Hence, this research will stimulate further study in the area of high-performance luminescence applications.In summary, the first stable FAPbBr


Rif1, Tk1, Fan1 and Mn1. These genes are primarily related, among other less well characterized functions, to nucleic acid biology. We also associated distinct motor learning patterns with groups of functionally related genes. These functional sets included preferentially histone H3 methyltransferase activity for mice that learnt at an accelerated rate relative to the rest of mutant mice. The results allow for an estimation of the fraction of mutations that can modify a behavior influential for evolution such as locomotion. They may also enable, once the loci are further validated and the mechanisms elucidated, the harnessing of the activity of the newly identified genes to enhance motor ability or to counterbalance disability or disease.Evolution depends upon genetic variations that influence physiology. As defined in a genetic screen, phenotypic performance may be enhanced or degraded by such mutations. We set out to detect mutations that influence motor function, including motor learning. Thus, we tested the motor effects of 36,444 non-synonymous coding/splicing mutations induced in the germline of C57BL/6J mice with N-ethyl-N-nitrosourea by measuring changes in the performance of repetitive rotarod trials while blinded to genotype. Automated meiotic mapping was used to implicate individual mutations in causation. 32,726 mice bearing all the variant alleles were screened. This was complemented with the simultaneous testing of 1,408 normal mice for reference. 16.3% of autosomal genes were thus rendered detectably hypomorphic or nullified by mutations in homozygosity and motor tested in at least 3 mice. This approach allowed us to identify superperformance mutations in  Rather, experimental evidence is required to establish, in a function-specific manner, when the characteristics of the organism are faciliated or degraded by individual genes or when they are subject to more numerous genetic influences. We set out to experimentally elucidate the relative contribution of these two possibilities to locomotion and motor learning. The motivation for the study of movement is that it represents a trainable, complex or high-order organism function where both individual genes and groups of genes may be expected to be influential. Ample precedent exists for both kinds of influence: On the reductionist end are laboratory mice subject to spontaneous or intended single-gene mutations that degrade motor phenotype. This is also the case for some human diseases that deteriorate motor ability where the causal links between gene product, cellular and organism function are few or stereotyped. In contrast, on the opposite, more complex end, most of the behaviors considered indispensable in the natural environment require adaptation to changes in initial and intervening conditions. This adaptation is often enabled by the concerted interaction of gene products that subserve the activity of the whole cell, organ or organism rather than one of their constitutent parts. The disease counterpart of this concerted action is exemplified by the dependence of mitochondrial disease manifestations on the transcriptome of a functionally related gene family rather than on individual genes  behaviour , locomotThus, we performed a forward genomic screening of mutations that may impact locomotion. Given the mutability of the laboratory mouse, we examined whether definable mouse mutations artificially induced and tested for effect in both homozygous and heterozygous states, might augment or degrade motor performance as assayed on a standard  rotarod over several consecutive testing trials performed while blinded to genotype and compared with control mice tested simultaneously. Our work was faciliated by the fact that, originally, the C57BL/6J laboratory mouse underwent a process of stringent inbreeding, fixing alleles that might impede motor performance in the homozygous state. Indeed, many wild mice and some laboratory strains exhibit motor performance inferior or superior to C57BL/6J mice. We reasoned that most randomly induced point mutations are inconsequential, but some, by causing loss or gain of function, may alter and even augment performance at the organism level. We were particularly interested in motor superperformance mutants given the far reaching therapeutic development potential of any gain of function thus identified, since genetic gain of motor function might ultimately prove amenable to replication via the development of mechanism-related pharmacological or other interventions.This study was approved by the Institutional Animal Care and Use Committee of UT Southwestern Medical Center. All other relevant institutional regulations were followed, in addition to ARRIVE  guidelines 2.0.Mice were generated as previously . In brieWhile some of the mutations were homozygous lethal prior to weaning age and therefore were tested only in the heterozygous state, the majority were homozygous viable. To compute genome saturation, or the fraction of all genes destroyed or severely damaged and examined three or more times in the homozygous state, we used a method based on observation of a curated set of essential genes: those known to be essential for life based on the results of robust knockout mutations on the C57BL/6J background. We also used a larger set of ENU-induced mutations and checked the effect of mutations falling into genes of this type. This allowed us to correlate “severe damage or destruction” manifested as a lethal effect with PolyPhen-2  scores  was ~99.69% to a depth ≥10 nucleotides . Since we give consideration to two or more identical aberrant calls among 10 reads , fewer than one in 10,000 mutations within the target sequence will be missed in sequencing. False positive mutations, while exceedingly rare, are eliminated by resequencing every non-synonymous coding/splicing change identified using Ion Torrent sequencing technology in the course of genotyping G1, G2, and G3 mice (described below).On average, 60 coding/splicing changes were identified per G1 male. Each G1 male with ≥30 mutations and adequate breeding capacity was used to generate a large pedigree. G1 males were crossed to C57BL/6J females to yield 10 to 15 G2 daughters. Each G1 male was then crossed to its own daughters to yield ~40 to 60 G3 mice , in which most of the mutations were brought to homozygosity. G1, G2, and G3 mice were genotyped at all mutation sites identified in the G1 founder, this time using Ion Torrent sequencing. In rare instances, mutations identified by Illumina sequencing were not confirmed, and were eliminated from further consideration. All validated mutations were assessed for zygosity in G2 mothers and G3 offspring. This allowed detection of probable lethal effects, evidenced by distortion of the expected Mendelian ratio. It also allowed subsequent mapping of each observed phenotype to a causative mutation, using the statistical computation method we have described .p value; the magnitude of the phenotypic effect; the number of homozygotes assayed; the variance of the data; the degree of overlap between phenotypic performance of homozygous mutant and homozygous reference allele populations; the existence of multiple alleles supporting causation; the predicted damage caused by each allele (assessed by another machine learning program); the predicted essentiality of the candidate gene ; and others. Candidate Explorer is trained on the basis of CRISPR/Cas9 validation data.Candidate Explorer, a machine learning program developed by us , determiRif1I107T/I107T this translates into an increase in maximum linear mouse speed of approximately from 1.0 m/ min (control) to 1.4 m/ min.We used a standard rotarod  in several consecutive testing trials performed over 7 days while blinded to genotype and simultaneously testing control littermate mice. The rotarod accelerates; hence, a prolonged time to fall (or raw score) at elevated angular velocities indicates much greater motor performance than a similar prolongation at lower speeds. For example, in the case of the superpeforming mutant We conducted a rigor and effectiveness analysis by applying rotarod screening to mice mutated as above at a gradually increasing rate from 100 to 900 mice per week by a team of six testers who tested an approximately equal number of mice per week. Testing more than 900 mice per week nearly exceeded testing capacity and was thus considered prohibitive for efficient screening. Conversely, low mouse numbers also negatively impacted phenotypic mutation detection rate. th trial in these mice tested over time showed stability . 32,075 of these mutations resided in a total of 32,726 G3 mice, which were screened in the homozygous state at least once. By computing gene damage or destruction, we estimate that 16.3% of all autosomal genes were severely damaged or destroyed by mutations that were brought to homozygosity in five or more G3 mice, each tested six times using the rotarod. Note that the X chromosome is not mutagenized in our G3 mice: it was derived from a wild type C57BL/6J female mated to a mutagenized G0 male, which contributed a Y chromosome to the G1 founder. However, spontaneous mutations of the X chromosome did occur and were detected. If all chromosomes are included in the saturation estimate, at least 11.4% of all genes were damaged or destroyed by mutations that were brought to homozygosity in five or more G3 mice, each tested six times using the rotarod. The average percent of each mutation effect was: Probably benign: 38.3%, possibly damaging: 15.1%, probably damaging: 38.8%, and probably null: 4.5%. The remaining 3.3% can be accounted for by mutations classified as "unknown".By the criterion of “good or excellent” candidate ranking in Candidate Explorer , we notestruggles, an allele of Nfatc3 that is also associated with reduced survival. All are unambiguously assigned to a single mutation except even-steven, a phenotype that results from one of two co-segregating mutations, in Ak8 or Ntng2 (Candidate Explorer favored the former). Two separate mutations in the leptin receptor (Lepr) gene, both leading to obesity, caused poor rotarod performance. Although Lepr is expressed in the brain, obesity mutations  can cause poor rotarod performance. Among the mutant genes that enhance rotarod performance, we unambiguously assigned  a single recessive ENU-induced mutation, Rif1I107T/I107T (nietzsche) that increases rotarod performance , Fan1I618N/I618N  and Mn1  mutations in relation to motor superperformance  is comparable to other reports that also use ENU mutagenesis and behavioral screening for sleep  2 behav  motor leaning rate.We identified Rif1, Tk1, Fan1 or Mn1 is uneven. For example, whereas these genes are differentially expressed in brain areas potentially relevant to locomotion and whereas all have been implicated in nucleoside phosphorylation, more significant evidence is available regarding the function of Rif1. Rif1 regulates several DNA damage response processes, particularly in relation to double stranded break (DSB) repair, as it influences repair pathway selection between non-homologous end-joining (NHEJ) versus homologous recombination repair (HRR)  . DNA repir (HRR) , which mir (HRR) , whereasir (HRR) . Additioir (HRR) . Rif1 renous DSB  relevantnous DSB . Thus, tA salient association of the 6,096 genes related, when mutated, to relative accelerated motor leaning is with histone H3 methyltransferase activity. Whereas numerous additional activities may be relevant given the large number of these genes, mouse histone regulation via methylation has been implicated in certain forms of synaptic plasticity , includia priori that ion channel and other membrane function genes would be best situated to exert the changes of neuronal function necessary for superperformance or increased motor learning, we find that the most robust associations point to DNA regulatory aspects.In summary, whereas we might have entertained −9) × (7.9×10−9) or 6.24×10−17. Taking into account our results indicating that 0.15% and 0.27% of all the homozygous ENU mutations studied modify locomotor performance and motor learning phenotype, about (7.9×10−9) × (7.9×10−9)×0.15) mutations, or 9.36×10−18 (1.19×10−7%) of mutations per replication may endow a superior phenotypic impact. Similarly, 2.13×10−7 % of mutations per replication may endow superior motor learning.Given the rate of locomotor-impacting mutations we have identified and the estimated spontaneous mutation rate, it is possible to conjecture the probability of appearance of a modification in locomotor behavior over time. Despite the ever-increasing diversity of neurological disease mechanisms elucidated, motor impairment remains a common and poorly treatable outcome. For example, the motor consequences of stroke alone account for over one half of all the neurological disability in the world . In contSupplement 1Supplement 2Supplement 3Supplement 4