A density functional theory study of a water gas shift reaction on Ag(111): potassium effect.

Density functional theory (DFT) calculations are executed to investigate the effect of a potassium (K) promoter on the activity of the water gas shift reaction (WGSR) over an Ag(111) surface. It is found that the WGSR proceeds mainly through the OH(O)-assisted carboxy pathway in which H2O dehydrogenation is the rate-controlling step on both Ag(111) and K/Ag(111) surfaces. Energetic span model analysis shows that K addition can enhance the activity of the WGSR by reducing the apparent activation energy of the whole reaction since it can promote H2O dissociation and stabilize the adsorption of the reactants (CO and H2O). Importantly, the K adatom can stabilize the binding of all oxygenates by direct K-O bonding and the stabilizing effect of K on OH adsorption of the transition state (TS) plays a leading role in promoting H2O dissociation. Moreover, the K-O distance and K coverage are two key factors affecting H2O activation, that is, the shorter the K-O distance (2-3 Å) the more the K coverage (25%) contributes to the stronger promotion effect. For various metals catalyzing the WGSR, K promotes H2O dissociation on inert metals like Ag, Au and Cu better than those on reactive metals (Pd and Ni) since the more inert metal surfaces would weaken the K and O binding and accordingly strengthen the interaction between them, resulting in a higher promotion effect.

Genetic and molecular characterization of determinant of six-rowed spike of barley carrying vrs1.a4.

KEY MESSAGE: Decisive role of reduced vrs1 transcript abundance in six-rowed spike of barley carrying vrs1.a4 was genetically proved and its potential causes were preliminarily analyzed. Six-rowed spike 1 (vrs1) is the major determinant of the six-rowed spike phenotype of barley (Hordeum vulgare L.). Alleles of Vrs1 have been extensively investigated. Allele vrs1.a4 in six-rowed barley is unique in that it has the same coding sequence as Vrs1.b4 in two-rowed barley. The determinant of row-type in vrs1.a4 carriers has not been experimentally identified. Here, we identified Vrs1.b4 in two-rowed accessions and vrs1.a4 in six-rowed accessions from the Qinghai-Tibet Plateau at high frequency. Genetic analyses revealed a single nuclear gene accounting for row-type alteration in these accessions. Physical mapping identified a 0.08-cM (~ 554-kb) target interval on chromosome 2H, wherein Vrs1 was the most likely candidate gene. Further analysis of Vrs1 expression in offspring of the mapping populations or different Vrs1.b4 and vrs1.a4 lines confirmed that downregulated expression of vrs1.a4 causes six-rowed spike. Regulatory sequence analysis found a single 'TA' dinucleotide deletion in vrs1.a4 carriers within a 'TA' tandem-repeat-enriched region ~ 1 kb upstream of the coding region. DNA methylation levels did not correspond to the expression difference and therefore did not affect Vrs1 expression. More evidence is needed to verify the causal link between the 'TA' deletion and the downregulated Vrs1 expression and hence the six-rowed spike phenotype.

Relationship Between Serum ET-1, HDL-C, and sVCAM-1 and Hearing Loss in Patients with Sudden Deafness.

Vascular causes are most commonly associated with sudden sensorineural hearing loss (SSHL). This study was performed to determine the relationship between serum endothelin-1 (ET-1), high-density lipoprotein cholesterol (HDL-C), soluble vascular cell adhesion molecule-1 (sVCAM-1) levels, and the degree of hearing loss in patients with SSHL. Firstly, 60 SSHL patients were admitted to The First Hospital of Shanxi Medical University. In the same period, 60 healthy subjects matching the age and gender of SSHL patients were selected as the control group. Then, serum levels of ET-1, HDL-C, and sVCAM-1 were measured by enzyme-linked immunosorbent assay (ELISA). Next, the relationship between serum levels of ET-1, HDL-C, and sVCAM-1 with clinicopathological factors and their diagnostic and prognostic values were analyzed and evaluated. Serum ET-1 and sVCAM-1 were increased, and HDL-C was decreased in patients with SSHL. Serum ET-1 and sVCAM-1 were higher and HDL-C was lower in patients aged ≥ 45 years, or severe hearing loss patients (P < 0.05). ROC analysis determined that ET-1 (AUC = 0.839), HDL-C (AUC = 0.830), and sVCAM-1 (AUC = 0.865) had excellent diagnostic values. In addition, patients with low levels of ET-1 and sVCAM-1 and high levels of HDL-C had better hearing prognosis (P < 0.05). Abnormal serum ET-1, HDL-C, and sVCAM-1 in patients with SSHL are closely related to age, and degree of hearing loss, and perform diagnostic and prognostic values.

Revolutionizing agriculture with nanotechnology: Innovative approaches in fungal disease management and plant health monitoring.

Nanotechnology has emerged as a transformative force in modern agriculture, offering innovative solutions to address challenges related to fungal plant diseases and overall agricultural productivity. Specifically, the antifungal activities of metal, metal oxide, bio-nanoparticles, and polymer nanoparticles were examined, highlighting their unique mechanisms of action against fungal pathogens. Nanoparticles can be used as carriers for fungicides, offering advantages in controlled release, targeted delivery, and reduced environmental toxicity. Nano-pesticides and nano-fertilizers can enhance nutrient uptake, plant health, and disease resistance were explored. The development of nanosensors, especially those utilizing quantum dots and plasmonic nanoparticles, promises early and accurate detection of fungal pathogens, a crucial step in timely disease management. However, concerns about their potential toxic effects on non-target organisms, environmental impacts, and regulatory hurdles underscore the importance of rigorous research and impact assessments. The review concludes by emphasizing the significant prospects of nanotechnology in reshaping the future of agriculture but advocates for a balanced approach that prioritizes safety, sustainability, and environmental stewardship.

Assessing genetic diversity in critically endangered Chieniodendron hainanense populations within fragmented habitats in Hainan.

Habitat fragmentation has led to a reduction in the geographic distribution of species, making small populations vulnerable to extinction due to environmental, demographic, and genetic factors. The wild plant Chieniodendron hainanense, a species with extremely small populations, is currently facing endangerment and thus requires urgent conservation efforts. Understanding its genetic diversity is essential for uncovering the underlying mechanisms of its vulnerability and for developing effective conservation strategies. In our study, we analyzed 35 specimens from six different populations of C. hainanense using genotyping-by-sequencing (GBS) and single nucleotide polymorphism (SNP) methodologies. Our findings indicate that C. hainanense has limited genetic diversity. The observed heterozygosity across the populations ranged from 10.79 to 14.55%, with an average of 13.15%. We categorized the six populations of C. hainanense into two distinct groups: (1) Diaoluoshan and Baishaling, and (2) Wuzhishan, Huishan, Bawangling, and Jianfengling. The genetic differentiation among these populations was found to be relatively weak. The observed loss of diversity is likely a result of the effects of natural selection.

The N545S and K717N substitution at the N-glycosylation sites of the S2 subunit of avian infectious bronchitis virus can significantly enhance viral pathogenicity.

The S2 subunit of infectious bronchitis virus (IBV) is a heavily glycosylated protein that can impact various characteristics of the virus. It is currently known that N-glycosylation modifications are predominantly located on the S2 subunit. However, the exact role of their N-glycosylation modification remains undisclosed. To elucidate the function of these N-glycosylation sites, we identified 14 common sites distributed on the S2 subunit of the 5 genotypes of IBV in present study. Subsequently, we selected 7 sites to generate mutants and assessed their impact on viral virulence, replication ability, and antigenicity. Our finding revealed that only 2 substitutions, N545S and K717N, increased the viral replication titer and antigenicity, and ultimately the pathogenicity in chicks. To delve into the mechanisms underlying this increased pathogenicity, we discovered that K717N can change the structure of antigenic epitopes. The N545S substitution not only influenced antigenic epitope structure, but also enhanced the ability of the virus to enter CEKs during the early stages of viral replication. These results suggest that the enhanced viral pathogenicity associated with N545S and K717N substitutions is multifaceted, with acceleration of the viral membrane fusion process and alterations in epitope structure representing crucial factors in the capability of N-glycosylation modifications to boost viral virulence. These insights provide valuable guidance for the efficient development of live attenuated vaccines.

Distribution patterns and drivers of urban green space and plant diversity in Haikou, China.

Investigating historical and ongoing changes in urban green space (UGS) and urban plant diversity (UPD) provides critical insights into urban ecology and urban planning development. The present study illuminates some of the transformations which can occur in rapidly developing urban landscapes. In this work, we used 30 m resolution images from the Landsat 5 satellite from 2015 to investigate UGS patterns in Haikou City, China. Metrics of UPD were obtained using field surveys, allowing the proportion of UGS and UPD to be determined in each urban functional unit (UFU) of Haikou. The results revealed that leisure and entertainment areas (such as parks) had the highest diversity, whereas roads and transportation hubs had the lowest. More frequent anthropogenic maintenance had a positive effect on the total number of species, including cultivated, tree, and herb species. Similarly, increased watering frequency had a positive impact on the diversity of cultivated and shrub species. By providing demonstrating a crucial link between UGS and UPD, the results provide valuable information for planning sustainable urban development in Haikou City and other tropical regions. They highlight the important role of UGS in maintaining biodiversity and providing a range of ecosystem services. This research will inform policymakers and urban planners about the need to consider UGS and UPD in urban planning and management process, in order to promote sustainability and conservation of biodiversity.

The air we breathe: An In-depth analysis of PM2.5 pollution in 1312 cities from 2000 to 2020.

In recent decades, the phenomenon of rapid urbanization in various parts of the world has led to a significant increase in PM2.5 concentration, which has emerged as a growing social concern. In order to achieve the objective of sustainable development, the United Nations Global Sustainable Development Goals (SDGs) have established the goal of creating inclusive, safe, resilient, and sustainable cities and human habitats (SDG 11). Goal 11.6 aims to decrease the negative environmental impact per capita in cities, with an emphasis on urban air quality and waste management. However, the global distribution of PM2.5 pollution varies due to disparities in urbanization development in different regions. The purpose of this paper is to explore the global spatial distribution and temporal variation of PM2.5 in cities with populations greater than 300,000 from 2000 to 2020, to gain insight into the issue. The findings indicate that PM2.5 concentrations are expected to continue increasing as urbanization progresses, but the rate of evolution of PM2.5 concentration varies depending on the continent, country, and city. From 2000 to 2020, PM2.5 concentration increased significantly in Asia and Africa, with the majority of the increased concentrations located in Asian countries and some African countries. On the other hand, most European and American countries had lower PM2.5 concentrations. The results of this study have the potential to inform urbanization policy formulation by providing knowledge about the spatial distribution of PM2.5 pollution during global urbanization. Addressing the issue of PM2.5 pollution is critical in achieving SDG 11.6 and promoting sustainable and coordinated development in cities worldwide.

Corrigendum: Genetic diversity assessment of Hopea hainanensis in Hainan Island.

[This corrects the article DOI: 10.3389/fpls.2022.1075102.].

Amphioxus CaVPT and creatine kinase are crucial immune-related molecules in response to bacterial infection and immunization.

Although a great progress has been made, our understanding of innate immunity is incomplete. Here, we hypothesize that the innate immune response to pathogens is attributed into a group of functional proteins. The group contains information on host status post bacterial entry (infection or immunity) and bacterial species (Gram-positive or Gram-negative bacteria). Investigation of the group of proteins may result in disclosing of biomarkers identifying the status and species. For this regard, differential proteomics approach coupled with the pattern recognition methods are used to identify biomarkers from the proteins that being specifically regulated during the innate immune response of amphioxus to Gram-positive and Gram-negative bacteria with live or dead status. Four proteins, Calcium vector protein (CaVP), sarcoplasmic calcium-binding protein (SCP), CaVP-target protein (CaVPT) and creatine kinase (CK), are selected as the key biomarkers. Since immunoprotection of CaVP and SCP has been reported, the role of CaVPT and CK are further investigated. Gut CaVPT appears in dying amphioxus, whereas humoral fluid CK downregulates and gut CK keep no change in animals with immunity. The responses are stronger in Gram-negative than Gram-positive bacteria. These results indicate that CaVPT, CK, CaVP and SCP are the most important biomarkers to uncover amphioxus innate immunity to bacteria, and the approach is an efficient way to identify key biomarkers.

Inhibition of autophagy-potentiated chemosensitivity to cisplatin in laryngeal cancer Hep-2 cells.

OBJECTIVE: The purposes of this study were to determine whether autophagy was involved in cisplatin (CDDP) resistance and to investigate the role of the autophagy in the regulation of chemosensitivity to CDDP in laryngeal cancer Hep-2 cells. METHODS: A WST-1 assay was performed to determine cell viability and cell proliferation. Autophagy activation and proapoptotic effects were characterized using monodansylcadaverine labeling and Hoechest staining, respectively. Western blot analysis was used to detect the expression of apoptotic and autophagy-related genes. Flow cytometry was used to assess cell apoptosis ratio. RESULTS: Exposure to CDDP induced the aggregation of autophagosomes in the cytoplasms of Hep-2 cells and up-regulated the expression of Beclin 1 and LC3II. However, CDDP treatment could not lead to obvious inhibition of cell proliferation, which implies that the autophagy may protect CDDP-treated cells from undergoing cell death. Meanwhile, the WST-1 assay indicated that knockdown of the autophagic gene Beclin 1 sensitized Hep-2 cells to CDDP. Furthermore, CDDP-mediated apoptotic cell death was further potentiated by pretreatment with autophagy inhibitor 3-methyladenine or small interfering RNA against Beclin 1. For the definite mechanism of Beclin 1-enhancing chemosensitivity to CDDP, we found that Beclin1 augmented CDDP-induced apoptotic signaling via enhancing caspase-9 and caspase-3 activity but not caspase-8. CONCLUSION: Our results suggest that functional autophagy in response to CDDP may lead to cell survival in Hep-2 cells, whereas defective autophagy may contribute to CDDP-induced apoptosis in Hep-2 cells. Thus, modulators of autophagy may be used beneficially as adjunctive therapeutic agents during the treatment of laryngeal cancer with CDDP therapy.

Promising natural lysine specific demethylase 1 inhibitors for cancer treatment: advances and outlooks.

Lysine specific demethylase 1 (LSD1), a transcriptional corepressor or coactivator that serves as a demethylase of histone 3 lysine 4 and 9, has become a potential therapeutic target for cancer therapy. LSD1 mediates many cellular signaling pathways and regulates cancer cell proliferation, invasion, migration, and differentiation. Recent research has focused on the exploration of its pharmacological inhibitors. Natural products are a major source of compounds with abundant scaffold diversity and structural complexity, which have made a major contribution to drug discovery, particularly anticancer agents. In this review, we briefly highlight recent advances in natural LSD1 inhibitors over the past decade. We present a comprehensive review on their discovery and identification process, natural plant sources, chemical structures, anticancer effects, and structure-activity relationships, and finally provide our perspective on the development of novel natural LSD1 inhibitors for cancer therapy.

Genetic diversity assessment of Hopea hainanensis in Hainan Island.

Hopea hainanensis (Dipterocarpaceae) is an endangered tree species restricted to Hainan Island, China, and a small part of Northern Vietnam. On Hainan Island, it is an important indicator species for tropical forests. The wood of Hopea hainanensis has a very high utilization value in nature since it is compact in structure, hard in texture, not easily deformed after drying, durable, and resistant to sunlight and water. As a result of its high quality, it has been felled and mined by humans without restraint, resulting in a reduction of its population size, severe habitat fragmentation, and a sharp decline in its population. Therefore, its conservation biology needs to be researched urgently. Researchers are currently focusing on the ecological factors and seed germination in the habitat of Hopea hainanensis to determine its endangered status. In the literature, there are no systematic analyses of the endangered mechanism of Hopea hainanensis in terms of genetic diversity. It focuses especially on the systematic genetic diversity of Hopea hainanensis in fragmented habitats. Using single nucleotide polymorphism (SNP) and genotyping-by-sequencing (GBS) technology, 42 samples from seven different cohabitation groups were genotyped. The results showed that the average heterozygosity of the six populations of Hopea hainanensis was 19.77%, which indicated that the genetic diversity of Hopea hainanensis was low. Genetic diversity research is essential for rare and endangered plant protection research. We can find a scientific basis for protecting endangered plants on slope bases by analyzing genetic differences and relationships among populations.

Four-year experience with more than 1000 cases of total laparoscopic liver resection in a single center.

BACKGROUND: Laparoscopic liver resection (LLR) has become a safe surgical procedure that needs additional summarization. AIM: To review 4 years of total LLR surgeries, exceeding 1000 cases, which were performed at a single center. METHODS: Patients who underwent LLR at West China Hospital of Sichuan University between January 2015 and December 2018 were identified. Surgical details, including the interventional year, category of liver disease, and malignant liver tumors prognosis, were evaluated. The learning curve for LLR was evaluated using the cumulative sum method. The Kaplan-Meier method was used to perform survival analysis. RESULTS: Ultimately, 1098 patients were identified. Hepatocellular carcinoma (HCC) was the most common disease that led to the need for LLR at the center (n = 462, 42.08%). The average operation time was 216.94 ± 98.51 min. The conversion rate was 1.82% (20/1098). The complication rate was 9.20% (from grade II to V). The 1-year and 3-year overall survival rates of HCC patients were 89.7% and 81.9%, respectively. The learning curve was grouped into two phases for local resection (cases 1-106 and 107-373), three phases for anatomical segmentectomy (cases 1-44, 45-74 and 75-120), and three phases for hemihepatectomy (cases 1-17, 18-48 and 49-88). CONCLUSION: LLR may be considered a first-line surgical intervention for liver resection that can be performed safely for a variety of primary, secondary, and recurrent liver tumors and for benign diseases once technical competence is proficiently attained.

Spatiotemporal Variation of Urban Plant Diversity and above Ground Biomass in Haikou, China.

Understanding the drivers of urban plant diversity (UPD) and above ground biomass (AGB) in urbanized areas is critical for urban ecosystem services and biodiversity protection. The relationships between UPD and AGB have been investigated simultaneously. However, the drivers of UPD and AGB have been explored independently in tropical coastal areas at different time points. To fill this gap, we conducted a remote sensing interpretation, field plant plot surveys, and compiled socioeconomic and urban greening management survey data. We conducted spatial analyses to investigate the relationships among UPD and socioeconomic variables across different primary and secondary urban functional units (UFUs) in the tropical urban ecosystems of the coastal city of Haikou, China. The primary UFUs with the highest AGB were the recreation and leisure districts in 2015 and 2021. In 2015, AGB was mainly correlated with the number of herb species in undeveloped land and the districts of industry, business, recreation, and leisure. In 2021, AGB was affected primarily by the frequency of fertilizing, maintenance, and watering. Our study found that the relationship between UPD and AGB varied across time and space in Haikou. The plant diversity and AGB's response to human activities and socioeconomics appear to have a time-lag effect. These results provide new insights in understanding how management decisions affect urban vegetation and could be used to guide future urban green space planning in Haikou.

Distance-dependent metal-enhanced quantum dots fluorescence analysis in solution by capillary electrophoresis and its application to DNA detection.

Here the distance dependence of metal-enhanced quantum dots (QDs) fluorescence in solution is studied systematically by capillary electrophoresis (CE). Complementary DNA oligonucleotides-modified CdSe/ZnS QDs and gold nanoparticles (Au NPs) were connected together in solution by the hybridization of complementary oligonucleotides, and a model system (QD-Au) for the study of metal-enhanced QDs fluorescence was constructed, in which the distance between the QDs and Au NPs was controlled by adjusting the base number of the oligonucleotide. In our CE experiments, the metal-enhanced fluorescence of the QDs solution was only observed when the distance between the QDs and Au NPs ranged from 6.8 to 18.7 nm, and the maximum enhancement by a factor of 2.3 was achieved at 11.9 nm. Furthermore, a minimum of 19.6 pg of target DNA was identified in CE based on its specific competition with the QD-DNA in the QD-Au system. This work provides an important reference for future study of metal-enhanced QDs fluorescence in solution and exhibits potential capability in nucleic acid hybridization analysis and high-sensitivity DNA detection.

[Selection of surgical modalities for T3 glottic carcinoma].

OBJECTIVE: To determine the optimal surgical modality for T3 glottic carcinoma. METHODS: Clinical data of 57 cases of T3 glottic carcinoma were retrospectively reviewed. Their clinical characteristics, surgical procedures and prognosis were analyzed. At different ages and by surgical procedures performed, the 3-year disease-free survival rate of the patients were analyzed. RESULTS: All cases underwent surgical procedures including total laryngectomy, near total laryngectomy and partial laryngectomy, and the 3-year disease-free survival rate was 63.2% (36/57). The 3-year disease-free survival rate of patients who received total laryngectomy was 66.7% (16/24), near total laryngectomy 50.0% (4/8), and partial laryngectomy 64.0% (16/25, P = 0.694). The 3-year survival rate of the cases ≥ 70.0 years old was 70.0% (7/10), and that of < 70 years old was 61.7% (29/47, P = 0.621). Thirty-six cases had neck dissection, including 2 cases with radical neck dissection, 6 cases with modified neck dissection, and 28 cases with selective neck dissection. The lymph node metastasis rate of all cases was 17.5%. Ten cases were diagnosed as postoperative local recurrence, including 1 cases treated with total laryngectomy, 2 cases treated with near total laryngectomy and 7 cases treated with partial laryngectomy. CONCLUSIONS: Both total laryngectomy and partial laryngectomy are important surgical procedures for treating patients with T3 glottic carcinoma. The optimal individual surgical procedure for the patient with T3 glottic carcinoma should be determined on the basis of the local lesions and physical status. Total laryngectomy is prior to partial laryngectomy for the patients with T3 glottic carcinoma ≥ 70 years old.

Field-amplified sample injection for the determination of albumin and transferrin in human urines by MEKC.

An MEKC method combining field-amplified sample injection (FASI) has been developed for the analysis of albumin (Alb) and transferrin (TRF) in human urines. Various parameters affecting FASI preconcentration were optimized systematically. Under the optimum conditions, using electrokinetic sample injection for 90 s, the sensitivity was improved 77.6- and 45.3-fold for TRF and Alb, respectively, without loss of separation efficiency when compared with hydrodynamical sample injection for 5 s (0.5 psi). The detection limits of TRF and Alb were found to be 0.31 and 0.14 mg/L, respectively. The proposed method has been successfully applied to the determination of TRF and Alb in desalted urines from patients with kidney diseases, and the recoveries of TRF and Alb were 91.1-101.7%.

Calix[4]arene crown-4 ether modified glassy carbon electrode for electrochemical determination of norepinephrine.

A sensitive and selective electrochemical method for the determination of norepinephrine (NE) was developed by using a calix[4]arene crown-4 ether (CACE) film modified glassy carbon electrode (GCE). The fabrication of the CACE film and its electrocatalytic effect for electrochemical oxidation of NE were investigated by electrochemical impedance spectroscopy (EIS) and voltammetric methods. It was found that the electrochemical behavior of NE on the CACE film modified electrode depended on the film thickness of CACE. Based on the electrochemical data, the CACE film surface coverage (theta), the charge transfer number (z) and the amount of the surface-bound NE (Gamma) were calculated. The effects of the experimental variables such as the solution pH and the applied potential were investigated for optimum analytical performance. Excellent linear relationships were obtained in two concentration ranges of NE: one was from 0.55 to 9.7 microM and the other was 9.7 to 230 microM. The detection limit (S/N = 3) of NE obtained by steady-state amperometry was 0.28 microM. The relative standard deviation (RSD) for 10 successive measurements of 0.05 mM NE was 2.2%. This method could be applied for the direct determination of NE in injection samples. In addition, effects of possible interferences (e.g. ascorbic acid, AA) were investigated. The present work provides an effective method for amperometric determination of NE by using a CACE modified electrode.

An amperometric hydrogen peroxide biosensor based on immobilization of horseradish peroxidase on an electrode modified with magnetic dextran microspheres.

A new kind of magnetic dextran microsphere (MDMS) with uniform shape and narrow diameter distribution has been prepared from magnetic iron nanoparticles and dextran. Horseradish peroxidase (HRP) was successfully immobilized on the surface of an MDMS-modified glassy-carbon electrode (GCE), and the immobilized HRP displayed excellent electrocatalytic activity in the reduction of H(2)O(2) in the presence of the mediator hydroquinone (HQ). The effects of experimental variables such as the concentration of HQ, solution pH, and the working potential were investigated for optimum analytical performance. This biosensor had a fast response to H(2)O(2) of less than 10 s and an excellent linear relationship was obtained in the concentration range 0.20 micromol L(-1)-0.68 mmol L(-1), with a detection limit of 0.078 micromol L(-1) (S/N = 3) under the optimum conditions. The response showed Michaelis-Menten behavior at larger H(2)O(2) concentrations, and the apparent Michaelis-Menten constant K(M)(app) was estimated to be 1.38 mmol L(-1). Moreover, the selectivity, stability, and reproducibility of the biosensor were evaluated, with satisfactory results.