Impact of European Union label changes to avoid inadvertent use of medicinal products containing methotrexate for once-weekly administration: A survey amongst prescribers, pharmacists and patients on awareness, knowledge, and behaviour.

PURPOSE: To determine the effectiveness of risk minimisation measures (RMM) to avoid inadvertent daily instead of weekly methotrexate (MTX) use, introduced by the European Medicines Agency (EMA) from 2019 onwards. METHODS: Using a cross-sectional online survey in France, Greece, Germany, Poland, and Sweden in 2022, we assessed awareness, knowledge, and self-declared behaviour for respondents who prescribed, dispensed, or used once-weekly MTX in the last 3 months. Respondents' answers were considered as 'successful' with regards to RMM effectiveness (vs. unsuccessful) if they provided correct ('desirable') responses to 100% of questions regarding awareness and self-declared behaviour and correct responses to ≥80% of questions about knowledge. Per target population, an outcome was considered successful if achieved by ≥80% of respondents. Effectiveness of RMM was defined by ≥80% being successful on all outcomes. RESULTS: One-hundred-fifty-one prescribers, 150 pharmacists, and 150 patients completed the survey. Success rates were 56% (95% CI 48.0%-64.3%) for awareness, 42% (95% CI 34.4%-50.7%) for knowledge, and 31% (95% CI 23.8%-39.2%) for self-declared behaviour among prescribers, 18% (95% CI 12.8%-25.8%) for awareness, 7% (95% CI 3.7%-12.7%) for knowledge, and 50% (95% CI 41.7%-58.3%) for self-declared behaviour among pharmacists, and 29% (95% CI 21.6%-36.6%) for awareness, and 3% (95% CI 1.1%-7.6%) for knowledge among patients. Overall success was not attained by any target population. CONCLUSIONS: RMM were evaluated as not effective across outcomes of awareness, knowledge, and self-declared behaviour in prescribers, pharmacists, and patients. Findings suggested we need continued efforts for RMM across all target populations and across all outcomes.

Method validation, residue behaviour and dietary risk assessment of insecticides (cyantraniliprole, acetamiprid, flubendiamide and its metabolite, des-iodo flubendiamide) in or on broccoli using LC-MS/MS.

Residue behaviour and dietary risk assessment of cyantraniliprole, flubendiamide and acetamiprid in broccoli were carried out using the QuEChERS (quick, easy, cheap, effective, rugged and safe) technique coupled with LC-MS/MS. The QuEChERS technique was validated on parameters such as linearity, accuracy, precision, robustness, matrix effects, limit of quantification (LOQ), specificity, retention time and ion ratio as per SANTE (Directorate General for Health and Food Safety) guidelines to attest to the specificity, accuracy and precision of the analytical method in estimating insecticide residues in and on broccoli heads and cropped soil. The LOQ of the method for all three insecticides was 0.01 mg/kg. The initial deposits of cyantraniliprole, flubendiamide and acetamiprid reduced to half of its concentration in 1.873-2.354, 1.975-2.484 and 1.371-1.620 days, respectively. No residues were detected in broccoli-cropped soil at harvest time (30 days after last spray). The proposed maximum residue limits (MRLs) of 1.5, 0.5-0.9 and 2.0-3 mg/kg for cyantraniliprole, flubendiamide and acetamiprid were calculated using the Organisation for Economic Co-operation and Development MRL calculator. The acute and chronic dietary risk assessment of the tested insecticides identified no appreciable dietary risk to the Indian population from the consumption of broccoli heads. The findings of no dietary risk highlight the importance of informed pesticide usage in broccoli and the proposed MRL derived from this study offers crucial guidelines for the regulatory authorities, ensuring the safety of broccoli consumption.

Dissipation kinetics and the evaluation of dietary risks associated with deltamethrin, ethion, fenazaquin, and fenpropathrin on bell pepper (Solanum annuum L.).

Bell peppers, a globally significant crop, face infestations from various pests. In a study, bell peppers were treated with deltamethrin, ethion, fenazaquin, and fenpropathrin at recommended and double the doses, repeated twice with a 10-day interval. The QuEChERS method underwent validation for linearity, matrix match, accuracy, and precision in bell pepper matrices for residue analysis. The limit of detection for the tested pesticides on bell peppers was 0.01 mg/L, with a quantification limit of 0.05 mg/L. Recovery studies showed a range of 94.80% to 102.80%. Initial deposits of deltamethrin, ethion, fenazaquin, and fenpropathrin on bell peppers at recommended doses were 0.371, 1.237, 0.617, and 0.640 mg/L, respectively, and at double doses were 0.712, 1.945, 1.221, and 1.189 mg/L, respectively. Safe waiting periods of 10, 11, 10, and 8 days were suggested for deltamethrin, ethion, fenazaquin, and fenpropathrin, respectively. The corresponding half-lives for the pesticides were 1.96, 1.79, 2.06, and 1.69 days, all following first-order dissipation kinetics. Dietary risk assessment indicated Hazard Quotients (HQ) below 1 and Theoretical Maximum Daily Intake (TMDI) below Acceptable Daily Intake (ADI) and Maximum Permissible Intake (MPI) levels. Therefore, at their recommended doses, the pesticides were deemed safe for bell pepper cultivation.

IL-1α Mediates Innate and Acquired Resistance to Immunotherapy in Melanoma.

Inflammation has long been associated with cancer initiation and progression; however, how inflammation causes immune suppression in the tumor microenvironment and resistance to immunotherapy is not well understood. In this study, we show that both innate proinflammatory cytokine IL-1α and immunotherapy-induced IL-1α make melanoma resistant to immunotherapy. In a mouse melanoma model, we found that tumor size was inversely correlated with response to immunotherapy. Large tumors had higher levels of IL-1α, Th2 cytokines, polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs), and regulatory T cells but lower levels of IL-12, Th1 cytokines, and activated T cells. We found that therapy with adenovirus-encoded CD40L (rAd.CD40L) increased tumor levels of IL-1α and PMN-MDSCs. Blocking the IL-1 signaling pathway significantly decreased rAd.CD40L-induced PMN-MDSCs and their associated PD-L1 expression in the tumor microenvironment and enhanced tumor-specific immunity. Similarly, blocking the IL-1 signaling pathway improved the antimelanoma activity of anti-PD-L1 Ab therapy. Our study suggests that blocking the IL-1α signaling pathway may increase the efficacy of immunotherapies against melanoma.

Residue behaviour and health risk assessment of chlorpyrifos and mancozeb in apple fruits and soil.

Mancozeb residue estimation was done using second derivative ultraviolet spectroscopy by Shimadzu ultraviolet-visible spectrophotometer, and chlorpyrifos was estimated by QuEChERS technique using GC-FPD. The persistence for chlorpyrifos was carried out at two locations, and for mancozeb, persistence studies were carried out at four locations. Initial deposits of mancozeb on apple fruits ranged from 1.33 to 1.63 mg/kg at the recommended dose and from 2.55 to 3.26 mg/kg at double the recommended dose at all four locations. Chlorpyrifos residues in apple fruits had an initial deposit of 0.94-0.99 mg/kg at recommended dose and 1.75-1.92 mg/kg at double the recommended dose. Mancozeb residues in apple fruit were below the detection limit (BDL) after 20 days at recommended dose and after 25 days at double the recommended dose at two locations, while mancozeb residue at the other two locations and the residues of chlorpyrifos at all locations reached BDL after 15 and 20 days at recommended and double the recommended doses, respectively. Half-life of mancozeb varied from 3.07 to 4.02 days at recommended dose and from 3.30 to 4.32 days at double the recommended dose, whereas chlorpyrifos residues dissipated to half their initial concentration on 2.33-2.35 days at recommended dose and 2.89-2.90 days at double the recommended dose. The soil samples showed no presence of residues of chlorpyrifos and mancozeb at harvest. The risk assessment revealed that hazard quotient for the intake of mancozeb was in the range of 0.06-0.13% and 0.20-0.44% for rural and urban population, while for the intake of chlorpyrifos, hazard quotient was in the range of 0.10-0.12% for rural population and 0.33-0.38% for urban population, and theoretical maximum dietary intake (9.67 × 10-5 mg/person and 3.18 × 10-4 mg/person for rural population and urban population in case of mancozeb and 3.22 × 10-5 mg/person and 1.06 × 10-4 mg/person for rural population and urban population in case of chlorpyrifos) was also found to be less than maximum permissible intake (1.38 mg/kg for mancozeb and 0.60 mg/kg for chlorpyrifos). The results of risk assessment thereby indicated that apple consumption does not pose a risk to human health.

Dissipation pattern and dietary risk assessment of some commonly used insecticides on tomato (Solanum lycopersicum L.).

Pesticide residue in food commodities is a serious concern in relation to consumer safety and also the most significant barrier in the trade of food commodities. The dissipation pattern of four insecticides, namely novaluron, λ-cyhalothrin, imidacloprid, and fenazaquin, was evaluated on tomato fruits and cropped soil. The residues were extracted using the QuEChERS analytical method and quantized using a gas chromatograph with electron capture detector, gas chromatograph mass spectrometer, and high-performance liquid chromatography with photo diode array detector. The analytical method was validated using parameters like recovery, linearity, accuracy, matrix effect, and specificity, with limit of detection and limit of quantitation established to be 0.01 and 0.05 mg/kg, respectively, for all the pesticides. The average initial deposits (samples collected after 2 h of application) at the recommended dose of novaluron, λ-cyhalothrin, imidacloprid, and fenazaquin were 0.593, 0.293, 0.227, and 0.431 mg/kg on tomato fruits, respectively, and were below the limit of quantification in soil. The pre-harvest interval of 17, 8, 1, and 13 days was suggested for novaluron, λ-cyhalothrin, imidacloprid, and fenazaquin on tomato, respectively. Risk assessment studies revealed that all pesticides under study are safe and do not pose any threat to humans as theoretical maximum dietary intake is less than the maximum permissible intake and acceptable daily intake.

Anti-Depressant Properties of Crocin Molecules in Saffron.

Saffron is a valued herb, obtained from the stigmas of the C. sativus Linn (Iridaceae), with therapeutic effects. It has been described in pharmacopoeias to be variously acting, including as an anti-depressant, anti-carcinogen, and stimulant agent. The therapeutic effects of saffron are harbored in its bioactive molecules, notably crocins, the subject of this paper. Crocins have been demonstrated to act as a monoamine oxidase type A and B inhibitor. Furthermore, saffron petal extracts have experimentally been shown to impact contractile response in electrical field stimulation. Other research suggests that saffron also inhibits the reuptake of monoamines, exhibits N-methyl-d-aspartate antagonism, and improves brain-derived neurotrophic factor signaling. A host of experimental studies found saffron/crocin to be similarly effective as fluoxetine and imipramine in the treatment of depression disorders. Saffron and crocins propose a natural solution to combat depressive disorders. However, some hurdles, such as stability and delivery, need to be overcome.

Dissipation kinetics and dietary risk assessment of spiromesifen on major summer vegetables using good agricultural practices.

Dissipation kinetics and dietary risk assessment of spiromesifen is worked out on four summer vegetables, viz. okra, chilli, capsicum and brinjal (eggplant or aubergine) during March-April 2015 at the experimental farm of the Department of Entomology, Dr. Yashwant Singh Parmar University of Horticulture and Forestry Nauni, Solan using good agricultutral practices. Two foliar applications of spiromesifen @ 144.0 g.a.i./ha each were given at 10 days interval with a knapsack sprayer with the first application at the fruit initiation stage. Sample were collected up to 15 days after pesticide application and processed using a modified QuEChERS method, which was validated by doing recovery studies having recovery range and RSD within established guidelines of SANCO. Estimation of spiromesifen residues was conducted on GC-MS. The initial deposits after spraying of spiromesifen on okra, capsicum, chilli and brinjal fruit after 2 h of treatment were 1.327, 0.727, 0.800 and 0.738 mg/kg, respectively. The residues persisted up to 7 days and further dissipated and declined below the limit of quantification of <0.025 mg/kg at 10 days after treatment in all of the crops under investigation. Dissipation of spiromesifen followed first-order kinetics. The spiromesifen residues dissipated to half in 1.6, 1.8, 1.9 and 1.7 days with the suggested safe waiting period of 8.9, 5.2, 6.0 and 7.0 in the respective crops. The hazard quotient was <1 and theoretical maximum dietary intake was less than the maximum permissible intake, which was less than the maximum residue limit in all of the vegetable crops under investigation.

A unique PE_PGRS protein inhibiting host cell cytosolic defenses and sustaining full virulence of Mycobacterium marinum in multiple hosts.

Despite intense research, PE_PGRS proteins still represent an intriguing aspect of mycobacterial pathogenesis. These cell surface proteins influence virulence in several pathogenic species, but their diverse and exact functions remain unclear. Herein, we focussed on a PE_PGRS member from Mycobacterium marinum, MMAR_0242, characterized by an extended and unique C-terminal domain. We demonstrate that an M. marinum mutant carrying a transposon insertion in MMAR_0242 is highly impaired in its ability to replicate in macrophages and amoebae, because of its inability to inhibit lysosomal fusion. As a consequence, this mutant failed to survive intracellularly as evidenced by a reduced number of cytosolic actin tail-forming bacteria and by quantitative electron microscopy, which mainly localized MMAR_0242::Tn within membrane-defined vacuoles. Functional complementation studies indicated that the C-terminus, but not the N-terminal PE_PGRS domain, is required for intracellular growth/survival. In line with these findings, disruption of MMAR_0242 resulted in a highly attenuated virulence phenotype in zebrafish embryos, characterized by restricted bacterial loads and a failure to produce granulomas. Furthermore, expression of MMAR_0242 in Mycobacterium smegmatis, a non-pathogenic species naturally deficient in PE_PGRS production, resulted in increased survival in amoebae with enhanced cytotoxic cell death and increased survival in infected mice with splenomegaly. Overall, these results indicate that MMAR_0242 is required for full virulence of M. marinum and sufficient to confer pathogenic properties to M. smegmatis.

Panacea to lost gingival tissue architecture and spacing: Silicone gingival prosthesis.

The positive psychological effects of improving patients' smile often contribute to an improved self-image and enhanced self-esteem. Periodontal disease may lead to tooth and tissue loss that can result in esthetic problems. Combined periodontal/prosthodontic treatment for patients with advanced disease is well documented. This case report illustrates a method of treatment for an advanced tissue loss in an esthetic area using a removable silicone gingival prosthesis/mask.

Environmental risk of heavy metal pollution and contamination sources using multivariate analysis in the soils of Varanasi environs, India.

This study assessed soil pollution in the Varanasi environs of Uttar Pradesh in India. Assessing the concentration of potentially harmful heavy metals in the soils is imperative in order to evaluate the potential risks to human. To identify the concentration and sources of heavy metals and assess the soil environmental quality, 23 samples were collected from different locations covering dumping, road and agricultural area. The average concentrations of the heavy metals were all below the permissible limits according to soil quality guidelines except Cu (copper) and Pb (lead) in dumping and road soils. Soil heavy metal contamination was assessed on the basis of geoaccumulation index (Igeo), pollution index (PI) and integrated pollution index (IPI). The IPI of the metals ranged from 0.59 to 9.94, with the highest IPI observed in the dumping and road soils. A very significant correlation was found between Pb and Cu. The result of principal component analysis suggested that PC1 was mainly affected by the use of agrochemicals, PC2 was affected by vehicular emission and PC3 was affected by dumping waste. Meanwhile, PC4 was mainly controlled by parent material along with anthropogenic activities. Appropriate measures should be taken to minimize the heavy metal levels in soils and thus protect human health.

Perspective on utilization of Bacillus species as plant probiotics for different crops in adverse conditions.

Plant probiotic bacteria are a versatile group of bacteria isolated from different environmental sources to improve plant productivity and immunity. The potential of plant probiotic-based formulations is successfully seen as growth enhancement in economically important plants. For instance, endophytic Bacillus species acted as plant growth-promoting bacteria, influenced crops such as cowpea and lady's finger, and increased phytochemicals in crops such as high antioxidant content in tomato fruits. The present review aims to summarize the studies of Bacillus species retaining probiotic properties and compare them with the conventional fertilizers on the market. Plant probiotics aim to take over the world since it is the time to rejuvenate and restore the soil and achieve sustainable development goals for the future. Comprehensive coverage of all the Bacillus species used to maintain plant health, promote plant growth, and fight against pathogens is crucial for establishing sustainable agriculture to face global change. Additionally, it will give the latest insight into this multifunctional agent with a detailed biocontrol mechanism and explore the antagonistic effects of Bacillus species in different crops.

Applying innovative technological interventions in the preservation and packaging of fresh seafood products to minimize spoilage - A systematic review and meta-analysis.

Seafood, being highly perishable, faces rapid deterioration in freshness, posing spoilage risks and potential health concerns without proper preservation. To combat this, various innovative preservation and packaging technologies have emerged. This review delves into these cutting-edge interventions designed to minimize spoilage and effectively prolong the shelf life of fresh seafood products. Techniques like High-Pressure Processing (HPP), Modified Atmosphere Packaging (MAP), bio-preservation, and active and vacuum packaging have demonstrated the capability to extend the shelf life of seafood products by up to 50%. However, the efficacy of these technologies relies on factors such as the specific type of seafood product and the storage temperature. Hence, careful consideration of these factors is essential in choosing an appropriate preservation and packaging technology.

Residue estimation and dietary risk assessment of fenvalerate, novaluron, and profenofos in bell pepper grown under protected and open field conditions.

Residue studies were conducted in bell pepper crops (green and yellow bell pepper) to ensure the safe use of fenvalerate, profenofos, and novaluron (under open field and protected conditions) in randomized block design (RBD) following three applications at a 10-day interval over two consecutive years, 2021 and 2022. A robust analytical method was developed using quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction and gas chromatography-tandem mass spectrometry (GC-MS/MS) for the determination of pesticide residues in bell pepper samples. The half-lives for fenvalerate were 2.47-2.87 and 2.50-3.03 days on bell pepper under open field conditions, whereas the corresponding values for bell pepper under protected conditions were 3.84-4.58 and 4.17-4.71 days, during 2021 and 2022, respectively. Profenofos displayed half-lives of 2.03-2.65 and 2.15-2.77 days in open field conditions and 3.05-3.89 and 3.16-3.78 days in protected conditions during 2021 and 2022, respectively. Similarly, novaluron had half-lives of 2.87-3.49 and 3.24-3.75 days under protected conditions in 2021 and 2022, respectively. The maximum residue limits (MRLs) were calculated to be 0.6 mg/kg for fenvalerate, while for profenofos it was 0.7 mg/kg on bell pepper under open field conditions at double doses, at the proposed pre-harvest interval (PHI) of 3 days. Likewise, for bell peppers grown under protected conditions, MRLs at the PHI of 3 days were determined to be 0.8 mg/kg for fenvalerate, 0.3 mg/kg for novaluron, and 1.5 mg/kg for profenofos. A dietary risk assessment study indicated that the percentage of acute hazard index (% aHI) was significantly lower than 100, and hazard quotient (HQ) values were below 1, signifying no acute or chronic risk to consumers. These findings underscore the safety of consuming bell peppers treated with fenvalerate, profenofos, and novaluron under the protected and open field conditions.

Residue behavior and consumer risk assessment of spirotetramat and chlorpyrifos on cabbage heads and cropped soil.

A field experiment following good agricultural practices was laid out to study the dissipation of spirotetramat (90 g a.i. ha-1 and 180 g a.i. ha-1) and chlorpyrifos (400 g a.i. ha-1 and 800 g a.i. ha-1) on cabbage heads and soil. Samples were processed using quick, easy, cheap, effective, rugged, and safe (QuEChERS) method for residue estimation of spirotetramat and chlorpyrifos, which were further detected using HPLC-PDA and GC-FPD respectively. The residues of spirotetramat on cabbage heads reached below detection limit (BDL) (< 0.05 mg kg-1) on 7th and 10th day and for chlorpyrifos, BDL (< 0.01 mg kg-1) was achieved on 10th and 15th day for X and 2X dose, respectively. On 20th day after second spray, residues in soil were found to be BDL for both the pesticides. Half-life of spirotetramat and chlorpyrifos was found to be 3 and 2 days, respectively while a safe pre-harvest interval (PHI) of 9 days for spirotetramat and 10 days for chlorpyrifos is suggested on cabbage. The dietary risk assessment studies for various age groups of Indian population, ascertained safety of treated cabbage heads for consumption, as current study revealed that hazard quotient (HQ) < 1 and theoretical maximum dietary intake (TMDI) < maximum permissible intake (MPI) for both the pesticides at respective PHI.

Assessing the Global Impact of Ambient Air Pollution on Cancer Incidence and Mortality: A Comprehensive Meta-Analysis.

PURPOSE: This study aims to examine the association between exposure to major ambient air pollutants and the incidence and mortality of lung cancer and some nonlung cancers. METHODS: This meta-analysis used PubMed and EMBASE databases to access published studies that met the eligibility criteria. Primary analysis investigated the association between exposure to air pollutants and cancer incidence and mortality. Study quality was assessed using the Newcastle Ottawa Scale. Meta-analysis was conducted using R software. RESULTS: The meta-analysis included 61 studies, of which 53 were cohort studies and eight were case-control studies. Particulate matter 2.5 mm or less in diameter (PM2.5) was the exposure pollutant in half (55.5%), and lung cancer was the most frequently studied cancer in 59% of the studies. A pooled analysis of exposure reported in cohort and case-control studies and cancer incidence demonstrated a significant relationship (relative risk [RR], 1.04 [95% CI, 1.02 to 1.05]; I2, 88.93%; P < .05). A significant association was observed between exposure to pollutants such as PM2.5 (RR, 1.08 [95% CI, 1.04 to 1.12]; I2, 68.52%) and nitrogen dioxide (NO2) (RR, 1.03 [95% CI, 1.01 to 1.05]; I2, 73.52%) and lung cancer incidence. The relationship between exposure to the air pollutants and cancer mortality demonstrated a significant relationship (RR, 1.08 [95% CI, 1.07 to 1.10]; I2, 94.77%; P < .001). Among the four pollutants, PM2.5 (RR, 1.15 [95% CI, 1.08 to 1.22]; I2, 95.33%) and NO2 (RR, 1.05 [95% CI, 1.02 to 1.08]; I2, 89.98%) were associated with lung cancer mortality. CONCLUSION: The study confirms the association between air pollution exposure and lung cancer incidence and mortality. The meta-analysis results could contribute to community cancer prevention and diagnosis and help inform stakeholders and policymakers in decision making.

Polystyrene microplastic particles in the food chain: Characteristics and toxicity - A review.

Polystyrene (PS) is a crucial material for modern plastic manufacturers, but its widespread use and direct discard in the environment severely affect the food chain. This review provides a detailed study on the impact of PS microplastics (PS-MPs) on the food chain and the environment, including information on their mechanism, degradation process, and toxicity. The accumulation of PS-MPs in organisms' different organs leads to various adverse reactions, such as reduced body weight, premature deaths, pulmonary diseases, neurotoxicity, transgenerational issues, oxidative stress, metabolic alterations, ecotoxicity, immunotoxicity, and other dysfunctions. These consequences affect diverse elements in the food chain, spanning from aquatic species to mammals and humans. The review also addresses the need for sustainable plastic waste management policies and technological developments to prevent the adverse impacts of PS-MPs on the food chain. Additionally, it emphasizes the importance of developing a precise, flexible, and effective methodology for extracting and quantifying PS-MPs in food, considering their characteristics like particle size, polymer types, and forms. While several studies have focused on the toxicity of polystyrene microplastics (PS-MPs) in aquatic species, further investigation is required to understand the mechanisms by which they are transferred across multiple trophic levels. Therefore, this article serves as the first comprehensive review, examining the mechanism, degradation process, and toxicity of PS-MPs. It presents an analysis of the current research landscape of PS-MPs in the global food chain, providing insights for future researchers and governing organizations to adopt better approaches to managing PS-MPs and preventing their adverse impacts on the food chain. As far as we know this is the first article on this specific and impactant topic.

Influence of vehicles used for oral dosing of test molecules on the progression of Mycobacterium tuberculosis infection in mice.

Preclinical evaluation of drug-like molecules requires their oral administration to experimental animals using suitable vehicles. We studied the effect of oral dosing with corn oil, carboxymethyl cellulose, dimethyl sulfoxide, and polysorbate-80 on the progression of Mycobacterium tuberculosis infection in mice. Infection was monitored by physical (survival time and body weight) and bacteriological (viable counts in lungs) parameters. Compared with water, corn oil significantly improved both sets of parameters, whereas the other vehicles affected only physical parameters.

B Cells Are Required to Generate Optimal Anti-Melanoma Immunity in Response to Checkpoint Blockade.

Immunotherapies such as checkpoint blockade therapies are known to enhance anti-melanoma CD8+ T cell immunity, but only a fraction of patients treated with these therapies achieve durable immune response and disease control. It may be that CD8+ T cells need help from other immune cells to generate effective and long-lasting anti-tumor immunity or that CD8+ T cells alone are insufficient for complete tumor regression and cure. Melanoma contains significant numbers of B cells; however, the role of B cells in anti-melanoma immunity is controversial. In this study, B16 melanoma mouse models were used to determine the role of B cells in anti-melanoma immunity. C57BL/6 mice, B cell knockout (KO) C57BL/6 mice, anti-CD19, and anti-CXCL13 antibody-treated C57BL/6 mice were used to determine treatment efficacy and generation of tumor-specific CD8+ T cells in response to PD-L1 blockade alone or combination with TLR-7/8 activation. Whole transcriptome analysis was performed on the tumors from B cell depleted and WT mice, untreated or treated with anti-PD-L1. Both CD40-positive and CD40-negative B cells were isolated from tumors of TLR-7/8 agonist-treated wild-type mice and adoptively transferred into tumor-bearing B cell KO mice, which were treated with anti-PD-L1 and TLR-7/8 agonist. Therapeutic efficacy was determined in the presence of activated or inactivated B cells. Microarray analysis was performed on TLR-7/8-treated tumors to look for the B cell signatures. We found B cells were required to enhance the therapeutic efficacy of monotherapy with anti-PD-L1 antibody and combination therapy with anti-PD-L1 antibody plus TLR-7/8 agonist. However, B cells were not essential for anti-CTLA-4 antibody activity. Interestingly, CD40-positive but not CD40-negative B cells contributed to anti-melanoma immunity. In addition, melanoma patients' TCGA data showed that the presence of B cell chemokine CXCL13 and B cells together with CD8+ T cells in tumors were strongly associated with improved overall survival. Our transcriptome data suggest that the absence of B cells enhances immune checkpoints expression in the tumors microenvironment. These results revealed the importance of B cells in the generation of effective anti-melanoma immunity in response to PD-1-PD-L1 blockade immunotherapy. Our findings may facilitate the design of more effective anti-melanoma immunotherapy.