RESUMO
We present a rare case of a patient with toluene exposure manifesting as anti-glomerular basement membrane (GBM) disease on a background of phospholipase A2 receptor (PLA2R)-associated membranous nephropathy. A 23-year-old man presented to the emergency department with hypertension, headache, hemoptysis, anemia, acute kidney injury, glomerular hematuria, and proteinuria. He endorsed repeated exposure to toluene-containing products while repairing dirt bikes. Serologies were positive for anti-GBM antibodies. Kidney biopsy showed crescentic glomerulonephritis with linear immunoglobulin G and granular PLA2R staining by immunofluorescence. He was initially treated with high-dose steroids, plasmapheresis, and hemodialysis for pulmonary-renal syndrome followed by oral cyclophosphamide and prednisone, which were discontinued after 3 months when follow-up biopsies confirmed little chance for renal recovery. He remained on dialysis 1 year later. This case exhibits a unique presentation of anti-GBM syndrome and underlying membranous nephropathy following repeated hydrocarbon exposure. Inhaled toxins promote recurrent localized inflammation, unmasking previously hidden epitopes. Early diagnosis and appropriate use of immunosuppressive and extracorporeal therapies are necessary to prevent morbidity and to improve survival in this rare condition.
Assuntos
Doença Antimembrana Basal Glomerular , Glomerulonefrite Membranosa , Humanos , Masculino , Adulto Jovem , Doença Antimembrana Basal Glomerular/induzido quimicamente , Doença Antimembrana Basal Glomerular/complicações , Doença Antimembrana Basal Glomerular/diagnóstico , Autoanticorpos , Ciclofosfamida/uso terapêutico , Glomerulonefrite Membranosa/induzido quimicamente , Glomerulonefrite Membranosa/diagnóstico , Glomerulonefrite Membranosa/tratamento farmacológico , Fosfolipases/uso terapêutico , Poliésteres/uso terapêutico , Receptores da Fosfolipase A2 , Tolueno/uso terapêuticoRESUMO
RATIONALE & OBJECTIVE: Rituximab is the first-choice therapy for patients with primary membranous nephropathy (MN) and nephrotic syndrome. However, approximately 30% of patients are treatment-resistant or become treatment-intolerant with hypersensitivity reactions upon repeated drug exposures. We aimed to assess whether ofatumumab, a fully human second-generation anti-CD20 antibody, could be a valuable alternative to rituximab in this population. STUDY DESIGN: Case series. SETTING & PARTICIPANTS: 7 rituximab-intolerant and 10 rituximab-resistant patients with MN who consented to receive ofatumumab (50-300mg, single intravenous infusion) and were followed at the nephrology unit of Azienda Socio-Sanitaria Territoriale Papa Giovanni XXIII (Bergamo, Italy) between September 2015 and January 2019. FINDINGS: Over a median (IQR) follow-up of 5.0 (3.0-9.8) months, all 7 rituximab-intolerant and 3 of the 10 rituximab-resistant patients exhibited complete (proteinuria<0.3g/d) or partial (proteinuria<3.5g/d with≥50% reduction vs baseline) remission of nephrotic syndrome. Circulating B cells were similarly depleted in all patients by 1 week, and serum anti-phospholipase A2 receptor antibody concentrations decreased to<2.7 relative units/mL in 3 of 4 rituximab-intolerant and 4 of 8 rituximab-resistant patients with phospholipase A2 receptor-related disease. Ofatumumab significantly reduced 24-hour urinary protein and immunoglobulin G excretion and increased serum albumin and immunoglobulin G levels. These effects were greater in rituximab-intolerant than in rituximab-resistant patients. Measured glomerular filtration rate significantly increased by an average of 13.4% at 24 months compared with baseline (P=0.036) among all patients in the series. There were 14 nonserious infusion-related adverse events in 9 patients that recovered with temporary infusion interruption. LIMITATIONS: Retrospective design, limited number of patients. CONCLUSIONS: Ofatumumab may represent an effective and safe treatment for rituximab-intolerant cases of MN. Larger prospective studies will be needed to validate these preliminary findings and explore the effectiveness of other second-generation anti-CD20 antibodies in this clinical setting. PLAIN-LANGUAGE SUMMARY: Primary membranous nephropathy (MN) is one of the most frequent causes of nephrotic syndrome (NS) in adults. In this case series, we explored the efficacy of ofatumumab, a fully human second-generation anti-CD20 antibody, in 17 patients with MN and NS who were intolerant or unresponsive to rituximab. All 7 rituximab-intolerant patients exhibited complete or partial clinical remission, compared with only 3 of the 10 rituximab-resistant patients. Autoantibody levels decreased in all patients with phospholipase A2 receptor-related disease. Ofatumumab achieved a significant reduction in urinary protein and immunoglobulin G excretion while increasing serum albumin and immunoglobulin G levels. Ofatumumab may be a promising option for patients with MN who are rituximab-intolerant. Further investigations are warranted to validate these preliminary findings.
Assuntos
Glomerulonefrite Membranosa , Síndrome Nefrótica , Adulto , Humanos , Rituximab/uso terapêutico , Síndrome Nefrótica/tratamento farmacológico , Estudos Retrospectivos , Estudos Prospectivos , Anticorpos Monoclonais Humanizados/uso terapêutico , Imunoglobulina G , Proteinúria/tratamento farmacológico , Albumina Sérica/uso terapêutico , Fosfolipases/uso terapêutico , Imunossupressores/uso terapêutico , Receptores da Fosfolipase A2RESUMO
BACKGROUND: Choosing the appropriate antipsychotic drug (APD) treatment for patients with schizophrenia (SCZ) can be challenging, as the treatment response to APD is highly variable and difficult to predict due to the lack of effective biomarkers. Previous studies have indicated the association between treatment response and genetic and epigenetic factors, but no effective biomarkers have been identified. Hence, further research is imperative to enhance precision medicine in SCZ treatment. METHODS: Participants with SCZ were recruited from two randomized trials. The discovery cohort was recruited from the CAPOC trial (n = 2307) involved 6 weeks of treatment and equally randomized the participants to the Olanzapine, Risperidone, Quetiapine, Aripiprazole, Ziprasidone, and Haloperidol/Perphenazine (subsequently equally assigned to one or the other) groups. The external validation cohort was recruited from the CAPEC trial (n = 1379), which involved 8 weeks of treatment and equally randomized the participants to the Olanzapine, Risperidone, and Aripiprazole groups. Additionally, healthy controls (n = 275) from the local community were utilized as a genetic/epigenetic reference. The genetic and epigenetic (DNA methylation) risks of SCZ were assessed using the polygenic risk score (PRS) and polymethylation score, respectively. The study also examined the genetic-epigenetic interactions with treatment response through differential methylation analysis, methylation quantitative trait loci, colocalization, and promoter-anchored chromatin interaction. Machine learning was used to develop a prediction model for treatment response, which was evaluated for accuracy and clinical benefit using the area under curve (AUC) for classification, R2 for regression, and decision curve analysis. RESULTS: Six risk genes for SCZ (LINC01795, DDHD2, SBNO1, KCNG2, SEMA7A, and RUFY1) involved in cortical morphology were identified as having a genetic-epigenetic interaction associated with treatment response. The developed and externally validated prediction model, which incorporated clinical information, PRS, genetic risk score (GRS), and proxy methylation level (proxyDNAm), demonstrated positive benefits for a wide range of patients receiving different APDs, regardless of sex [discovery cohort: AUC = 0.874 (95% CI 0.867-0.881), R2 = 0.478; external validation cohort: AUC = 0.851 (95% CI 0.841-0.861), R2 = 0.507]. CONCLUSIONS: This study presents a promising precision medicine approach to evaluate treatment response, which has the potential to aid clinicians in making informed decisions about APD treatment for patients with SCZ. Trial registration Chinese Clinical Trial Registry ( https://www.chictr.org.cn/ ), 18. Aug 2009 retrospectively registered: CAPOC-ChiCTR-RNC-09000521 ( https://www.chictr.org.cn/showproj.aspx?proj=9014 ), CAPEC-ChiCTR-RNC-09000522 ( https://www.chictr.org.cn/showproj.aspx?proj=9013 ).
Assuntos
Antipsicóticos , Esquizofrenia , Humanos , Antipsicóticos/efeitos adversos , Esquizofrenia/tratamento farmacológico , Esquizofrenia/genética , Esquizofrenia/induzido quimicamente , Olanzapina/farmacologia , Olanzapina/uso terapêutico , Risperidona/efeitos adversos , Aripiprazol/farmacologia , Aripiprazol/uso terapêutico , Medicina de Precisão , Multiômica , Benzodiazepinas/efeitos adversos , Ensaios Clínicos Controlados Aleatórios como Assunto , Fosfolipases/uso terapêuticoRESUMO
Vaginal candidiasis is a medical condition characterized by the overgrowth of Candida spp. in the vaginal cavity with complex recurrent pathogenicity as well as tolerance to antifungal therapy and hence is awaiting more safe and effective treatments. This work aimed to assess the potential antifungal activity of galloylquinic acid compounds (GQAs) from Copaifera lucens leaves against vaginal Candida albicans. The antifungal susceptibility test was performed against 20 isolates of multidrug-resistant (MDR) C. albicans using agar diffusion and broth microdilution assays. The results showed that GQAs exhibited strong antagonistic activity against the test isolates, with inhibition zone diameters ranging from 26 to 38 mm and low MICs (1 to 16 µg/mL) as well as minimum fungicidal concentrations (2 to 32 µg/mL). The MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] assay confirmed the safety of GQAs against the Vero cell line, showing a 50% inhibitory concentration (IC50) of 168.17 mg/mL. A marked difference in the growth pattern of the treated and untreated pathogens was also observed, where a concentration-dependent reduction in the growth rate occurred. Moreover, a pronounced fungicidal effect was demonstrated 6 h after treatment with 1× the minimum fungicidal concentration (MFC), as evidenced by time-kill assays, where the number of survivors was decreased a 6-fold. GQAs effectively inhibited and eradicated about 80% of C. albicans biofilm at 6 µg/mL and 32 µg/mL, respectively. Interestingly, GQAs disturbed the fungal membrane integrity, induced cell lysis, and reduced the virulence factors (proteinase and phospholipase) as well as the catalase activity. Moreover, the ergosterol content in the plasma membrane decreased in a concentration-dependent manner. Additionally, the altered mitochondrial membrane potential was associated with an increased release of cytochrome c from mitochondria to the cytosol, suggesting the initiation of early apoptosis in GQA-treated cells. Transcriptional analysis revealed that all test genes encoding virulence traits, including SAP1, PLB1, LIP1, HWP1, and ALS1, were markedly downregulated in GQA-treated cells compared to the control. The in vivo murine model of vaginal candidiasis further confirmed the therapeutic activity of GQAs (4 mg/kg of body weight) against C. albicans. This work comprehensively evaluated the antifungal, antivirulence, and antibiofilm activities of GQAs against C. albicans isolates using in vitro and in vivo models, providing molecular-level insights into the antifungal mechanism of action and experimental evidence that supports the potential use of GQAs for the treatment of vaginal candidiasis. IMPORTANCE Our work presents a new perspective on the potential use of GQAs as safe and highly effective phytochemicals against MDR C. albicans. This microorganism colonizes the human vaginal epithelium, causing vaginal candidiasis, a condition characterized by recurrent pathogenicity and tolerance to traditional antifungal therapy. Based on the results of in vitro tests, our study reports GQAs antifungal modes of action. These compounds exhibited an anticandidal effect by deactivating the fungal hydrolytic enzymes, reducing ergosterol content in the plasma membrane, altering the potential of the mitochondrial membrane, and inducing apoptosis. Additionally, GQAs showed high activity in eradicating the biofilm formed by the fungus via the downregulation of HWP1, ALS, SAP, PLB, and LIP genes, which are constitutively expressed in the biofilm. In an in vivo murine model of vaginal candidiasis, GQAs further demonstrated strong evidence of their effectiveness as an antifungal therapy. In this regard, our findings provide novel insights into the potential therapeutic use of these phytoactive molecules for vaginal candidiasis treatment.
Assuntos
Candidíase Vulvovaginal , Candidíase , Fabaceae , Feminino , Camundongos , Humanos , Animais , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Modelos Animais de Doenças , Citocromos c/farmacologia , Citocromos c/uso terapêutico , Ágar/farmacologia , Ágar/uso terapêutico , Catalase/farmacologia , Catalase/uso terapêutico , Candidíase Vulvovaginal/tratamento farmacológico , Candidíase Vulvovaginal/microbiologia , Candida albicans , Candidíase/tratamento farmacológico , Biofilmes , Testes de Sensibilidade Microbiana , Fatores de Virulência , Ergosterol , Fosfolipases/farmacologia , Fosfolipases/uso terapêutico , Peptídeo Hidrolases/farmacologia , Peptídeo Hidrolases/uso terapêuticoRESUMO
We studied an amorphous solid dispersion of berberine with absorption enhancer sodium caprate (Huang-Gui solid dispersion preparations, HGSD). A therapeutic effect of HGSD was revealed in mice with type 2 diabetes mellitus and palmitate-induced injury to MIN6 ß-cells. HGSD treatment (150 mg/kg) improved glucose metabolism and decreased ß-cell apoptosis in diabetic mice. Furthermore, the effective component of HGSD berberine significantly attenuated the palmitate-induced decrease in MIN6 ß-cells viability and insulin secretion. Moreover, molecular docking analysis and Western blotting showed that berberine decreased cell apoptosis and expression of group VIA phospholipase A2 (iPLA2), p38 mitogen-activated protein kinase (p38 MAPK), and caspase-3. These data suggest that HGSD treatment protected ß-cells via inhibiting the iPLA2/p38 MAPK pathway.
Assuntos
Berberina , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Células Secretoras de Insulina , Ilhotas Pancreáticas , Animais , Apoptose , Berberina/farmacologia , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/tratamento farmacológico , Células Secretoras de Insulina/metabolismo , Camundongos , Simulação de Acoplamento Molecular , Palmitatos/metabolismo , Palmitatos/farmacologia , Palmitatos/uso terapêutico , Fosfolipases/metabolismo , Fosfolipases/farmacologia , Fosfolipases/uso terapêutico , Fosfolipases A2 Independentes de Cálcio/metabolismo , Fosfolipases A2 Independentes de Cálcio/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Pseudomonas aeruginosa is an important nosocomial pathogen with a capacity of resistance to multiple antibiotics and production of various extracellular and cell-associated virulence factors that clearly contribute to its pathogenicity. The objective of this study was to investigate the antibiotic susceptibility, virulence factors, and clonal relationship among clinical isolates of P. aeruginosa. Different clinical specimens from hospitalized patients were investigated for P. aeruginosa. Susceptibility of the isolates was evaluated by disc diffusion and broth microdilution methods, as described by the Clinical and Laboratory Standards Institute (CLSI) guideline. A total of 97 P. aeruginosa isolates were recovered from clinical specimens. The percentage of isolates resistant to antimicrobials was imipenem 25.77%, meropenem 15.46%, gentamicin 16.49%, tobramycin 15.46%, amikacin 16.49%, ciprofloxacin 20.61%, levofloxacin 24.74, ceftazidime 20.61%, piperacillin 15.46%, piperacillin/tazobactam 12.37%, colistin 9.27%, and polymyxin B 11.34%. Of isolates, 87.62% possessed ß-hemolytic activity, 78.35% lecithinase, 59.8% elastase, 37.11% DNase, and 28.86% twitching motility. The frequency of virulence genes in isolates was lasB 82.47%, plcH 82.47%, exoA 58.76%, exoS 56.7%, and pilA 10.3%. ERIC-PCR typing clustered P. aeruginosa isolates to 19 common types (CT1-CT19) containing isolates from different hospitals and 43 single types (ST1-ST43). Colistin and polymyxin B were the most effective agents against the majority of P. aeruginosa isolates, emphasizing the effort to maintain their antibacterial activity as last-line therapy. The frequency of some virulence factors and genes was noticeably high, which is alarming. In addition, more effective strategies and surveillance are necessary to confine and prevent the inter-hospital and/or intra-hospital dissemination of P. aeruginosa between therapeutic centers.
Assuntos
Infecções por Pseudomonas , Pseudomonas aeruginosa , Amicacina/farmacologia , Amicacina/uso terapêutico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Ceftazidima/farmacologia , Ceftazidima/uso terapêutico , Ciprofloxacina/farmacologia , Ciprofloxacina/uso terapêutico , Colistina/farmacologia , Desoxirribonucleases/genética , Desoxirribonucleases/farmacologia , Desoxirribonucleases/uso terapêutico , Farmacorresistência Bacteriana/genética , Genótipo , Gentamicinas/farmacologia , Gentamicinas/uso terapêutico , Humanos , Imipenem/farmacologia , Imipenem/uso terapêutico , Irã (Geográfico) , Levofloxacino/farmacologia , Levofloxacino/uso terapêutico , Meropeném/farmacologia , Meropeném/uso terapêutico , Testes de Sensibilidade Microbiana , Elastase Pancreática/genética , Elastase Pancreática/farmacologia , Elastase Pancreática/uso terapêutico , Fosfolipases/genética , Fosfolipases/farmacologia , Fosfolipases/uso terapêutico , Piperacilina/farmacologia , Piperacilina/uso terapêutico , Combinação Piperacilina e Tazobactam/farmacologia , Combinação Piperacilina e Tazobactam/uso terapêutico , Polimixina B/farmacologia , Infecções por Pseudomonas/microbiologia , Tobramicina/farmacologia , Tobramicina/uso terapêutico , Fatores de Virulência/genéticaRESUMO
Robust biofilms and the complex airway environment with thick sputum, local hypoxia and persistent inflammation induce the intractability of chronic pulmonary infections caused by Pseudomonas aeruginosa (P. aeruginosa). Herein, we proposed a type of antibiotic-adjuvant liposomes (NANO@PS-LPs), co-incorporating azithromycin (AZI), adjuvant (2-nitroimidazole derivative, 6-NIH) and biofilm dispersant (nitric oxide donor, DETA NONOate). NANO@PS-LPs possessing negatively-charged surface and good hydrophilicity could easily penetrate through the sputum layer, then disassembled triggered by overexpressed phospholipase A2 (PLA2) in the microenvironment around biofilms. Nitric oxide produced by DETA NONOate promoted P. aeruginosa biofilms dispersal. 6-NIH was reduced to 2-aminomidazole derivative (6-AIH) under a hypoxic condition, and hence acted as an AZI adjuvant to enhance the antibacterial activity of AZI. It was found that NANO@PS-LPs could significantly eliminate mature P. aeruginosa biofilms, effectively kill dispersed bacteria, inhibit the metabolism of survivors and prevent P. aeruginosa adherence to airway epithelial cells, accordingly restrain recurrent infections. Additionally, NANO@PS-LPs performed a remarkable advantage in killing AZI-resistant P. aeruginosa and removing their biofilms. In summary, NANO@PS-LPs present a potential nano-strategy to treat stubborn pseudomonal pulmonary infections and overcome correlative drug resistance.
Assuntos
Antibacterianos , Azitromicina , Biofilmes , Pseudomonas aeruginosa , Antibacterianos/farmacologia , Azitromicina/farmacologia , Biofilmes/efeitos dos fármacos , Humanos , Hipóxia , Lipopolissacarídeos , Lipossomos/farmacologia , Testes de Sensibilidade Microbiana , Fosfolipases/farmacologia , Fosfolipases/uso terapêutico , Infecções por Pseudomonas , Pseudomonas aeruginosa/efeitos dos fármacosRESUMO
Psoriasis is an inflammatory disorder characterized by keratinocyte hyper-proliferation and Th17-type immune responses. However, the roles of bioactive lipids and the regulation of their biosynthesis in this chronic skin disease are not fully understood. Herein, we show that group IVE cytosolic phospholipase A2 (cPLA2 ε/PLA2G4E) plays a counterregulatory role against psoriatic inflammation by producing the anti-inflammatory lipid N-acylethanolamine (NAE). Lipidomics analysis of mouse skin revealed that NAE species and their precursors (N-acyl-phosphatidylethanolamine and glycerophospho-N-acylethanolamine) were robustly increased in parallel with the ongoing process of imiquimod (IMQ)-induced psoriasis, accompanied by a marked upregulation of cPLA2 ε in epidermal keratinocytes. Genetic deletion of cPLA2 ε exacerbated IMQ-induced ear swelling and psoriatic marker expression, with a dramatic reduction of NAE-related lipids in IMQ-treated, and even normal, skin. Stimulation of cultured human keratinocytes with psoriatic cytokines concomitantly increased PLA2G4E expression and NAE production, and supplementation with NAEs significantly attenuated the cytokine-induced upregulation of the psoriatic marker S100A9. Increased expression of cPLA2 ε was also evident in the epidermis of psoriatic patients. These findings reveal for the first time the in vivo role of cPLA2 ε, which is highly induced in the keratinocytes of the psoriatic skin, promotes the biosynthesis of NAE-related lipids, and contributes to limiting psoriatic inflammation.
Assuntos
Psoríase , Animais , Anti-Inflamatórios/uso terapêutico , Anticorpos , Citocinas/metabolismo , Etanolaminas , Humanos , Imiquimode , Inflamação , Lipídeos/efeitos adversos , Camundongos , Fosfolipases/uso terapêutico , Psoríase/induzido quimicamente , Psoríase/tratamento farmacológicoRESUMO
Background: B cells can contribute to immune-mediated disorders. Targeting CD20 has proved to be efficacious in several B cell-mediated immunopathologies, as illustrated by the use of rituximab, the first anti-CD20 monoclonal antibody (mAb). Following rituximab, second- and third-generation anti-CD20 mAbs have been developed and tried in immune-mediated diseases, including obinutuzumab, ocrelizumab, ofatumumab, ublituximab, and veltuzumab. However, their safety and efficacy has not been systematically reviewed. Objective: To evaluate safety and efficacy of obinutuzumab, ocrelizumab, ofatumumab, ublituximab, and veltuzumab for the treatment of immune-mediated disorders compared to placebo, conventional treatment or other biologics. Methods: The PRISMA checklist guided the reporting of the data. We searched the PubMed database between 4 October 2016 and 22 July 2021 concentrating on immune-mediated disorders. Results: The literature search identified 2220 articles. After screening titles and abstracts against the inclusion and exclusion criteria and assessing full texts, 27 articles were finally included in a narrative synthesis. Conclusions: Obinutuzumab has shown promising results in a case series of patients with phospholipase A2 receptor-associated membranous nephropathy and mixed results in systemic lupus erythematosus. Ocrelizumab has been approved for the use in patients with relapsing-remitting multiple sclerosis and primary progressive multiple sclerosis. Ocrelizumab was also tested in patients with rheumatoid arthritis, demonstrating promising results, and in systemic lupus erythematosus, revealing mixed results; however, in these conditions, its use was associated with increased risk of serious infections. Ofatumumab received approval for treating patients with relapsing-remitting multiple sclerosis. Moreover, ofatumumab showed promising results in patients with anti-neutrophil cytoplasmic antibody-associated vasculitis, rheumatoid arthritis, and systemic lupus erythematosus, as well as mixed results in phospholipase A2 receptor-associated membranous nephropathy. Ublituximab was assessed in relapsing-remitting multiple sclerosis and neuromyelitis optica spectrum disorder, with promising results, however, the included number of patients was too small to conclude. Veltuzumab was tested in patients with immune thrombocytopenia resulting in improved platelet counts. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/, identifier CRD4201913421.
Assuntos
Artrite Reumatoide , Produtos Biológicos , Glomerulonefrite Membranosa , Doenças do Sistema Imunitário , Lúpus Eritematoso Sistêmico , Esclerose Múltipla Recidivante-Remitente , Esclerose Múltipla , Antígenos CD20 , Artrite Reumatoide/tratamento farmacológico , Produtos Biológicos/uso terapêutico , Glomerulonefrite Membranosa/tratamento farmacológico , Humanos , Doenças do Sistema Imunitário/tratamento farmacológico , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Fosfolipases/uso terapêutico , Rituximab/efeitos adversosRESUMO
The apicomplexan parasite Cryptosporidium parvum is an important cause of diarrhea in humans and cattle, and it can persistently infect immunocompromised hosts. No consistently effective parasite-specific pharmaceuticals or immunotherapies for control of cryptosporidiosis are presently available. The innate immune system represents the first line of host defense against a range of infectious agents, including parasitic protozoa. Several types of antimicrobial peptides and proteins, collectively referred to herein as biocides, constitute a major effector component of this system. In the present study, we evaluated lactoferrin, lactoferrin hydrolysate, 5 cationic peptides (lactoferricin B, cathelicidin LL37, indolicidin, ß-defensin 1, ß-defensin 2), lysozyme, and 2 phospholipases (phospholipase A2, and phosphatidylinositol-specific phospholipase C) for anti-cryptosporidial activity. The biocides were evaluated either alone or in combination with 3E2, a monoclonal antibody (MAb) against C. parvum that inhibits sporozoite attachment and invasion. Sporozoite viability and infectivity were used as indices of anti-cryptosporidial activity in vitro. All biocides except lactoferrin had a significant effect on sporozoite viability and infectivity. Lactoferrin hydrolysate and each of the 5 cationic peptides were highly parasiticidal and strongly reduced sporozoite infectivity. While each phospholipase also had parasiticidal activity, it was significantly less than that of lactoferrin hydrolysate and each of the cationic peptides. However, each phospholipase reduced sporozoite infectivity comparably to that observed with lactoferrin hydrolysate and the cationic peptides. Moreover, when 3 of the cationic peptides (cathelicidin LL37, ß-defensin 1, and ß-defensin 2) were individually combined with MAb 3E2, a significantly greater reduction of sporozoite infectivity was observed over that by 3E2 alone. In contrast, reduction of sporozoite infectivity by a combination of either phospholipase with MAb 3E2 was no greater than that by 3E2 alone. These collective observations suggest that cationic peptides and phospholipases neutralize C. parvum by mechanisms that are predominantly either parasiticidal or non-parasiticidal, respectively.
Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Criptosporidiose/tratamento farmacológico , Cryptosporidium parvum/efeitos dos fármacos , Fosfolipases/farmacologia , Animais , Animais Recém-Nascidos , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/farmacologia , Peptídeos Catiônicos Antimicrobianos/uso terapêutico , Células CACO-2 , Bovinos , Cryptosporidium parvum/imunologia , Cryptosporidium parvum/fisiologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Quimioterapia Combinada , Humanos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Fosfolipases/uso terapêutico , Organismos Livres de Patógenos Específicos , Esporozoítos/efeitos dos fármacos , Esporozoítos/fisiologiaRESUMO
Antibacterial, antiparasitidal and antiviral properties have recently been attributed to members of the secreted phospholipases A(2) (sPLA(2)s) superfamily. Seven sPLA(2)s from groups IA, IB, IIA and III, were tested here in different culture conditions for inhibition of the in vitro intraerythrocytic development of Plasmodium falciparum, the causative agent of the most severe form of human malaria. In the presence of human serum, all sPLA(2)s were inhibitory, with three out of seven exhibiting IC(50)<0.1 nM. In all cases, inhibition could be induced by enzymatic pre-treatment of the serum. By contrast, no effect was observed when parasites were grown in a semi-defined medium (AlbuMAX II) devoid of lipoproteins and containing 10 times less phospholipids than the medium with human serum, strongly suggesting that hydrolysis of serum generating toxic lipid by-products, rather than a direct interaction of the sPLA(2) with the infected erythrocyte, is a general feature of the anti-Plasmodium properties of sPLA(2)s. Furthermore, in serum, six out of the seven sPLA(2)s were toxic against both trophozoite and schizont stages of the parasite development, contrasting with the trophozoite-selective bee venom enzyme's toxicity. Deciphering the molecular mechanisms at play in the phenotypic singularity of the bee venom enzyme toxicity might offer new prospects in antimalarial fight.
Assuntos
Antimaláricos/farmacologia , Fosfolipases/farmacologia , Plasmodium falciparum/efeitos dos fármacos , Animais , Antimaláricos/metabolismo , Antimaláricos/uso terapêutico , Meios de Cultura , Humanos , Malária Falciparum/tratamento farmacológico , Testes de Sensibilidade Parasitária , Fosfolipases/metabolismo , Fosfolipases/uso terapêutico , Plasmodium falciparum/crescimento & desenvolvimento , Escorpiões , Soro , Venenos de Serpentes/química , SerpentesRESUMO
Hepatic lipase (HL) is an endothelial-bound lipolytic enzyme which functions as a phospholipase as well as a triacylglycerol hydrolase and is necessary for the metabolism of IDL and HDL. To evaluate the feasibility of replacing an enzyme whose in vivo physiologic function depends on its localization on the vascular endothelium, we have infused recombinant replication-deficient adenovirus vectors expressing either human HL (HL-rAdV; n = 7) or luciferase cDNA (Lucif-rAdV; n = 4) into HL-deficient mice with pretreatment plasma cholesterol, phospholipid, and HDL cholesterol values of 176 +/- 9, 314 +/- 12, and 129 +/- 9, respectively. After infusion of HL-rAdV, HL could be detected in the postheparin plasma of HL-deficient mice by immunoblotting and postheparin plasma HL activities were 25,700 +/- 4,810 and 1,510 +/- 688 nmol/min/ml on days 5 and 15, respectively. Unlike the mouse HL, 97% of the newly synthesized human HL was heparin releasable, indicating that the human enzyme was virtually totally bound to the mouse vascular endothelium. Infusion of HL-rAdV in HL-deficient mice was associated with a 50-80% decrease in total cholesterol, triglyceride, phospholipids, cholesteryl ester, and HDL cholesterol (P < 0.001) as well as normalization of the plasma fast protein liquid chromatography lipoprotein profile by day 8. These studies demonstrate successful expression and delivery of a lipolytic enzyme to the vascular endothelium for ultimate correction of the HL gene defect in HL-deficient mice and indicate that recombinant adenovirus vectors may be useful in the replacement of endothelial-bound lipolytic enzymes in human lipolytic deficiency states.
Assuntos
Endotélio Vascular/enzimologia , Terapia Genética/métodos , Hiperlipidemias/terapia , Lipase/uso terapêutico , Fosfolipases/uso terapêutico , Adenoviridae/genética , Animais , Colesterol/sangue , Humanos , Lipase/sangue , Lipase/deficiência , Lipase/genética , Lipídeos/sangue , Lipoproteínas/sangue , Masculino , Camundongos , Camundongos Mutantes , Fosfolipases/sangue , Fosfolipases/deficiência , Fosfolipases/genética , Proteínas Recombinantes/uso terapêuticoRESUMO
In order to evaluate the efficacy and safety of gabexate mesilate (FOY = [ethyl-4-(6-guanidinohexanoyloxy) benzoate methane sulfonate]), a synthetic inhibitor of various proteases as well as phospholipase A 2, 17 patients with acute pancreatitis were treated with FOY in doses of 3 X 150 mg/day per i.v. perfusor. The clinical course of the disease and the laboratory findings were compared with the results of a control group of 21 patients also suffering from acute pancreatitis. 30% of the patients in the FOY-group and 40% in the control group had different complications during the first week after admission to the hospital. The laboratory findings in the two groups revealed that alpha-amylase levels in serum and urine in the FOY-group were significantly lower than in the control group. Side effects of the drug were not observed. FOY may be considered a promising drug for treatment of acute pancreatitis in view of its few side effects and especially with regard to its mode of action.
Assuntos
Guanidinas/uso terapêutico , Pancreatite/tratamento farmacológico , Fosfolipases A/uso terapêutico , Fosfolipases/uso terapêutico , Inibidores de Proteases/uso terapêutico , Doença Aguda , Adulto , Idoso , Amilases/sangue , Feminino , Gabexato , Humanos , Masculino , Pessoa de Meia-Idade , Pancreatite/enzimologia , Fosfolipases A2RESUMO
The activities of amylase, lipase, phospholipase, and trypsin in four commercially available preparations of pancreatin with different galenic and adjunctive protective properties were estimated in vitro, using human small-intestinal juice as the incubation medium. These preparations were administered to healthy subjects and to patients with severe pancreatic insufficiency, and their ability to increase the intestinal concentrations of pancreatic enzymes was evaluated. The relations between in vitro and in vivo activities were also studied. In vitro testing showed that the preparations contained high but varying activities of enzymes, with the greatest variations in lipase and trypsin. Pancreatin in the form of tablets, with or without protective measures against acid, did not cause any apparent increase in the activities of pancreatic enzymes in the upper part of the gut in patients with pancreatic insufficiency. Granulated pancreatin, on the other hand, brought about an increase in the activities of amylase, phospholipase, lipase, and trypsin. Relatively higher activities of the enzymes in granulated form reached the small intestine as compared with those of the tablets.
Assuntos
Secreções Intestinais/enzimologia , Pâncreas/enzimologia , Pancreatopatias/tratamento farmacológico , Pancreatina/uso terapêutico , Amilases/metabolismo , Amilases/uso terapêutico , Humanos , Lipase/metabolismo , Lipase/uso terapêutico , Pancreatopatias/enzimologia , Pancreatina/administração & dosagem , Fosfolipases/metabolismo , Fosfolipases/uso terapêutico , Tecnologia Farmacêutica , Tripsina/metabolismo , Tripsina/uso terapêuticoRESUMO
Injection of bacterial lipopolysaccharide into pregnant mice resulted in fibrinogen accumulation, thrombosis and haemorrhage in the placental tissue and foetal death. Depletion of circulating fibrinogen by a thrombin-like enzyme from the venom of Malayan pit viper, Arvin, prevents foetal death. Foetal protection was also obtained by treating the mothers with a preparation of phospholipase C from Bacillus cereus known to inactivate tissue thromboplastin. It is suggested the lipopolysaccharide causes foetal death by inducing thrombosis as a consequence of activation of placental thromboplastin.
Assuntos
Ancrod/uso terapêutico , Morte Fetal/prevenção & controle , Fosfolipases/uso terapêutico , Animais , Feminino , Morte Fetal/etiologia , Fibrinogênio/antagonistas & inibidores , Lipopolissacarídeos/toxicidade , Camundongos , Placenta/patologia , Gravidez , Trombose/induzido quimicamenteRESUMO
Infusions of purified tissue thromboplastin in rats cause the accumulation of fibrin and platelets in the lungs and produce marked changes in the platelet count and in the coagulation factors V, VII and VIII. Tissue thromboplastin in a dose corresponding to less than 2 mug of protein per rat is lethal when given as a bolus injection. Simultaneous i.v. administration purified phospholipase C effectively prevents all these changes and protects rats from otherwise lethal doses of tissue thromboplastin. The necessary doses of phospholipase C are well below the toxic level for phospholipase C alone.