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PURPOSE: The incidence of pneumonia caused by multidrug-resistant gram-negative bacteria (MDR GNB) is increasing, which imposes significant burden on public health. Inhalation combined with intravenous polymyxins has emerged as a viable treatment option. However, pharmacokinetic studies focusing on intravenous and inhaled polymyxin B (PMB) are limited. METHODS: This study included seven patients with MDR GNB-induced pneumonia who were treated with intravenous plus inhaled PMB from March 1 to November 30, 2022, in the intensive care unit of the First Affiliated Hospital of Zhejiang University School of Medicine. Clinical outcomes and therapeutic drug monitoring data of PMB in both plasma and epithelial lining fluid (ELF) were retrospectively reviewed. RESULTS: Median PMB concentrations in the ELF were 7.83 (0.72-66.5), 116.72 (17.37-571.26), 41.1 (3.69-133.78), and 33.82 (0.83-126.68) mg/L at 0, 2, 6, and 12 h, respectively, and were much higher than those detected in the serum. ELF concentrations of PMB at 0, 2, 6, and 12 h were higher than the minimum inhibitory concentrations of pathogens isolated from the patients. Steady-state concentrations of PMB in the plasma were > 2 mg/L in most patients. Of the patients, 57.14 % were cured and 71.43 % showed a favorable microbiological response. The incidence of side effects with PMB was low. CONCLUSIONS: Inhaled plus intravenous PMB can achieve high ELF concentrations and favorable clinical outcomes without an increased adverse effect profile. This treatment approach appears promising for the treatment of patients with pneumonia caused by MDR-GNB.
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Aggregation of aberrant proteins is a common pathological hallmark in neurodegeneration such as polyglutamine (polyQ) and other repeat-expansion diseases. Here through overexpression of ataxin3 C-terminal polyQ expansion in Drosophila gut enterocytes, we generated an intestinal obstruction model of spinocerebellar ataxia type3 (SCA3) and reported a new role of nuclear-associated endosomes (NAEs)-the delivery of polyQ to the nucleoplasm. In this model, accompanied by the prominently increased RAB5-positive NAEs are abundant nucleoplasmic reticulum enriched with polyQ, abnormal nuclear envelope invagination, significantly reduced endoplasmic reticulum, indicating dysfunctional nucleocytoplasmic trafficking and impaired endomembrane organization. Consistently, Rab5 but not Rab7 RNAi further decreased polyQ-related NAEs, inhibited endomembrane disorganization, and alleviated disease model. Interestingly, autophagic proteins were enriched in polyQ-related NAEs and played non-canonical autophagic roles as genetic manipulation of autophagic molecules exhibited differential impacts on NAEs and SCA3 toxicity. Namely, the down-regulation of Atg1 or Atg12 mitigated while Atg5 RNAi aggravated the disease phenotypes both in Drosophila intestines and compound eyes. Our findings, therefore, provide new mechanistic insights and underscore the fundamental roles of endosome-centered nucleocytoplasmic trafficking and homeostatic endomembrane allocation in the pathogenesis of polyQ diseases.
Assuntos
Autofagia , Endossomos , Peptídeos , Animais , Peptídeos/metabolismo , Endossomos/metabolismo , Núcleo Celular/metabolismo , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Transporte Ativo do Núcleo Celular , Drosophila melanogaster/metabolismo , Drosophila melanogaster/genética , Doença de Machado-Joseph/metabolismo , Doença de Machado-Joseph/genética , Doença de Machado-Joseph/patologia , Enterócitos/metabolismo , Modelos Animais de Doenças , Ataxina-3/metabolismo , Ataxina-3/genética , Drosophila/metabolismoRESUMO
INTRODUCTION: Moyamoya syndrome is commonly associated with sickle cell anemia, neurofibromatosis type 1, cranial therapeutic irradiation, and Down syndrome. However, it is rare for Moyamoya syndrome associated with Graves' disease. CASE REPORT: Here we report a case of Moyamoya syndrome associated with Graves' disease in a 19-year-old girl with sudden weakness of the right arm, progressive caries, and alopecia for 4 years. Brain magnetic resonance imaging revealed acute intraventricular hemorrhage and cerebral infarction of left middle cerebral artery territory and narrowing of the proximal portion of bilateral anterior and middle cerebral arteries. CONCLUSION: Acute cerebral infarction and intraventricular hemorrhage can occur simultaneously in Moyamoya syndrome associated with Graves' disease. Hydrocortisone, combined with prothiouracil medication, can correct thyroid dysfunction and improve neurological function. Caries may be the first symptom of Graves' disease.
Assuntos
Doença de Graves , AVC Isquêmico , Doença de Moyamoya , Feminino , Humanos , Adulto Jovem , Adulto , AVC Isquêmico/complicações , Doença de Moyamoya/complicações , Doença de Moyamoya/diagnóstico por imagem , Hemorragia Cerebral/complicações , Hemorragia Cerebral/diagnóstico por imagem , Infarto Cerebral/etiologia , Hemorragias Intracranianas/complicações , Doença de Graves/complicações , Doença de Graves/diagnóstico por imagemRESUMO
Polyglutamine (polyQ)-mediated mitochondria damage is one of the prime causes of polyQ toxicity, which leads to the loss of neurons and the injury of non-neuronal cells. With the discovery of the crucial role of the gut-brain axis and gut microbes in neurological diseases, the relationship between visceral damage and neurological disorders has also received extensive attention. This study successfully simulated the polyQ mitochondrial damage model by expressing 78 or 84 polyglutamine-containing Ataxin3 proteins in Drosophila intestinal enterocytes. In vivo, polyQ expression can reduce mitochondrial membrane potential, mitochondrial DNA damage, abnormal mitochondrial morphology, and loose mitochondrial cristae. Expression profiles evaluated by RNA-seq showed that mitochondrial structural genes and functional genes (oxidative phosphorylation and tricarboxylic acid cycle-related) were significantly down-regulated. More importantly, Bioinformatic analyses demonstrated that pathological polyQ expression induced vitamin B6 metabolic pathways abnormality. Active vitamin B6 participates in hundreds of enzymatic reactions and is very important for maintaining mitochondria's activities. In the SCA3 Drosophila model, Vitamin B6 supplementation significantly suppressed ECs mitochondria damage in guts and inhibited cellular polyQ aggregates in fat bodies, indicating a promising therapeutic strategy for the treatment of polyQ. Taken together, our results reveal a crucial role for the Vitamin B6-mediated mitochondrial protection in polyQ-induced cellular toxicity, which provides strong evidence for this process as a drug target in polyQ diseases treatment.
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Ataxina-3/genética , Modelos Animais de Doenças , Doença de Machado-Joseph/genética , Mitocôndrias/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Vitamina B 6/farmacologia , Animais , Animais Geneticamente Modificados , Ataxina-3/metabolismo , Drosophila , Redes Reguladoras de Genes/fisiologia , Humanos , Doença de Machado-Joseph/tratamento farmacológico , Doença de Machado-Joseph/metabolismo , Mitocôndrias/genética , Fármacos Neuroprotetores/uso terapêutico , Vitamina B 6/uso terapêuticoRESUMO
Calcium treatment effects on malate metabolism and the GABA pathway in 'Cripps Pink' apple fruit during storage were investigated. Postharvest apple fruit treated with 1% and 4% calcium chloride solutions were stored at 25 ± 1 °C. The 4% calcium treatment suppressed declines in titratable acidity and malate content and increased succinate and oxalate concentrations. Calcium treatment also reduced the respiration rate and decreased ethylene production peak during storage. Moreover, 4% calcium treatment significantly enhanced cyNAD-MDH and PEPC activities and upregulated MdMDH1, MdMDH2, MdPEPC1 and MdPEPC2 expression while inhibiting cyNADP-ME and PEPCK activities and downregulating MdME1, MdME4 and MdPEPCK2 expression. Surprisingly, calcium treatment changed the content of some free amino acids (GABA, proline, alanine, aspartic acid and glutamate), two of which (glutamate and GABA) are primary metabolites of the GABA pathway. Furthermore, calcium application enhanced GABA pathway activity by increasing MdGAD1, MdGAD2, MdGABA-T1/2 and MdSSADH transcript levels.
Assuntos
Cálcio/farmacologia , Frutas/efeitos dos fármacos , Malatos/metabolismo , Malus/efeitos dos fármacos , Malus/metabolismo , Ácido gama-Aminobutírico/metabolismo , Aminoácidos/análise , Aminoácidos/metabolismo , Etilenos/metabolismo , Qualidade dos Alimentos , Frutas/química , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Malus/química , Malus/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
PTEN-induced putative kinase 1 (PINK1) mutation induces autosomal recessive Parkinson's Disease (PD), mitochondrial dysfunction is the central pathogenic process. However, more and more studies presented the bulk of the damage to neurons with mitochondrial dysfunction stems from the endoplasmic reticulum (ER) stress. In mitochondria damaged PINK1B9 fly model how protein kinase RNA-like ER kinase (PERK) arm of ER stress functions remains a mystery. Thus, we generated both PERK overexpressed (PEK OE) and down expressed (PEK RNAi) PINK1B9 flies and monitored their motor activity. We found PEK OE decreased the abnormal wing posture rate and rescued PINK1B9 flies' motor activity. Furthermore, we observed the increased number of dopaminergic neurons of protocerebral posterior lateral 1 (PPL1) and the tyrosine hydroxylase (TH) protein levels in PINK1B9 flies. When testing the mitochondrial morphology in flight muscle with TEM, we found that the shape of the mitochondria became normal. The ATP levels of flight muscle tissues were significantly elevated in PEK OE PINK1B9 flies with the increased function of mitochondrial Electron Transport Chain (ETC) Complex I (CI) but not Complex â ¡ (Câ ¡) which is further confirmed by oxygen consumption experiments, Western Blot, and RT-PCR to examine the corresponding subunits. We suggest that overexpression of PERK can rescue PINK1B9 PD flies' pathogenic phenotypes and it is linked with the improved mitochondrial function especially CI of ETC but not Câ ¡. Our findings may pave a way for the target of the drug for alleviating the suffering of PINK1 mutant autosomal recessive PD patients.
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Proteínas de Drosophila/biossíntese , Mitocôndrias/metabolismo , Doenças Neurodegenerativas/metabolismo , Doenças Neurodegenerativas/prevenção & controle , Fenótipo , Proteínas Serina-Treonina Quinases/biossíntese , Animais , Animais Geneticamente Modificados , Drosophila , Proteínas de Drosophila/genética , Expressão Gênica , Masculino , Mitocôndrias/genética , Doenças Neurodegenerativas/genética , Proteínas Serina-Treonina Quinases/genéticaRESUMO
Organic acid is an important indicator of fruit quality, and malate is the predominant organic acid in apple fruit. However, the regulation of malate metabolism in postharvest fruit is rarely reported. Here, we found that, compared with a control treatment, a 10 mM γ-aminobutyric acid (GABA) treatment remarkably delayed the loss of tiftratable acidity and malate and increased the succinate and oxalate contents in "Cripps Pink" fruit stored in polyethylene bags at room temperature. The higher malate levels in GABA-treated fruit were accompanied by higher activities of cytosolic nicotinamide adenine dinucleotide-dependent malate dehydrogenase (cyNAD-MDH) and phosphoenolpyruvate carboxylase (PEPC) but lower cytosolic NAD phosphate-dependent malic enzyme (cyNADP-ME) and phosphoenolpyruvate carboxykinase (PEPCK) activities than those seen in control fruit. Notably, ethylene production was significantly reduced by GABA treatment, paralleling the downregulation of MdACS, MdACO, and MdERF expression. Meanwhile, GABA treatment also enhanced the activity of the GABA shunt and promoted the accumulation of GABA. This study provides new insights into the regulation of malate metabolism and reports for the first time the possible interplay between GABA and ethylene signaling pathways in apple fruit during postharvest storage.