Melatonin enhances L‐DOPA therapeutic effects,helps to reduce its dose,and protects dopaminergic neurons in 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine‐induced parkinsonism in mice

L‐3,4‐dihydroxyphenylalanine (L‐DOPA) reduces symptoms of Parkinson's disease (PD), but suffers from serious side effects on long‐term use. Melatonin (10–30 mg/kg, 6 doses at 10 hr intervals) was investigated to potentiate L‐DOPA therapeutic effects in 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP)‐induced parkinsonism in mice. Striatal tyrosine hydroxylase (TH) immunoreactivity, TH, and phosphorylated ser 40 TH (p‐TH) protein levels were assayed on 7th day. Nigral TH‐positive neurons stereology was conducted on serial sections 2.8 mm from bregma rostrally to 3.74 mm caudally. MPTP caused 39% and 58% decrease, respectively, in striatal fibers and TH protein levels, but 2.5‐fold increase in p‐TH levels. About 35% TH neurons were lost between 360 and 600 μm from 940 μm of the entire nigra analyzed, but no neurons were lost between 250 μm rostrally and 220 μm caudally. When L‐DOPA in small doses (5–8 mg/kg) failed to affect MPTP‐induced akinesia or catalepsy, co‐administration of melatonin with L‐DOPA attenuated these behaviors. Melatonin administration significantly attenuated MPTP‐induced loss in striatal TH fibers (82%), TH (62%) and p‐TH protein (100%) levels, and nigral neurons (87–100%). Melatonin failed to attenuate MPTP‐induced striatal dopamine depletion. L‐DOPA administration (5 mg/kg, once 40 min prior to sacrifice, p.o.) in MPTP‐ and melatonin‐treated mice caused significant increase in striatal dopamine (31%), as compared to L‐DOPA and MPTP‐treated mice. This was equivalent to 8 mg/kg L‐DOPA administration in parkinsonian mouse. Therefore, prolonged, effective use of L‐DOPA in PD with lesser side effects could be achieved by treating with 60% lower doses of L‐DOPA along with melatonin.     

CCR7 facilitates the pro‐inflammatory function of dendritic cells in experimental leishmaniasis

Cutaneous leishmaniasis, caused by the parasite Leishmania major, results in lesions at the site of infection, which are self‐healing in resistant hosts. However, in the absence of the chemokine receptor CCR7, mice are unable to heal the lesion and develop chronic disease. These B6.CCR7^?/? mice display an increased number of Th2 cells and immunosuppressive cytokine levels, as well as more regulatory T cells. As CCR7 is expressed on activated dendritic cells (DCs), and these cells require CCR7 to migrate to the draining lymph node, we expected decreased migration of DCs into the lymph node in the absence of CCR7 during cutaneous leishmaniasis. Consequently, in an attempt to initiate a self‐healing response, we adoptively transferred CCR7⁺ (B6.WT) DCs into the site of infection of B6.CCR7^?/? mice. Surprisingly, instead of healing the lesion, B6.CCR7^?/? mice inoculated with B6.WT DCs developed augmented lesions and showed increased immunosuppression compared to control B6.CCR7^?/? mice transferred with B6.CCR7^?/? DCs or B6.WT mice with B6.WT DCs. Finally, B6.WT mice injected with B6.CCR7^?/? DCs also presented delayed healing of the lesion. These results indicate that CCR7 must be expressed on DCs, as well as peripheral cells, to allow an efficient immune response to L. major.     

Safety and immunogenicity of hepatitis B surface antigen‐pulsed dendritic cells in patients with chronic hepatitis B

Summary. The immune modulator capacity of antigen‐pulsed dendritic cells (DC) has been documented in patients with cancers and in animal models of chronic viral infections. Cancer antigen‐pulsed DC are now used for treating patients with cancer. But viral antigen‐pulsed DC are not used in chronic viral‐infected patients because safety of antigen‐pulsed DC has not been evaluated in these patients. DC were isolated from human peripheral blood mononuclear cells by culturing with human‐grade granulocyte‐macrophage colony stimulating factor and interleukin‐4. Human blood DC were cultured with hepatitis B surface antigen (HBsAg) for 8 h to prepare HBsAg‐pulsed DC. After immunogenicity assessment of HBsAg‐pulsed DC in vitro, five million HBsAg‐pulsed DC were administered intradermally to five patients with chronic hepatitis B (CHB) 1–3 times. HBsAg‐pulsed DC were immunogenic in nature because they produced significantly higher levels of interleukin‐12 and interferon‐γ compared to unpulsed DC (P < 0.05). Also, HBsAg‐pulsed DC induced proliferation of HBsAg‐specific T lymphocytes in vitro. CHB patients injected with HBsAg‐pulsed DC did not exhibit generalized inflammation, exacerbation of liver damage, abnormal kidney function, or features of autoimmunity. Administration of HBsAg‐pulsed DC induced anti‐HBs in two patients and HBsAg‐specific cellular immunity in 1 patient. This is the first study about preparation of antigen‐pulsed DC using human consumable materials for treating patients with CHB. Because HBsAg‐pulsed DC were safe for all patients with CHB and had immune modulation capacity in some patients, phase I and phase II clinical trials with antigen‐pulsed DC in CHB and other chronic infections are warranted.     

Proteomic diversity of high‐density lipoprotein explains its association with clinical outcome in patients with heart failure

Aims

Previously, low high‐density lipoprotein (HDL) cholesterol was found to be one of the strongest predictors of mortality and/or heart failure (HF) hospitalisation in patients with HF. We therefore performed in‐depth investigation of the multifunctional HDL proteome to reveal underlying pathophysiological mechanisms explaining the association between HDL and clinical outcome.

Methods and results

We selected a cohort of 90 HF patients with 1:1 cardiovascular death/survivor ratio from BIOSTAT‐CHF. A novel optimised protocol for selective enrichment of lipoproteins was used to prepare plasma. Enriched lipoprotein content of samples was analysed using high resolution nanoscale liquid chromatography‐mass spectrometry‐based proteomics, utilising a label free approach. Within the HDL proteome, 49 proteins significantly differed between deaths and survivors. An optimised model of 12 proteins predicted death with 76% accuracy (Nagelkerke R²=0.37, P < 0.001). The strongest contributors to this model were filamin‐A (related to crosslinking of actin filaments) [odds ratio (OR) 0.31, 95% confidence interval (CI) 0.15–0.61, P = 0.001] and pulmonary surfactant‐associated protein B (related to alveolar capillary membrane function) (OR 2.50, 95% CI 1.57–3.98, P < 0.001). The model predicted mortality with an area under the curve of 0.82 (95% CI 0.77–0.87, P < 0.001). Internal cross validation resulted in 73.3 ± 7.2% accuracy.

Conclusion

This study shows marked differences in composition of the HDL proteome between HF survivors and deaths. The strongest differences were seen in proteins reflecting crosslinking of actin filaments and alveolar capillary membrane function, posing potential pathophysiological mechanisms underlying the association between HDL and clinical outcome in HF.

     

Promoting dementia‐friendly communities to improve the well‐being of individuals with and without dementia

A community is the totality of human relationships, in which individuals live with and for others. This article discusses promoting dementia‐friendly community from the perspective of human relationships within a community. Improving the social well‐being of aging people is important; it is especially important for people with dementia, as dementia is a life‐changing syndrome that affects all aspects of one's life, including human relationships. Dementia‐friendly community requires support from the community in improving the social adaptation of people with dementia, as they experience greater difficulties in social interaction as a result of cognitive decline, especially deterioration of social cognition. They need to be empowered to stay motivated for the opportunity to maintain an active and meaningful life, despite dementia. Flexible provisions of such tailored support can be effective to improve the support network in the community to meet the individual's needs. As social and human resources are limited, it is also necessary to discuss how to share socioeconomic burdens to meet both social sustainability demands and the welfare of aging adults. More discussions that include people with dementia and their family members are required to achieve the purpose of dementia‐friendly community. This is important to enhance the well‐being of individuals with and without dementia, as well as the society as a whole, through dementia support and dementia prevention activities. Geriatr Gerontol Int 2020; ??: ??–?? .     

Bone density and health‐related quality of life in adult patients with severe haemophilia

Summary. Severe haemophilia and reduced bone density can negatively influence perception of patient’s health‐related quality of life (HRQoL), especially considering future aspects, the risk of losing independence or pain suffering. The aim of this study was to assess levels of HRQoL in severe haemophilia patients and to compare HRQoL to those of the general population as well as to determine whether reduced bone density is correlated to the perceived HRQoL. Patients were divided into two groups based on timing of being treated with prophylaxis: Group A (started prophylaxis at age of ≤3 years; n = 22); Group B (at age of >3 years; n = 15). The bone mineral density (BMD g cm^?2) of different measured sites was measured by dual energy X‐ray absorptiometry (DXA). HRQoL was assessed using SF‐36 questionnaire. Group A have mean BMD T‐score >?1.0 (i.e. normal score) at all measured sites, and have almost similar scores in the SF‐36 domains compared with the reference population. Group B have mean BMD T‐score ?1.0 at lumbar spine and total body, and their scores in the SF‐36 domains were lower compared with the reference population. Moreover, significant correlations were found between BMD at femoral neck and total body with physical domains. With adequate long‐term prophylaxis since early childhood, adult patients with haemophilia report a comparable BMD and HRQoL to the Swedish reference population. Reduced BMD in group B correlated with impaired physical health, which underscores the importance of early onset of adequate prophylactic treatment.     

Melatonin synergizes the chemotherapeutic effect of 5‐fluorouracil in colon cancer by suppressing PI3K/AKT and NF‐κB/iNOS signaling pathways

5‐Fluorouracil (5‐FU) is one of the most commonly used chemotherapeutic agents in colon cancer treatment, but has a narrow therapeutic index limited by its toxicity. Melatonin exerts antitumor activity in various cancers, but it has never been combined with 5‐FU as an anticolon cancer treatment to improve the chemotherapeutic effect of 5‐FU. In this study, we assessed such combinational use in colon cancer and investigated whether melatonin could synergize the antitumor effect of 5‐FU. We found that melatonin significantly enhanced the 5‐FU‐mediated inhibition of cell proliferation, colony formation, cell migration and invasion in colon cancer cells. We also found that melatonin synergized with 5‐FU to promote the activation of the caspase/PARP‐dependent apoptosis pathway and induce cell cycle arrest. Further mechanism study demonstrated that melatonin synergized the antitumor effect of 5‐FU by targeting the PI3K/AKT and NF‐κB/inducible nitric oxide synthase (iNOS) signaling. Melatonin in combination with 5‐FU markedly suppressed the phosphorylation of PI3K, AKT, IKKα, IκBα, and p65 proteins, promoted the translocation of NF‐κB p50/p65 from the nuclei to cytoplasm, abrogated their binding to the iNOS promoter, and thereby enhanced the inhibition of iNOS signaling. In addition, pretreatment with a PI3K‐ or iNOS‐specific inhibitor synergized the antitumor effects of 5‐FU and melatonin. Finally, we verified in a xenograft mouse model that melatonin and 5‐FU exerted synergistic antitumor effect by inhibiting the AKT and iNOS signaling pathways. Collectively, our study demonstrated that melatonin synergized the chemotherapeutic effect of 5‐FU in colon cancer through simultaneous suppression of multiple signaling pathways.     

不同剂量阿托伐他汀治疗急性冠状动脉综合征合并基线低水平低密度脂蛋白的疗效分析

目的 探讨不同剂量阿托伐他汀治疗急性冠状动脉综合征(ACS)合并基线低水平低密度脂蛋白患者的疗效。方法 抽取本院于2016年1月—2017年12月收治的ACS合并基线低水平低密度脂蛋白患者共102例,以随机综合序贯法分为低剂量短期组、高剂量短期组及低剂量长期组,每组各34例。低剂量短期组和低剂量长期组患者每天口服阿托伐他汀10 mg,治疗时间分别为9个月和12个月;高剂量短期组患者每天口服阿托伐他汀40 mg,治疗时间为9个月。全自动生化分析仪检测甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDLC)、低密度脂蛋白胆固醇(LDLC)血脂4项指标。比较三组患者治疗终点的血脂水平和主要不良心脑血管事件发生率。结果 治疗前三组患者基线资料、血脂4项指标比较差异无统计学意义(P＞0.05)。治疗第3、9个月后高剂量短期组TC、LDLC和TG水平比低剂量短期组、低剂量长期组低,HDLC水平比低剂量短期组、低剂量长期组高(P均＜0.05);低剂量长期组治疗12个月后血脂4项指标较低剂量短期组治疗9个月的疗效更佳,低剂量长期组治疗12个月后血脂4项指标与高剂量短期组治疗9个月后的疗效无明显差异(P＞0.05)。三组治疗终点均未出现死亡病例,高剂量短期组、低剂量长期组的主要不良心脑血管事件发生率均为11.76%,稍低于低剂量短期组17.65%,但各组间比较差异均无统计学意义(P＞0.05)。结论 使用阿托伐他汀治疗ACS合并基线低水平低密度脂蛋白具有剂量和时间依赖性,因此,在治疗此类患者时建议适当加大用药剂量、延长治疗时间。     

Melatonin protects embryonic development and maintains sleep/wake behaviors from the deleterious effects of fluorene‐9‐bisphenol in zebrafish (Danio rerio)

Environmental endocrine chemicals have various adverse effects on the development of vertebrates. Fluorene‐9‐bisphenol (BHPF), a substitute of bisphenol A (BPA), is widely used in commercial production. The effects of BHPF on development and behavior are unclear. Melatonin plays a protective role under many unfavorable conditions. In this study, we investigated the effects of BHPF on the development and behaviors of zebrafish and whether melatonin reverses effects induced by BHPF. Zebrafish embryos were exposed to 0.1, 10, or 1000 nmol/L BHPF with or without 1 μmol/L melatonin from 2 hours postfertilization to 6 days postfertilization. The results showed that 0.1 and 10 nmol/L BHPF had little effect on development. High‐dose BHPF (1000 nmol/L) delayed the development, increased mortality and surface tension of embryonic chorions, caused aberrant expression of the key genes (ntl, shh, krox20, pax2, cmlc2) in early development detected by in situ hybridization, and damaged the CaP motor neurons, which were associated with locomotion ability detected by immunofluorescence. Melatonin addition reversed or weakened these adverse effects of BHPF on development, and melatonin alone increased surface tension as the effects of high‐dose BHPF. However, all groups of BHPF exposure triggered insomnia‐like behaviors, with increased waking activity and decreased rest behaviors. BHPF acted on the hypocretin (hcrt) system and upregulated the expression of sleep/wake regulators such as hcrt, hcrt receptor (hcrtr), arylalkylamine N‐acetyltransferase‐2 (aanat2). Melatonin recovered the alternation of sleep/wake behaviors induced by BHPF and restored abnormal gene expression to normal levels. This study showed that high‐dose BHPF had adverse effects on early development and induced behavioral alternations. However, melatonin prevented BHPF‐induced aberrant development and sleep/wake behaviors.