Neuronal types in the spinal dorsal gray of the turtle Chrysemys d'orbigny: a Golgi study

At thoracic and lumbar levels the spinal dorsal gray of young specimens of the turtle Chrysemys d'orbigny consists of a cell-free neuropil and an aggregation of perikarya termed here the lateral column of the dorsal horn (LCDH). Nerve cell clusters also occur in the dorsal commissure. The main neuropil area can be divided into a thin superficial layer containing some myelinated fibers (neuropil area Ib) and a compact core composed of unmyelinated axon terminals, dendritic branches, and thin glial processes (neuropil area II). A looser neuropil area is located at the horn base (neuropil area III). The so-called marginal zone of de Lange represents a fourth synaptic field termed here neuropil area Ia. The LCDH consists of neurons of different size and shape. Two peculiar nerve cell types have been recognized in the dorsal horn: giant and bitufted neurons. The former exhibits a large dendritic arbor, which after passing through neuropil areas II and Ib projects into neuropil area Ia and the adjacent white matter. Most frequently Golgi-stained giant neurons have perikarya and dendritic domains on the same side (ipsilateral giant neurons). There are also heterolateral giant neurons whose dendritic branches invade the opposite horn. Bitufted neurons are characterized by the presence of two main dendritic shafts connecting neuropil area II of both dorsal horns. At neuropil levels the major dendritic branches ramify profusely giving rise to short tortuous terminal processes. Perikarya of bitufted neurons occur in the dorsal commissure. The LCDH also contains many small and medium-sized neurons. These are oriented in two main directions: parallel or radial with respect to the dorsal horn surface. The population of horizontally oriented neurons comprises two subtypes termed here alpha and beta. Radially oriented neurons are pleomorphic, defying precise, unequivocal classification.     

Development of chicken intrafusal muscle fibers

The first sign of developing intrafusal fibers in chicken leg muscles appeared on embryonic day (E) 13 when sensory axons contacted undifferentiated myotubes. In sections incubated with monoclonal antibodies against myosin heavy chains (MHC) diverse immunostaining was observed within the developing intrafusal fiber bundle. Large primary intrafusal myotubes immunostained moderately to strongly for embryonic and neonatal MHC, but they were unreactive or reacted only weakly with antibodies against slow MHC. Smaller, secondary intrafusal myotubes reacted only weakly to moderately for embryonic and neonatal MHC, but 1–2 days after their formation they reacted strongly for slow and slow-tonic MHC. In contrast to mammals, slow-tonic MHC was also observed in extrafusal fibers. Intrafusal fibers derived from primary myotubes acquired fast MHC and retained at least a moderate level of embryonic MHC. On the other hand, intrafusal fibers developing from secondary myotubes lost the embryonic and neonatal isoforms prior to hatching and became slow. Based on relative amounts of embryonic, neonatal and slow MHC future fast and slow intrafusal fibers could be first identified at E14. At the polar regions of intrafusal fibers positions of nerve endings and acetylcholinesterase activity were seen to match as early as E16. Approximately equal numbers of slow and fast intrafusal fibers formed prenatally; however, in postnatal muscle spindles fast fibers were usually in the majority, suggesting that some fibers transformed from slow to fast.     

Discrimination of folding types of globular proteins based on average distance maps constructed from their sequences

It has been shown that probable portions which form contacts in a protein can be predicted by means of an average distance map (ADM) as well as regular structures (-helices and -turns) defined as short-range compact regions (Kikuchiet al., 1988a,c). In this paper, we analyze the occurrence of those portions and short-range compact regions on ADMs for various proteins regarding their folding types. We have found out that each folding type of proteins shows characteristic distribution of such parts on ADMS. We also discuss the possibility of the prediction of folding types of proteins by ADMs.     

Subcellular Distribution and Chemical Forms of Cadmium in the Medicine Food Homology Plant <i>Platycodon grandiflorum</i> (Jacq.) A.DC.

Although Platycodon grandiflorum (Jacq.) A.DC. is a renowned medicine food homology plant, reports of excessive cadmium (Cd) levels are common, which affects its safety for clinical use and food consumption. To enable its Cd levels to be regulated or reduced, it is necessary to first elucidate the mechanism of Cd uptake and accumulation in the plant, in addition to its detoxification mechanisms. This present study used inductively couple plasma-mass-spectrometry to analyze the subcellular distribution and chemical forms of Cd in different tissues of P. grandiflorum. The experimental results showed that Cd was mainly accumulated in the roots [predominantly in the cell wall (50.96%–61.42%)], and it was found primarily in hypomobile and hypotoxic forms. The proportion of Cd in the soluble fraction increased after Cd exposure, and the proportion of insoluble phosphate Cd and oxalate Cd increased in roots and leaves, with a higher increase in oxalate Cd. Therefore, it is likely that root retention mechanisms, cell wall deposition, vacuole sequestration, and the formation of low mobility and low toxicity forms are tolerance strategies for Cd detoxification used by P. grandiflorum. The results of this study provide a theoretical grounding for the study of Cd accumulation and detoxification mechanisms in P. grandiflorum, and they can be used as a reference for developing Cd limits and standards for other medicine food homology plants.     

珍稀濒危植物蒜头果种子萌发特征与幼苗类型的研究

以去除果皮后阴干的珍稀濒危植物蒜头果种子为材料,在温室大棚沙池内进行层积处理,从层积处理开始至子叶出土的不同萌发阶段,考察蒜头果种子发育形态、贮藏物质积累、酶活性以及幼苗类型等变化特征,初步探讨其种子休眠形成原因。结果显示：(1)蒜头果种子从解除休眠开始至萌发形成幼苗的过程约需195 d,其中幼胚的形态发育后熟约需75 d,随后30 d内是种子集中萌发的时期,其发芽率达到最高(53.33%);依据种胚发育形态的标志特征将此过程划分为7个阶段(S1～S7阶段):S1阶段种子未萌发,S2阶段种胚“露白”,S3阶段胚根突破种皮长超过1 cm, S4阶段下胚轴与胚根连接处形成弯钩结构,S5阶段“S”型胚形成及胚根前端膨大,S6阶段种子不仅具有膨大的胚根且已有侧根的分化,S7阶段子叶脱落,胚芽出土,真叶出现。(2)蒜头果种子在湿沙层积过程中,种胚胚长和胚率从S1阶段的(5.49±1.57)mm和(19.48±5.72)%分别增加至S6阶段的(67.92±2.94)mm和(240.75±15.29)%,胚率平均增加了12.4倍,显示蒜头果种子的胚需要经历后熟过程才能萌发,属于胚后熟型种子。(3)从...     

Exosomes for angiogenesis induction in ischemic disorders

Ischaemic disorders are leading causes of morbidity and mortality worldwide. While the current therapeutic approaches have improved life expectancy and quality of life, they are unable to “cure” ischemic diseases and instate regeneration of damaged tissues. Exosomes are a class of extracellular vesicles with an average size of 100–150 nm, secreted by many cell types and considered a potent factor of cells for paracrine effects. Since exosomes contain multiple bioactive components such as growth factors, molecular intermediates of different intracellular pathways, microRNAs and nucleic acids, they are considered as cell-free therapeutics. Besides, exosomes do not rise cell therapy concerns such as teratoma formation, alloreactivity and thrombotic events. In addition, exosomes are stored and utilized more convenient. Interestingly, exosomes could be an ideal complementary therapeutic tool for ischemic disorders. In this review, we discussed therapeutic functions of exosomes in ischemic disorders including angiogenesis induction through various mechanisms with specific attention to vascular endothelial growth factor pathway. Furthermore, different delivery routes of exosomes and different modification strategies including cell preconditioning, gene modification and bioconjugation, were highlighted. Finally, pre-clinical and clinical investigations in which exosomes were used were discussed.     

COVID-19 plasma proteome reveals novel temporal and cell-specific signatures for disease severity and high-precision disease management

Coronavirus disease 2019 (COVID-19) is a systemic inflammatory condition with high mortality that may benefit from personalized medicine and high-precision approaches. COVID-19 patient plasma was analysed with targeted proteomics of 1161 proteins. Patients were monitored from Days 1 to 10 of their intensive care unit (ICU) stay. Age- and gender-matched COVID-19-negative sepsis ICU patients and healthy subjects were examined as controls. Proteomic data were resolved using both cell-specific annotation and deep-analysis for functional enrichment. COVID-19 caused extensive remodelling of the plasma microenvironment associated with a relative immunosuppressive milieu between ICU Days 3–7, and characterized by extensive organ damage. COVID-19 resulted in (1) reduced antigen presentation and B/T-cell function, (2) increased repurposed neutrophils and M1-type macrophages, (3) relatively immature or disrupted endothelia and fibroblasts with a defined secretome, and (4) reactive myeloid lines. Extracellular matrix changes identified in COVID-19 plasma could represent impaired immune cell homing and programmed cell death. The major functional modules disrupted in COVID-19 were exaggerated in patients with fatal outcome. Taken together, these findings provide systems-level insight into the mechanisms of COVID-19 inflammation and identify potential prognostic biomarkers. Therapeutic strategies could be tailored to the immune response of severely ill patients.     

Treatment-resistant depression: definition,prevalence, detection,management, and investigational interventions

Treatment-resistant depression (TRD) is common and associated with multiple serious public health implications. A consensus definition of TRD with demonstrated predictive utility in terms of clinical decision-making and health outcomes does not currently exist. Instead, a plethora of definitions have been proposed, which vary significantly in their conceptual framework. The absence of a consensus definition hampers precise estimates of the prevalence of TRD, and also belies efforts to identify risk factors, prevention opportunities, and effective interventions. In addition, it results in heterogeneity in clinical practice decision-making, adversely affecting quality of care. The US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) have adopted the most used definition of TRD (i.e., inadequate response to a minimum of two antidepressants despite adequacy of the treatment trial and adherence to treatment). It is currently estimated that at least 30% of persons with depression meet this definition. A significant percentage of persons with TRD are actually pseudo-resistant (e.g., due to inadequacy of treatment trials or non-adherence to treatment). Although multiple sociodemographic, clinical, treatment and contextual factors are known to negatively moderate response in persons with depression, very few factors are regarded as predictive of non-response across multiple modalities of treatment. Intravenous ketamine and intranasal esketamine (co-administered with an antidepressant) are established as efficacious in the management of TRD. Some second-generation antipsychotics (e.g., aripiprazole, brexpiprazole, cariprazine, quetiapine XR) are proven effective as adjunctive treatments to antidepressants in partial responders, but only the olanzapine-fluoxetine combination has been studied in FDA-defined TRD. Repetitive transcranial magnetic stimulation (TMS) is established as effective and FDA-approved for individuals with TRD, with accelerated theta-burst TMS also recently showing efficacy. Electroconvulsive therapy is regarded as an effective acute and maintenance intervention in TRD, with preliminary evidence suggesting non-inferiority to acute intravenous ketamine. Evidence for extending antidepressant trial, medication switching and combining antidepressants is mixed. Manual-based psychotherapies are not established as efficacious on their own in TRD, but offer significant symptomatic relief when added to conventional antidepressants. Digital therapeutics are under study and represent a potential future clinical vista in this population.     

Evidence for constraint on species coexistence in vegetation of the Park Grass experiment

Repeated patterns, of a type that would be expected to result from limitations to species coexistence (i.e. assembly rules) were sought in the Park Grass experiment. This classical grassland experiment was sampled in two years, using replicated biomass samples. Variance in a number of measures was examined, and compared to the variance expected under appropriate null models, the latter based on assumptions of no interactions between species. In each case, an assembly rule would result in low variance. Examining variance in species richness between quadrats within a treatment, there was no indication of constraint on species co-occurrences; variance in richness was actually greater than expected under the null model, attributable to environmental variation or perhaps positive interactions between species. However, there was control on biomass, evidenced by variance in total biomass (i.e. over all species) within a treatment being significantly lower than expected under the null model. There was no indication of community structure based on guilds (i.e. functional types). Although there was in 1991 some, non-significant, indication of a constant proportion of species from the legume guild, there was no sign of such an effect in 1992. Searches for intrinsic guilds failed to converge. There was no indication at all of constancy in the proportional representation of guilds by biomass. Thus, there is good evidence for competitive control on plant growth, but none for control of species occurrences. There is no convincing evidence for guild structure in this community at the scale sampled. Possible conflict is discussed between the existence of evidence for temporal stability but the absence of evidence for spatial uniformity. It is concluded that most of the mechanisms proposed for temporal stability will not necessarily lead to control on spatial variation. For many mechanisms, this would depend on the spatial scale examined.     

红头豆芫菁成虫芫菁素含量的研究

芫菁素在红头豆芫菁体内主要贮存在雄虫的生殖腺和卵内。与野生群体两性芫菁素的平均含量相比较,刚交配过的雄虫失去体内７０％的芫菁素,其雌配偶体内芫菁素含量相应升高４３％。红头豆芫菁可用作中药材。经１１０℃烘干后的雄虫,用酸水解后提取的芫菁素含量比直接提取的含量增高４倍。