Cytochemical investigation of genic male-sterility in Chinese cabbage

A genic male sterile Chinese cabbage, Brassica campestris L. ssp. chinensis Makino, was examined using cytological and cytochemical methods to characterize the process of pollen abortion in this plant. Thick sections of both fertile and sterile anthers at different developmental stages were stained using Toluidine Blue O, Periodic Acid-Schiff’s (PAS) reaction and Sudan Black B to detect cytochemical changes that may occur in the distribution of insoluble polysaccharide and lipid storage bodies. Pollen abortion in sterile anthers occurs at an early stage of microspore development. During early microspore development, reductions in the number of starch grains in the connective tissue of fertile anthers coincide with the accumulation of starch grains in cells of the anther wall. In the late microspore stage, a large vacuole forms in the microspore, and tapetal cells synthesize and accumulate lipid droplets. The cellular organization of tapetal cells in sterile anthers appears similar to that in fertile anthers, except for the absence of lipid droplets in cells of sterile anthers and diffusely labeled tapetal polysaccharides, suggesting defects in nutrient storage. Supported by National Natural Science Foundation of CHINA (30170060)     

The diaphragm is better protected from oxidative stress than hindlimb skeletal muscle during CLP-induced sepsis

Objectives: The aim of this study was to determine whether non-lethal sepsis induced by cecal ligation and puncture (CLP) modulates oxidative damage and enzymatic antioxidant defenses in diaphragm and hindlimb skeletal muscles (soleus and Extensor Digitorus Longus (EDL)).

Methods: Female Wistar rats were divided into four experimental groups: (1) control animals, (2) animals sacrificed 2?hours or (3) 7 days after CLP, and (4) sham-operated animals. At the end of the experimental procedure, EDL, soleus, and diaphragm muscles were harvested and 4-hydroxynonenal (HNE)-protein adducts and protein carbonyl contents were examined in relation to superoxide dismutase and catalase expression and activities.

Results: We observed that both non-respiratory oxidative (i.e. soleus) and glycolytic skeletal muscles (i.e. EDL) are more susceptible to sepsis-induced oxidative stress than diaphragm, as attested by an increase in 4-HNE protein adducts and carbonylated proteins after 2?hours of CLP only in soleus and EDL.

Discussion: These differences could be explained by higher basal enzymatic antioxidant activities in diaphragm compared to hindlimb skeletal muscles. Together, these results demonstrate that diaphragm is better protected from oxidative stress than hindlimb skeletal muscles during CLP-induced sepsis.     

CT-based semi-automatic quantification of vertebral fracture restoration

Minimally invasive surgeries aiming to restore fractured vertebral body are increasing; therefore, our goals were to create a 3D vertebra reconstruction process and design clinical indices to assess the vertebral restoration in terms of heights, angles and volumes. Based on computed tomography (CT)-scan of the vertebral spine, a 3D reconstruction method as well as relevant clinical indices were developed. First, a vertebra initial solution requiring 5 min of manual adjustments is built. Then an image processing algorithm places this solution in the CT-scan images volume to adjust the model's nodes. On the vertebral body's anterior and posterior parts, nine robust heights, volume and endplate angle measurement methods were developed. These parameters were evaluated by reproducibility and accuracy studies. The vertebral body reconstruction accuracy was 1.0 mm; heights and volume accuracy were, respectively, 1.2 and 179 mm³. In conclusion, a 3D vertebra reconstruction process requiring little user time was proposed as well as 3D clinical indices assessing fractured and restored vertebra.     

红背山麻杆叶的化学成分研究(Ⅱ)——黄酮和苯乙醇苷类化合物

采用80％丙酮提取物的水萃取部位,利用凝胶、MCI、反相碳18、及 Toyopearl Butyl-650C 柱色谱进行分离纯化得到7个黄酮和3个苯乙醇苷类化合物。根据化合物的波谱数据分析鉴定为槲皮素(1)、槲皮苷(2)、异懈皮苷(3)、芦丁(4)、异牡荆素(5)、牡荆素(6)、木犀草素-7-O-α-L-鼠李糖(1→6)-β-D-葡萄糖苷(7)、2-phenethylβ-D-glucoside(8)、icariside D1(9)、2-苯乙基-D-芸香甙(10)。其中化合物1-3、5-6、8-10为首次从本属植物中分离得到。     

Proteomic analysis of human osteoarthritis synovial fluid

Background

Osteoarthritis is a chronic musculoskeletal disorder characterized mainly by progressive degradation of the hyaline cartilage. Patients with osteoarthritis often postpone seeking medical help, which results in the diagnosis being made at an advanced stage of cartilage destruction. Sustained efforts are needed to identify specific markers that might help in early diagnosis, monitoring disease progression and in improving therapeutic outcomes. We employed a multipronged proteomic approach, which included multiple fractionation strategies followed by high resolution mass spectrometry analysis to explore the proteome of synovial fluid obtained from osteoarthritis patients. In addition to the total proteome, we also enriched glycoproteins from synovial fluid using lectin affinity chromatography.

Results

We identified 677 proteins from synovial fluid of patients with osteoarthritis of which 545 proteins have not been previously reported. These novel proteins included ADAM-like decysin 1 (ADAMDEC1), alanyl (membrane) aminopeptidase (ANPEP), CD84, fibulin 1 (FBLN1), matrix remodelling associated 5 (MXRA5), secreted phosphoprotein 2 (SPP2) and spondin 2 (SPON2). We identified 300 proteins using lectin affinity chromatography, including the glycoproteins afamin (AFM), attractin (ATRN), fibrillin 1 (FBN1), transferrin (TF), tissue inhibitor of metalloproteinase 1 (TIMP1) and vasorin (VSN). Gene ontology analysis confirmed that a majority of the identified proteins were extracellular and are mostly involved in cell communication and signaling. We also confirmed the expression of ANPEP, dickkopf WNT signaling pathway inhibitor 3 (DKK3) and osteoglycin (OGN) by multiple reaction monitoring (MRM) analysis of osteoarthritis synovial fluid samples.

Conclusions

We present an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis. We believe that the catalog of proteins generated in this study will further enhance our knowledge regarding the pathophysiology of osteoarthritis and should assist in identifying better biomarkers for early diagnosis.     

Multidimensional analysis of landscape dynamics in a Central African forest-savannah mosaic

Most landscape cover assessments for conservation programmes rely largely on remote sensing analyses. These analyses, however, neglect how people inhabiting protected zones perceive and structure land cover. Using socio-ecological systems (SES) analysis in a forest-savannah mosaic on the Congo Basin forest edge (Democratic Republic of Congo), we investigated how human practices, landscape perceptions and land use patterns affected forest cover. We conducted remote sensing analysis using a Sentinel-2 satellite image and 187 GPS landmarks, producing a land cover map with 11 classes. Our results yielded an 81.85% correspondence with additional 164 GPS landmarks, a robust score in tropical areas. We conducted 40 individual interviews, eighteen focus group discussion workshops and nine months of participant observation of human practices to identify 19 land units and elaborate a granular SES system structuring the landscape. Integrating local knowledge and practices with general ecological and agronomic processes, we developed a landscape dynamics model revealing progressive forest colonisation of savannahs. Our methods demonstrate that a forest-edge landscape cover evaluation through remote sensing and local knowledge can contribute to finer-grained assessment of land cover and ecosystem services. This assessment can assist conservation efforts by considering local populations' practices on and perceptions of landscape change.     

Establishment of functional primary cultures of heart cells from the clam <Emphasis Type="Italic">Ruditapes decussatus</Emphasis>

Heart cells from the clam Ruditapes decussatus were routinely cultured with a high level of reproducibility in sea water based medium. Three cell types attached to the plastic after 2 days and could be maintained in vitro for at least 1 month: epithelial-like cells, round cells and fibroblastic cells. Fibroblastic cells were identified as functional cardiomyocytes due to their spontaneous beating, their ultrastructural characteristics and their reactivity with antibodies against sarcomeric α-actinin, sarcomeric tropomyosin, myosin and troponin T-C. Patch clamp measurements allowed the identification of ionic currents characteristic of cardiomyocytes: a delayed potassium current (I _K slow) strongly suppressed (95%) by tetraethylammonium (1 mM), a fast inactivating potassium current (I _K fast) inhibited (50%) by 4 amino-pyridine at 1 mM and, at a lower level (34%) by TEA, a calcium dependent potassium current (I _KCa) activated by strong depolarization. Three inward voltage activated currents were also characterized in some cardiomyocytes: L-type calcium current (I _Ca) inhibited by verapamil at 5 × 10⁻⁴ M, T-type Ca²⁺ current, rapidly activated and inactivated, and sodium current (I _Na) observed in only a few cells after strong hyperpolarization. These two currents did not seem to be physiologically essential in the initiation of the beatings of cardiomyocytes. Potassium currents were partially inhibited by tributyltin (TBT) (1 μM) but not by okadaic acid (two marine pollutants). DNA synthesis was also demonstrated in few cultured cells using BrdU (bromo-2′-deoxyuridine). Observed effects of okadaic acid and TBT demonstrated that cultured heart cells from clam Ruditapes decussatus can be used as an experimental model in marine toxicology.