A Protocol for Genetic Induction and Visualization of Benign and Invasive Tumors in Cephalic Complexes of Drosophila melanogaster

Drosophila has illuminated our understanding of the genetic basis of normal development and disease for the past several decades and today it continues to contribute immensely to our understanding of complex diseases ^1-7. Progression of tumors from a benign to a metastatic state is a complex event ⁸ and has been modeled in Drosophila to help us better understand the genetic basis of this disease ⁹. Here I present a simple protocol to genetically induce, observe and then analyze the progression of tumors in Drosophila larvae. The tumor induction technique is based on the MARCM system ¹⁰ and exploits the cooperation between an activated oncogene, Ras^V12 and loss of cell polarity genes (scribbled, discs large and lethal giant larvae) to generate invasive tumors ⁹. I demonstrate how these tumors can be visualized in the intact larvae and then how these can be dissected out for further analysis. The simplified protocol presented here should make it possible for this technique to be utilized by investigators interested in understanding the role of a gene in tumor invasion.     

Orcadian Variation in Human Stature

Thyroxine (T₄) and triiodothyronine (T₃) plasma concentrations have been determined during 24-hr sampling periods in six mongrels (age 12-36 months), six beagles (age 35-37 months), three labradors (age 3.5 months) and three beagles (age 5 months). The mean T₄ levels of the labradors were significantly lower than the values found for mongrels or older beagles (P < 0.05), whereas T₃ was higher in the 5 month old beagles compared to the mongrels (P < 0.001), young beagles (P < 0.05) or labradors (P < 0.01).

Circadian and ultradian rhythmicities have been evaluated by cosinor and Fourier analysis. Mongrels and older beagles did have a 12-hr rhythmicity in plasma T₄ (P < 0.05), whereas 5 month old beagles had a circadian one (P < 0.01). A 12-hr rhythmicity was also found for T₃ in the older Beagles (P < 0.05). However, Fourier analysis indicated that the daily variation in T₄ and T₃ plasma levels was inadequately mathematically described by single sinusoidal rhythm and that more harmonic components are to be taken into account.

The obtained data during a 24-hr period indicate that T₄ and T₃ concentrations in plasma may vary according to breed, age and sampling hour.     

Characterization of Cre recombinase mouse lines enabling cell type-specific targeting of postnatal intervertebral discs

Cre/loxP technology is an important tool for studying cell type-specific gene functions. Cre recombinase mouse lines, including Agc1-CreER^T2, Col2a1-Cre; Col2a1-CreER^T2, Shh-Cre, Shh-CreER^T2, and Osx-Cre, have been proven to be valuable tools to elucidate the biology of long bones, yet the information for their activity in postnatal intervertebral disc (IVD) tissues was very limited. In this study, we used R26-mTmG fluorescent reporter to systematically analyze cell specificity and targeting efficiency of these six mouse lines in IVD tissues at postnatal growing and adult stages. We found that Agc1-CreER^T2 is effective to direct recombination in all components of IVDs, including annulus fibrosus (AF), nucleus pulposus (NP), and cartilaginous endplate (CEP), upon tamoxifen induction at either 2 weeks or 2 months of ages. Moreover, Col2a1-Cre targets most of the cells in IVDs, except for some cells in the outer AF (OAF) and NP. In contrast, the activity of Col2a1-CreER^T2 is mainly limited to the IAF of IVD tissues at either stage of tamoxifen injection. Similarly, Shh-Cre directs recombination specifically in all NP cells, whereas Shh-CreER^T2 is active only in a few NP cells when tamoxifen is administered at either stage. Finally, Osx-Cre targets cells in the CEP, but not in the NP or AF of IVDs tissues at these two stages. Thus, our data demonstrated that all these Cre lines can direct recombination in IVD tissues at postnatal stages with different cell type specificity and/or targeting efficiency, and can, therefore, serve as valuable tools to dissect cell type-specific gene functions in IVD development and homeostasis.     

Computation models simulating notochordal cell extinction during early ageing of an intervertebral disc

Lower back pain due to intervertebral disc (IVD) degeneration is a prevalent problem which drastically affects the quality of life of millions of sufferers. Healthy IVDs begin with high populations of notochordal cells in the nucleus pulposus, while by the second stage of degeneration, these cells will be replaced by chondrocyte-like cells. Because the IVD is avascular, these cells rely on passive diffusion of nutrients to survive. It is thought that this transition in cell phenotype causes the shift of the IVD's physical properties, which impede the flow of nutrients. Our computational model of the IVD illustrates its ability to simulate the evolving chemical and mechanical environments occurring during the early ageing process. We demonstrate that, due to the insufficient nutrient supply and accompanying changes in physical properties of the IVD, there was a resultant exponential decay in the number of notochordal cells over time.     

Expression of serglycin in human disc is increased in degenerated discs and up-regulated in vitro by exposure to IL-1ß or TNF-α

We investigated whether the multifunctional intercellular proteoglycan, serglycin, is expressed in human intervertebral disc cells and assessed its localization. We also investigated expression levels of serglycin in human annulus fibrosus (annulus) cells exposed to IL-1ß and TNF-α, which are two proinflammatory cytokines that are expressed during disc degeneration. Immunolocalization of serglycin was common in many cells of the human annulus, but less common in the nucleus pulposus (nucleus). Both intracellular and cell membrane localization were observed. Annulus cells from Thompson grades III, IV and V degenerated discs exhibited a 4.69 fold up-regulation in serglycin expression vs. cells from healthier grades I and II discs. In monolayer annulus cell culture, cells from more degenerated discs exhibited a 9.4 fold up-regulation of serglycin expression compared to cells from healthier discs. Exposure of cultured cells to IL-1ß or TNF-α caused significant up-regulation of serglycin expression. We found that serglycin expression increased with increasing disc degeneration both in vivo and in vitro, and also increased with exposure in vitro to IL-1ß and TNF-α.     

放射线引导下经椎间盘入路内脏大小神经毁损术治疗上腹部癌痛的临床分析

目的:观察放射线引导下经椎间盘入路毁损内脏神经治疗上腹部癌痛的疗效及安全性。方法:选择上腹部癌痛患者26例,在放射线引导下经T11~12椎间盘入路穿刺,注射无水乙醇5.0 mL毁损内脏大、小神经,观察穿刺成功率,并记录术前、术后1天、1周、2周、1月、2月的疼痛强度(NRS评分)、生活质量评分(QOL),阿片类药物的用量以及手术不良反应的发生情况。结果:所有患者均穿刺到位,无严重并发症出现。和术前相比,术后各时点的NRS评分、每日吗啡消耗量下降,QOL评分增加(P0.05)。其中,6例患者发生暂时性腹泻,一周内恢复;5例患者出现不同程度的腹背部烧灼感,未经特殊处理24 h后症状消失。结论:在放射线引导下经椎间盘入路毁损内脏神经治疗上腹部癌痛的操作简单,疗效好,可显著提高患者的生活质量,且安全性高。     

winged eye Induces Transdetermination of Drosophila Imaginal Disc by Acting in Concert with a Histone Methyltransferase,Su(var)3-9

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Plasma N-glycome composition associates with chronic low back pain

Background

Low back pain (LBP) is the symptom of a group of syndromes with heterogeneous underlying mechanisms and molecular pathologies, making treatment selection and patient prognosis very challenging. Moreover, symptoms and prognosis of LBP are influenced by age, gender, occupation, habits, and psychological factors. LBP may be characterized by an underlying inflammatory process. Previous studies indicated a connection between inflammatory response and total plasma N-glycosylation. We wanted to identify potential changes in total plasma N-glycosylation pattern connected with chronic low back pain (CLBP), which could give an insight into the pathogenic mechanisms of the disease.

Prey consumption and survival of the predatory mite,Amblydromalus limonicus,on different prey and host plants

Amblydromalus limonicus consumed fewer first instar Frankliniella occidentalis thrips than Bactericera cockerelli psyllids per day when on the same (1.7 thrips, 3.7 psyllids) or separate (2.9 thrips, 4.4 psyllids) capsicum leaf discs. Mites ate fewer psyllids per day on tomato (1.9) than on capsicum (3.1). Mite survival was similar on both prey and plants.