The co-distribution of Plasmodium falciparum and hookworm among African schoolchildren

Background

On the island of Bioko (Equatorial Guinea), insecticide-treated nets (ITNs) have been the main tool used to control malaria over the last 13 years. In 2004, started an indoor residual spraying (IRS) campaign to control malaria. The purpose of this study is to asses the impact of the two control strategies on the island of Bioko (Equatorial Guinea), with regards to Plasmodium infection and anaemia in the children under five years of age.

Methods

Two transversal studies, the first one prior to the start of the IRS campaign and the second one year later. Sampling was carried out by stratified clusters. Malaria infection was measured by means of thick and thin film, and the packed cell volume (PCV) percentage. Data related to ITN use and information regarding IRS were collected. The Pearson's chi-square and logistic regression statistical tests were used to calculate odds ratios (OR)

Results

In the first survey, 168 children were sampled and 433 children in the second one. The prevalence of infection was 40% in 2004, and significantly lower at 21.7% in 2005. PCV was 41% and 39%, respectively. 58% of the children surveyed in 2004 and 44.3% in 2005 had slept under an ITN. 78% of the dwellings studied in 2005 had been sprayed. In the 2005 survey, sleeping without a mosquito net meant a risk of infection 3 times greater than sleeping protected with a net hanged correctly and with no holes (p < 0.05).

Conclusion

IRS and ITNs have proven to be effective control strategies on the island of Bioko. The choice of one or other strategy is, above all, a question of operational feasibility and availability of local resources.     

Plant Vascular Biology and Agriculture

The evolution of animal and plant vascular systems played a pivotal role in the advancement from simple to complex organisms, through the provision of a delivery system for the distribution     

不同术式的经内镜下行乳头括约肌切开术（EST）对肠胆反流的影响

目的：目前认为,十二指肠胆道反流是引起胆道反复感染,进而导致胆道结石再发和胆道狭窄的原因之一。近年来应用以内镜下逆行胆胰管造影术（endoscopicretrogradecholangiopancreatography,ERCP）为基础的微创治疗胆总管结束的技术开展颇为广泛。它主要包括ERCP、Oddi括约肌切开术（endoscopicsphincterotomy,EST）、十二指肠乳头球囊扩张术（endoscopicpapillarybal—Iondilation,EPBD）、胆管结石碎石取石术、胆总管支架植入术和鼻胆管引流术六大技术。本文主要研究了采用不同术式的EST,即EST中切口和EST小切口＋EPBD术,在术后早期对患者十二指肠胆道反流的影响。方法：63例胆总管结石患者,男30例,女33例,予行经内镜下逆行的胆胰管造影（ERCP）后分别采用不同术式EST,术后安放胆总管引流管。术后l周留取胆汁采用口服核素和测定胆汁中的胃蛋白酶I、II的浓度,对十二指肠胆道反流进行定量和定性的测定。结果：EST中切口术组、EST小切口＋球囊扩张（EPBD）组分别与无EST组相比,年龄和性别无统计学意义（P=0．07,P=0．416）。行EST中切开和小切开＋球囊扩张患者胆汁中的锝计数明显高于无EST组,且这两组不同术式的患者锝计数存在显著的统计学差异（P〈0．05）。行EST中切口者、EST小切口＋球囊扩张术者胆汁中的PGII质量浓度明显低于无EST组（P〈0．05）,但是EST中切口者和EST小切口＋球囊扩张术后两组间胆汁中PGII的质量浓度无统计学差异。结论：行EST中切口取胆总管结石的患者在手术早期较易发生十二指肠胆道的反流。因此,建议对于胆总管结石患者尽量选择行EST小切口＋球裳扩张术（EPBD）的手术方式。     

苎麻光合生理生态特性研究

以大田栽培的苎麻植株为材料,用TPS-2便携式光合作用测定系统测定自然条件下生长的苎麻叶片的光合气体交换参数,以及光响应曲线和CO2响应曲线,并通过回归和相关法分析探讨净光合速率与主要生理、生态因子间的关系.结果表明:(1)苎麻叶片的净光合速率(Pn)日变化曲线呈现双峰型,2个光合峰高度接近,其净光合速率具有典型的午休"现象;蒸腾速率(Tr)日变化曲线呈现单峰型,其走势与气孔导度(Gs)日变化一致.(2)苎麻叶片光合作用的光饱和点(LSP)为1 568.5μmol?m-2?s-1,光补偿点(LCP)为54.18μmol?m-2?s-1,表观量子效率(AQY)为0.025 8 mol?mol-1;而其CO2补偿点(CCP)、饱和点(CSP)和羧化效率(CE)分别为49.25、1 746.9μmol?mol-1和0.045;因此,苎麻属于喜光性阳生植物,且对强光有一定的耐受能力.(3)苎麻叶片Pn日变化的主要限制因子是胞间CO2浓度(Ci),主要决定生理因子是气孔导度(Gs).     

Serpina3n accelerates tissue repair in a diabetic mouse model of delayed wound healing

Chronic, non-healing wounds are a major complication of diabetes and are characterized by chronic inflammation and excessive protease activity. Although once thought to function primarily as a pro-apoptotic serine protease, granzyme B (GzmB) can also accumulate in the extracellular matrix (ECM) during chronic inflammation and cleave ECM proteins that are essential for proper wound healing, including fibronectin. We hypothesized that GzmB contributes to the pathogenesis of impaired diabetic wound healing through excessive ECM degradation. In the present study, the murine serine protease inhibitor, serpina3n (SA3N), was administered to excisional wounds created on the dorsum of genetically induced type-II diabetic mice. Wound closure was monitored and skin wound samples were collected for analyses. Wound closure, including both re-epithelialization and contraction, were significantly increased in SA3N-treated wounds. Histological and immunohistochemical analyses of SA3N-treated wounds revealed a more mature, proliferative granulation tissue phenotype as indicated by increased cell proliferation, vascularization, fibroblast maturation and differentiation, and collagen deposition. Skin homogenates from SA3N-treated wounds also exhibited greater levels of full-length intact fibronectin compared with that of vehicle wounds. In addition, GzmB-induced detachment of mouse embryonic fibroblasts correlated with a rounded and clustered phenotype that was prevented by SA3N. In summary, topical administration of SA3N accelerated wound healing. Our findings suggest that GzmB contributes to the pathogenesis of diabetic wound healing through the proteolytic cleavage of fibronectin that is essential for normal wound closure, and that SA3N promotes granulation tissue maturation and collagen deposition.Diabetic skin ulcers are non-healing, chronic wounds that pose a major health burden to society.¹ Up to a quarter of diabetic patients will develop these ulcers in their lifetime and as a result, nearly a fifth of these diabetic patients will require non-traumatic lower limb amputations.² Numerous advanced treatment options for the management of diabetic ulcers have been explored, including bioengineered skin substitutes, hyperbaric oxygen therapy and negative pressure dressings.^{3, 4, 5} However, they have largely been unsuccessful. As such, a combination of lifestyle modification, pressure off-loading, local surgical or larval debridement and infection control continue to be the standard recommended treatment strategy.⁶Wound healing is a complex process that involves overlapping and sequential phases involving haemostasis, inflammation, granulation tissue formation and tissue remodelling. For wounds to heal timely and properly, there must be a fine balance of interaction between various cell types, cytokines, growth factors, proteases and extracellular matrix (ECM) components. However, in diabetic patients, the normal continuum of wound healing is disrupted, and wounds enter a chronic, non-healing state characterized by persistent inflammation, enhanced proteolytic activity and impaired ECM deposition.⁷ The roles of various proteases, primarily matrix metalloproteinases (MMPs), have been extensively studied. Although MMPs were once believed to be the major culprits in impaired wound healing, it is now recognized that they are essential for normal wound healing by modulating inflammation, angiogenesis and tissue remodelling.^{8, 9, 10} Therefore, other proteases may also contribute to the pathogenesis of chronic wound healing.Granzyme B (GzmB) is a cytotoxic serine protease that is often viewed exclusively as a pro-apoptotic serine protease that is released from cytotoxic lymphocytes, along with the pore-forming, molecule perforin, to induce cell death.¹¹ However, because GzmB can be induced in other types of immune and non-immune cells that often do not express perforin and/or do not form immunological synapses with target cells, there is emerging evidence supporting the paradigm that GzmB can accumulate and function in the extracellular milieu.¹² In support, many ECM proteins are GzmB substrates and the consequences of such cleavage may be implicated in many diseases associated with aging and/or chronic inflammation such as abdominal aortic aneurysm (AAA), skin aging, atherosclerosis and wound healing using GzmB knockout mice.^{13, 14, 15, 16} Fibronectin is one such ECM glycoprotein that has an important role in cell attachment, differentiation and migration during wound healing and is cleaved by GzmB.¹⁷The serine protease inhibitors, also known as serpins, are the largest protease inhibitor super family and are divided into 16 clades.¹⁸ Serpina3n (SA3N), which is part of the SERPINA clade, is the mouse orthologue of the human anti-chymotrypsin (ACT) and has been identified as an inhibitor of both human and mouse GzmB in vitro.^{19, 20, 21} The human ACT is encoded by only one gene; however, extensive diversification and duplication in mice have resulted in 13 related serpina genes clustered at chromosome 12F1.¹⁹ Out of these, SA3N was identified to be the only extracellular inhibitor of GzmB.²¹ A previous study by our group had demonstrated efficacy for SA3N as an in vivo GzmB inhibitor in a murine model of AAA.²⁰The purpose of this study was to determine whether local inhibition of extracellular GzmB could accelerate wound closure in a genetically induced type-II diabetic mouse model of delayed wound healing. As many of the proteins in the ECM involved in wound healing are subject to cleavage by GzmB, we propose that SA3N accelerates wound healing by inhibiting protease-related ECM degradation.     

E. coli chaperones DnaK,Hsp33 and Spy inhibit bacterial functional amyloid assembly

Amyloid formation is an ordered aggregation process, where β-sheet rich polymers are assembled from unstructured or partially folded monomers. We examined how two Escherichia coli cytosolic chaperones, DnaK and Hsp33, and a more recently characterized periplasmic chaperone, Spy, modulate the aggregation of a functional amyloid protein, CsgA. We found that DnaK, the Hsp70 homolog in E. coli, and Hsp33, a redox-regulated holdase, potently inhibited CsgA amyloidogenesis. The Hsp33 anti-amyloidogenesis activity was oxidation dependent, as oxidized Hsp33 was significantly more efficient than reduced Hsp33 at preventing CsgA aggregation. When soluble CsgA was seeded with preformed amyloid fibers, neither Hsp33 nor DnaK were able to efficiently prevent soluble CsgA from adopting the amyloid conformation. Moreover, both DnaK and Hsp33 increased the time that CsgA was reactive with the amyloid oligomer conformation-specific A11 antibody. Since CsgA must also pass through the periplasm during secretion, we assessed the ability of the periplasmic chaperone Spy to inhibit CsgA polymerization. Like DnaK and Hsp33, Spy also inhibited CsgA polymerization in vitro. Overexpression of Spy resulted in increased chaperone activity in periplasmic extracts and in reduced curli biogenesis in vivo. We propose that DnaK, Hsp33 and Spy exert their effects during the nucleation stages of CsgA fibrillation. Thus, both housekeeping and stress induced cytosolic and periplasmic chaperones may be involved in discouraging premature CsgA interactions during curli biogenesis.Key words: chaperone, curli, functional amyloid, CsgA, DnaK, Hsp33, Spy     

核心组蛋白的融合表达和纯化

目的:克隆核心组蛋白H2A、H2B、H3和H4的基因,表达并纯化组蛋白与谷胱甘肽S-转移酶(GST)的融合蛋白。方法:用PCR方法从乳腺文库中扩增核心组蛋白H2A、H2B、H3和H4的编码序列,分别将其以正确相位与pGEX-KG载体中的GST编码序列融合,得到重组质粒pGST-H2A、pGST-H2B、pGST-H3和pGST-H4,分别转化大肠杆菌BL21,表达融合蛋白GST-H2A、GST-H2B、GST-H3和GST-H4;用谷胱甘肽-Sepharose 4B亲和纯化融合蛋白;用Western印迹检测融合蛋白的表达及纯化。结果:分别构建了核心组蛋白H2A、H2B、H3和H4的融合表达载体;Western印迹检测表明,融合蛋白GST-H2A、GST-H2B、GST-H3和GST-H4获得表达及纯化。结论:表达并纯化了H2A、H2B、H3和H4的融合蛋白,为进一步研究核心组蛋白的功能奠定了基础。     

Granzyme B cleaves decorin, biglycan and soluble betaglycan, releasing active transforming growth factor-β1

Objective

Granzyme B (GrB) is a pro-apoptotic serine protease that contributes to immune-mediated target cell apoptosis. However, during inflammation, GrB accumulates in the extracellular space, retains its activity, and is capable of cleaving extracellular matrix (ECM) proteins. Recent studies have implicated a pathogenic extracellular role for GrB in cardiovascular disease, yet the pathophysiological consequences of extracellular GrB activity remain largely unknown. The objective of this study was to identify proteoglycan (PG) substrates of GrB and examine the ability of GrB to release PG-sequestered TGF-β1 into the extracellular milieu.

Methods/Results

Three extracellular GrB PG substrates were identified; decorin, biglycan and betaglycan. As all of these PGs sequester active TGF-β1, cytokine release assays were conducted to establish if GrB-mediated PG cleavage induced TGF-β1 release. Our data confirmed that GrB liberated TGF-β1 from all three substrates as well as from endogenous ECM and this process was inhibited by the GrB inhibitor 3,4-dichloroisocoumarin. The released TGF-β1 retained its activity as indicated by the induction of SMAD-3 phosphorylation in human coronary artery smooth muscle cells.

Conclusion

In addition to contributing to ECM degradation and the loss of tissue structural integrity in vivo, increased extracellular GrB activity is also capable of inducing the release of active TGF-β1 from PGs.