Regrow or repair: Potential regenerative therapies for the kidney

Regenerative medicine is being heralded in a similar way as gene therapy was some 15 yr ago. it is an area of intense excitement and potential, as well as myth and disinformation. However, with the increasing rate of end-stage renal failure and limited alternatives for its treatment, we must begin to investigate seriously potential regenerative approaches for the kidney. This review defines which regenerative options there might be for renal disease, summarizes the progress that has been made to date, and investigates some of the unique obstacles to such treatments that the kidney presents. The options discussed include in situ organ repair via bone marrow recruitment or dedifferentiation; ex vivo stem cell therapies, including both autologous and nonautologous options; and bioengineering approaches for the creation of a replacement organ.

Reducing seed viability of flaxleaf fleabane, feathertop Rhodes grass and awnless barnyard grass

In the sub-tropical grain region of Australia, cotton and grains systems are now dominated by flaxleaf fleabane (Conyza bonariensis (L.) Cronquist), feathertop Rhodes grass (Chloris virgata Sw.) and awnless barnyard grass (Echinochloa colona (L.) Link). While control of these weed species is best achieved when they are young, previous studies have shown a potential for reducing seed viability and minimising seed bank replenishment by applying herbicides when plants are reproductive. Pot trials were established over two growing seasons to examine the effects of 2,4-D, 2,4-D + picloram, glyphosate and glufosinate which had been successful on other species, along with paraquat and haloxyfop (grasses only). Herbicides were applied at ¾ field rates in an attempt not to kill the plants. Flaxleaf fleabane plants were sprayed at two growth stages (budding and flowering) and the grasses were sprayed at two stages (late tillering/booting and flowering). Spraying flaxleaf fleabane at flowering reduced seed viability to 0% (of untreated) in all treatments except glyphosate (51%) and 2,4-D + picloram (8%). Seed viability was not reduced with the first and second regrowths with the exception of 2,4-D + picloram where viability was reduced to 20%. When sprayed at budding only 2,4-D + picloram reduced seed viability in both trials. Spraying the grasses at late tillering/booting did not reduce viability except for glufosinate on awnless barnyard grass (50%). Applying herbicides at flowering resulted in 0% seed viability in awnless barnyard grass from glufosinate, paraquat and glyphosate and 0% viability in feathertop Rhodes grass for glufosinate. These herbicides were less effective on heads that emerged and flowered after spraying, only slightly reducing seed viability. These trials have shown that attempts to reduce seed viability have potential, however flaxleaf fleabane and feathertop Rhodes grass are able to regrow and will need on-going monitoring and control measures.

不同环境紫茎泽兰幼苗动态和特征

紫茎泽兰（Eupatorium adenophorum Spreng.）入侵我国已有70余年的时间，目前已经对我国西南地区造成了严重危害，却缺少有效的防治方法加以控制。本研究着眼于紫茎泽兰治理效果，在两片大面积综合防治样地的基础上，针对综合防治后样地紫茎泽兰再生长情况展开调查研究，并在野外观测的基础上设计两项置于样地附近的盆栽实验，研究紫茎泽兰在不同环境条件下的存活情况，以及与不同物种混播的种间竞争情况。 对综合治理1年后的样地中4个紫茎泽兰种群调查、分析结果表明，在不同的生境，采取不同的方法治理紫茎泽兰会导致紫茎泽兰采取不同的生长和繁殖对策进行再生长。紫茎泽兰生长速度为：水分充足的环境大于水分不充足的环境，萌生植株大于实生植株。种植替代物种绞股蓝（Gynostmma pentaphyllum）能够抑制紫茎泽兰的高生长，但在绞股蓝的压迫下紫茎泽兰长成L型，反而增加了冠幅、分蘖数目和单位株高的分枝数。在繁殖方式方面：采用拔除方法治理的区域紫茎泽兰以克隆繁殖为主，而采用喷洒化学除草剂治理的区域则以有性繁殖为主。 固定样方调查显示，紫茎泽兰再生长的1年当中，群落随季节变化而变。群落的盖度、密度和物种丰富度均随着旱季的深入而达到最低，第二年雨季到来后上升，紫茎泽兰在群落中处于绝对优势，尤其是在旱季优势更为明显。一年生紫茎泽兰在旱季部分植株死亡（死亡率为53.38±1.55%），少部分（5.66±0.45%）开花，产生种子的密度约为50 000ind∙m-2。与本地优势灌木物种车桑子（Dodonaea viscosa）、台湾相思（Acacia confuse）和马桑（Coriaria nepalensis）相比，紫茎泽兰具有更快的生长速度，更大的高度、密度和盖度。 紫茎泽兰种子的萌发需要充足的水分和遮荫条件。在盆栽实验当中，浇水或遮荫条件下足量紫茎泽兰的种子能够萌发更多的幼苗，而且幼苗的死亡率也比无浇水或无遮荫的处理低。在遮荫或去除其它物种竞争的条件下，紫茎泽兰幼苗生长速度更快。只要水分充足紫茎泽兰幼苗不会因拥挤而死亡，但是个体平均生物量会因拥挤而减小。 紫茎泽兰幼苗与本地3种草本植物比较结果，在未遮荫条件下，紫茎泽兰幼苗数量极少，且个体也非常小，而相同条件下荩草（Arthraxon hispidus）和含羞草决明（Cassia mimosoides）生长良好;在遮荫条件下荩草、含羞草决明和戟叶酸模（Rumex haxtatus ）与紫茎泽兰相比则几乎没有优势。 成株紫茎泽兰生命力强，难以控制，幼苗时期是采用替代控制方法治理紫茎泽兰的最佳时期。在替代控制物种选择方面，可以在根除后播种本地灌木和草本植物，因为草本植物在早期可以抑制紫茎泽兰幼苗的萌发和生长，而同时播种的灌木种类则可以在植被恢复的后期起到控制紫茎泽兰再生长的重要作用。

REGROWTH RATES OF AMORPHOUS LAYERS IN SILICON-ON-SAPPHIRE FILMS.

The rate and direction of regrowth of amorphous layers, created by self-implantation, in silicon-on-sapphire (SOS) have been studied using time resolved reflectivity (TRR) experiments performed simultaneously at two wavelengths. Regrowth of an amorphous layer towards the surface was observed in specimens implanted with 3 multiplied by (times) 10**1**5Si** plus /cm**2 at 50keV and regrowth of a buried amorphous layer, from a surface seed towards the sapphire, was observed in specimens implanted with 1 multiplied by (times) 10**1**5Si** plus /cm**2 at 175keV. Rapid isothermal heating to regrow the layers was performed in an electron beam annealing system. The combination of 514. 5nm and 632. 8nm wavelengths was found to be particularly useful for TRR studies since the high absorption in amorphous silicon, at the shorter wavelength, means that the TRR trace is not complicated by reflection from the silicon-sapphire interface until regrowth is nearly complete.

STRUCTURAL DEFECTS AND THEIR ELECTRICAL-ACTIVITY IN GERMANIUM IMPLANTED SILICON

The electrical and structural characteristics of secondary defects in regrown amorphous layers formed in n-type Si(100) with a resistivity of 2 OMEGA cm and 6 OMEGA cm using Ge+ ions, has been studied. The amorphous layers with a thickness of 460 nm are formed by implantation of 1 x 10(15) Ge+ cm-2 at an energy of 400 keV. Both conventional furnace and rapid thermal annealing were used to regrow the amorphous layer and the residual defects have been characterised in terms of their concentration depth distribution and activation energies using C-V and DLTS. Structural information has been obtained from RBS and XTEM. By choosing suitable anneal conditions it is possible to eliminate extended defects, apart from a low concentration of end of range dislocation loops. However, a substantial population of electrically active point defects remain after simple low thermal budget anneals. In a sample implanted with 1 x 10(15) Ge+ cm-2 at 400 keV a region of deep donors approximately 460 nm from the surface is always present When the samples are annealed at higher temperatures (> 850-degrees the total deep donor concentration is reduced by one order of magnitude. Other electrically active defects not observable in the low (750-degrees-C) temperature annealed layers become apparent during anneals at intermediate temperatures.

Cysteine proteinase-1 and cut protein isoform control dendritic innervation of two distinct sensory fields by a single neuron.

Dendrites often exhibit structural changes in response to local inputs. Although mechanisms that pattern and maintain dendritic arbors are becoming clearer, processes regulating regrowth, during context-dependent plasticity or after injury, remain poorly understood. We found that a class of Drosophila sensory neurons, through complete pruning and regeneration, can elaborate two distinct dendritic trees, innervating independent sensory fields. An expression screen identified Cysteine proteinase-1 (Cp1) as a critical regulator of this process. Unlike known ecdysone effectors, Cp1-mutant ddaC neurons pruned larval dendrites normally but failed to regrow adult dendrites. Cp1 expression was upregulated/concentrated in the nucleus during metamorphosis, controlling production of a truncated Cut homeodomain transcription factor. This truncated Cut, but not the full-length protein, allowed Cp1-mutant ddaC neurons to regenerate higher-order adult dendrites. These results identify a molecular pathway needed for dendrite regrowth after pruning, which allows the same neuron to innervate distinct sensory fields.

Thresholds for irreversible decline of the Greenland ice sheet

The Greenland ice sheet will decline in volume in a warmer climate. If a sufficiently warm climate is maintained for a few thousand years, the ice sheet will be completely melted. This raises the question of whether the decline would be reversible: would the ice sheet regrow if the climate cooled down? To address this question, we conduct a number of experiments using a climate model and a high-resolution ice-sheet model. The experiments are initialised with ice sheet states obtained from various points during its decline as simulated in a high-CO2 scenario, and they are then forced with a climate simulated for pre-industrial greenhouse gas concentrations, to determine the possible trajectories of subsequent ice sheet evolution. These trajectories are not the reverse of the trajectory during decline. They converge on three different steady states. The original ice-sheet volume can be regained only if the volume has not fallen below a threshold of irreversibility, which lies between 80 and 90% of the original value. Depending on the degree of warming and the sensitivity of the climate and the ice-sheet, this point of no return could be reached within a few hundred years, sooner than CO2 and global climate could revert to a pre-industrial state, and in that case global sea level rise of at least 1.3 m would be irreversible. An even larger irreversible change to sea level rise of 5 m may occur if ice sheet volume drops below half of its current size. The set of steady states depends on the CO2 concentration. Since we expect the results to be quantitatively affected by resolution and other aspects of model formulation, we would encourage similar investigations with other models.

Enhanced antitumor efficacy of a herpes simplex virus mutant isolated by genetic selection in cancer cells

Replication-competent, attenuated herpes simplex virus-1 (HSV-1) derivatives that contain engineered mutations into the viral γ34.5 virulence gene have been used as oncolytic agents. However, as attenuated mutants often grow poorly, they may not completely destroy some tumors and surviving cancer cells simply regrow. Thus, although HSV-1 γ34.5 mutants can reduce the growth of human tumor xenografts in mice and have passed phase I safety studies, their efficacy is limited because they replicate poorly in many human tumor cells. Previously, we selected for a γ34.5 deletion mutant variant that regained the ability to replicate efficiently in tumor cells. Although this virus contains an extragenic suppressor mutation that confers enhanced growth in tumor cells, it remains attenuated. Here, we demonstrate that the suppressor virus replicates to greater levels in prostate carcinoma cells and, importantly, is a more potent inhibitor of tumor growth in an animal model of human prostate cancer than the γ34.5 parent virus. Thus, genetic selection in cancer cells can be used as a tool to enhance the antitumor activity of a replication-competent virus. The increased therapeutic potency of this oncolytic virus may be useful in the treatment of a wide variety of cancers.

Intrinsic changes in developing retinal neurons result in regenerative failure of their axons.

The failure of mature mammalian central nervous system axons to regenerate after transection is usually attributed to influences of the extraneuronal milieu. Using explant cocultures of retina and midbrain tectum from hamsters, we have found evidence that these influences account for failure of regrowth of only a small minority of retinal axons. For most of the axons, there is a programmed loss of ability to elongate in the central nervous system. We show that there is a precipitous decline in the ability of retinal axons to reinnervate tectal targets when the retina is derived from pups on or after postnatal day 2, even when the target is embryonic. By contrast, embryonic retinal axons can regrow into tectum of any age, overcoming growth-inhibiting influences of glial factors.

Pathways to improving skin regeneration

A significant proportion of the human population suffers from some form of skin disorder, whether it be from burn injury or inherited skin anomalies. The ideal treatment for skin disorders would be to regrow skin tissue from stem cells residing in the individual patient's skin. Locating these adult stem cells and elucidating the molecules involved in orchestrating the production of new skin cells are important steps in devising more-efficient methods of skin production and wound healing via the ex vivo expansion of patient keratinocytes in culture. This review focuses on the structure of the skin, the identification of skin stem cells, and the role of Notch, Wnt and Hedgehog signalling cascades in regulating the fate of epidermal stem cells. © 2005 Cambridge University Press.

Effects of high temperature on the growth and composition of sugarcane internodes

Sugarcane grown in the Ord River district of Western Australia has lower sucrose content than expected from earlier trials and experience in other irrigated districts. High temperatures have been hypothesised as a possible cause. The effects of high temperature (above 32 degrees C) on growth and carbon partitioning were investigated. A temperature regime of (25-38 degrees C) was compared with (23-33 degrees C). In one experiment, 7-month-old plants of cvv. Q117 and Q158 were subjected to the treatments for 2 months. In another experiment, the plants were allowed to regrow (ratoon) for 6 months. In both experiments, the higher temperature resulted in more, shorter internodes and higher moisture content. Most internodes from plants in the higher temperature treatment had lower sucrose content than internodes from the lower temperature. On a dry mass basis the internodes from the plants in the higher temperature had proportionately more fibre and hexoses but lower sucrose. Combined with an increased number of nodes in a stem of similar or shorter length this would result in higher stalk fibre and lower sucrose content. The data provided evidence that sugarcane partitions less carbon to stored sucrose when grown under high compared with low temperatures. The two cultivars partitioned carbon between soluble (sugars) and insoluble (fibre) fractions to different degrees. These experiments also indicate that the current models describing leaf appearance and perhaps sugarcane growth at temperatures above 32 degrees C, in general, need revision.