Absence of DICER in Monocytes and Its Regulation by HIV-1

MicroRNAs (miRNAs) are a class of small RNA molecules that function to control gene expression and restrict viral replication in host cells. The production of miRNAs is believed to be dependent upon the DICER enzyme. Available evidence suggests that in T lymphocytes, HIV-1 can both suppress and co-opt the host''s miRNA pathway for its own benefit. In this study, we examined the state of miRNA production in monocytes and macrophages as well as the consequences of viral infection upon the production of miRNA. Monocytes in general express low amounts of miRNA-related proteins, and DICER in particular could not be detected until after monocytes were differentiated into macrophages. In the case where HIV-1 was present prior to differentiation, the expression of DICER was suppressed. MicroRNA chip results for RNA isolated from transfected and treated cells indicated that a drop in miRNA production coincided with DICER protein suppression in macrophages. We found that the expression of DICER in monocytes is restricted by miR-106a, but HIV-1 suppressed DICER expression via the viral gene Vpr. Additionally, analysis of miRNA expression in monocytes and macrophages revealed evidence that some miRNAs can be processed by both DICER and PIWIL4. Results presented here have implications for both the pathology of viral infections in macrophages and the biogenesis of miRNAs. First, HIV-1 suppresses the expression and function of DICER in macrophages via a previously unknown mechanism. Second, the presence of miRNAs in monocytes lacking DICER indicates that some miRNAs can be generated by proteins other than DICER.     

Inactivation of TNF-α ameliorates diabetic neuropathy in mice

Tumor necrosis factor (TNF)-α is a potent proinflammatory cytokine involved in the pathogenesis of diabetic neuropathy. We inactivated TNF-α to determine if it is a valid therapeutic target for the treatment of diabetic neuropathy. We effected the inactivation in diabetic neuropathy using two approaches: by genetic inactivation of TNF-α (TNF-α(-/-) mice) or by neutralization of TNF-α protein using the monoclonal antibody infliximab. We induced diabetes using streptozotocin in wild-type and TNF-α(-/-) mice. We measured serum TNF-α concentration and the level of TNF-α mRNA in the dorsal root ganglion (DRG) and evaluated nerve function by a combination of motor (MNCV) and sensory (SNCV) nerve conduction velocities and tail flick test, as well as cytological analysis of intraepidermal nerve fiber density (IENFD) and immunostaining of DRG for NF-κB p65 serine-276 phosphorylated and cleaved caspase-3. Compared with nondiabetic mice, TNF-α(+/+) diabetic mice displayed significant impairments of MNCV, SNCV, tail flick test, and IENFD as well as increased expression of NF-κB p65 and cleaved caspase-3 in their DRG. In contrast, although nondiabetic TNF-α(-/-) mice showed mild abnormalities of IENFD under basal conditions, diabetic TNF-α(-/-) mice showed no evidence of abnormal nerve function tests compared with nondiabetic mice. A single injection of infliximab in diabetic TNF-α(+/+) mice led to suppression of the increased serum TNF-α and amelioration of the electrophysiological and biochemical deficits for at least 4 wk. Moreover, the increased TNF-α mRNA expression in diabetic DRG was also attenuated by infliximab, suggesting infliximab's effects may involve the local suppression of TNF-α. Infliximab, an agent currently in clinical use, is effective in targeting TNF-α action and expression and amelioration of diabetic neuropathy in mice.     

香港的生物多样性及其保育工作

香港位于热带,属海洋性气候。地势崎岖多山,山地约占全港总面积的3/4。城市发展多集中在沿海平坦地带。目前香港的城市和乡镇面积约占总面积的20%,农地约占5%(当中大部份已遭荒废),余下的均为郊野地区,这包括天然林和人工林(约占14%)、灌丛(约占36%)及草地(约占17%)。由于良好的气候和地理条件,形成了众多不同的生态环境,使总面积仅1090 km²的弹丸之地孕育出种类多样的动植物,生物多样性十分丰富。香港约有2500种原生植物,包括被子植物约1900种,裸子植物7种,蕨类植物220多种及苔藓植物300多种。动物方面,已记录的野生哺乳类动物有40多种,鸟类超过459种,两栖类23种,爬行类70多种。昆虫种类繁多,其中蜻蜓目100多种,鳞翅目2200多种(蝴蝶200多种,蛾类2000多种)。有很多是国家保护物种和特有种。植物方面属国家一级保护的有1种——刺桫椤(Alsophila spinulosa);国家二级保护的有6种,如四药门花(Tetrathryrium subcordatum);国家三级保护的有8种,如穗花杉(Amentotaxus argotaenia)。此外,香港特有种有16种,例如紫萁科(Osmundaceae)的粤紫萁(Osmunda mildei)、马兜铃科(Aristolochiaceae)的香港细辛(Asarum hongkongense)和兰科(Orchidaceae)的谢氏卷瓣兰(Bulbophyllum tseanum)。动物方面有9种属国家一级保护,例如中华白海豚(Sousa chinensis);79种属国家二级保护。特有种则有卢氏小树蛙(Philautus romeri)、包氏双足蜥(Dibamus bogadeki)及多种昆虫。为了保护丰富的野生动植物及其栖息的环境,香港特别行政区政府制定了一些法例并推行了不少保护措施,例如设立了21个郊野公园和14个特别地区,占全港陆地总面积约38%。此外,还成立了2个禁区、3个海岸公园和1个海洋保护区。另一方面,政府还设立了59个“具特殊科学价值地点”,以保护及研究各种动植物、生态系统和特殊的地质地貌。香港地少人多,总人口超过600万,是世界上人口最稠密的地区之一。多年来香港这个生物宝库不断地遭受人类活动的威胁,近年来由于人口急剧上升,对土地需求迫切,不少郊野地区被开发利用,环境污染亦日益严重。此外,一些野生植物因具有药用价值、观赏价值或其他用途而遭盗伐或采集。上述种种因素已使香港野生动植物及其生境受到严重损害,一些物种更濒于灭绝,进行生物多样性的保育工作刻不容缓。因此,香港确实需要制定整体的生物多样性保护策略。有鉴于此,香港大学生态及分类学系于1996年展开了一项为期3年的香港生物多样性调查,以增加对动植物资源现况的了解,为保护香港的珍稀濒危物种和日益恶化的自然环境提出补救方案,并为制订长远的保育策略奠定基础。     

Piwi-Interacting RNAs (piRNAs) Are Dysregulated in Renal Cell Carcinoma and Associated with Tumor Metastasis and Cancer-Specific Survival

Piwi-interacting RNAs (piRNAs) are a distinct group of small noncoding RNAs (sncRNAs) that silence transposable genetic elements to protect genome integrity. Because of their limited expression in gonads and sequence diversity, piRNAs remain the most mysterious class of small RNAs. Studies have shown piRNAs are present in somatic cells and dysregulated in gastric, breast and liver cancers. By deep sequencing 24 frozen benign kidney and clear cell renal cell carcinoma (ccRCC) specimens and using the publically available piRNA database, we found 26,991 piRNAs present in human kidney tissue. Among 920 piRNAs that had at least two copies in one specimen, 19 were differentially expressed in benign kidney and ccRCC tissues, and 46 were associated with metastasis. Among the metastasis-related piRNAs, we found three piRNAs (piR-32051, piR-39894 and piR-43607) to be derived from the same piRNA cluster at chromosome 17. We confirmed the three selected piRNAs not to be miRNAs or miRNA-like sncRNAs. We further validated the aberrant expression of the three piRNAs in a 68-case formalin-fixed and paraffin-embedded (FFPE) ccRCC tissue cohort and showed the up-regulation of the three piRNAs to be highly associated with ccRCC metastasis, late clinical stage and poor cancer-specific survival.     

EARLY STAGES IN THE DEVELOPMENT OF PLASTID FINE STRUCTURE IN RED AND FAR-RED LIGHT

Developmental changes in fine structure were studied in plastids of etiolated bean leaves during the time required for the protochlorophyllide-chlorophyllide transformation and the following lag phase prior to chlorophyll accumulation. In agreement with some other workers, two distinct stages of change in the fine structure of proplastids were found to occur upon illumination during this period. The first involves a dissociation of the previously fused units in the prolamellar bodies of the proplastids and occurs simultaneously with the protochlorophyllide-chlorophyllide conversion in light of 655 mµ, but not of 682, 700, or 730 mµ. The effect of the red light could not be reversed by a simultaneously supplied stronger far-red irradiation. The energy requirements for these structural changes parallel those for the pigment conversion. During the following step the vesicles which arose from the fused units of the prolamellar body were dispersed in rows through the stroma, and the prolamellar bodies themselves disappeared. For these changes to occur, higher light energies were required and the leaves had to be illuminated for longer periods. A red preillumination seemed to accelerate the development somewhat. The structural changes could be induced by light of 655 mµ, but also, to a lesser degree, of 730 mµ. No measurable additional chlorophyll accumulated during this period. Thus, the structural changes observed were independent of major changes in pigment content.     

Recovery of novel bacterial diversity from a forested wetland impacted by reject coal

Sulphide mineral mining together with improperly contained sulphur-rich coal represents a significant environmental problem caused by leaching of toxic material. The Savannah River Site's D-area harbours a 22-year-old exposed reject coal pile (RCP) from which acidic, metal rich, saline runoff has impacted an adjacent forested wetland. In order to assess the bacterial community composition of this region, composite sediment samples were collected at three points along a contamination gradient (high, middle and no contamination) and processed for generation of bacterial and archaeal 16S rDNA clone libraries. Little sequence overlap occurred between the contaminated (RCP samples) and unimpacted sites, indicating that the majority of 16S rDNAs retrieved from the former represent organisms selected by the acidic runoff. Archaeal diversity within the RCP samples consisted mainly of sequences related to the genus Thermoplasma and to sequences of a novel type. Bacterial RCP libraries contained 16S rRNA genes related to isolates (Acidiphilium sp., Acidobacterium capsulatum, Ferromicrobium acidophilium and Leptospirillum ferrooxidans) and environmental clones previously retrieved from acidic habitats, including ones phylogenetically associated with organisms capable of sulphur and iron metabolism. These libraries also exhibited particularly novel 16S rDNA types not retrieved from other acid mine drainage habitats, indicating that significant diversity remains to be detected in acid mine drainage-type systems.     

The roles of geological history and colonization abilities in genetic differentiation between mammalian populations in the Philippine archipelago

Aim To test hypotheses that: (1) late Pleistocene low sea‐level shorelines (rather than current shorelines) define patterns of genetic variation among mammals on oceanic Philippine islands; (2) species‐specific ecological attributes, especially forest fidelity and vagility, determine the extent to which common genetic patterns are exhibited among a set of species; (3) populations show reduced within‐population variation on small, isolated oceanic islands; (4) populations tend to be most highly differentiated on small, isolated islands; and (5) to assess the extent to which patterns of genetic differentiation among multiple species are determined by interactions of ecological traits and geological/geographic conditions. Location The Philippine Islands, a large group of oceanic islands in Southeast (SE) Asia with unusually high levels of endemism among mammals. Methods Starch‐gel electrophoresis of protein allozymes of six species of small fruit bats (Chiroptera, Pteropodidae) and one rodent (Rodentia, Muridae). Results Genetic distances between populations within all species are not correlated with distances between present‐day shorelines, but are positively correlated with distances between shorelines during the last Pleistocene period of low sea level; relatively little intraspecific variation was found within these ‘Pleistocene islands’. Island area and isolation of oceanic populations have only slight effects on standing genetic variation within populations, but populations on some isolated islands have heightened levels of genetic differentiation, and reduced levels of gene flow, relative to other islands. Species associated with disturbed habitat (all of which fly readily across open habitats) show more genetic variation within populations than species associated with primary rain forest (all of which avoid flying out from beneath forest canopy). Species associated with disturbed habitats, which tend to be widely distributed in SE Asia, also show higher rates of gene flow and less differentiation between populations than species associated with rain forest, which tend to be Philippine endemic species. One rain forest bat has levels of gene flow and heterozygosity similar to the forest‐living rodent in our study. Main conclusions The maximum limits of Philippine islands that were reached during Pleistocene periods of low sea level define areas of relative genetic homogeneity, whereas even narrow sea channels between adjacent but permanently isolated oceanic islands are associated with most genetic variation within the species. Moreover, the distance between ‘Pleistocene islands’ is correlated with the extent of genetic distances within species. The structure of genetic variation is strongly influenced by the ecology of the species, predominantly as a result of their varying levels of vagility and ability to tolerate open (non‐forested) habitat. Readily available information on ecology (habitat association and vagility) and geological circumstances (presence or absence of Pleistocene land‐bridges between islands, and distance between oceanic islands during periods of low sea level) are combined to produce a simple predictive model of likely patterns of genetic differentiation (and hence speciation) among these mammals, and probably among other organisms, in oceanic archipelagos.