昆虫差异蛋白质组: 进展和展望

黑腹果蝇Drosophila melanogaster、家蚕Bombyx mori、意大利蜜蜂Apis mellifera和冈比亚按蚊Anopheles gambiae等昆虫的基因组测序已经基本完成,蛋白质组学技术将是阐明这些基因组功能的重要工具。本文综述了应用差异蛋白质组学技术在昆虫诱导性免疫、抗性机制和分子病理研究方面取得的一些成果：（1） 细菌、真菌、脂多糖或机械损伤诱导的昆虫血淋巴或血细胞来源细胞系的蛋白质表达的改变; （2） Bt抗性昆虫中肠刷状缘膜和抗锥虫采采蝇Glossina morsitans唾液腺多种蛋白质表达的改变; （3）化学农药处理和姬蜂Chelonus inanitus携带的多分DNA病毒引起的昆虫蛋白质组的变化。最后讨论了昆虫差异蛋白质组学研究的瓶颈与对策。     

苜蓿假盘菌子囊孢子萌发特性及子实体超微结构观察

【背景】苜蓿假盘菌(Pseudopeziza medicaginis)侵染苜蓿叶片引起的褐斑病是苜蓿的重要病害之一。研究表明,该菌以子囊盘越冬,翌年释放子囊孢子完成初侵染,因此观察该菌子实体的超微结构特征,有利于进一步揭示子囊菌的侵染机制。【目的】明确P. medicaginis子囊孢子萌发、Ca²⁺信号途径是否参与调控附着胞形成;观察子囊孢子和子实体的超微结构特征。【方法】采用光学显微镜技术,研究不同碳、氮源和Ca²⁺/CaM信号途径对P. medicaginis子囊孢子萌发、附着胞形成的影响,并采用电镜技术观察子实体和子囊孢子的超微结构。【结果】碳、氮源有利于P. medicaginis子囊孢子的萌发,葡萄糖、麦芽糖、蔗糖、氨基乙酸、酵母粉、尿素、蛋白胨、硝酸铵、硝酸钠等碳、氮源诱导形成附着胞,以尿素诱导效果最明显;Ca²⁺不仅促进子囊孢子的萌发,显著诱导附着胞的形成,而且高于碳、氮源;10mmol/L的Ca²⁺诱导效果最明显,随其浓度的增加,子囊孢子的萌发率和附着胞的形成率下降;低浓度Ca     

The physiological effects of salmon lice infection on post-smolt of Atlantic salmon

The physiological effects of salmon lice infections on post-smolt of Atlantic salmon were examined by experimentally infecting hatchery reared post-smolts with infective copepodids. Even at high infection intensities, ranging from 30–250 lice per fish, early chalimus stages did not have severe, physiological effects on the fish. There was a sudden increase in fish mortality after the appearance of preadult I stages. Infected fish were then suffering due to lesions and osmoregulatory failure. Plasma chloride level increased significantly and total protein, albumin and haematocrit decreased significantly in infected compared to uninfected fish. All infected fish became moribund before adult lice appeared. Infection intensities above 30 salmon lice larvae per fish thus appear to cause death of Atlantic salmon post-smolt soon after the lice reach their pre-adult stage.     

Host age, sex, and pathogen exposure level as factors in the susceptibility of Musca domestica to Entomophthora muscae

Adult Musca domestica were exposed to conidial showers from cadavers that were killed by Entomophthora muscae. Of the 4 host ages (14 days, 7 days, 3 days and <1 day), older flies were more likely to die without postmortem signs of infection, while younger flies were more likely to die with signs and produce primary conidia. The youngest fly group also succumbed to infection significantly sooner than older flies. Total mortality did not differ for the 2 sexes, but females died significantly more often with signs in one trial. Males died significantly earlier than did females, but differences were not significant when the sexes were paired by weight. Increased levels of pathogen exposure significantly increased both total mortality and the number of flies dying with signs; it also decreased incubation period. The potential significance of these findings in the epidemiology of the disease is discussed.

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Zusammenfassung Die Mortalitätsrate von Musca domestica nach Behandlung mit Entomophthora muscae Sporen wurde in Laborversuchen in Abhängigkeit von Wirtsalter,-geschlecht und Expositionsdauer festgestellt. Fliegen verschiedenen Alters (14 Tage, 7 Tage, 3 Tage und jünger als 1 Tag) wurden gemeinsam in 237 ml Kartonbehältern gehalten. Die Behälter besassen Netzdeckel, auf denen frisch an einer Entomophthora Infektion gestorbene Fliegen ausgelegt wurden. Auf diese Weise wurden die Versuchstiere 8 Stunden (low exposure) bzw. 48 Stunden (high exposure) lang mit Entomophthora Sporen kontaminiert. Mit Hilfe von am Boden der Versuchsgefässe ausgelegten Objektträgern wurde die relative Menge Entomophthora Sporen festgestellt, der die Versuchstiere ausgesetzt waren. Der Totenfall wurde täglich für die Dauer von 10 Tagen registriert; dabei wurde zwischen Konidienträgern (external signs) und Leichen ohne Konidien (no external signs) unterschieden.Die Versuchsgruppen, die 48 Stunden lang mit Entomophthora Sporen behandelt worden waren, wiesen im Vergleich zu den kürzer behandelten Versuchsgruppen höhere Mortalität und einen höheren Anteil an Konidienträgern auf. Darüber hinaus wurde das Auftreten von Konidienträgern bei den länger behandelten Gruppen früher beobachtet. Im Vergleich zu unbehandelten Kontrollgruppen erhöhte die Behandlung mit Entomophthora Sporen auch die Rate nicht konidientragender Leichen.Bei gleicher Expositionsdauer zeigten Weibchen und Männchen ähnliche Mortalitätsraten. Ein höherer Anteil Konidienträger bei den Weibchen wurde bei einem von zwei entsprechenden Versuchen beobachtet. Konidienträger traten bei Weibchen später als bei Männchen auf. In Bezug auf die Inkubationszeit konnte kein geschlechsspezifischer Unterschied festgestellt werden, wenn Versuchsgruppen mit gleichem Gewicht verglichen wurden. Bei beiden Geschlechtern erfolgte die Bildung von Konidien bei grösseren Individuen später als bei kleineren Individuen.Das Alter der Versuchstiere beeinflusste sowohl die Mortalitätsrate als auch die Häufigkeit und das zeitliche Auftreten von Konidienträgern. Jüngere Fliegen wiesen einen höheren Anteil Konidienträger auf; bei Fliegen mit einem Alter von weniger als einem Tag wurde das Auftreten von Konidienträgern früher beobachtet als bei älteren Individuen.Die untersuchten Faktoren können auch für die Epidemiologie von Entomophthora freilebender Musca domestica Populationen eine wichtige Rolle spielen. In Populationen mit niedrigerem Durchschnittsalter wird sich eine Infektion rascher ausbreiten, da jüngere Fliegen eine kürzere Inkubationszeit aufweisen und auch nach kürzerer Kontaminationsdauer Konidien an den getöteten Individuen ausgebildet werden. Darüber hinaus wird sich eine Infektion dann rascher ausbreiten, wenn die Population mit einer grösseren Sporenmenge infiziert wird, da dann eine kürzere Inkubationzeit und ein höherer Anteil Konidienträger zu erwarten ist.

     

Renal FNA-based typing of renal masses remains a useful adjunctive modality: evaluation of 31 renal masses with correlative histology

Objectives:  Given the advances in renal imaging modalities in the recent years, a greater number of renal cell carcinomas (RCCs) with tumour size of <3 cm are being detected radiologically. Consequently, there is a pressing need for accurate typing of RCCs which, in turn, will aid in selection of cases of nephron-sparing surgery.
Methods:  A total of 31 cases of renal masses with available fine needle aspiration (FNA) material and concomitant histopathology details were retrieved. They included 27 RCCs (17 clear cells, eight papillary and two chromophobe), one oncocytoma, one liposarcoma and two benign lesions – one xanthogranulomatous pyelonephritis (XPN) and one benign cyst. Two investigators reviewed all FNA material. The degree of concordance between cytological typing and histological typing was assessed.
Results:  There was excellent agreement between the FNA typing and the final diagnosis, with correct classification in 28 of 31 cases. Among the three discordant cases, two were RCCs. The first was a papillary RCC (PRCC) that was misdiagnosed on FNA as clear cell RCC. Another case that was typed as a PRCC on final histopathology was diagnosed 'suspicious cells' on FNA. The third case was an XPN that was misdiagnosed on FNA as RCC with necrosis.
Conclusions:  There is an excellent concordance (90.3%) between the FNA diagnosis and the final histological diagnosis, especially in RCCs. There is a tendency for misdiagnosis with PRCC. Lesions with extensive necrosis and relatively insufficient diagnostic material on FNA specimens must be interpreted with caution. Better concordance might be observed with more extensive sampling.     

孤立性纤维性肿瘤15例CT影像病理特征分析

对比研究孤立性纤维性肿瘤(SFTs)的CT影像学特征及其与病理学的关系,评估CT在SFTs诊断中的应用价值。方法:收集15例孤立性纤维性肿瘤,回顾分析CT表现以及病理形态学和免疫表型变化。结果:15例中6例发生于胸腔内(3例发生于肺,3例位于壁层胸膜);此外鼻腔2例,上颌窦、腹腔、咽旁、腮腺、肾脏、眼眶、髂窝各1例。临床主要表现为局部肿块及其引起的压迫症状,CT扫描显示肿块边界清楚,增强扫描早期呈明显均质性强化。病理学改变显示瘤细胞呈梭形细胞为主,具有细胞密集区与细胞疏松区特征,间质可见瘢痕样的胶原和鹿角样厚壁血管,瘤细胞排列方式多样,免疫组化染色CD34阳性(91．67%), CD99(88．88%),Bcl-2(87．5%)和Vim(100%)。结论:SFT因有完整的包膜,故多为境界清楚的软组织密度肿瘤,如境界不清多提示侵袭性生长。因病理学上肿瘤细胞分布以及有无变性坏死而呈现不同的密度特征,因富血管表现而在增强CT扫描时有较为明显的强化;最终诊断需要免疫组织化学染色证实。     

Natural mortality and cause of death analysis of the captive chimpanzee (Pan troglodytes): A 35‐year review

We present the spontaneous causes of mortality for 137 chimpanzees (Pan troglodytes) over a 35‐year period. A record review of the pathology database was performed and a primary cause of mortality was determined for each chimpanzee. The most common causes of mortality were as follows: cardiomyopathy (40% of all mortalities), stillbirth/abortion, acute myocardial necrosis, chimpanzee‐induced trauma, amyloidosis, and pneumonia. Five morphologic diagnoses accounted for 61% of mortalities: cardiomyopathy, hemorrhage, acute myocardial necrosis, amyloidosis, and pneumonia. The most common etiologies were degenerative, undetermined, bacterial, traumatic, and neoplastic. The cardiovascular system was most frequently involved, followed by the gastrointestinal, respiratory, and multisystemic diseases. Degenerative diseases were the primary etiological cause of mortality of the adult captive chimpanzee population. Chimpanzee‐induced trauma was the major etiological cause of mortality among the perinatal and infant population. This information should be a useful resource for veterinarians and researchers working with chimpanzees.     

X-ray dark-field phase-contrast imaging: Origins of the concept to practical implementation and applications

The basic idea of X-ray dark-field imaging (XDFI), first presented in 2000, was based on the concepts used in an X-ray interferometer. In this article, we review 20 years of developments in our theoretical understanding, scientific instrumentation, and experimental demonstration of XDFI and its applications to medical imaging. We first describe the concepts underlying XDFI that are responsible for imparting phase contrast information in projection X-ray images. We then review the algorithms that can convert these projection phase images into three-dimensional tomographic slices. Various implementations of computed tomography reconstructions algorithms for XDFI data are discussed. The next four sections describe and illustrate potential applications of XDFI in pathology, musculoskeletal imaging, oncologic imaging, and neuroimaging. The sample applications that are presented illustrate potential use scenarios for XDFI in histopathology and other clinical applications. Finally, the last section presents future perspectives and potential technical developments that can make XDFI an even more powerful tool.     

Performance and mortality of farmed hares

Performance and mortality of hares were evaluated for 2 consecutive years in a large farm in Veneto Region (Northern Italy). On average, fertile reproductive pairs (n=318) gave birth 4.8 times and produced 11.4 live leverets, weaned 8.4 leverets and produced 7.0 growing hares (60 days) every year. Mean mortality was 3.6%, 22.9%, 9.7% and 2.5% in newborn (0 to 2 days of age), suckling (3 to 25 days), growing (26 to 60 days) and sub-adult (61 days until sale) hares, respectively. The main causes of mortality were enteric diseases (75.5%, 75.9% and 12.1% in suckling, growing and sub-adult hares, respectively), followed by respiratory diseases (3.4%, 8.0% and 36.2% in suckling, growing and sub-adult hares, respectively), starvation (11.3% and 8.8% in suckling and growing hares, respectively) and trauma (7.1%, 2.3% and 30.2% in suckling, growing and sub-adult hares, respectively). In reproducing hares, mortality was 24.7% and 15.4% in 2010 and 2011, respectively. Respiratory diseases (34.8%) and ulcerative pododermatitis (18.9%) were the most common pathological changes detected in reproducing hares. Farmed hares seem to be affected by diseases resembling those of rabbits reared under intensive conditions. It seems necessary to improve the husbandry of hares to reach satisfactory technical standards and to preserve their health.     

Analysis of the Beta‐Tubulin Gene and Morphological Changes of the Microsporidium Anncaliia algerae both Suggest Albendazole Sensitivity

The Microsporidium, Anncaliia algerae, an obligate intracellular parasite, has been identified as an opportunistic human pathogen, but treatment has not been evaluated for infections with this organism. Albendazole, an antitubulin polymerization drug used against parasitic worm infections, has been the medication of choice used to treat some microsporidial infections affecting humans, with varying results ranging from clearing infection (Encephalitozoon) to resistance (Enterocytozoon). This study illustrates the effect of albendazole treatment on A. algerae infection in Rabbit Kidney (RK13) cells and Human Fetal Lung (HFL‐1) fibroblasts. Albendazole appears to have an attenuating effect on A. algerae infection and albendazole's IC₅₀ in RK13 cells is 0.1 μg/ml. Long‐term treatment inhibits up to 98% of spore production, but interrupting treatment reestablishes the infection without new exposure to the parasite as supported by microscopic observations. The parasite's beta‐tubulin gene was purified, cloned, and sequenced. Five of the six specific amino acids, associated with benzimidazole sensitivity, are conserved in A. algerae. These findings suggest that A. algerae is sensitive to albendazole; however, the organism is not completely cleared from cultures.