Long-term neuroretinal full-thickness transplants in a large animal model of severe retinitis pigmentosa

Background The purpose of this study was to explore neuroretinal transplantation in a large animal model of severe retinitis pigmentosa and to establish graft development, long-term survival, graft-host integration, and effects on the host retina. Methods Rhodopsin transgenic pigs, aged 6 months, received in one eye a fetal full-thickness neuroretinal sheet in the subretinal space by means of vitrectomy and retinotomy. Six months postoperatively, eyes were studied in the light microscope and with immunohistochemical markers. Full-field electroretinography (ERG) was performed at 4 and 6 months. Results Laminated grafts with well-organized photoreceptors, rod bipolar cells, and Müller cells were found in five of six eyes. Neuronal connections between graft and host retina were not seen. In the five eyes containing a graft, the number of surviving rods in the host retina was significantly higher compared with unoperated eyes. The ERG did not reveal any significant difference in b-wave amplitude between operated and control eyes, but the cone-derived response in operated eyes increased significantly from 4 to 6 months while the rod response in control eyes decreased significantly. Conclusions Fetal full-thickness neuroretina can be transplanted safely to an eye with severe retinal degeneration. In their major part, the transplants develop a normal laminated morphology and survive for at least 6 months. Graft and host retinal neurons do not form connections. Retinal function in the host is reduced initially by the surgical trauma, but the presence of a well-laminated graft counteracts this effect and rescues rods from degeneration. Supported by The Foundation Fighting Blindness (grant# C-NC02-798-0078), The Faculty of Medicine, University of Lund, The Swedish Research Council, The Princess Margaretas Foundation for Blind Children, The 2nd ONCE International Award for New Technologies for the Blind.     

1020例不孕症的子宫及输卵管形态学分析

用复方泛影葡胺注射液对1,020例不孕症者作子宫输卵管造影。结果:子宫腔形态异常540例。双侧输卵管通畅但形态异常者268例。单侧输卵管通畅但形态异常者110例。结果表明:子宫输卵管形态异常与不孕有密切关系。     

Backspatter from experimental close-range shots to the head

Backspatter is the ejection of biological material from a gunshot entrance wound against the line of fire. This phenomenon was investigated experimentally in transverse gunshots to the heads of calves (n = 9) using two types of 9 mm Parabellum ammunition from shooting distances of 0–10 cm. The resulting bloodstains were documented on white paper placed horizontally 60 cm below the impact site. In this report the analysis was restricted to stains with a diameter > 0.5 mm. Backspatter was documented after every gunshot. The number of stains varied from 31–324 per gunshot and appeared to be independent of the shooting distance. The maximum distance droplets travelled varied from 72–119 cm. The majority of droplets accumulated between 0 and 50 cm. The number of droplets and the distances travelled should be higher in man for anatomical reasons. The direction a single droplet can take comprises every possible angle between the most tangential ones to the skin surface. This resulted in a semi-circle of 180° covered with stains. Skin ruptures of the entrance wound were not observed. The succession of events was documented on high speed film and started with the recoil of the firearm, immediately followed by a blow-out effect of the skin. Large droplets exited approximately 0.7–4 ms after the bullet impacted the skin. The calculated minimum initial velocity of these droplets was 13–61 m/s. Backspatter from gunshots to the head likely is caused by the hot gases expanding subcutaneously and by cavitation-related intracranial overpressure and tail splashing. In three out of nine gunshots, secondary backspatter additionally occurred as a result of droplets produced by a stream of blood from the entrance wound impacting the paper surface.     

The periodontal differentiation in the phylogeny of teeth–an overview

During the evolution of teeth, different types of periodontal attachment have been developed. On the basis of the comparative histology of periodontal tissues, protoacrodontal, acrodontal, acro-protothecodontal, pleurodontal and thecodontal structures can be distinguished that depend upon the area of attachment (crestal, marginal or socketed type) and the mode of attachment (ankylosis, fibrous or combined type). Due to "phylogenetic memory", changes of the periodontium in health and disease could be interpreted as copies of phylogenetically older patterns. The greatest variations in tooth attachment have originated in acrodont bony fishes and in pleurodont reptiles, whereas the selection for a single thecodont or socketed type was an important event in the evolution of mammals. The detailed structures of cementum, of the fiber apparatus and of the junctional epithelium vary from type to type and within one type. These principal structures are decisive for reaction patterns of degeneration and regeneration. Therefore, comparative periodontology could be an important adjunct to help interpret the natural history of periodontal diseases, to help in the selection of experimental animals and to help provide treatment strategies in both human and veterinary situations.     

New metrics for analysis of dendritic branching patterns demonstrating similarities and differences in ON and ON-OFF directionally selective retinal ganglion cells.

The morphology and dendritic branching patterns of retinal ganglion cells have been studied in Golgi-impregnated, whole-mount preparations of rabbit retina. Among a large number of morphological types identified, two have been found that correspond to the morphology of ON and ON-OFF directionally selective (DS) ganglion cells identified in other studies. These two kinds of DS ganglion cell are compared with each other, as well as with examples of class I, class II, and class III cells, defined here with reference to our previous studies. Cell body, dendritic field size and branching pattern are analyzed in this paper and levels of dendritic stratification are examined in the following paper. ON DS ganglion cells are about 10% larger in soma size and about 5 times the dendritic field area of ON-OFF DS ganglion cells, when compared at the same retinal location. These two morphological types of ganglion cell can be said to define the upper and lower bounds of an intermediate range of cell body and dendritic field sizes within the whole population of ganglion cells. Nevertheless, in previous physiological studies receptive field sizes of the two types were shown to be similar. This discrepancy between morphological and physiological evidence is considered in the Discussion in terms of a model of the excitatory receptive field of ON-OFF DS ganglion cells incorporating starburst amacrine cells. A new set of metrics is introduced here for the quantitative analysis and characterization of the branching pattern of neuronal arborizations. This method compares the lengths of terminal and preterminal dendritic branches (treated separately), as a function of the distances of their origins from the soma, viewed graphically in a two-dimensional scatter plot. These values are derived from computer-aided 3D logging of the dendritic trees, and distance from the soma is measured as the shortest distance tracked along the dendritic branches. From these metrics of the "branch length distributions," scale-independent branching statistics are derived. These make use of mean branch lengths and distances, slopes of lines fitted to the distributions, and elliptical indices of scatter in the distributions. By these measures, ON and ON-OFF DS ganglion cells have similar branching patterns, which they share to varying degrees with functionally unrelated class III.1 ganglion cells. The scale of the branching patterns of ON and ON-OFF DS cells and their degree of uniformity are different, however.(ABSTRACT TRUNCATED AT 400 WORDS)     

Sound-generating (sonic) motor system in a teleost fish (Porichthys notatus): sexual polymorphisms and general synaptology of sonic motor nucleus

The sonic motor nucleus of the plainfin midshipman, Porichthys notatus, is a midline nucleus located at the junction of the caudal medulla and rostral spinal cord. Its motoneurons innervate sonic "drumming" muscles that are attached to the lateral walls of the swimbladder. There are two classes of sexually mature males referred to as Type I and Type II. The Type I males are larger and generate sounds during the breeding season. The Type II males are smaller and, like adult females, have not yet been shown to generate sounds. This study examined possible sex differences in the size of sonic motoneurons, and the type and distribution of their afferent terminal boutons. The average soma diameter of motoneurons of Type I males is about 50% larger than that of Type II males and females. There is also a small but significant difference in soma diameter between Type II males and females; they are smaller in the former class. There were no sex differences in the presence or distribution of different classes of axosomatic and axodendritic terminal boutons, which included: (1) active zones with either clear, round, or pleomorphic vesicles, (2) active zones with both clear, round vesicles, and larger dense core vesicles, (3) "mixed synapses" with gap junctions and active zones usually associated with pleomorphic vesicles. The results are discussed within the context of sexual differentiation of vertebrate motor systems and the functional organization of the sonic motor system in fishes. Sex differences in soma diameter correlate with a number of sex differences in the gross and ultrastructural features that distinguish the sonic muscles of Type I males from those of Type II males and females, which are similar to each other. The absence of qualitative sex differences in synaptic morphology suggest that the central neuronal circuitry of the sonic motor system is similar among all three adult morphs.     

Sperm ubiquitination positively correlates to normal morphology in human semen

BACKGROUND: Global ubiquitination in human semen has been found to negatively correlate with standard semen parameters, indicating that ubiquitination can be considered a marker of poor semen quality. However, the inclusion of all semen components in the analysis may be misleading on the biological significance of ubiquitination of sperm cells. We have recently demonstrated the variable presence of bodies of different size, with the highest concentration in oligoasthenoteratozoospermia. The purpose of the present study was to evaluate the relationship between ubiquitination and standard semen parameters, after distinguishing between ubiquitinated sperm and bodies in each sample. METHODS: Ubiquitination was evaluated by flow cytometric sperm ubiquitin tag immunoassay (SUTI) in sperm samples from 45 subjects. Semen analysis was performed according to WHO (1999) guidelines. RESULTS: When only ubiquitinated sperm were considered, a positive correlation with number, motility and normal morphology was found. When correlation was evaluated considering the percentage of ubiquitinated bodies, a negative correlation was found with good semen quality. CONCLUSIONS: Results indicate that the negative correlations previously found between global semen ubiquitination and parameters of semen analysis are mainly driven by components other than sperm. The positive correlation between sperm ubiquitination and good quality parameters suggests a previously unrecognized role for sperm ubiquitination.     

Maternal origin of transferrin receptor positive cells in venous blood of pregnant women

We studied the origin of transferrin receptor (CD71) positive cells in blood from seven women pregnant with a male fetus in order to explore if fetal cells could be detected among them. We used a technique that allows direct chromosomal analysis by in situ hybridization on immunologically and morphologically classified cells. Enrichment was performed by magnetic activated cell sorting (miniMACS)® using an anti-CD71 monoclonal antibody. The cells were immunophenotyped by alkaline phosphatase anti-alkaline phosphatase immunostaining with the same antibody. The origin of the immunophenotyped cells was studied by in situ hybridization using an X cosmid Y repeat chromosome specific probe cocktail. CD71 positive cells were found in six of the seven women at the range of 4 to 43 in respective samples. Over 90% of the CD71 positive cells were nucleated erythrocytes. None of the detected positive cells were shown to be fetal. Thus, the use of transferrin receptor antigen alone in combination with the miniMACS® may not be sufficient for enrichment of fetal cells.     

Sperm morphological assessment based on strict criteria and in-vitro fertilization outcome.

One-hundred-and-twenty-three in-vitro fertilization (IVF) cycles were analysed in order to clarify the influence of strictly normal morphology (SNM) of spermatozoa on IVF outcome. SNM was defined using strict criteria according to Kruger with our modifications. The IVF cycles studied were divided into three groups: %SNM less than 12% (13 cycles), 12 less than 40% (68 cycles), greater than or equal to 40% (42 cycles). The cleavage rates per oocyte were higher in the groups with 12-40% and greater than or equal to 40% of %SNM than in the group with %SNM less than 12%. The embryo transfer rate per cycle increased with increasing %SNM. The overall pregnancy rate per cycle increased with increasing %SNM (7.7% in %SNM less than 12%, 22.1% in 12-40% of %SNM, and 40.5% in %SNM greater than or equal to 40%). The ongoing pregnancy rate per cycle also increased with increasing %SNM (7.7% in %SNM less than 12%, 14.7% in 12-40% of %SNM, and 31.0% in %SNM greater than or equal to 40%). The miscarriage rate was lower in %SNM greater than or equal to 40% (23.5%) than in 12-40% of %SNM (33.3%). It was suggested that %SNM is a good predictor of IVF outcome.     

大鼠脑组织细胞培养的形态学观察

为观察原代培养 SD大鼠大脑组织 ,包括神经干细胞在内的各种细胞的形态 ,取新生大鼠 ,无菌操作下取出整个大脑 ,制备成单细胞悬液 ,接种培养 ,免疫细胞化学鉴定各类细胞的同时用光镜和扫描电镜对不同阶段和种类的细胞进行形态学观察。结果显示 :在体外培养出神经元、胶质细胞和神经干细胞 ;神经细胞在体外的发育时程跟体内基本一致 ,神经干细胞在体外 2 0 d左右分化成熟 ,获取了神经细胞和干细胞体外培养的光镜和扫描电镜形态。结果提示 ,成体神经细胞体外分离、培养的条件关键点在于尽可能贴近自然 ,由此可保证细胞培养的质量 ,获得可靠的神经干细胞、神经元和胶质细胞。为各类神经细胞提供了形态学资料。