Splenic white pulp as a thymus-independent area in the African clawed toad,Xenopus laevis

Summary An indirect immunofluorescence study of the frozen sections of the spleen of an anuran amphibian, Xenopus laevis, showed that lymphocytes bearing a small amount of immunoglobulin (Ig) were localized mostly in the white pulp of non-immunized toads. There were fewer fluorescent cells in the red pulp. In the toads hyperimmunized with human gamma globulin (HGG), cells with strong cytoplasmic fluorescence increased significantly in the outer part of the white pulp. Electron microscopy of spleens from these toads showed that plasma cells at different stages of maturation were abundant in the white pulp, whereas in the red pulp, a smaller number of maturer plasma cells were observed. These results indicate that, in contrast with its mammalian counterpart, the splenic white pulp of this anuran is the site where thymusin-dependent lymphocytes commence blast formation and transformation into plasma cells.     

Strain Differences in the Immunogenicity of Aggregated Human IgG and the Adjuvant Action of Lipopolysaccharide on the Low-Responder Strain of Mice

Strain differences in the antibody response to human IgG (HGG) were observed when aggregated HGG was injected intravenously. Lipopolysaccharide (LPS) administered subsequently markedly enhanced the antibody response to HGG in low responder C57BL/6 mice as compared with that in high responder DDD, C3H/He or (C57BL/6 × DDD)F₁ mice. Aggregate-free preparation of HGG at a dose of 0.5 mg induced immunological tolerance in all strains of mice tested. LPS injected subsequently converted tolerogenic, aggregate-free HGG into immunogen in DDD mice but not in C57BL/6 mice. To determine the correlation between adjuvanticity and mitogenicity of LPS, spleen cells from normal mice were cultured in the presence of LPS and ³H-thymidine uptake was measured. Spleen cells of DDD mice incorporated three times as much ³H-thymidine as those of C57BL/6 mice. There seems no strong correlation between both activities of LPS. The data obtained are discussed in terms of strain differences in the macrophage function for processing the antigen.     

Effect of Pulp Cell Number and Assimilate Availability on Dry Matter Accumulation Rate in a Banana Fruit [ Musa sp. AAA group 'Grande Naine' (Cavendish subgroup)]

Fruit position on the bunch (inflorescence) is an importantpart of variability in banana fruit weight at harvest, as fruitsat the bottom of the bunch (distal fruits) are approx. 40% smallerthan those at the top (proximal fruits). In this study, therespective roles of cell number and cell filling rate in thedevelopment of pulp dry weight are estimated. To this end, thesource/sink ratio in the plant was altered at different stagesof fruit development. Leaf shading (reducing resource availability),bunch bagging (increasing sink activity by increasing fruittemperature), and bunch trimming (decreasing sink size by fruitpruning), applied once cell division had finished, showed thatthe pulp filling rate depends on resource availability. Bunchbagging and bunch trimming were also carried out before theend of cell division to estimate the role of pulp cell numberin the development of pulp dry weight. A sampling method wascalibrated to evaluate pulp cell number from the digestion ofa fixed portion of the pulp in a solution of chromic and nitricacids. A relationship was found between pulp cell number andfruit length at the end of cell division. It was observed thatpulp cell number is a determining factor in pulp dry weightvariability within a bunch. On the other hand, the cell fillingrate was identical for all fruits in the bunch and was influencedby the source/sink ratio. A Michaelis-Menten relationship wasinvoked to relate the cell filling rate in a bunch to the source/sinkratio during bunch filling. Copyright 2001 Annals of BotanyCompany Banana fruit, Musa sp., fruit growth, cell number, cell filling rate, source/sink ratio, temperature     

Lymphoid organ development in Xenopus thymectomized at eight days of age

This paper examines the effect of early thymectomy on the subsequent development of lymphoid tissues in the toad, Xenopus laevis. At the time of thymic removal (8 days post-fertilization) all the lymphoid organ anlagen are at a rudimentary state of differentiation and contain few, if any, small lymphocytes. Despite the absence of any thymic tissue all thymectomized animals grew normally. Thymectomized larvae developed relatively normal lymphoid organs. However, lymphoid depletion was apparent in the splenic red pulp and in the pharyngeal ventral cavity bodies. Examination of the lymphoid organs of post-metamorphic Xenopus revealed reduction in spleen size following thymectomy. Lymphoid depletion was evident in the splenic red pulp of many thymectomized toadlets and reduction in proportion of white to red pulp was also noted in a few of these animals. Absence of the thymus had no apparent effect on the histology of the other lymphoid organs examined.     

小菜蛾烟碱型乙酰胆碱受体α亚基cDNA的克隆、序列分析与不同发育阶段表达分析

烟碱型乙酰胆碱受体(nAChR)介导昆虫中枢神经系统中胆碱能突触兴奋性神经递质的快速传递,也是新烟碱类杀虫剂和多杀菌素的作用靶标。本研究利用RT-PCR和RACE技术,克隆了小菜蛾Plutella xylostella nAChR α亚基的一个新基因(Pxα8)的全长cDNA(GenBank登录号为EU914853)。Pxα8的cDNA序列全长1 744 bp,开放阅读框为1 602 bp,编码534个氨基酸,具有nAChR α亚基的典型特征,与其他昆虫nAChR α8亚基具有77%～96%的相似性,与果蝇nAChR β2亚基具有76%的相似性。Pxα8的开放阅读框存在单核苷酸多态性位点,导致多个位点氨基酸的替换。雌性4龄幼虫的多态性位点多于雄性4龄幼虫,而且雌、雄4龄幼虫的多态性位点均不相同。半定量RT-PCR研究结果表明,Pxα8 mRNA在成虫期表达量高于蛹期和4龄幼虫期。本研究结果为进一步研究小菜蛾nAChR 亚基的多样性和对多杀菌素的靶标抗性机制提供重要基础。     

中国黄粉蝶亚科六属间基于COⅡ和EF-1α基因部分序列的系统发育关系（鳞翅目：粉蝶科）

为了探讨中国黄粉蝶亚科属间的系统发育关系,我们对其中6属9种的细胞色素氧化酶Ⅱ（COⅡ）的部分序列和延伸因子基因（EF-1α）部分序列进行了分析。分别采用最大简约法（maximum parsimony, MP）、最大似然法（maximum likelihood, ML）和贝叶斯推论法（bayesian inference, BI）构建黄粉蝶亚科分子系统树。结果表明：在测得的COⅡ基因的648 bp序列和EF-1α基因的504 bp序列中,有261个变异位点,151个简约信息位点,黄粉蝶亚科内各属COⅡ基因A+T含量（77.3%）均明显偏高。系统发育分析显示黄粉蝶属为亚科中较为原始的类群,分化较早,豆粉蝶属和迁粉蝶属亲缘关系较近,但钩粉蝶属与豆粉蝶属、迁粉蝶属之间的亲缘关系还不能确定。本研究结果和传统的基于形态学的黄粉蝶亚科的分类体系有所不同,最显著的分歧是本研究支持内群中分化最早的属应为黄粉蝶属,而不是豆粉蝶属和迁粉蝶属。     

Vegetative Parthenocarpy in the Cactus Pear Opuntia ficus-indica (L.) Mill.

The dependence of fruit development on fertilization was studiedin two clones of Opuntia ficus-indica, Ofer and BSI. Fruitsof the clone Ofer bear fully developed seeds, whereas fruitsof the clone BSI contain only degenerated seeds and it was suspectedthat BSI is parthenocarpic. The two clones differed in theirpattern of fruit development. The increase in fruit fresh weightin Ofer was a result of both peel and pulp growth, whereas inBSI fruit growth was mainly due to pulp growth and the peelhad reached its final weight almost at anthesis. Pulp growthin BSI commenced earlier and was faster than in Ofer, but thefinal pulp weight in BSI was only 64% of that in Ofer. Seedgrowth in BSI was limited to the development of semilignifiedand lignified seed coats, whereas in Ofer 43·5% of theseeds were completely developed, the remainder being similarto those found in BSI. Fruits bearing well-developed seeds werealso found in a 'mutant' of BSI that, like Ofer, contained smallerovules at anthesis than flower of the regular BSI genotype.Germination in vivo and in vitro were similar in BSI and Ofer,but pollen tubes failed to reach the ovules in the regular BSIgenotype, while penetrating the ovules in Ofer and the small-ovuledgenotype of BSI. Good-quality fruits similar to Ofer fruitsin weight but with a higher peel to pulp ratio developed inBSI after flower-sterilization in the spring and in the autumn.In Ofer, sterilized flowers failed to develop fruits in springand partially set fruit in the autumn; the fruits consistedalmost exclusively of peel tissue. It was concluded that BSIis a vegetative parthenocarpic clone, i.e., that pollinationis not required for fruit set and development.Copyright 1993,1999 Academic Press Opuntia ficus-indica, cactus pear, fruit development, parthenocarpy, pollen-tube-growth, fluorescence microscopy     

Endogenous Hormones in Seeds, Germination Behaviour and Early Seedling Characteristics in a Normal and ogura Cytoplasmic Male Sterile Line of Rapeseed (Brassica napus L.)

Endogenous hormones, namely cytokinins (CKs), indole-3-aceticacid (IAA) and abscisic acid (ABA) were quantified by specificenzyme-linked immunosorbent assay (ELISA) in the mature seedof normal (cv. Westar) and ogura (ogu) cytoplasmic male sterile(CMS) lines of rapeseed (Brassica napus L.). Dihydrozeatin (DZ)and dihydrozeatin riboside (DZR) were the major CK base andriboside, respectively, in seeds of both the normal and oguCMS lines. The normal seed had more than 4-fold DZ levels incomparison to that of ogu CMS. On the other hand, the ogu CMSseed had higher levels of CK o-glucosides and CK. nucleotidesthan normal seed. Seeds of the normal line contained 5-foldmore IAA but had one-quarter the level of ABA in comparisonto those of the ogu CMS line. The normal line also had greaterseed diameter and weight than the ogu CMS line and the normalseed germinated earlier than the male sterile seed. DZ (10^–6M) promoted the germination of ogu CMS seeds, but it was notcomparable to that of the normal line. ABA (10^–6 M) inhibitedseed germination of ogu CMS but had little effect on the normalline. The normal seedlings had shorter primary roots, more lateralroots, longer hypocotyls, greater cotyledon fresh weight andhigher chlorophyll levels in comparison to ogu CMS seedlings.Exogenously supplied DZ, IAA and ABA affected the various parametersof both the normal and ogu CMS seedlings, but in most casesdid not fully restore the differences in the two lines. The results presented show that in the ogura cytoplasmic malesterile line of B. napus (1) a number of seed and seedling characteristicsare affected, and (2) the altered seed morphology is accompaniedby changes in the levels of various hormones. Key words: Brassica napus, cytoplasmic male sterility, enzyme-linked immunosorbent assay, hormone, seed germination     

Phenotypic differences between red pulp capillary and sinusoidal endothelia help localizing the open splenic circulation in humans

The distribution of capillaries, sinuses and larger vessels was investigated by immunohistology in paraffin sections of 12 adult human spleens using a panel of antibodies. Double staining for CD34 and CD141 (thrombomodulin) revealed that capillary endothelia in the cords of the splenic red pulp and at the surface of follicles were CD34⁺CD141⁻, while red pulp sinus endothelia had the phenotype CD34⁻CD141⁺. Only in the direct vicinity of splenic follicles did sinus endothelial cells exhibit both antigens. Thus, splenic sinuses do not replace conventional capillaries, but exist in addition to such vessels. The endothelium in arterioles, venules and larger arteries and veins was uniformly CD34⁺CD141⁺. Anti-CD34 and anti-CD141 both additionally reacted with different types of splenic stromal cells. Differential staining of capillaries and sinuses may permit a three-dimensional reconstruction of serial sections to unequivocally delineate the “open” and “closed” splenic circulation in humans.     

Macrophage colony-stimulating factor is indispensable for repopulation and differentiation of Kupffer cells but not for splenic red pulp macrophages in osteopetrotic (<Emphasis Type="Italic">op</Emphasis>/<Emphasis Type="Italic">op</Emphasis>) mice after macrophage depletion

We previously reported that macrophage colony-stimulating factor (M-CSF, CSF-1) played important roles in the process of the repopulation of Kupffer cells after their elimination by administration of liposome-entrapped dichloromethylene diphosphonate (lipo-MDP). In this study, we examined the repopulation of Kupffer cells and splenic red pulp macrophages in osteopetrotic (op/op) mice defective in the production of functional M-CSF and their littermate mice by using the lipo-MDP model. In untreated op/op mice, numbers of F4/80-positive Kupffer cells in the liver and F4/80-positive splenic red pulp macrophages were reduced. Repopulation of Kupffer cells and splenic macrophages was observed in littermate (op/+) mice liver by 14 days after depletion. However, in op/op mice, repopulation of Kupffer cells was not observed in Kupffer-cell-depleted op/op mice until 56 days after depletion, whereas splenic red pulp macrophages repopulated and recovered to the level of control op/op mice by 10 days after depletion. Single injection of M-CSF was effective for the induction of the repopulation of Kupffer cells, and daily administration of M-CSF induced remarkable repopulation and maturation of Kupffer cells and proliferation of macrophage precursor cells in the liver of Kupffer-cell-depleted op/op mice. These results suggest that Kupffer cells are completely M-CSF-dependent tissue macrophages, whereas splenic red pulp macrophages are composed of M-CSF-dependent macrophages and M-CSF-independent macrophages. This mouse model provides a useful tool for the study of effects of growth factor on Kupffer cell differentiation in vivo. This study was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, and Culture of Japan, NIH grant CA20408, and a Tsukada Memorial Grant (2000).     

Selective inhibition of antimycin A-insensitive respiration in Ustilago maydis and Ceratocystis ulmi

Sporidia of Ustilago maydis and conidia of Ceratocystis ulmipossess an antimycin A and azide-tolerant electron transportpathway which apparently diverts electrons to O₂ from some pointon the substrate side of the antimycin A block. The alternatepathway (induced by 0.5 µg/ml antimycin A or 5x10^–4M sodium azide) supports a respiratory rate 1.5–2 timesthat of the normal system, but has a terminal oxidase with alower than normal affinity for O₂. A similarly high respiratoryrate in U. maydis is supported by the normal pathway when uncoupledby 4 µg/ml of 4,5-dichloro-2-trifluoromethylbenzimidazole,but a high affinity for O₂ in this case indicates that the normalterminal oxidase is utilized. Respiration by the normal pathway in both fungi is only slightlyor moderately inhibited by 1.5x10^–3 M benzohydroxamicacid (BHAM) and 5x10^–4 M 8-hydroxyquinoline. The alternatepathway in U. maydis, however, is inhibited as much as 84 and92% respectively by these two compounds, while alternate respirationin C. ulmi can be inhibited as much as 86 and 76% respectively.BHAM, 8-hydroxyquinoline, 2-pyridinethiol-1-oxide, a,a'-dipyridyl,carboxin, and diphenylamine inhibit alternate respiration ata site on the alternate pathway which is not part of the normalelectron transport system. Antimycin A and azide-insensitiverespiration found in U. maydis and C. ulmi closely resemblesinhibitor insensitivity noted in several fungi and some higherplants. Such an alternate respiratory pathway may be an earlystep in the evolution of oxidative phosphorylation. (Received June 27, 1972; )     

Microvascularization of the spleen in larval and adult Xenopus laevis: Histomorphology and scanning electron microscopy of vascular corrosion casts

Microvascular anatomy and histomorphology of larval and adult spleens of the Clawed Toad, Xenopus laevis were studied by light microscopy of paraplast embedded serial tissue sections and scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Histology showed i) that white and red pulp are present at the onset of metamorphic climax (stage 57) and ii) that splenic vessels penetrated deeply into the splenic parenchyma at the height of metamorphic climax (stage 64). Scanning electron microscopy of VCCs demonstrated gross arterial supply and venous drainage, splenic microvascular patterns as well as the structure of the interstitial (extravasal) spaces representing the “open circulation routes.” These spaces identified themselves as interconnected resin masses of two distinct forms, namely “broccoli‐shaped” forms and highly interconnected small resin structures. Arterial and venous trees were clearly identified, as were transitions from capillaries to interstitial spaces and from interstitial spaces to pulp venules. Venous sinuses were not diagnosed (nonsinusal spleen). The splenic circulation in Xenopus laevis is “open.” It is hypothesized that red blood cells circulate via splenic artery, central arteries, penicillar arteries, and red pulp capillaries primarily via “broccoli‐shaped” interstitial spaces, pulp venules and veins into subcapsular veins to splenic veins while lymphocytes circulate also via the interstitial spaces represented by the highly interconnected small resin structures in vascular corrosion casts. In physiological terms, the former most likely represent the fast route for blood circulation, while the latter represent the slow route. J. Morphol. 277:1559–1569, 2016. © 2016 Wiley Periodicals, Inc.     

Influence of localized auxiliary heating on hand comfort during cold exposure

There is a needfor a hand-heating system that will keep the hands warm during coldexposure without hampering finger dexterity. The purpose of this studywas to examine the effects of torso heating on the vasodilativeresponses and comfort levels of cooled extremities during a 3-hexposure to 15°C air. Subjects were insulated, but theirupper extremities were left exposed to the cold ambient air. The effectof heating the torso [torso-heating test (THT)] on handcomfort was compared with a control condition in which no torso heatingwas applied, but Arctic mitts were worn [control test(CT)]. The results indicate that mean finger temperature, meanfinger blood flow, mean toe temperature, mean body skin temperature, body thermal comfort, mean finger thermal comfort, and rate of bodyheat storage were all significantly (P < 0.05) higher on average (n = 6)during THT. Mean body heat flow was significantly (P < 0.05) lower during THT. Therewere no significant differences (P  0.05) in rectal temperature between CT and THT. Mean unheated body skintemperature and mean unheated body heat flow (both of which did notinclude the torso area in the calculation of mean body skin temperatureand mean body heat flow) were also calculated. There were nosignificant differences (P  0.05) inmean unheated body skin temperature and mean unheated body heat flowbetween CT and THT. It is concluded that the application of heat to the torso can maintain finger and toe comfort for an extended period oftime during cold exposure.

     

Expression of insulin-like growth factor II (IGF-II) and histological changes in the thymus and spleen of transgenic mice overexpressing IGF-II

 Previously, transgenic mice were constructed overexpressing human insulin-like growth factor II (IGF-II) under control of the H₂k^b promoter. The IGF-II transgene was highly expressed in thymus and spleen, and these organs showed an increase in weight. In the current study we have analyzed the sites of IGF-II mRNA expression, the distribution of IGF-II, IGF-I, and both IGF receptors, and histomorphometrical changes in thymus and spleen. With in situ mRNA hybridization, expression of the IGF-II transgene is found with high intensity in the thymic medulla and in the white pulp/marginal zone of the spleen, whereas there were scattered positive cells in the thymic cortex and in the splenic red pulp. Hybridization was restricted to non-lymphocytic cells. Immunohistochemistry revealed intense IGF-II peptide staining with the same distribution as IGF-II mRNA. There was additional intense IGF-II staining of all elements in the splenic red pulp (including trabeculae) and diffuse, low level staining in the thymic cortex. These findings were not observed in control mice. In the thymic medulla, most IGF-II producing cells co-labelled with keratin, whereas a minor population also stained for the monocyte/macrophage marker MOMA-2. In the spleen, co-labelling of IGF-II producing cells was found with MOMA-1 (marginal zone), or with the dendritic cell marker NLDC-145 (red pulp). IGF-I and both IGF receptors were found in these organs in nearly all cell types, with a similar pattern in transgenic mice and in control animals. Histomorphometric analysis revealed a marked increase of thymus cortex size and an increased trabecular size in the spleen. This suggests that IGF-II overproduction induces local effects (auto/paracrine) in the thymic cortex, but not in the thymic medulla. Trabecular growth in the spleen most likely is a distant effect (paracrine or endocrine) of IGF-II overproduction. Accepted: 5 September 1996     

Loss of Function of the Mouse Sharpin Gene Results in Peyer’s Patch Regression

Peyer’s patches (PP) are an important component in the immune response against intestinal pathogens. Two independent, spontaneous mutations in the mouse Sharpin gene (Sharpin^cpdm and Sharpin^cpdm-Dem) result in the absence of PP and disrupted splenic white pulp in adult mice, although a full complement of lymph nodes is present. Here we report that rudimentary PP begin to develop in Sharpin^cpdm mice during embryogenesis, but lack the organizational patterns that are typical of this tissue. In the present study, small intestines examined at weekly intervals from birth to maturity showed spontaneous regression of PP in mutant mice with concurrent infiltration of granulocytes. At 5 to 6 weeks of age, only indistinct remnants of granulocytic accumulations remain. Transplantation of normal bone marrow into Sharpin^cpdm mice at 7 days of age did not prevent regression of PP in bone marrow chimeras examined at 7 to 8 weeks of age. These findings indicate that SHARPIN expression is required for the normal development and maintenance, but not initiation, of PP.     

Stomatal Pattern in Four Species of Monocotyledons

The distribution of stomata has been investigated in the leafepidermis of Chlorophytum comosum, Galanthus nivalis, Schizostyliscoccinea and Scilla lancifolia. The epidermis was consideredto consist of units of construction of two kinds: type A, along epidermal cell with a stoma at its distal end, and typeB, a long epidermal cell without an associated stoma. Exceptin Scilla, the probability of an epidermal unit being type Aincreases approximately with its length. Considering the epidermisas rows of units, alternating sequences of type A and type Bdo not occur randomly along the rows. In Chlorophytum, Galanthusand Schizostylis, both type A and B units tend to be aggregatedinto longer sequences than would be expected on a random basis.It is suggested that homoeogenetic induction (i.e. of like bylike) may be occurring during development. No case can be madefor homoeogenetic induction of units in Scilla. There is a slighttendency to periodicity of distribution of type A units in Galanthus,Schizostylis and Scilla, but this does not seem to representa primary element of pattern. There is interaction between rowsin the sense that unit ends (transverse walls) tend to avoidthose in neighbour rows; this affects the relative distributionof stomata, but there is no evidence of any direct interactionbetween stomata in different rows. Chlorophytum comosum, Galanthus nivalis, Schizostylis coccinea, Scilla lancifolia, leaf, epidermis, stomata, pattern     

Crocodilians and Their Helminth Parasites: Macroevolutionary Considerations

Crocodilian relationships supported by the phylogenetic relationshipsof digenean and nematode parasites are compared with currentestimates of crocodilian phylogeny. The parasite data support(1) the placement of Gavialis as the sister-group of the alligatoridsand the crocodylids, (2) the monophyly of alligators and caimans,(3) the placement of Caiman (as a monophyletic group) as thesister-group of Melanosuchus plus Paleosuchus, and (4) ancientorigins of Crocodylus consistent with patterns of continentaldrift. The parasite data do not support the monophyly of Crocodylus,but the "misplaced" species (C. palustris and Osteolaemus) havehad few parasites reported from them. There is evidence of widespreadhost-switching, but most of the ambiguity appears to resultfrom uneven representation of parasite groups in host species.This is probably due both to uneven sampling by parasitologistsand to parasite extinctions associated with crocodilian extinctions.     

Membrane State and Pollen Viability

The relationship between germinability and fluorochromasia (FCR)has been studied in pollen of eight genera, Secale, Iris, Carex,Eleocharis, Cytisus, Digitalis, Plantago and Lonicera. The FCRtests two properties of the pollen, (a) the integrity of theplasmalemma of the vegetative cell and (b) the presence of anesterase capable of cleaving the fluorogenic ester, fluoresceindiacetate. In general, the correlations between FCR and germinabilitywere found to be very highly significant. This is interpretedas meaning that the primary determinant of pollen viabilityin short-term storage is the state of the vegetative cell membranes.It is suggested that in the partly dehydrated grain at the timeof dispersal the membranes are largely dissociated and do notform an osmotic barrier, but that normal properties are recoveredduring controlled hydration which would normally take placeon the stigma. According to this view, the decay of apparentviability is related to the progressive loss of the capacityof the vegetative cell membranes to regain a normal structureon rehydration. The genera investigated varied in their longevityin storage in low and high humidity conditions. After low-humiditystorage, most showed some enhancement of germination with 1h exposure to a humid atmosphere before transfer to the germinationmedium. During this ‘conditioning’ period the membranesrecover their fluorescein retentivity in step with the increasein germinability. pollen testing, pollen membranes, pollen fluorochromasia, Secale cereale L., Iris pseudacorus L., Carex ovalis Good., Carex nigra (L.) Reichard, Eleocharis palustris (L.) R.Br., Cytisus battandieri Maire, Plantago lanceolata L., Digitalis purpurea L., Lonicera periclymenum L.