The development of the human lymph node

Summary Lymph nodes of human fetuses from the 11th to the 20th gestational week (g.w.) were investigated by light- and electron microscopy under particular consideration of the development of the T-cell and the B-cell regions and their specific reticulum cells. Lymph node development begins as a mesenchymal condensation, containing capillaries and mesenchymal cells; this primordium bulges into a lymph sac. Within the primordium of the lymph node granulopoiesis and erythropoiesis occur temporarily from the 12th to the 14th g.w. The first lymphoid cells and undifferentiated blast cells are seen in the 12th g.w.; monocytes and macrophages can be found from the 13th g.w. onward.The development of the T-cell regions begins during the 13th g.w., before differentiation into cortex and medulla becomes obvious in the 14th g.w. Near the marginal sinus, cells displaying features of interdigitating reticulum cells (IDC) show similarities to monocytes. The morphological differentiation of the IDC is complete in the 17th g.w. when they are found in the paracortical region. Among the IDC, lymphoid cells with features of thymocytes are arranged in small groups.The first indication of the development of B-cell regions can be recognized in the 14th g.w. when precursors of dendritic reticulum cells (DRC) are seen near the marginal sinus; this area also displays lymphoblasts, immunoblasts, and plasmoblasts. During the 20th g.w. small primary follicles are discernible in the outer cortex; in addition to blast cells they contain small lymphocytes, none of which show features of thymocytes. The morphological development of DRCs is not entirely complete until the 20th g.w.; however, some cells already show a characteristic network of interwoven processes.The probable origin of (i) the IDC from monocytes, and (ii) the DRC and fibroblastic reticulum cells from a common type of mesenchymal precursor cells, as well as their significance for a specific micromilieu in the T-cell and the B-cell regions, are discussed.This investigation was supported by grants from the Deutsche Forschungsgemeinschaft and the Sonderforschungsbereich 111 of the Deutsche ForschungsgemeinschaftThe authors appreciate the contribution of human fetal material from Dr. J. von Hollweg and Dr. J. Körner, Hospital Heidberg, Hamburg, and the excellent technical assistance of Mrs. O.M. Bracker, Mrs. H. Hansen, Mrs. R. Köpke, Mrs. I. Knauer, Mrs. F. Müller, Mrs. H. Siebke, and Mrs. H. Waluk     

β-Arrestin Promotes Wnt-induced Low Density Lipoprotein Receptor-related Protein 6 (Lrp6) Phosphorylation via Increased Membrane Recruitment of Amer1 Protein

β-Arrestin is a scaffold protein that regulates signal transduction by seven transmembrane-spanning receptors. Among other functions it is also critically required for Wnt/β-catenin signal transduction. In the present study we provide for the first time a mechanistic basis for the β-arrestin function in Wnt/β-catenin signaling. We demonstrate that β-arrestin is required for efficient Wnt3a-induced Lrp6 phosphorylation, a key event in downstream signaling. β-Arrestin regulates Lrp6 phosphorylation via a novel interaction with phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P₂)-binding protein Amer1/WTX/Fam123b. Amer1 has been shown very recently to bridge Wnt-induced and Dishevelled-associated PtdIns(4,5)P₂ production to the phosphorylation of Lrp6. Using fluorescence recovery after photobleaching we show here that β-arrestin is required for the Wnt3a-induced Amer1 membrane dynamics and downstream signaling. Finally, we show that β-arrestin interacts with PtdIns kinases PI4KIIα and PIP5KIβ. Importantly, cells lacking β-arrestin showed higher steady-state levels of the relevant PtdInsP and were unable to increase levels of these PtdInsP in response to Wnt3a. In summary, our data show that β-arrestins regulate Wnt3a-induced Lrp6 phosphorylation by the regulation of the membrane dynamics of Amer1. We propose that β-arrestins via their scaffolding function facilitate Amer1 interaction with PtdIns(4,5)P₂, which is produced locally upon Wnt3a stimulation by β-arrestin- and Dishevelled-associated kinases.     

Dobrava-Belgrade hantavirus from Germany shows receptor usage and innate immunity induction consistent with the pathogenicity of the virus in humans

Background

Dobrava-Belgrade virus (DOBV) is a European hantavirus causing hemorrhagic fever with renal syndrome (HFRS) in humans with fatality rates of up to 12%. DOBV-associated clinical cases typically occur also in the northern part of Germany where the virus is carried by the striped field mouse (Apodemus agrarius). However, the causative agent responsible for human illness has not been previously isolated.

Methodology/Principal Findings

Here we report on characterization of a novel cell culture isolate from Germany obtained from a lung tissue of “spillover” infected yellow necked mouse (A. flavicollis) trapped near the city of Greifswald. Phylogenetic analyses demonstrated close clustering of the new strain, designated Greifswald/Aa (GRW/Aa) with the nucleotide sequence obtained from a northern German HFRS patient. The virus was effectively blocked by specific antibodies directed against β3 integrins and Decay Accelerating Factor (DAF) indicating that the virus uses same receptors as the highly pathogenic Hantaan virus (HTNV). In addition, activation of selected innate immunity markers as interferon β and λ and antiviral protein MxA after viral infection of A549 cells was investigated and showed that the virus modulates the first-line antiviral response in a similar way as HTNV.

Conclusions/Significance

In summary, our study reveals novel data on DOBV receptor usage and innate immunity induction in relationship to virus pathogenicity and underlines the potency of German DOBV strains to act as human pathogen.     

A characteristic sequence of trisomies starting with trisomy 7 in benign thyroid tumors

The cytogenetic results from a series of 113 thyroid hyperplasias and adenomas are reported; 15 showed clonal karyotypic alterations. In addition to a group showing translocations involving 19q13, another subset of lesions characterized by polysomies can be found. Based on our own cases belonging to this subset and a review of the cases reported in the literature, we conclude that the characteristic feature of this group is a sequence that always starts with trisomy 7, but that sometimes even leads to chromosome numbers in the hypertriploid range. This subset of thyroid tumors may be an example of a more common genetic pathway in human solid tumors.     

Nitric oxide synthase and interferon-gamma receptor immunoreactivities in relation to ascending spinal pathways to thalamus,hypothalamus, and the periaqueductal grey in the rat

The retrograde tracer fluoro-gold was injected into the periaqueductal grey, thalamus or hypothalamus, and spinal cord sections were processed for neuronal nitric oxide synthase (nNOS) immunohistochemistry to investigate the relationships of nNOS immunoreactive, and spinomesencephalic, spinothalamic and spinohypothalamic projection neurones. In addition, in the lateral spinal nucleus the relationship between spinomesencephalic, -thalamic and -hypothalamic projection neurones, and nNOS and interferon-gamma receptor immunoreactive structures was investigated at the lumbar level. No single retrogradely labelled spinomesencephalic, -thalamic or -hypothalamic neurone showed nNOS immunoreactivity. In the lateral spinal nucleus, however, many fluoro-gold-labelled neurones were closely apposed by both nNOS and interferon-gamma receptor immunoreactive structures, especially prominent in the hypothalamic injection cases. This study gave no evidence for nNOS immunoreactivity in spinal neurones projecting to the periaqueductal grey, thalamus or hypothalamus, but suggests that in the lateral spinal nucleus such neurones are contacted by both nNOS- and interferon-gamma receptor-containing axon terminals.     

How can we protect rare hemiparasitic plants? Early-flowering taxa ofEuphrasia andRhinanthus on the Baltic island of Gotland

Euphrasia stricta var.suecica, E. stricta var.tenuis, andRhinanthus serotinus subsp.vernalis are three endangered, hemiparasitic annual herbs found in traditionally managed hay meadows on Gotland, Sweden. We have studied — experimentally and in the field — how some features in the present and traditional management cycle affect their fitness. We set up permanent 50 × 50-cm plots in a coastal hay meadow where bothE. stricta var.suecica andRhinanthus are still quite common. The survival, growth and fecundity of cohorts ofEuphrasia andRhinanthus were followed throughout the growing season for three years. The length of the growing season was vital forEuphrasia andRhinanthus performance. In cooler summers, the percentage ofEuphrasia individuals that produced mature seeds was decreased by 20% at the time of mowing. This indicates the need to individually adjust the time of mowing, as was formerly done, to the actual phenological development in order to maintain healthy populations. It is also important to consider the effect of time of mowing on total species richness, asEuphrasia more successfully established in 10 × 10-cm squares with high species richness. Also, the time in spring when hemiparasite growth started was crucial. Cohorts of bothEuphrasia stricta var.suecica andRhinanthus that connected to hosts and started growing early in the season had a strong advantage over later cohorts by having fitness values four to eight times higher. From this we conclude that the traditional practice of spring raking is important for the long-term persistence of these hemiparasites, since raking promotes an earlier onset of both host and hemiparasite growth. Another important issue is the traditional, but nowadays often neglected practice of letting the hay dry in the meadow after mowing. Hay that was left to dry in the meadow contributed significantly moreRhinanthus seed to the meadow than hay that was taken away immediately after mowing. We also found that seedling emergence and subsequent growth ofEuphrasia stricta var.suecica was greatly enhanced by a second hay cut in September. A second cutting mimics some of the positive effects of the traditional practice of aftermath grazing, which is nowadays often abandoned.     

Periodontal disease status of pregnant women with diabetes mellitus

The aim of the present study was to evaluate the association between type I diabetes mellitus (DM) and periodontal disease in pregnant women. Fifty-two pregnant women aged 27.9 +/- 6.9 years with type I DM participated in the present study. Forty-two non-pregnant type I female diabetics (mean age: 27.9 +/- 6.1 years) and 121 healthy non-pregnant women (mean age: 29.1 +/- 5.7 years) without diabetes formed the control group. All subjects were given a clinical periodontal examination including probing pocket depth (PPD), probing attachment level (PAL), assessment of plaque and gingivitis scores (SBI). Blood parameters included levels of hemoglobin, glycosylated hemoglobin, total cholesterol, triglyceride and leukocytes. The pregnant diabetic subjects showed despite a good metabolic control significantly higher values for the SBI compared to the controls. Pregnant diabetic subjects displayed a significant correlation between the dose of insulin per day and PPD (p < or = 0.05) as well as the PAL (p < or = 0.05). In conclusion, the results of the study indicate that pregnant diabetics demonstrate a higher degree of periodontal inflammation and destruction compared to non-pregnant diabetics and healthy non-pregnant patients.     

Simultaneous Hypoxia and Low Extracellular pH Suppress Overall Metabolic Rate and Protein Synthesis In Vitro

Background

The tumor microenvironment is characterized by regions of hypoxia and acidosis which are linked to poor prognosis. This occurs due to an aberrant vasculature as well as high rates of glycolysis and lactate production in tumor cells even in the presence of oxygen (the Warburg effect), which weakens the spatial linkage between hypoxia and acidosis.

Methods

Five different human squamous cell carcinoma cell lines (SiHa, FaDu_DD, UTSCC5, UTSCC14 and UTSCC15) were treated with hypoxia, acidosis (pH 6.3), or a combination, and gene expression analyzed using microarray. SiHa and FaDu_DD were chosen for further characterization of cell energetics and protein synthesis. Total cellular ATP turnover and relative glycolytic dependency was determined by simultaneous measurements of oxygen consumption and lactate synthesis rates and total protein synthesis was determined by autoradiographic quantification of the incorporation of ³⁵S-labelled methionine and cysteine into protein.

Results

Microarray analysis allowed differentiation between genes induced at low oxygen only at normal extracellular pH (pH_e), genes induced at low oxygen at both normal and low pH_e, and genes induced at low pH_e independent of oxygen concentration. Several genes were found to be upregulated by acidosis independent of oxygenation. Acidosis resulted in a more wide-scale change in gene expression profiles than hypoxia including upregulation of genes involved in the translation process, for example Eukaryotic translation initiation factor 4A, isoform 2 (EIF4A2), and Ribosomal protein L37 (RPL37). Acidosis suppressed overall ATP turnover and protein synthesis by 50%. Protein synthesis, but not total ATP production, was also suppressed under hypoxic conditions. A dramatic decrease in ATP turnover (SiHa) and protein synthesis (both cell lines) was observed when hypoxia and low pH_e were combined.

Conclusions

We demonstrate here that the influence of hypoxia and acidosis causes different responses, both in gene expression and in de novo protein synthesis, depending on whether the two factors induced alone or overlapping, and as such it is important for in vivo studies to take this into account.