Expression of the T cell receptor αβ on a CD123+ BDCA2+ HLA-DR+ subpopulation in head and neck squamous cell carcinoma

Human Plasmacytoid Dendritic Cells (PDCs) infiltrating solid tumor tissues and draining lymph nodes of Head and Neck Squamous Cell Carcinoma (HNSCC) show an impaired immune response. In addition to an attenuated secretion of IFN-α little is known about other HNSCC-induced functional alterations in PDCs. Particular objectives in this project were to gain new insights regarding tumor-induced phenotypical and functional alterations in the PDC population. We showed by FACS analysis and RT-PCR that HNSCC orchestrates an as yet unknown subpopulation exhibiting functional autonomy in-vitro and in-vivo besides bearing phenotypical resemblance to PDCs and T cells. A subset, positive for the PDC markers CD123, BDCA-2, HLA-DR and the T cell receptor αβ (TCR-αβ) was significantly induced subsequent to stimulation with HNSCC in-vitro (p = 0.009) and also present in metastatic lymph nodes in-vivo. This subgroup could be functionally distinguished due to an enhanced production of IL-2 (p = 0.02), IL-6 (p = 0.0007) and TGF-β (not significant). Furthermore, after exposure to HNSCC cells, mRNA levels revealed a D-J-beta rearrangement of the TCR-beta chain besides a strong enhancement of the CD3ε chain in the PDC population. Our data indicate an interface between the PDC and T cell lineage. These findings will improve our understanding of phenotypical and functional intricacies concerning the very heterogeneous PDC population in-vivo.     

A genome-wide CRISPR screen identifies interactors of the autophagy pathway as conserved coronavirus targets

Over the past 20 years, 3 highly pathogenic human coronaviruses (HCoVs) have emerged—Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), and, most recently, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)—demonstrating that coronaviruses (CoVs) pose a serious threat to human health and highlighting the importance of developing effective therapies against them. Similar to other viruses, CoVs are dependent on host factors for their survival and replication. We hypothesized that evolutionarily distinct CoVs may exploit similar host factors and pathways to support their replication cycles. Herein, we conducted 2 independent genome-wide CRISPR/Cas-9 knockout (KO) screens to identify MERS-CoV and HCoV-229E host dependency factors (HDFs) required for HCoV replication in the human Huh7 cell line. Top scoring genes were further validated and assessed in the context of MERS-CoV and HCoV-229E infection as well as SARS-CoV and SARS-CoV-2 infection. Strikingly, we found that several autophagy-related genes, including TMEM41B, MINAR1, and the immunophilin FKBP8, were common host factors required for pan-CoV replication. Importantly, inhibition of the immunophilin protein family with the compounds cyclosporine A, and the nonimmunosuppressive derivative alisporivir, resulted in dose-dependent inhibition of CoV replication in primary human nasal epithelial cell cultures, which recapitulate the natural site of virus replication. Overall, we identified host factors that are crucial for CoV replication and demonstrated that these factors constitute potential targets for therapeutic intervention by clinically approved drugs.

This study identifies essential host dependency factors for human coronavirus replication, showing that these can be directly targeted by clinically approved inhibitors and that treatment leads to effective inhibition of coronavirus replication in primary human nasal epithelial cell cultures.     

Micro- and Nanoforaminifera from Abyssal Northeast Atlantic Sediments: A Preliminary Report

Rose Bengal stained benthic foraminifera which pass through a 63 μm mesh (microforaminifera and nanoforaminifera) have been extracted by handsorting the fine sieve residues (> 45 μm, 31 μm, 28 μm, 20 μm, 15 μm) of abyssal sediment samples. The samples were collected using a multiple corer in four areas of the northeast Atlantic between 31° N and 59° N. The abundance of these minute foraminifera varied from 2 specimens per 1 cm² (Madeira Abyssal Plain) to > 110 per 1 cm² (BIOTRANS area). They include a variety of taxa, the most common being certain rotaliid species, hormosinaceans and other multilocular agglutinated forms, the unilocular agglutinated genus Lagenammina, soft-bodied agglutinated sphaeres and flasks (saccamminids and psammosphaerids) and allogromiids. Some specimens are < 63 μm in maximum dimension but others belonging to elongate taxa are longer. Two samples taken 10 cm apart on the Porcupine Abyssal Plain suggest that minute foraminifera may be patchily distributed on a small scale. One sample, which was overlain by substantial amounts of phytodetritus, contained > 100 stained specimens (> 30 per 1 cm²) while the other, in which much less phytodetritus was present, yielded only 10 specimens (2.9 per 1 cm²). This observation suggests that some micro- and nanoforaminifera may flourish in the presence of decaying organic matter, perhaps consuming the associated bacteria. The presence of phytodetritus may also explain why two of our samples from the Madeira Abyssal Plain (MAP) contained an order of magnitude more stained tiny foraminifera than two other MAP samples in which phytodetritus was absent.     

Highly Predictive Model for a Protective Immune Response to the A(H1N1)pdm2009 Influenza Strain after Seasonal Vaccination

Understanding the immune response after vaccination against new influenza strains is highly important in case of an imminent influenza pandemic and for optimization of seasonal vaccination strategies in high risk population groups, especially the elderly. Models predicting the best sero-conversion response among the three strains in the seasonal vaccine were recently suggested. However, these models use a large number of variables and/or information post- vaccination. Here in an exploratory pilot study, we analyzed the baseline immune status in young (<31 years, N = 17) versus elderly (≥50 years, N = 20) donors sero-negative to the newly emerged A(H1N1)pdm09 influenza virus strain and correlated it with the serological response to that specific strain after seasonal influenza vaccination. Extensive multi-chromatic FACS analysis (36 lymphocyte sub-populations measured) was used to quantitatively assess the cellular immune status before vaccination. We identified CD4⁺ T cells, and amongst them particularly naive CD4⁺ T cells, as the best correlates for a successful A(H1N1)pdm09 immune response. Moreover, the number of influenza strains a donor was sero-negative to at baseline (NSSN) in addition to age, as expected, were important predictive factors. Age, NSSN and CD4⁺ T cell count at baseline together predicted sero-protection (HAI≥40) to A(H1N1)pdm09 with a high accuracy of 89% (p-value = 0.00002). An additional validation study (N = 43 vaccinees sero-negative to A(H1N1)pdm09) has confirmed the predictive value of age, NSSN and baseline CD4⁺ counts (accuracy = 85%, p-value = 0.0000004). Furthermore, the inclusion of donors at ages 31–50 had shown that the age predictive function is not linear with age but rather a sigmoid with a midpoint at about 50 years. Using these results we suggest a clinically relevant prediction model that gives the probability for non-protection to A(H1N1)pdm09 influenza strain after seasonal multi-valent vaccination as a continuous function of age, NSSN and baseline CD4 count.     

Guard cells undergo constitutive and pressure-driven membrane turnover

Summary. During stomatal movement, guard cells undergo large and reversible changes in cell volume and consequently surface area. These alterations in surface area require addition and removal of plasma membrane material. How this is achieved is largely unknown. Here we summarize recent studies of membrane turnover in guard cells using electrophysiology and fluorescent imaging techniques. The results implicate that membrane turnover in guard cells and most likely in plant cells in general is sensitive to changes in membrane tension. We suggest that this provides a mechanism for the adaptation of surface area of guard cells to osmotically driven changes in cell volume. In addition, guard cells also exhibit constitutive membrane turnover. Constitutive and pressure-driven membrane turnover were found to be associated with addition and removal of K⁺ channels. This implies that some of the exo- and endocytic vesicles carry K⁺ channels. Together the results demonstrate that exo- and endocytosis is an essential process in guard cell functioning. Correspondence and reprints: Institute of Botany, Darmstadt University of Technology, Schnittspahnstrasse 3, 64287 Darmstadt, Federal Republic of Germany.     

Arbuscular mycorrhiza enhances auxin levels and alters auxin biosynthesis in Tropaeolum majus during early stages of colonization

Plant hormones, including auxins, might be signals during the establishment of an arbuscular mycorrhizal (AM) symbiosis. Here, we report on the concentrations of three auxins native to nasturtium ( Tropaeolum majus L.) during early AM development. Indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and phenylacetic acid (PAA) were previously identified as endogenous compounds in this species by full-scan gas chromatography–mass spectrometry. All auxinic compounds were influenced by AM colonization but showed completely different patterns. At very early stage, free IAA and IBA were lower in infected than in control roots, whereas PAA concentration was higher in infected roots than in controls. At later stages, PAA was reduced in colonized roots, whereas, especially, IBA was increased in colonized roots compared with controls. Measurement of total auxins confirmed a complex regulation pattern for the three compounds. In hyphae of Glomus intraradices , none of the auxins was detectable. Biosynthesis of the three auxins was measured using heavy labeled isotopes as precursors in control and AM-inoculated roots. While not much difference was found in the IAA labeling pattern between controls and AM-inoculated roots at both time points, IBA synthesis was slightly higher in AM-inoculated roots. Double labeling experiments showed that two distinct pathways, a tryptophan-dependent and a tryptophan-independent biosynthetic pathway contribute to the synthesis of IAA in T. majus roots. Because T. majus is difficult to genetically manipulate, we have used tobacco plants transformed with the auxin-inducible promoter GH3 fused to the β-glucuronidase (GUS) reporter gene to investigate whether AM structures would co-localize to cells harboring the auxin-inducible promoter. Although the GUS activity increased significantly in AM-inoculated roots, there was no obvious correlation between GH3::GUS expression and fungal structures.     

Phosphate transport and arsenate resistance in the cyanobacterium Anabaena variabilis.

Cells of the cyanobacterium Anabaena variabilis starved for phosphate for 3 days took up phosphate at about 100 times the rate of unstarved cells. Kinetic data suggested that a new transport system had been induced by starvation for phosphate. The inducible phosphate transport system was quickly repressed by addition of Pi. Phosphate-starved cells were more sensitive to the toxic effects of arsenate than were unstarved cells, but phosphate could alleviate some of the toxicity. Arsenate was a noncompetitive inhibitor of phosphate transport; however, the apparent Ki values were high, particularly for phosphate-replete cells. Preincubation of phosphate-starved cells with arsenate caused subsequent inhibition of phosphate transport, suggesting that intracellular arsenate inhibited phosphate transport. This effect was not seen in phosphate-replete cells.     

International Commission for Protection against Environmental Mutagens and Carcinogens. ICPEMC Working Paper No. 15/7. Reproductive consequences of alcohol abuse: males and females compared and contrasted

The adverse effects of ethyl alcohol on the hypothalamic pituitary gonadal axes of men and women are discussed with particular attention being given to effects of alcohol upon reproduction. Data obtained from acute and chronic alcohol exposure studies are presented. The putative pathophysiologic mechanisms responsible for disturbed reproductive performances in alcohol abusing individuals are discussed where sufficient data are available.     

The quantitative separation of meiofauna

Summary 1. This paper presents the results of a meeting on the evaluation of quantitative procedures for the separation of meiofauna, held at the Marine Station of the Biologische Anstalt Helgoland, in May 1972. Close co-operation on the part of the participants (p. 194) has allowed assessment of advantages and disadvantages of the separation methods applied. The time needed for preserved methods can be reduced by changing to scanning rather than picking out for sorting and counting.2. Sorting without concentrations: This method is suitable for very fine-grained sediments, oozes; it is generally applied for preserved samples; adequate for hard fauna; insufficient for soft fauna; very time consuming.3. Decantation: Suitable for sandy sediments; generally applied for preserved samples; adequate for hard fauna; insufficient for soft fauna; time consuming.4. Elutriation: Suitable for sandy sediments; for unpreserved samples only with anaesthetization, more effective with preserved samples; limited for total live fauna. With preserved samples, adequate for hard fauna; insufficient for soft fauna; quick method.5. Warm-water elutriation: Suitable for sandy sediments; designed for live hard fauna (especially nematodes, ostracods); quick method.6. Sea-water ice treatment: Suitable for sandy sediments with microporal structure; only for live extraction; limited for hard fauna; well suited for soft fauna (including ciliates and flagellates); time consuming.

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Die quantitative Isolierung der Meiofauna. Ein Methodenvergleich

Kurzfassung Im Mai 1972 fand an der Meeresstation der Biologischen Anstalt Helgoland ein Arbeitstreffen mit dem Ziel statt, die verschiedenen Methoden zur quantitativen Isolierung der Meiofauna aus dem Sediment in ihrer Effektivität miteinander zu vergleichen. Die Verfahren zur Anreicherung der Meiofauna in der Probe, des Sortierens und Auszählens werden beschrieben und die durch die Teilnehmer am Arbeitstreffen gemeinsam erzielten Ergebnisse diskutiert. Die Wertung der einzelnen Methoden zur Erfassung der Gesamtfauna, der harten und der weichen Meiofauna, von konservierten und nichtkonservierten Organismen sowie die Anwendung der Methoden auf verschiedenen Sedimenttypen werden durch statistische Analysen abgesichert. Eine Zusammenfassung der Arbeitsverfahren sowie deren Charakterisierung und Leistungsfähigkeit beschließt die Darstellung.