Risk assessment of SARS-CoV-2 transmission in hospitality employees in a highly frequented tourist area.

Right from the start of the COVID pandemic in January 2020, the entire tourism sector was put under immense pressure because of its assumed role in SARS-CoV-2 transmission and infection dynamics. Based on reports of single superspreading events in the early days of the pandemic, the hotel industry appeared in a bad light that impaired a strategic risk-assessment of existing transmission risks between tourists and employees. We prospectively analysed samples of 679 employees of 21 hotels and restaurants from July 2020 to December 2020, a time during which more than 1.5 million tourists visited the Lübeck/Ostholstein Baltic Sea vacation area in Northern Germany. Employees were tested up to three times for an acute SARS-CoV-2 infection (PCR from nasopharyngeal swabs) and the presence of SARS-CoV-2 specific antibodies, and were asked to complete a short questionnaire. Despite the massive increase in tourist influx, no significant increase in SARS-CoV-2 cases was observed amongst employees of the tourism sector from July to September 2020. In a cluster-outbreak analysis of 104 study participants of one single hotel in the Lübeck/Ostholstein region in October 2020 being employed in the low-wage sector "housekeeping" could be determined as major risk factor for becoming infected. In conclusion, in a low incidence setting, touristic activities are safe under COVID-related hygiene measures for both the local population and employees of the tourism sector. Whereas, the field of work is a potential risk factor for increased infection dynamics.

Osteogenic induction from marmoset embryonic stem cells cultured in feeder-dependent and feeder-independent conditions.

UNLABELLED: Embryonic stem cells (ESCs) have become increasingly attractive for cell replacement therapies of osteodegenerative diseases; however, pre-clinical studies in large animal models to repair diseased or injured bone are lacking. As a first step into this direction, we describe here the feeder-free cultivation and directed osteogenic differentiation of marmoset ESCs. INTRODUCTION: Owing to their potential to self-renew and their enormous differentiation capability, ESCs are an adequate cell source for cell replacement therapies. To implement stem cell technology clinically, standardized cultivation and differentiation protocols and appropriate animal models are needed. Here, we describe the feeder-free cultivation of Callithrix jacchus ESCs (cESCs) in a chemically defined medium and their subsequent osteogenic differentiation. METHODS: cESCs were maintained on mouse embryonic fibroblast feeder layers or in feeder-free conditions with activin A and basic fibroblast growth factor. Differentiation into mature osteoblasts was steered with ascorbic acid, ß-glycerophosphate and 1α,25-(OH)2 vitamin D3 employing various induction strategies. RESULTS: In feeder-free conditions, cESCs maintained pluripotency as indicated by Oct-4 and Nanog expression, positive immunostaining for typical primate ESC markers and high telomerase activity. Cells also remained karyotypically normal after 40 passages without feeder cells. The hanging drop protocol as well as omitting the embryoid body step proved unsuccessful to initiate osteogenic differentiation. The highest degree of osteogenesis was achieved by formation of embryoid bodies employing the cell cluster technique as indicated by the amount of deposited calcium and bone marker gene expression. Early addition of retinoic acid further improved the yield of osteoblasts and led to an increase in calcium deposition. CONCLUSIONS: The osteogenic differentiation potential of feeder-free cESCs was equal if not higher compared to cells grown on feeders. These findings open the field for near clinical transplantation studies in primate models to evaluate the effectiveness of ESC-derived osteoblasts.

A functional toll-like receptor 8 variant is associated with HIV disease restriction.

BACKGROUND: Toll-like receptors (TLRs) play an important role in the innate immune response to pathogens. TLR8 has been found to recognize RNA derived from various viruses, including human immunodeficiency virus (HIV). Presently, very little is known about the influence of TLR8 genetic variation on susceptibility to and progression of HIV disease. METHODS AND RESULTS: We genotyped a population of 782 HIV-positive adults and 550 healthy control subjects for 3 nonsynonymous TLR8 single-nucleotide polymorphisms. We found that the presence of the most frequent TLR8 polymorphism, TLR8 A1G (rs3764880), confers a significantly protective effect regarding progression of the disease. In overexpression assays, we demonstrated that this receptor variant displays impaired NF-kappaB activation in vitro. Furthermore, we analyzed different cell types obtained from individuals differing in their TLR8 genotype and assessed their response to TLR8 ligands in vitro. The presence of the mutated receptor variant was associated with modulation of cytokine secretion profiles and lipid mediator synthesis patterns in monocytes and neutrophils. CONCLUSIONS: This first report of a functional TLR8 variant associated with a different clinical course of an RNA viral disease may have implications for the individual risk assessment of patients infected with HIV and other RNA viruses as well as for future HIV vaccine development.

Reporting recommendations for tumor marker prognostic studies (remark).

Despite years of research and hundreds of reports on tumor markers in oncology, the number of markers that have emerged as clinically useful is pitifully small. Often, initially reported studies of a marker show great promise, but subsequent studies on the same or related markers yield inconsistent conclusions or stand in direct contradiction to the promising results. It is imperative that we attempt to understand the reasons that multiple studies of the same marker lead to differing conclusions. A variety of methodologic problems have been cited to explain these discrepancies. Unfortunately, many tumor marker studies have not been reported in a rigorous fashion, and published articles often lack sufficient information to allow adequate assessment of the quality of the study or the generalizability of study results. The development of guidelines for the reporting of tumor marker studies was a major recommendation of the National Cancer Institute - European Organisation for Research and Treatment of Cancer (NCI - EORTC) First International Meeting on Cancer Diagnostics in 2000. As for the successful CONSORT initiative for randomized trials and for the STARD statement for diagnostic studies, we suggest guidelines to provide relevant information about the study design, preplanned hypotheses, patient and specimen characteristics, assay methods, and statistical analysis methods. In addition, the guidelines suggest helpful presentations of data and important elements to include in discussions. The goal of these guidelines is to encourage transparent and complete reporting so that the relevant information will be available to others to help them to judge the usefulness of the data and understand the context in which the conclusions apply.

REporting recommendations for tumour MARKer prognostic studies (REMARK).

Despite years of research and hundreds of reports on tumour markers in oncology, the number of markers that have emerged as clinically useful is pitifully small. Often initially reported studies of a marker show great promise, but subsequent studies on the same or related markers yield inconsistent conclusions or stand in direct contradiction to the promising results. It is imperative that we attempt to understand the reasons that multiple studies of the same marker lead to differing conclusions. A variety of methodological problems have been cited to explain these discrepancies. Unfortunately, many tumour marker studies have not been reported in a rigorous fashion, and published articles often lack sufficient information to allow adequate assessment of the quality of the study or the generalisability of the study results. The development of guidelines for the reporting of tumour marker studies was a major recommendation of the US National Cancer Institute and the European Organisation for Research and Treatment of Cancer (NCI-EORTC) First International Meeting on Cancer Diagnostics in 2000. Similar to the successful CONSORT initiative for randomised trials and the STARD statement for diagnostic studies, we suggest guidelines to provide relevant information about the study design, preplanned hypotheses, patient and specimen characteristics, assay methods, and statistical analysis methods. In addition, the guidelines suggest helpful presentations of data and important elements to include in discussions. The goal of these guidelines is to encourage transparent and complete reporting so that the relevant information will be available to others to help them to judge the usefulness of the data and understand the context in which the conclusions apply.

Generation of recombinant norovirus-like particles (VLP) in the human endothelial kidney cell line 293T.

Noroviruses (NVs) are the major cause of non-bacterial outbreaks of gastroenteritis. Here we report a new alternative system to generate recombinant NV virus-like particles (rNV-VLP) in a human endothelial kidney cell line (HEK). Transfecting HEK-293T cells with an expression vector coding for the ORF-2 gene lead to the expression of the viral structural protein VP1 which spontaneously assembled into virus-like particles (VLP), as shown by electron microscopy. The transfected cells did not show a cytopathic effect and released rNV-VLP into the culture medium. The HEK-293T cell derived particles were morphologically indistinguishable to the rNV-VLP produced from baculovirus and the Venezuelan equine encephalitis virus (VEE)-replicon. The produced particles were stable for at least 2.5 months at 4 degrees C.

The Cooperative Breast Cancer Tissue Resource: archival tissue for the investigation of tumor markers.

Investigators continue to search for reliable markers of prognosis of breast cancer. For many analyses, laboratory techniques permit the use of archival paraffin-embedded tissue collected years previously and readily linked to clinical and follow-up information. Laboratory investigators have often expressed the need for such a tissue resource. We have developed a publicly available resource of archival breast cancer specimens. The pathological material has been collected and reviewed by investigators at four institutions and currently includes breast cancer specimens from more than 9300 cases. Institutional pathologists reviewed slides and blocks using a common protocol and coding scheme. Clinical information and details of follow-up came from data routinely collected by the institutions' cancer registries. Coded data are maintained centrally in a single database. A subset of the data may be searched on the World Wide Web to determine the availability of cases with specified characteristics. The material collected by this Cooperative Breast Cancer Tissue Resource is generally representative of breast cancer diagnosed in community hospital settings in the United States. Seventy-two percent of the living cases have been followed for at least 5 years, and follow-up status is updated regularly. Interested laboratory investigators may apply to the Resource for the use of these tissues. This Resource is proving valuable to laboratory investigators who require large numbers of specimens for validation studies of prognostic markers of breast cancer.

Inhibition of beta(2) integrin-mediated leukocyte cell adhesion by leucine-leucine-glycine motif-containing peptides.

Many integrins mediate cell attachment to the extracellular matrix by recognizing short tripeptide sequences such as arginine-glycine-aspartic acid and leucine-aspartate-valine. Using phage display, we have now found that the leukocyte-specific beta(2) integrins bind sequences containing a leucine-leucine-glycine (LLG) tripeptide motif. An LLG motif is present on intercellular adhesion molecule (ICAM)-1, the major beta(2) integrin ligand, but also on several matrix proteins, including von Willebrand factor. We developed a novel beta(2) integrin antagonist peptide CPCFLLGCC (called LLG-C4), the structure of which was determined by nuclear magnetic resonance. The LLG-C4 peptide inhibited leukocyte adhesion to ICAM-1, and, interestingly, also to von Willebrand factor. When immobilized on plastic, the LLG-C4 sequence supported the beta(2) integrin-mediated leukocyte adhesion, but not beta(1) or beta(3) integrin-mediated cell adhesion. These results suggest that LLG sequences exposed on ICAM-1 and on von Willebrand factor at sites of vascular injury play a role in the binding of leukocytes, and LLG-C4 and peptidomimetics derived from it could provide a therapeutic approach to inflammatory reactions.

Reproducibility of p53 immunohistochemistry in bladder tumors. National Cancer Institute, Bladder Tumor Marker Network.

The National Cancer Institute Bladder Tumor Marker Network conducted a study to evaluate the reproducibility of immunohistochemistry for measuring p53 expression in bladder tumors. Fifty paraffin blocks (10 from each of the five network institutions) were chosen at random from among high-grade invasive primary bladder tumors. Two sections from each block were sent to each laboratory for staining and scoring, and then all sections were randomly redistributed among the laboratories for a second scoring. Intra- and interlaboratory reproducibility was assessed with regard to both staining and scoring. For overall assessments of p53 positivity, the results demonstrated that intralaboratory reproducibility was quite good. Concordance across the five participating laboratories was high for specimens exhibiting no or minimal nuclear immunostaining of tumor cells or high percentages of tumor cells with nuclear immunoreactivities. However, there was a reduced level of concordance on specimens with percentages of stained tumor cells in an intermediate range. The discordancies were due mainly to staining differences in one of the five laboratories and scoring differences in another laboratory. These results indicate that some caution must be used in comparing results across studies from different groups. Standardization of staining protocols and selection of a uniform threshold for binary interpretation of results may improve assay reproducibility between laboratories.

Tumor marker utility grading system: a framework to evaluate clinical utility of tumor markers.

Introduction of tumor markers into routine clinical practice has been poorly controlled, with few criteria or guidelines as to how such markers should be used. We propose a Tumor Marker Utility Grading System (TMUGS) to evaluate the clinical utility of tumor markers and to establish an investigational agenda for evaluation of new tumor markers. A Tumor Marker Utility Grading Worksheet has been designed. The initial portion of this worksheet is used to clarify the precise characteristics of the marker in question. These characteristics include the marker designation, the molecule and/or substance and the relevant alteration from normalcy, the assay format and reagents, the specimen type, and the neoplastic disease for which the marker is being evaluated. To determine the clinical utility of each marker, one of several potential uses must be designated, including risk assessment, screening, differential diagnosis, prognosis, and monitoring clinical course. For each of these uses, associations between marker assay results and expected biologic process and end points must be determined. However, knowledge of tumor marker data should contribute to a decision in practice that results in a more favorable clinical outcome for the patient, including increased overall survival, increased disease-free survival, improvement in quality of life, or reduction in cost of care. Semiquantitative utility scales have been developed for each end point. The only markers recommended for use in routine clinical practice are those that are assigned utility scores of "++" or " " on a 6-point scale (ranging from 0 to ) in the categories relative to more favorable clinical outcomes. Each utility score assignment should be supported by documentation of the level of evidence used to evaluate the marker. TMUGS will establish a standardized analytic technique to evaluate clinical utility of known and future tumor markers. It should result in improved patient outcomes and more cost-efficient investigation and application of tumor markers.

The psychotherapy of patients with complex partial seizures.

The direct neurological effect of complex partial seizures (CPS) is described, as is the psychological and interpersonal impact of the disorder, with emphasis on the iterative nature of the process. A model for the psychotherapy of CPS patients is presented that is based on an understanding of the neurological changes and continuous interaction between these changes and the psychological demands of adaptation.

Helium-neon laser therapy in the prevention of postoperative swelling and pain after wisdom tooth extraction.

Seventeen patients with symmetrically embedded lower wisdom teeth were selected for this study, intended to evaluate the local effect of soft laser therapy on postoperative pain. Both lower third molars were removed in the same operation. The test side, chosen by lot, was treated using a helium-neon laser (632,8 nm, 8 mW, 50 Hz) for 2 minutes. The other side served as the untreated control side. Facial swelling was measured using a modification of the face-bow technique. Postoperative pain was estimated using a visual analogue scale (VAS). When it became apparent that conventional statistical analysis was revealing no difference in postoperative swelling and pain between the test and the control groups, the study was discontinued for ethical reasons.

The science base underlying research on acquired immune deficiency syndrome.

In order to define the clinical syndrome of AIDS and begin to deal with it effectively, scientists needed to understand how the immune system works. Fortunately, considerable knowledge was available: research in immunology over the last two decades had provided the technological advances and basic information about cell-mediated immunity that were necessary for identification of the syndrome. Without this knowledge base, immune suppression would not have been recognized as the common link among AIDS patients manifesting a variety of infections and unusual neoplasms. Similarly, research on infectious diseases, and in particular on the role of viruses as etiologic agents, has had an important bearing on understanding of AIDS. The epidemiologic data to date indicate that an infectious agent most likely is involved and that transmission of the disease requires intimate contact and perhaps some passage of blood. Among the candidates for viral agents are Epstein-Barr virus, cytomegalovirus, and human T-cell leukemia virus. All have been isolated from the cells of AIDS victims, but whether they are etiologic agents or opportunistic pathogens remains unresolved. Knowledge gained from the study of any of these viruses will contribute to understanding of AIDS, and vice versa. In this paper, we have attempted to show the integral relationship between specific research on AIDS and the ongoing research effort in related disciplines. It is important to recognize that effective research is the result of careful consideration of which questions can and should be addressed and the development of innovative approaches to gain answers to those questions. Research on AIDS is proceeding as rapidly as it is only because of the solid foundations that have been developed in the areas of immunology and virology. It is this base of research that ultimately will provide the rationale and the tools for solving new problems.

Surgical treatment of congenital esotropia.

We conducted a retrospective study of 133 children (69 boys and 64 girls) who underwent bilateral medial rectus muscle recession (most by the augmented or en-bloc technique) for congenital esotropia. Esotropia was diagnosed before the age of 6 months in 84 patients and after the age of 6 months in the other 49. A total of 27 children underwent surgery before the age of 12 months; of these, three required second procedures. A total of 106 children underwent surgery after the age of 12 months; of these, eight required second procedures. The mean preoperative deviation was 40 prism diopters. Two patients had significant A pattern deviations and 17 had significant V pattern deviations. Six patients had dissociated vertical deviations. Five to 60 days after surgery, 52 patients had no deviation and 99 were within +/- 10 prism diopters of no deviation. Two months after surgery, 67 patients had no deviation and 114 were within +/- 10 prism diopters of no deviation. Final alignments (five months to seven years postoperatively) showed that 51 patients had no deviation and 109 were within +/- 10 prism diopters of no deviation. Despite adequate alignment, none of 13 patients whose esotropia was diagnosed before the age of 6 months, who underwent surgery before the age of 12 months, and who cooperated with testing achieved stereopsis. This suggested that there may be two types of congenital esotropia--one without fusion potential and one in which fusion is possible but lost secondarily because of peripheral esotropic factors.