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To monitor infection by the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) and successful vaccination against coronavirus disease 2019 (COVID‐19), the kinetics of neutralizing or blocking anti‐SARS‐CoV‐2 antibody titers need to be assessed. Here, we report the development of a quick and inexpensive surrogate SARS‐CoV‐2 blocking assay (SUBA) using immobilized recombinant human angiotensin‐converting enzyme 2 (hACE2) and human cells expressing the native form of surface SARS‐CoV‐2 spike protein. Spike protein‐expressing cells bound to hACE2 in the absence or presence of blocking antibodies were quantified by measuring the optical density of cell‐associated crystal violet in a spectrophotometer. The advantages are that SUBA is a fast and inexpensive assay, which does not require biosafety level 2‐ or 3‐approved laboratories. Most importantly, SUBA detects blocking antibodies against the native trimeric cell‐bound SARS‐CoV‐2 spike protein and can be rapidly adjusted to quickly pre‐screen already approved therapeutic antibodies or sera from vaccinated individuals for their ACE2 blocking activities against any emerging SARS‐CoV‐2 variants., A simple, inexpensive surrogate SARS‐CoV‐2 neutralization assay is introduced. Ectopic expression of the SARS‐CoV‐2 spike protein confers binding of Ramos‐Ig ‐ or HEK293 cells to plate‐bound recombinant human angiotensin‐converting‐enzyme‐2. This interaction can be blocked with reconvalescent or immune serum or with anti‐receptor‐binding domain antibodies. Bound spike‐expressing cells are quantitated with crystal violet.

Receptor occupancy, the ratio between amount of drug bound and amount of total receptor on single cells, is a biomarker for treatment response to therapeutic monoclonal antibodies. Receptor occupancy is traditionally measured by flow cytometry. However, spectral overlap in flow cytometry limits the number of markers that can be measured simultaneously. This restricts receptor occupancy assays to the analysis of major cell types, although rare cell populations are of potential therapeutic relevance. We therefore developed a receptor occupancy assay suitable for mass cytometry. Measuring more markers than currently available in flow cytometry allows simultaneous receptor occupancy assessment and high‐parameter immune phenotyping in whole blood, which should yield new insights into disease activity and therapeutic effects. However, varying sensitivity across the mass cytometer detection range may lead to misinterpretation of the receptor occupancy when drug and receptor are detected in different channels. In this report, we describe a method for optimization of mass cytometry receptor occupancy measurements by using antibody‐binding quantum simply cellular (QSC) beads for standardization across channels with different sensitivities. We evaluated the method in a mass cytometry‐based receptor occupancy assay for natalizumab, a therapeutic antibody used in multiple sclerosis treatment that binds to α4‐integrin, which is expressed on leukocyte cell surfaces. Peripheral blood leukocytes from a treated patient were stained with a panel containing metal‐conjugated antibodies for detection of natalizumab and α4‐integrin. QSC beads with known antibody binding capacity were stained with the same metal‐conjugated antibodies and were used to standardize the signal intensity in the leukocyte sample before calculating receptor occupancy. We found that QSC bead standardization across channels corrected for sensitivity differences for detection of drug and receptor and generated more accurate results than observed without standardization. © 2019 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

© 2021 The Authors., The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers., Hyun-Dong Chang is supported by the Dr. Rolf M. Schwiete Foundation. Susanne Melzer and Attila Tarnok thank De Novo Software for providing FCS Express. Enrico Lugli is supported by a grant from the Associazione Italiana per la Ricerca sul Cancro (AIRC IG20676). Gabriele De Simone and Giovanni Galletti were supported by Fellowships from the Fondazione Italiana per la Ricerca sul Cancro-Associazione Italiana per la Ricerca sul Cancro (FIRC-AIRC). Jun Dong is supported by Deutsche Forschungsgemeinschft (DFG, German Research Foundation) Projektnummer 389687267 and Chinesisch-Deutsches Zentrum für Wissenschaftsförderung [Sino-German Center for Research Promotion (SGC)] grant C-0072. Nicola Gagliani, Samuel Huber and Franziska Muscate are supported by DFG fundings: SFB841,GA 2441/3-1, HU 1714/10-1. The tetramer APC-conjugated H-2K (d) HIV-1 gag197-205 AMQMLKETI used in TDS assay for mouse blood T cells was obtained through the NIH Tetramer Facility. Larissa Nogueira Almeida was supported by DFG research grant MA 2273/14-1. Supported by the following grants: AIRC 5X1000 2018 id. 21147 (Lorenzo Moretta); AIRC IG 2017 id. 19920 (Lorenzo Moretta); RC-2020 OPBG (Lorenzo Moretta); AIRC and European Union's Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement No 800924 (Linda Quatrini). Dirk Baumjohann was supported by Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Emmy Noether Programme BA 5132/1-2 (252623821) and Germany's Excellence Strategy EXC2151 (390873048).

Hepatitis B virus (HBV) from 76 adult immigrants in Australia from Myanmar was characterized to determine the prevalence of different HBV genotypes and subgenotypes. A mutational analysis was then performed to determine the presence of clinically significant mutations and correlate them to clinical outcomes. Initial genotyping revealed 68 patients with genotype C (89.5%) and eight patients with genotype B (10.5%). Phylogenetic analysis revealed the large majority of the genotype C infections were of subgenotype C1 (67/68). Sequencing of the HBV polymerase gene (and overlapping surface gene) revealed no mutations associated with antiviral resistance. HBV surface gene mutations were detected in 10 patients with subgenotype C1. HBV BCP/PC sequencing was obtained for 71/76 (93%) patients. BCP and/or PC mutations were identified in 57/71 (80%) of PCR positive patients. Treatment had been commenced for 15/76 (18%) patients, a further 26 untreated patients were in a stage of disease where HBV treatment would be considered standard of care. It was identified that genotype C1 is the predominant sub-genotype in this population. Genotype C is known to be associated with increased risk of development of HCC. This highlights the need for screening for HCC given the potential for the development of liver cancer. It was also identified that people with HBV were potentially not receiving optimal therapy in a timely fashion.

Patient-derived cancer xenografts (PDX) are widely used to identify and evaluate novel therapeutic targets, and to test therapeutic approaches in preclinical mouse avatar trials. Despite their widespread use, potential caveats of PDX models remain considerably underappreciated. Here, we demonstrate that EBV-associated B-lymphoproliferations frequently develop following xenotransplantation of human colorectal and pancreatic carcinomas in highly immunodeficient NOD.Cg-Prkdcscid Il2rgtm1Wjl /SzJ (NSG) mice (18/47 and 4/37 mice, respectively), and in derived cell cultures in vitro. Strikingly, even PDX with carcinoma histology can host scarce EBV-infected B-lymphocytes that can fully overgrow carcinoma cells during serial passaging in vitro and in vivo. As serial xenografting is crucial to expand primary tumor tissue for biobanks and cohorts for preclinical mouse avatar trials, the emerging dominance of B-lymphoproliferations in serial PDX represents a serious confounding factor in these models. Consequently, repeated phenotypic assessments of serial PDX are mandatory at each expansion step to verify "bona fide" carcinoma xenografts.

Mass cytometry has pioneered >40-parameter single-cell analyses that allow for the characterization of complex cellular networks at unprecedented depth. Up to 135 parameters can be simultaneously detected, but limited availability of metal tags suitable for labeling of specific probes prevents optimal exploitation of the analytical capacity of mass cytometers. To this end, we here establish the application of elemental silver nanoparticles (AgNP) of different size for reporting cell surface antigens on human leukocytes in mass cytometry assays. The mass channels at 107 Da and 109 Da are uniquely occupied by silver isotopes and do not interfere with other mass cytometry reagents. Streptavidin-coated AgNP (SA-AgNP) facilitated distinct and specific detection of various antigens, such as CD8, CD244 and CD294 on peripheral blood leukocytes pre-incubated with respective biotinylated primary antibodies. Signal intensities elicited by 40 nm-sized AgNP allowed specific detection of the low abundance antigen CD25 on both, peripheral blood regulatory T cells and CD25lo CD127+ CD4+ T cells, enabling their distinct clustering in viSNE plots. SA-AgNP were of high elemental purity, showed minor background binding to cells in immunoassays, and were compatible with previously established staining protocols for PBMC and leukocytes, facilitating their use in complex mass cytometry panels. Considering the synthesis of AgNP from isotopically purified silver, the usage of AgNP extends the analytical capacity of mass cytometry panels by one, prospectively two, additional parameters, suitable for the detection of cellular targets of low abundance. © 2016 International Society for Advancement of Cytometry.

BACKGROUND: Evaluation of an outreach programme using a mobile transient elastography (TE) device (FibroScan) to improve liver disease assessment in different clinical settings. AIMS: To evaluate a programme of liver fibrosis assessment by TE and to compare fibrosis scores between different sites and patient groups. METHODS: Prospective cohort study. TE was conducted at a tertiary hospital and during outreach clinics in three different settings: community clinics, clinics for people who use drugs (PWUD) and regional clinics in rural Victoria. All patients referred for TE at the participating locations were eligible during the study period. RESULTS: A total of 200 of 623 patients was assessed and evaluated during outreach sessions (regional 100; PWUD 18; community 82). While the majority of patients in community centres were infected with hepatitis B (68%), most patients in regional clinics and in PWUD settings had hepatitis C virus (HCV) (81 and 100%, respectively). Significantly more patients assessed at regional clinics and PWUD settings presented with severe fibrosis (F3-F4, F4): regional clinics 39%; PWUD 31%; tertiary 11%; community 7%, (P

Background In 2011, the Australian Government introduced Medicare item numbers for telehealth consultations. This is a rapidly expanding method of healthcare provision. Aims We assessed the demographic and disease profile of refugee patients attending a new telehealth clinic, and calculated the patient travel avoided. We examined technical challenges and assessed the performance of two videoconferencing solutions using different bandwidth and latencies. Methods We audited the first 120 patients attending the telehealth clinic. During consultations, the patient was with the general practitioner (GP) and linked by internet videoconference using VIDYO, GoToMeeting or Skype, to the specialist at a tertiary referral hospital. Travel avoided was calculated and technical problems were assessed by the participating specialist. Bandwidth and latency variations were examined within a university broadband testing facility. Results The two most frequently managed conditions were hepatitis C and latent tuberculosis. Twenty-nine different GP were included and 42 consultations required an interpreter. Nearly 500 km of travel and 127 kg of CO2 production was avoided per consultation. Technical issues were faced in 25% of consultations, most frequently sound problems and connections dropping out. A bandwidth of at least 512 kbps and latency of no more than 300 ms was necessary to conduct an adequate multipoint videoconference. Conclusions Telehealth using videoconferencing adds a new component to care of refugee and immigrant patients settling in regional areas. Telehealth will be improved by changes to improve simplicity and standardisation of videoconferencing, but requires ongoing Medicare funding to allow sufficient administrative support.

Objective To determine the incidence of carpal tunnel syndrome (CTS) over 1 year in Latino poultry processing workers. Methods Symptoms and nerve conduction studies were used to identify Latino poultry processing workers (106 wrists) and Latinos in other manual labor occupations (257 wrists) that did not have CTS at baseline, and these individuals were then evaluated in the same manner 1 year later. Results Based on wrists, the 1-year incidence of CTS was higher in poultry processing workers than non-poultry manual workers (19.8% vs. 11.7%, P = 0.022). Poultry workers had a higher odds (1.89; P = 0.089) of developing CTS over 1 year compared to non-poultry manual workers. Discussion Latino poultry processing workers have an incidence of CTS that is possibly higher than Latinos in other manual labor positions. Latino poultry workers' high absolute and relative risk of CTS likely results from the repetitive and strenuous nature of poultry processing work. Am. J. Ind. Med. 57:362–369, 2014. © 2013 Wiley Periodicals, Inc.

Introduction: The prevalence of bifid median nerves and persistent median arteries, their co-occurrence, and their relationship to carpal tunnel syndrome (CTS) are only understood partially. Methods: We screened 1026 wrists of 513 Latino manual laborers in North Carolina for bifid median nerves and persistent median arteries using electrodiagnosis and ultrasound. Results: A total of 8.6% of wrists had a bifid median nerve, and 3.7% of wrists had a persistent median artery independent of subgroup ethnicity, age, gender, or type of work. An association with definite carpal tunnel syndrome was not found. The presence of either anatomic variant was associated with a high likelihood of co-occurrence of another variant in the same or the contralateral wrist. Conclusions: The occurrence of median anatomic variants can be determined in field studies using ultrasound. Persistent median arteries and bifid median nerves tend to co-occur but do not put manual laborers at additional risk of developing CTS. Abbreviations BMI: body mass index BMN: bifid median nerve CTS: carpal tunnel syndrome

Inadequate control of high systolic blood pressure in older adults has been largely attributable to poor control of overall hypertension (HTN). The Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) guidelines emphasize the importance of controlling isolated systolic HTN in older adults. The study examined demographics, self-reported health information, and clinical measures as predictors of uncontrolled HTN among individuals taking antihypertensive medications. The Community Initiative to Eliminate Stroke, a stroke risk factor screening and prevention project, collected data in two North Carolina counties. Statistical modeling of predictors included odds ratios (ORs) and logistic regression analyses. Of the 2663 participants, 43.5% and 22.8% had uncontrolled systolic and diastolic HTN, respectively. African Americans were more likely to have uncontrolled systolic (60%) or diastolic HTN (70.9%) compared with whites (40% and 29.1%, respectively). Participants 55 years and older were more likely to have uncontrolled systolic HTN compared with younger individuals. Regression analyses showed that race (OR, 1.239; P=.00), age (OR, 1.683; P=.00), and nonadherence with medications (OR, 2.593; P=.00) were significant predictors of uncontrolled systolic HTN. Future interventions should focus on improving management of isolated systolic HTN in older adults and African Americans to increase overall control of HTN.

Introduction: Manual laborers are at increased risk for carpal tunnel syndrome (CTS), and a combination of history, physical examination, and nerve conduction studies is often used to screen for CTS in this population. Neuromuscular ultrasound may be a better screening tool, because it is painless. In this study we compare the accuracy of nerve conduction studies and ultrasound for CTS screening. Methods: Five hundred thirteen manual laborers were screened prospectively for CTS using nerve conduction studies and neuromuscular ultrasound, and the accuracy of the 2 techniques was compared using the Katz hand diagram as the diagnostic standard. Results: The ROC curves for the 2 techniques were not significantly different (P = 0.542), indicating that the approaches had similar diagnostic accuracy. Conclusions: Neuromuscular ultrasound is a painless technique that has diagnostic accuracy similar to nerve conduction studies and can be used to screen large populations at risk for CTS. Abbreviations CTS: carpal tunnel syndrome ROC: receiver operating characteristic

Background Immigrant Latino workers represent an expanding workforce in rural areas of the USA, where their employment is concentrated in occupations such as poultry processing that entail chemical, infectious, and mechanical skin exposures. Occupation-related skin illnesses in this vulnerable population are not well characterized. Objectives This study was designed to describe the prevalences of skin diseases among immigrant Latino poultry processors and other manual workers in North Carolina. Methods Community-based sampling was used to recruit 742 immigrant Latino workers, 518 of whom underwent a physical examination supervised by a board-certified dermatologist. The presence or absence of skin disease on the face, neck, arms, hands, and feet was recorded. Results Workers ranged in age from 18 years to 68 years. Slightly over half of the sample were male (52.6%). Poultry workers represented 55.8% of the study sample. Infectious skin diseases were the most common diagnosis, present in 52.3% of workers. Inflammatory skin diseases were present in 28.2% and pigmentary disorders in 21.8% of workers. The most common skin conditions were tinea pedis (37.6%), onychomycosis (31.9%), scars (13.7%), acne (11.8%), and melasma (9.3%). Age, sex, first language, and work as a poultry processor accounted in part for the prevalence of these diseases. Conclusions Several skin diseases are highly prevalent in immigrant Latino workers and may relate to work environment. These may impair the quality of life of these workers and predispose them to further illness.

Herein, we report on the first enantioselective synthesis of the sparteine-type alkaloids virgidivarine (1) and virgiboidine (2). The stereoselective construction of the challenging dipiperidine core within 1 and 2 was realized by applying an intramolecular ene-ene-yne ring-rearrangement metathesis (RRM) protocol. In combination with this sequence, an unprecedented tandem metathesis/oxidation reaction was established. Subsequent to the RRM event, the Ru-alkylidene species was converted into the highly potent dihydroxylation catalyst RuO4 by the addition of NaIO4. This transformation rendered five sequential reactions that include an ene-ene-yne RRM/dihydroxylation/glycol cleavage in a one-pot procedure. This approach provides practical and general access to dipiperidines and piperidino-quinolizidines. Despite their wide use in organic and organometallic chemistry as well as their occurrence in biologically active natural products, methods are still lacking for the stereoselective synthesis of these motifs.

Background: The goal of this study is to improve understanding of immigrant Latino manual workers' occupational health, focusing on upper body musculoskeletal injury. Methods: Physical exams were conducted with a representative sample of 516 Latino poultry workers and manual laborers in western North Carolina; outcome measures were prevalence of epicondylitis, rotator cuff syndrome, and low back pain. Results: Low back pain (n?=?89; 17.2%) and rotator cuff syndrome (n?=?76; 14.7%) indicated by physical exam was common. Epicondylitis was less common, but still frequent (n?=?30; 5.8%). Prevalence of each outcome did not differ between poultry processing workers and other manual workers. Workers >40 years old had greater incidence of rotator cuff syndrome and epicondylitis. Conclusions: Epicondylitis, rotator cuff syndrome, and low back pain are common in immigrant Latino workers, and may negatively impact long-term health and contribute to occupational health disparities.

Zusammenfassung 1 Am 6. Tag nach dem Schlupfen ist im Dunndarminhalt von Kuken, die Futter mit 150 ppm Chlortetracyclin bekamen, etwa 50% weniger Cholin vorhanden als bei Kontrolltieren, die kein Antibiotikum erhielten. 2 Aus den Messungen der Blutkonzentrationen nach enteraler Applikation von radiomarkiertem Cholin ist zu schliesen, das Cholin bei vier Wochen alten Kuken schnell und vollstandig aus dem Duodenum und Jejunum resorbiert wird. Im Ileum wird ein betrachtlicher Anteil des Cholins abgebaut. 3 Bei acht Tage alten Hahnchen, deren Futter 20 ppm Chlortetracyclin enthielt, wurde Cholin schneller aus dem Darm resorbiert als bei den Kontrollen. Zwei Minuten nach der intraduodenalen Injektion von 1 μCi Cholin waren bei den mit dem Antibiotikum behandelten Tieren etwa 50%, bei den Kontrollen dagegen nur 25% der Radioaktivitat aus dem Darm aufgenommen. Die gleichen Verhaltnisse wurden bei Tieren im Alter von 5, 12 und 20 Tagen angetroffen. 4 Aus den Ergebnissen wird gefolgert, das der wachstumsfordernde Effekt von Chlortetracyclin auf einer gesteigerten Resorption essentieller Nahrungbestandteile beruht.

Context: Skin diseases are common occupational illnesses for migrant farmworkers. Farmworkers face many barriers in accessing health care resources. Purpose: Framed by the Health Behavior Model, the purpose of this study was to assess health care utilization for skin disease by migrant Latino farmworkers. Methods: Three hundred and four migrant and seasonal Latino farmworkers in North Carolina were enrolled in a longitudinal study of skin disease and health care utilization over a single agricultural season. Self-reported and dermatologist-diagnosed skin condition data were collected at baseline and at up to 4 follow-up assessments. Medical visit rates were compared to national norms. Findings: Self-reported skin problems and diagnosed skin disease were common among farmworkers. However, only 34 health care visits were reported across the entire agricultural season, and none of the visits were for skin diseases. Nevertheless, self-treatment for skin conditions was common, including use of non-prescription preparations (63%), prescription products (9%), and home remedies (6%). General medical office visits were reported in 3.2% of the assessments, corresponding to 1.6 office visits per person year. Conclusions: The migrant farmworker population consists largely of young men who make little use of clinic services. Skin conditions are very common among these workers, but use of medical services for these conditions is not common. Instead, farmworkers rely primarily on self-treatment. Clinic-based studies of farmworker skin conditions will not account for most injury or disease in this population and have the potential for biased estimates.

Background: Melasma is a common condition of Latino women that detracts from their quality of life (QOL). The prevalence and impact of melasma in Latino men is not well characterized. Aim: To assess the prevalence of melasma and its association with QOL in Latino men from Mexico and Central America working in the USA. Methods: The prevalence of melasma was assessed in three studies of Latino men: by direct examination in a study of 25 Latino poultry workers; by direct examination in a study of 54 Latino farm workers; and by examination of store-and-forward teledermatology images in a study of 300 Latino farm workers. QOL was assessed with a Spanish version of the Dermatology Life Quality Index (DLQI). Results: The prevalence of melasma was 36.0%, 7.4%, and 14.0% in the three studies. The prevalence of melasma was greatest amongst those aged 31 years and older, who were from Guatemala, and who spoke an indigenous language. The presence of melasma was associated with higher DLQI scores, indicating a poorer QOL, in the poultry worker population. Conclusions: Melasma is a common condition in Latino men and is associated with a poor QOL in some affected individuals. Clinicians should be aware that melasma may be a concern for their male Latino patients. Research on the association of skin conditions with QOL amongst minority men is needed.

Background: This study estimates the prevalence of self-reported skin problems among Latino farmworkers and identifies associated risk factors. Methods: The study used a longitudinal surveillance design. Participants were interviewed up to five times and reported skin problems and personal, work, and environment characteristics. Frequencies and counts were calculated for 13 skin problems. Adjusted odds ratios were obtained for six skin problems. Results: More than one-third of participants reported skin problems, including skin and nail fungus; sunburn; bumps, pimples, or acne; calluses; itching; rash; and insect bite. A variety of work and environment factors were associated with higher rates of skin problems. One of the strongest predictors was working in wet clothes or shoes. Conclusions: Programs are needed to educate farmworkers about measures they can take to decrease their risk of skin problems. Changes in work practices and personal protective equipment provided could help decrease the prevalence of skin problems.

The 5500T allele variant of the C5500T single nucleotide polymorphism in the human G protein beta3 subunit (GNB3) has been reported to be associated with primary hypertension. In this study, the GNB3 gene of primary hypertensive and normotensive dogs was examined for an analogous nucleotide polymorphism associated with hypertension. The genomic GNB3 dna, with 10 exons and nine introns coding for 340 amino acids, is described. PCR product sequencing of the GNB3 exon 9 from 25 dogs (including five hypertensive animals) failed to detect any nucleotide polymorphism. In contrast to human beings, there was no polymorphism at either the analogous nucleotide or in the respective exon. Only the human hypertension-associated thymine was detected, regardless of whether the dogs were hypertensive or normotensive. Furthermore, examinations of 565 dogs of 85 distinct breeds for the presence of the human 5500C nucleotide at the analogous nucleotide side failed to detect a cytosine that is present with high allele frequency in normotensive man. Owing to the lack of allele variance, it is concluded that canine primary hypertension is not associated with a polymorphism at either the respective human hypertension-associated nucleotide site or in the entire exon.

Agarose gel electrophoresis, which separates and sizes linear DNA and RNA fragments, is arguably the most basic and essential technique in molecular biology. It is commonly employed for analysis of PCR products, plasmid DNA, and products of restriction enzyme digestion. It is the first step for analysis of specific DNA and RNA fragments by northern and Southern blots. In this unit, we provide both written instructions and photographic images to take the reader from preparing a first agarose gel to analyzing results and determining the size of sample DNA. We include two protocols: agarose gel electrophoresis (commonly used to analyze DNA) and denaturing gel electrophoresis (for analyzing RNA). We have divided each protocol into four basic steps: (1) preparing and pouring the agarose gel; (2) preparing and loading samples; (3) running the agarose gel; and (4) staining the gel using the fluorescent stain ethidium bromide to visualize DNA and RNA. © 2015 by John Wiley & Sons, Inc. Keywords: DNA; RNA; separation; sizing; ethidium bromide; voltage; TAE; TBE; bromphenol blue; xylene cyanol; northern blotting; Southern blotting

Ultraviolet (UV) disinfection is being considered for inactivation of pathogens in filtered surface waters across North America. MS2 coliphage is the most commonly used test microbe for UV reactor validation in North America, and the development of UV dose-response data for MS2 at bench scale is an integral part of validation testing. This research evaluated the effect of water quality (e.g., turbidity, particle count, particle size, and absorbance) and sample depth on inactivation of MS2 coliphage in 17 filtered waters. In addition, the inactivation performance of low-pressure (LP) and medium-pressure (MP) lamp types was compared. Results indicated that turbidity, particle count, and absorbance, when factored into the bench-scale dose measurement, did not affect UV inactivation of MS2 coliphage in filtered waters meeting federal regulations. UV light from MP lamps appeared more effective than LP UV for inactivating MS2. These results suggest that water quality should not be considered a major factor in properly designed UV bench-scale tests or in reactor validation challenges that use inactivation kinetics of MS2 coliphage as a dose-measurement tool in filtered surface waters.

BACKGROUND: Within an ongoing birth cohort study (LISA) the cytokine production of cord blood T cells was compared between neonates from Leipzig (East Germany) and Munich (West Germany). The aim of this study was to analyse regional differences and influencing factors of the immune status. METHODS: Cytokine production was measured in a randomly selected subgroup of 158 children from the LISA (Life style - Immune system - Allergy) cohort by intracellular cytokine staining. Information on family "atopy" history (FAH) and home characteristics was obtained from questionnaires. RESULTS: Reduced numbers of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) producing T cells were found in association with biparental FAH and housing renovation during pregnancy. In addition, cytokine production was influenced by season. In Munich, the frequency of biparental FAH and of renovation measures during pregnancy was significantly higher as compared to Leipzig. Neonates from Munich showed significantly decreased amounts of IFN-gamma and TNF-alpha and elevated levels of interleukin-4 (IL-4) producing T cells. Differences in cytokine production between Munich and Leipzig were influenced by season (IL-4) and housing renovation (IFN-gamma, TNF-alpha). CONCLUSIONS: Since differences in the T cell cytokine production of neonates in Munich and Leipzig are independent from FAH our findings may provide evidence for the impact of environmental factors upon the fetal immune system. &nbsp

Indoor VOC (volatile organic compound) exposure has been shown to be correlated with airway symptoms and allergic manifestations in children. An investigation was conducted within an ongoing birth cohort study (LISA: Lifestyle-Immune System-Allergy) of the association between maternal exposure to VOCs and immune status at birth, in particular the cytokine secretion profile of cord-blood T cells. In a randomly selected group of 85 neonates, cytokine-producing cord-blood T cells were analyzed using intracellular cytokine detection. VOC exposure was measured in children's dwellings by passive sampling, while parents were asked to complete questionnaires about possible sources of VOC exposure. Adjusted odds ratios (ORs) were calculated by logistic regression based on categorized quartiles. A positive association was found between elevated percentages of interleukin-4-producing (IL-4) type 2 T cells and exposure to naphthalene (OR = 2.9) and methylcyclopentane (OR = 3.3). Exposure to tetrachloroethylene was associated with reduced percentages of interferon-gamma-producing (IFN-gamma) type 1 T cells (OR = 2.9). In addition, smoking during pregnancy was correlated with a higher indoor air concentration of naphthalene (OR = 3.8), new carpets in infants' bedrooms with elevated methylcyclopentane concentrations (OR = 4.1), and home renovation with a higher trichloroethylene burden (OR = 4.9). Our data suggest that maternal exposure to VOC may have an influence on the immune status of the newborn child.

Background: Ethnic minorities have been omitted from many occupational health studies, despite their substantial contributions to the labor force and documented cases of high exposures in some settings. To describe the occurrence of potentially work-related cancers among African American and Latino Workers, we conducted an epidemiologic study based on death certificate data. Methods: Data were obtained from 21 states during 1985-1992. Directly adjusted proportionate mortality ratios (PMRs) and standardized mortality rate ratios (SRRs) for ages 20-64 years were computed for cancers of the lung, nasal cavity, pleura, and peritoneum, malignant melanoma and leukemia using the mortality data and population counts from the 1990 census. Results: Proportionate mortality was generally low for both groups, but African Americans had higher than expected mortality rates for leukemia and cancers of the lung, nasal cavity, and peritoneum. Industry-specific analyses indicate excess leukemia among African American men in the rubber industry (PMR 2.08, 95% confidence interval (CI) 1.29-3.35), Latino men in textile (PMR 2.31, 95% CI 0.81-5.13) and wood industries (PMR 2.03, 95% CI0. 81-5.13), and Latino women in the chemical industry (PMR 2.18, 95% CI 0.59-8.10), among other findings. Excess cancer of the pleura and peritoneum was observed among workers with a variety of usual occupations, consistent with widespread exposure to asbestos. Conclusions: This study demonstrates the utility of surveillance data bases for generating basic epidemiologic information on historically neglected workers. Leads about specific workplace exposure can be followed up in more detailed studies.

Background: Occupational bladder cancer mortality among minority racial/ethnic groups is not well described compared to occupational bladder cancer mortality among non-minority males in the United States. Methods: Race/ethnicity- and sex-specific bladder cancer mortality (1985-1992) of workers employed in 21 states was examined using a proportionate mortality study design. Mortality of specific racial/ethnic/ occupational groups was compared separately with workers in the specific occupation and with members of the specific racial/ethnic group. Results: This study identified elevated bladder cancer mortality among African American males and females and Latino males in several occupational groups with exposure to suspected bladder carcinogens as well as among Asian males in sales (PMR = 2.13) and Asian females in the personal services industry (PMR = 5.25; CI: 1.64-16.75). Conclusion: Surveillance of occupational cancer risks among racial/ethnic minorities using regularly available death certificate data is facilitated when states code both usual occupation/industry and race/ethnicity

Human corticotropin-releasing hormone (hCRH) and thyrotropin-releasing hormone (TRH) are known to stimulate ventilation after i.v. administration in humans. In a placebo-controlled, single-blind study we aimed to clarify if both peptides act by altering central chemosensitivity. Two subsequent CO2-rebreathing tests were performed in healthy young volunteers. During the first test 0.9% NaCl was given i.v.; during the second test 200 micrograms of hCRH (n = 12) or 400 micrograms of TRH (n = 6) was administered i.v. Nine subjects received 0.9% NaCl i.v. during both rebreathing manoeuvres. The CO2-response curves for the two tests were compared within the same subject. In the hCRH group a marked parallel shift of the CO2-response curve to the left was observed after hCRH (P < 0.01). The same effect occurred following TRH but was less striking (P = 0.05). hCRH and TRH caused a reduction in the CO2 threshold. The CO2-response curves in the control group were nearly identical. The results indicate an additive effect of both releasing hormones on the hypercapnic ventilatory response in humans, presumably independent of central chemosensitivity.

A new type of ozone contactor, called the SVI-DT (sidestream venturi injection with downflow tube) contactor, was evaluated at pilot scale and compared with the performance of a traditional countercurrent flow, fine-bubble diffusion contactor. The specially designed downflow tube, equipped with one or more pressurized sidestream venturi injectors and installed inside an ozone reactor, facilitated the dissolution of ozone into water. The results showed that the SVI-DT contactor achieved high mass transfer rates and outperformed the fine-bubble diffusion contactor in removing color and yielding higher and longer-lasting dissolved ozone residuals. The hydraulic behavior of the new contactor was also evaluated, and operating limits were established for maintaining stable two-phase flow conditions in the downflow tube.

[PPh(4)][EI(4)] (E=As, Sb, Bi) salts were reacted with four and five equivalents of AgN(3) to form tetraazidopnictates and pentaazidopnictates of the type [PPh(4)][E(N(3))(4)] and [PPh(4)](2)[E(N(3))(5)], respectively. The synthesis of [PPh(4)][P(N(3))(4)] was also attempted from the reaction of P(N(3))(3) with [PPh(4)]N(3), but it yielded only the starting materials. Herein, we report the synthesis and structure elucidation of [PPh(4)][E(N(3))](4) (E=As, Sb) and pentaazidobismuthate, stabilized as the dimethyl sulfoxide (DMSO) anion adduct, [PPh(4)](2)[Bi(N(3))(5)(dmso)]. Successive anion formation along the series E(N(3))(3)+nN(3)(-) (n=1-3) and E(N(3))(5)+N(3)(-) was studied by density functional theory.

This protocol describes methods for monitoring intracellular phosphorylation-dependent signaling events on a single-cell basis. This approach measures cell signaling by treating cells with exogenous stimuli, fixing cells with formaldehyde, permeabilizing with methanol, and then staining with phospho-specific antibodies. Thus, cell signaling states can be determined as a measure of how cells interact with their environment. This method has applications in clinical research as well as mechanistic studies of basic biology. In clinical research, diagnostic or drug efficacy information can be retrieved by discovering how a disease affects the ability of cells to respond to growth factors. Basic scientists can use this technique to analyze signaling events in cell lines and human or murine primary cells, including rare populations, like B1 cells or stem cells. This technique has broad applications bringing standard biochemical analysis into primary cells in order to garner valuable information about signaling events in physiologic settings. Curr. Protoc. Immunol. 96:8.17.1-8.17.20. © 2012 by John Wiley & Sons, Inc. Keywords: phosphorylation; signaling; flow cytometry; FACS; intracellular staining

A new type of coarse media flocculator, using buoyant media instead of the heavier gravel in order to facilitate cleaning, was evaluated in laboratory and field studies. The results indicate that a two-stage buoyant coarse madia (BCM) flocculator provided effective treatment at higher loading rates and shorter residence times than are typically employed in designing mechanical flocculatores. Source-water quality and coagulant chemistry directly influenced flocculator configuration selected

We present a flux calibrated spectrum of the star listed as CY Tau in the `General Catalog of Variable Stars 4th ed.' in the spectral range 3700 A less than or equal to lambda less than or equal to 6400 A with a resolution of approximately equals 15 A showing the Balmer-Series from H(sub beta) to H(sub 10) as well as the CaII H (in blend with H(sub epsilon) and K lines in emission. Apart from the emission lines the spectrum is composed of a continuum equivalent to that of an ordinary pre-main sequence star (i.e. a `naked' T Tau) of spectral type M2 V with emission lines plus a `blue' veiling continuum that can be described as black body radiation of temperature T(sub BL) approximately equals 7000K due to accretion onto a boundary layer at a rate of M-dot(sub acc) greater than or approximately = 2.18 10(exp -8) solar mass/a.

Recently we reported on the stimulating effect of a cold dark pretreatment (cold-dp) on the chlorophyll accumulation (Schonbohm et al., 1988) and on the CO2- and O2-gas exchange (Schonbohm and Czeska-Werner, 1990) during de-etiolation of wheat leaves under white light. The goal of the present paper was to study the effect of a cold-dp on the accumulation of chloroplast pigments, on the kinetics of fluorescence quenching, on the synthesis of Cab-proteins and on the amounts of cab-transcripts during de-etiolation under white light. The following results were obtained: 1 Not only the chlorophyll synthesis but also the synthesis of xanthophylls and of β-carotene is highly increased by a cold-dp. 2 The kinetics of the PAM-fluorescence curves demonstrate that the activity and co-operation of the two photosystems start earlier and with higher efficiency during de-etiolation in cold-dp than in warm-dp wheat leaves. 3 The amount of cab-transcripts can be elevated by a cold-dp. 4 The amount of Cab-proteins during de-etiolation is much higher in cold-dp than in warm-dp wheat leaves. The presented data are in accordance with our earlier published results. Consequences of these results on seedlings growing under natural conditions with fluctuating temperatures during day and night are pointed out.

Compounds (II) and (III) are characterized by IR and Raman spectroscopy, and compounds (IV), (V), and (VII) by single crystal XRD, 14N NMR, IR, and Raman spectroscopy.

The principle of chemical amplification has proven to be successful for the design of highly sensitive, high resolution resist material. In many cases a strong Broensted acid, generated by photolysis of onium salt precursors, has been used to catalytically cleave an acid labile blocking moiety. A new approach to negative tone resist is based on acid catalyzed cleavage of acetal blocked aromatic aldehydes, which act as «latent electrophiles» and, under the influence of strong acid, react with the surrounding phenolic resin