Live-cell labeling of endogenous proteins with nanometer precision by transduced nanobodies.

Accurate labeling of endogenous proteins for advanced light microscopy in living cells remains challenging. Nanobodies have been widely used for antigen labeling, visualization of subcellular protein localization and interactions. To facilitate an expanded application, we present a scalable and high-throughput strategy to simultaneously target multiple endogenous proteins in living cells with micro- to nanometer resolution. For intracellular protein labeling, we advanced nanobodies by site-specific and stoichiometric attachment of bright organic fluorophores. Their fast and fine-tuned intracellular transfer by microfluidic cell squeezing enabled high-throughput delivery with less than 10% dead cells. This strategy allowed for the dual-color imaging of distinct endogenous cellular structures, and culminated in super-resolution imaging of native protein networks in genetically non-modified living cells. The simultaneous delivery of multiple engineered nanobodies does not only offer exciting prospects for multiplexed imaging of endogenous protein, but also holds potential for visualizing native cellular structures with unprecedented accuracy.

Real-time computation of subdiffraction-resolution fluorescence images.

In the recent past, single-molecule based localization or photoswitching microscopy methods such as stochastic optical reconstruction microscopy (STORM) or photoactivated localization microscopy (PALM) have been successfully implemented for subdiffraction-resolution fluorescence imaging. However, the computational effort needed to localize numerous fluorophores is tremendous, causing long data processing times and thereby limiting the applicability of the technique. Here we present a new computational scheme for data processing consisting of noise reduction, detection of likely fluorophore positions, high-precision fluorophore localization and subsequent visualization of found fluorophore positions in a super-resolution image. We present and benchmark different algorithms for noise reduction and demonstrate the use of non-maximum suppression to quickly find likely fluorophore positions in high depth and very noisy images. The algorithm is evaluated and compared in terms of speed, accuracy and robustness by means of simulated data. On real biological samples, we find that real-time data processing is possible and that super-resolution imaging with organic fluorophores of cellular structures with approximately 20 nm optical resolution can be completed in less than 10 s.

[Disposal of animal byproducts, dead and slaughtered animals]. / Entsorgung von Schlachtnebenprodukten, verendeten und getöteten Tieren.

The current state of epidemiological knowledge about BSE clearly indicates that certain practices in carcass rendering had a significant impact on maintenance and spreading of BSE in the cattle population. This awareness did not come up spontaneously. As a reflection community legislation continuously developed and still does. The decisive move was done in the year 2000 by eliminating ruminant tissues with a high infectious potential with regard to BSE (specified risk material--SRM) from the human and animal feed chains. This step as well as the subsequent feed ban for all farm animals dramatically changed the logistical as well as the economical preconditions of the rendering industry. In fact the basic treatment (pressure cooking) remained almost unchanged. But instead of physically recycling the products they are nowadays predominantly used as an energy source in industry. In case of products that originate from the treatment of SRM burning is mandatory. These changes require a well adapted and intensified official supervision.

Mastery of postprostatectomy incontinence and impotence: his work, her work, our work.

PURPOSE/OBJECTIVES: To describe couples' experiences of postprostatectomy incontinence and impotence. DESIGN: Descriptive, qualitative. SETTING: Northeastern U.S. metropolitan area. SAMPLE: Subsample of 20 (10 control and 10 intervention) couples from a large quantitative clinical trial of a Standardized Nursing Intervention Protocol (SNIP) postprostatectomy. METHODS: Interviews were conducted using a semistructured guide. Data were analyzed using grounded theory techniques. MAIN RESEARCH VARIABLE: Couples' experiences of coping with postprostatectomy incontinence and impotence. FINDINGS: Managing postprostatectomy incontinence and impotence required work. Men's work focused on regaining mastery and encompassed understanding incontinence as healing, mastering incontinence, networking, confronting impotence and putting it into perspective, and prioritizing. Wives were supportive by managing anxiety, encouraging mastery, putting impotence into perspective, and reassuring their spouses. Established routines brought couples through the experience together while strengthening intimacy. SNIP couples found the nurses to be sources of information, support, and affirmation. CONCLUSIONS: Couples worked to deal with postprostatectomy incontinence and impotence within the context of surviving cancer and maintaining a loving relationship. This gave unique meaning to their symptoms and led the couples to value the fact that the men were alive and work toward regaining mastery. Mastery emerged as a key concept from the findings. IMPLICATIONS FOR NURSING PRACTICE: Nurses can gain from an enhanced understanding of postprostatectomy incontinence and impotence as meaningful within the greater context of patients having had cancer. Nurses can hasten couples' abilities to regain a sense of mastery by providing information, supporting couples' work, providing positive affirmation, and being available.

Acculturation and perinatal health outcomes among rural women of Mexican descent.

Research has shown that health outcomes for urban women of Mexican descent are related to acculturation. The purpose of this research was to compare perinatal outcomes of 773 women of Mexican descent who gave birth in three rural northern California hospitals, in relation to acculturation measured three different ways: by place of birth, by language spoken, and by the two factors combined as an Acculturation Index (AI). The prenatal and birth records of 773 Mexico-born or U.S.-born women of Mexican descent were reviewed. Results showed that language spoken was a less useful indicator of acculturation associated with perinatal complications than place of birth or the AI. The categorization of acculturation with the AI enhanced understanding of more specific groups of rural women and their particular health outcomes.

The Hispanic teen mother's origin of birth, use of prenatal care, and maternal and neonatal complications.

The purpose of this study was to describe and compare differences in demographics, prenatal care use, and pregnancy, labor, postpartum, and neonatal complications for 129 pregnant Mexican-American adolescents who were either born in the United States or born in Mexico. A significant finding was that 25% of the teens, regardless of origin of birth, received either inadequate or no prenatal care. The findings of this study will be useful to clinicians providing health care for Hispanic teens and for administrators and policy makers who affect the types of health care and education available to these young mothers.

PIP: Currently comprising 9% of the US population, Hispanics are the fastest growing ethnic minority population in the country. 26% of California's population is Hispanic. The proportion of Hispanic youth is growing rapidly, with the Hispanic youth population projected to grow by 80% by 2030. The authors describe and compare differences in demographics, the use of prenatal care, and pregnancy, labor, postpartum, and neonatal complications among 129 pregnant Mexican-American adolescents who were either born in the US or in Mexico. The mean age of the 48 young women born in Mexico was 17.9 years, while the 81 born in the US were of mean age 17.2 years. More US teens were enrolled in school than were the teens born in Mexico, Mexico-born teens were more likely to be married, and the US-born teens reported a higher incidence of cigarette smoking. US-born and Mexico-born women had an average of 6.5 and 5.5 recorded prenatal visits, respectively. 25% of the teens, regardless of the country of origin, received either inadequate or no prenatal care. 14 prenatal complications, 17 labor complications, 7 postpartum complications, and 7 neonatal complications were identified.

More does not mean better: prenatal visits and pregnancy outcome in the Hispanic population.

Early and consistent prenatal care (PNC) is thought to play an important role in the reduction of low birthweight (LBW) in the United States. It has been reported that LBW and delayed PNC are common to the Hispanic woman. In California, this cultural group comprises approximately 26% of the population, and much debate concerning health care reform has been targeted at this problem. A comparative study was conducted in California to examine the number of prenatal visits and the outcomes of Mexico-born Hispanics and U.S.-born Hispanics. Obstetric and medical record review for 783 women was done. The results show that more prenatal visits did not improve the outcome during pregnancy, labor, or the postpartum period. Because a large portion of PNC is now delivered by the advanced practice nurse, implications for practice include exploring alternatives for the delivery of culturally relevant care, addressing the barriers to caring for this population, and finding alternative models of care that have the potential to produce positive outcomes.