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Christian S Mayer,1,* Isabella Baur,1,* Julia Storr,2 Alexander Markard,1 Ramin Khoramnia1 1Department of Ophthalmology, University Hospital Heidelberg, University of Heidelberg, Heidelberg, Germany; 2Department of Ophthalmology, Technical University Munich, Munich, Germany*These authors contributed equally to this workCorrespondence:Christian S MayerDepartment of Ophthalmology, University Hospital Heidelberg, University of Heidelberg, Im Neuenheimer Feld 400, Heidelberg 69120, GermanyEmail Christian.Mayer@med.uni-heidelberg.dePurpose: Patients with traumatic aniridia, aphakia and retinal complications can require silicone oil endotamponade. In the absence of compartmentalization, there is a risk of silicone oil migrating to the anterior chamber which can cause long-term complications. We report a two-step procedure in trauma cases, using sutures for silicone oil retention in primary care and subsequently prior to secondary artificial iris (AI) and intraocular lens (IOL) implantation, to achieve a reconstruction of the anterior and posterior chamber.Material and Methods: Seven patients with loss of the iris-lens-diaphragm after an ocular trauma and the need for silicone oil endotamponade underwent a primary intervention including wound closure, placement of silicone oil retention sutures and silicone oil filling. Four of those underwent secondary reconstruction with silicone oil removal and AI and IOL implantation and could be included in this retrospective observational study. All main outcome measures were evaluated after the first and after the second surgery. The main outcome measures were morphological findings, subjective impairment from glare, subjective cosmetic disfigurement, patient satisfaction and intraocular pressure (IOP) as well as best-corrected distance visual acuity and objective refraction.Results: The retention sutures could effectively prevent silicone oil migration into the anterior chamber in the first surgery. Silicone oil emulsifications in the anterior chamber were observed in one patient. In two patients, pigment deposition at the sutures was seen. Subjective impairment from glare and subjective cosmetic disfigurement could be reduced after the second surgery compared to the evaluation after the first surgery. IOP and best-corrected distance visual acuity remained stable. No eye needed silicone oil refilling after the secondary reconstruction surgery.Conclusion: Our two-step approach is viable and provides good functional and aesthetic results. We observed a high patient satisfaction.Keywords: aniridia, aphakia, trauma, open globe injury, retinal detachment

Alizé A Rogge,1 Isabel Baur,2 Gabriele Blettner,3 Ulrike Holtkamp,4 Markus Horneber,5 Patrick Jahn,6 Stefanie Joos,7 Silva Keberle,8 Anita Kettelgerdes,9 David Klemperer,10 Alfred Längler,11 Petra Voiß,12,13 Joachim Weis,14 Claudia M Witt1,15,16 1Institute for Social Medicine, Epidemiology, and Health Economics, Charité – Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität Zu Berlin, and Berlin Institute of Health, Berlin, Germany; 2Legal Institute, Competence Center Medicine - Ethics - Law Helvetiae, University of Zurich, Zurich, Switzerland; 3Deutsche Krebshilfe, INFONETZ KREBS, Bonn, Germany; 4German Leukemia & Lymphoma Patients‘ Association, Bonn, Germany; 5Department of Internal Medicine, Division of Oncology and Hematology, Paracelsus Medical University, Klinikum Nuremberg, Nuremberg, Germany; 6Department for Nursing Science, Institute for Health Science, Medical Faculty, Eberhard Karls University Tübingen, Tübingen, Germany; 7Institute for General Practice and Interprofessional Health Care, University Clinic Tübingen, Tübingen 72076, Germany; 8Eskamed AG, Basel, Switzerland; 9TK – Techniker Krankenkasse, Hamburg, Germany; 10Faculty of Social and Health Sciences, Ostbayerische Technische Hochschule Regensburg, Regensburg., Germany; 11Gemeinschaftskrankenhaus Herdecke, Institute of Integrative Medicine, Witten/Herdecke University, Herdecke, Germany; 12Department of Internal and Integrative Medicine, Evang. Kliniken Essen-Mitte, Faculty of Medicine, University of Duisburg-Essen, Essen, Germany; 13Gynaecological Clinic, Städtisches Klinikum Karlsruhe gGmbH, Karlsruhe, Germany; 14Department of Self-Help Research, Comprehensive Cancer Center University Clinic Freiburg, Freiburg i.Br., Germany; 15Institute for Complementary and Integrative Medicine, University of Zurich and University Hospital Zurich, Zurich, Switzerland; 16Center for Integrative Medicine, University of Maryland School of Medicine Baltimore, Baltimore, MarylandCorrespondence: Claudia M WittInstitute for Complementary and Integrative Medicine, University Hospital Zurich and University Zurich, Sonneggstrasse 6, Zurich 8091, SwitzerlandEmail claudia.witt@uzh.chPurpose: Even in cases of positive evidence for complementary medicine (CM) therapies, it is still difficult for cancer patients to identify reputable providers. The aim of this study was to develop and evaluate a criteria list to provide guidance to cancer patients seeking a reputable CM provider.Methods: The design combined a literature review, an expert consensus procedure (n=15) and an assessment from three stakeholder perspectives (patients (n=18), CM providers (n=26) and oncology physicians (n=20)).Results: A total of 30 existing CM criteria were extracted from the literature, and 12 more were added by the experts. The main challenge was to define criteria that could easily be applied by the patients. A final comprehensive list of 8 criteria guiding cancer patients to find a reputable CM provider was developed.Conclusion: Health professionals and cancer information services might find the criteria list helpful when aiming to strengthen patients’ awareness of quality-related factors associated with CM providers. The criteria developed might be helpful when standards are established for quality assurance in CM in oncology.Keywords: neoplasms, complementary medicine, patient-centered care, standards, healthcare quality assurance

Dairy farmers face various challenges in developing their businesses. The current literature identifies several constraints towards a more efficient dairy sector that relate mostly to on-farm management practices. The available studies analyze constraints mostly from the objective viewpoint of the researcher, whereas very little is known about what farmers themselves perceive to be challenging. To better understand the farmers' perspective and what they perceive to be challenging, and how these challenges differ across regions, this paper builds on a survey of 529 dairy farmers in Malawi. In the survey, respondents were asked in an open question to name their three main challenges. Based on the responses, a three-level coding scheme was elaborated. Data were then coded by two researchers to ensure reliability of coding. Codes were then used to calculate relative frequencies for the different challenges and to perform chi-square tests to check for regional differences in frequencies. Results suggest that farmers perceive low milk price and milk yield, animal health, availability of feedstock, and the costs for drugs as the main challenges. The analysis also revealed that the challenges vary strongly between the three main regions in the country--Blantyre (south), Lilongwe (central), and Mzuzu (north). The perceived challenges reflect the different production systems in the regions. In Blantyre, dairy farming is low in intensity, and farmers thus perceive inputs, in particular the availability of fodder, as well as outputs, in particular milk yields more frequently as a challenge than farmers in Lilongwe and Mzuzu. In contrast, dairy farming in Lilongwe is the most intense. Accordingly, farmers in Lilongwe perceive costs, in particular for inputs such as mash and concentrates as major constraints. Farmers in Mzuzu, which is a rather remote region, consider a lack of assistance, in particular concerning extension services significantly more challenging than the farmers in the other two regions do. Considering these insights, development work and extensions services may be able to increase outcomes in the dairy sector by targeting the regional challenges. Key words: Challenges, dairy production, smallholders, Malawi, qualitative study, regions, INTRODUCTION Dairy cattle production has the potential to contribute to the livelihood of many smallholders in Malawi through income generation, increased resilience during food crises, and by strengthening social security [...]

The distribution of type VII collagen was examined in the normal human nervous system, in brain tumour biopsies and in glioma cell lines by immunohistochemistry and western blotting. In normal tissue, positivity was observed beneath choroid plexus epithelial cells and around pineal gland and pituitary gland cell nests, while other brain regions and peripheral nerves were negative. Expression was preserved in most related tumours (choroid plexus papilloma, pineoblastoma, pituitary adenoma). Scattered abnormal vessels showed neo-expression of type VII collagen in about half of the astrocytic and ependymal tumours. Glioma cells in situ were consistently negative for type VII collagen, whereas the glioblastoma cell lines were positive. Our results suggest that anchoring fibrils or at least epitopes of their major structural component are present in normal and pathological cerebral structures, indicating a unique distribution of type VII collagen in the nervous system. [ABSTRACT FROM AUTHOR]

This commentary focuses on the challenges and possibilities that adolescents and young adults with cancer (AYA) desiring parenthood face under Swiss law. The regulation of reproductive medicine procedures is stricter in Switzerland than in some other countries. Health insurance is compulsory, but the interventions that are covered are in constant flux. Recent changes pertain to the possibilities of future AYA parenthood and keeping up to date with practical and legal ramifications is taxing even for health professionals. AYA facing treatment decisions are uniquely vulnerable and dependent on comprehensive, clear, current, and country-specific information regarding risks and options pertaining to their fertility. This commentary provides a short overview of the Swiss legal framework related to reproductive medicine, highlighting its access restrictions and prohibitions, as well as recent changes. While the importance of patient, peer, caregiver, and interest groups supporting people affected by health conditions has long been recognized in many countries, an AYA organization was only recently established in Switzerland. Such organizations are vital for providing accurate, country-specific information and support, while individualized medical guidance, informed by the most current legal framework and its consequences, remains essential in addressing AYAs' specific needs in connection with the desire to have children.

(1) Background: Laparoscopic resection for colon and rectal cancer was introduced in the early 1990s; the aim of this analysis was to show possible advantages of minimal-invasive approaches in rectal cancer surgery. (2) Methods: From 2016 to 2020, all patients undergoing open, laparoscopic or robotic-assisted rectal cancer surgery in Germany were retrospectively analyzed regarding sex distribution, conversion rates and in-hospital mortality rates according to nationwide hospital billing data based on diagnosis-related groups (DRGs). (3) Results: In total, 68,112 patients were analyzed, and most commonly, low anterior rectal resections with primary anastomosis ( n = 25,824) were performed with an increase of minimal-invasive procedures over the years (open: 51% to 27%; laparoscopic: 47% to 63% and robotic: 2% to 10%). In-hospital mortality rate was 2.95% ( n = 2012). In total, 4.61%, 1.77%, 1.14% and 3.95% of patients with open, laparoscopic, robotic and converted-to-open surgery died during hospital stay, respectively (open vs. laparoscopic p < 0.0001; open vs. robotic p < 0.00001; laparoscopic vs. robotic p = 0.001). Conversion rates were significantly more favorable in the robotic compared to the laparoscopic group. (11.94% vs. 2.53%; p < 0.0001). (4) Conclusion: Minimal-invasive rectal cancer surgery might have some advantages in terms of a reduced in-hospital mortality, and an improved conversion rate for the robotic approach.

Objective: Tailored approaches in gastrointestinal oncology have been more frequently introduced in past years and for patients with peritoneal metastases. This article attempts to overview the current strategies in surgical gastrointestinal oncology, with a focus on gastrointestinal peritoneal metastases., Methods: In 2019, all patients undergoing PIPAC therapy in Germany were retrospectively analyzed regarding morbidity and in-hospital mortality rates. Furthermore, patients with chemotherapy-refractory peritoneal metastases from gastric cancer undergoing PIPAC-therapy at our institution were analyzed., Results: In 2019, 534 patients received PIPAC treatment in german hospitals. The in-hospital mortality rate was 0%. In total, 36 patients suffered from postoperative complications (8%). From April 2016 to September 2021, a total of 44 patients underwent 93 PIPAC applications at our institution. The non-access-rate was 0%. The median PRGS was two (range, 1-4). Eleven patients (44%) showed histologically stable disease, whereas six patients (24%) showed histological regression. Median survival, calculated from the date of the first PIPAC application, was 181 days (range, 43-636 days)., Conclusions: PIPAC is a safe and feasible procedure with a low in-hospital morbidity and mortality. Furthermore, PIPAC in the palliative and chemorefractory setting and is an appealing approach for patient management in the future.

Purpose: To analyze the tomographically non-affected second eyes of keratoconus patients using the Corvis ST to detect any biomechanical abnormalities or subclinical keratoconus., Methods: In this retrospective, single-center, consecutive case series 244 eyes of 122 keratoconus patients were analyzed between November 2020 and February 2021. Fourteen fellow eyes fulfilled the inclusion criteria and showed no clinical or tomographic signs of keratoconus. Main outcome measures included best-corrected visual acuity, tomographic and biomechanical analyses using Scheimpflug imaging: Pentacam and Corvis ST (Oculus, Wetzlar, Germany). Tomographic analyses included anterior and posterior simulated keratometry, K-Max, central corneal thickness, thinnest corneal thickness, Belin/Ambrosio Ectasia Display, and the ABCD grading system. For biomechanical analyses, the corneal biomechanical index (CBI) and tomographic biomechanical index were used., Results: The mean best-corrected visual acuity was 0.01 ± 0.10 logMAR. Mean K-Max was 43.79 ± 1.12 D, mean central corneal thickness 529 ± 25 µm, mean thinnest corneal thickness 524 ± 23 µm, and mean Belin/Ambrosio Ectasia Display 1.0 ± 0.32. The mean CBI was 0.30 ± 0.21. Regular CBI values were found in six of 14 patients. The mean tomographic biomechanical index was 0.47 ± 0.22 with regular values observed in only two of 14 patients. No signs of tomographic or biomechanical abnormalities were shown in only one of 14 keratoconus fellow eyes, with regular ABCD, Belin/Ambrosio Ectasia Display, CBI and tomographic biomechanical index values., Conclusions: Tomographically normal fellow eyes of keratoconus patients are rare. In these cases, a biomechanical analysis of the cornea may help detect a subclinical keratoconus. The tomographic biomechanical index was the most sensitive index to verify a mild ectasia.

Background: Invasiveness is a major factor contributing to metastasis of tumour cells. Given the broad variety and plasticity of invasion mechanisms, assessing potential metastasis-promoting effects of irradiation for specific mechanisms is important for further understanding of potential adverse effects of radiotherapy. In fibroblast-led invasion mechanisms, fibroblasts produce tracks in the extracellular matrix in which cancer cells with epithelial traits can follow. So far, the influence of irradiation on this type of invasion mechanisms has not been assessed., Methods: By matrix-embedding coculture spheroids consisting of breast cancer cells (MCF-7, BT474) and normal fibroblasts, we established a model for fibroblast-led invasion. To demonstrate applicability of this model, spheroid growth and invasion behaviour after irradiation with 5 Gy were investigated by microscopy and image analysis., Results: When not embedded, irradiation caused a significant growth delay in the spheroids. When irradiating the spheroids with 5 Gy before embedding, we find comparable maximum migration distance in fibroblast monoculture and in coculture samples as seen in unirradiated samples. Depending on the fibroblast strain, the number of invading cells remained constant or was reduced., Conclusion: In this spheroid model and with the cell lines and fibroblast strains used, irradiation does not have a major invasion-promoting effect. 3D analysis of invasiveness allows to uncouple effects on invading cell number and maximum invasion distance when assessing radiation effects., (© 2021. The Author(s).)

Purpose: We present the case of a 36-year old Curschmann-Steinert myotonic dystrophy patient with posterior subcapsular cataract that we treated with unilateral implantation of an extended depth of focus intraocular lens to address his wish for spectacle independence at far and intermediate distance., Observations: The patient underwent phacoemulsification with subsequent implantation of the AcrySof IQ Vivity IOL (Alcon, Fort Worth, TX, USA) in his left eye. Uncorrected distance visual acuity (UDVA) on the left eye increased from +0.40 logMAR preoperatively to -0.12 logMAR at 3 months postoperatively. At the three months follow-up distance corrected intermediate visual acuity (DCIVA) at 80 cm distance was -0.08 logMAR and DCIVA at 66 cm distance was 0.14 logMAR for the left eye. The defocus curve showed a functional defocus of 2.0 diopters at 0.2 logMAR or better, corresponding to the extended depth of focus. Dysphotopsia evaluation with a Halo & Glare simulator (Eyeland-Design Network GmbH, Vreden, Germany) revealed a very low level of photic phenomena., Conclusions and Importance: Unilateral implantation of a new generation, non-diffractive extended depth of focus IOL was well tolerated and provided good functional results for far and intermediate distances. The patient reported a very low level of photic phenomena., Competing Interests: G. Auffarth and R. Khoramnia report grants, personal fees and non-financial support from Alcon, Oculus and Johnson&Johnson. This work was supported by an Investigator Initiated Trial Grant IIT# 62791973 from Alcon., (© 2021 The Authors.)

Competing Interests: G. Auffarth and R. Khoramnia report grants, personal fees and non-financial support from Rayner. T. Yildirim and I. Baur have nothing to disclose.

The interaction between residential preferences and dwellings is a complex system whose function thus far remains insufficiently explored. In this paper, we investigate housing functions as orchestrators of households' residential mobility in the context of Swiss rental housing. We propose a theoretical multi-step model and use survey data from 878 Swiss tenants to inspect the model's linkages. From the statistical analysis, we firstly observe that tenants' residential satisfaction is more likely to increase when the gap between ideal housing functions and those actually fulfilled by the current dwelling decreases. Secondly, results show that the effectiveness of an event (e.g. a job opportunity) in triggering the move is significantly related to both residential satisfaction and the functions the dwelling fulfils prior to the trigger. Thirdly, findings show that these trigger events can be grouped into three types: radical change, problem-solving and opportunity. With a medium effect size, a radical change was found to bring about the strongest change in housing functions between past and current dwellings. Lastly, in line with the hypothesis that residential preferences vary over the life course, socio-demographic characteristics and tenancy types are found to be significant explanatory variables for households' ideal housing functions. By disentangling the complexity of the housing system, the proposed multi-step model can be used to integrate households' preferences with supply-side constraints in agent-based model simulations, thereby contributing to fostering the provision of quality housing, i.e. dwellings able to meet the needs of current and future occupants., Competing Interests: Conflict of interestThe authors have no relevant financial or non-financial interests to disclose., (© The Author(s) 2021.)

Purpose: To present the case of a 60-year-old patient with severe nanophthalmic eyes, who underwent cataract surgery with a bilateral implantation of custom-made high-power intraocular lenses (IOLs)., Observations: The axial length was 14.94 and 15.05 mm of the right and the left eye, respectively. The preoperative corrected distance visual acuity (CDVA) was +0.46 logMAR (20/63) in the right eye and +0.58 logMAR (20/80) in the left eye with rigid contact lenses of +17.5 D bilaterally. The calculated IOL power for emmetropia with different formulas ranged from +55.28 to +70.09 D. The IOL power selection was based on the average value from four formulas (Haigis, Holladay 1, Holladay 2, SRK/T) with the target refraction of emmetropia. Custom-made +56.0 and + 58.0 D Aspira-aAY IOLs (HumanOptics AG, Erlangen, Germany) were implanted without any complications. The postoperative CDVA was +0.40 logMAR (20/50) and +0.60 logMAR (20/80). The manifest refraction spherical equivalents were +0.625 D and -0.375 D., Conclusions and Importance: Even in eyes with the axial length of only 15 mm, cataract surgery can be successfully performed after adequate preparation. High-power customized IOLs allow complete correction of hyperopia but caution is required with the results from different IOL power calculation formulas, which can be misleading., Competing Interests: No conflict of interest (TN, DS, IB, RK, GUA)., (© 2020 Published by Elsevier Inc.)

Nucleoli have attracted interest for their role as cellular stress sensors and as potential targets for cancer treatment. The effect of DNA double-strand breaks (DSBs) in nucleoli on rRNA transcription and nucleolar organisation appears to depend on the agent used to introduce DSBs, DSB frequency and the presence (or not) of DSBs outside the nucleoli. To address the controversy, we targeted nucleoli with carbon ions at the ion microbeam SNAKE. Localized ion irradiation with 1-100 carbon ions per point (about 0.3-30 Gy per nucleus) did not lead to overall reduced ribonucleotide incorporation in the targeted nucleolus or other nucleoli of the same cell. However, both 5-ethynyluridine incorporation and Parp1 protein levels were locally decreased at the damaged nucleolar chromatin regions marked by γH2AX, suggesting localized inhibition of rRNA transcription. This locally restricted transcriptional inhibition was not accompanied by nucleolar segregation, a structural reorganisation observed after inhibition of rRNA transcription by treatment with actinomycin D or UV irradiation. The presented data indicate that even multiple complex DSBs do not lead to a pan-nucleolar response if they affect only a subnucleolar region., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)

Purpose: Clinical evaluation of the visual outcomes after implantation of a small-aperture extended depth of focus (EDOF) intraocular lens (IOL) and a segmental refractive bifocal lens., Methods: In this prospective study, 13 patients with cataract received a small-aperture IC-8 IOL (AcuFocus, Irvine, CA) in one eye and a segmental-refractive Lentis Mplus LS-313 MF20 IOL (Oculentis, Berlin, Germany) in the fellow eye. The clinical examination included measurements of uncorrected (UDVA) and corrected (CDVA) distance visual acuity, uncorrected (UIVA) and distance-corrected (DCIVA) intermediate visual acuity, and uncorrected (UNVA) and distance-corrected (DCNVA) near visual acuity. Additionally, a defocus curve was obtained from +2.00 to -5.00 diopters (D) and a Salzburg Reading Desk was used to assess the patients' reading acuity at intermediate and near distances. A halo and glare simulator was used to evaluate the postoperative perception of dysphotopsia., Results: At 5 months postoperatively, the mean binocular visual results demonstrated UDVA, UIVA, and UNVA values of -0.04 ± 0.11, 0.00 ± 0.10, and 0.11 ± 0.08 logMAR, respectively. The binocular distance-corrected reading performance test also confirmed the improved visual function, with an intermediate reading acuity of 0.12 logMAR at 69.21 cm and a near reading acuity of 0.19 logMAR at 41.63 cm. The mean halo size was 32.54 ± 22.38, mean halo intensity was 34.46 ± 21.95, mean glare size was 9.00 ± 17.47, and mean glare intensity was 9.92 ± 16.84., Conclusions: The new concept of a combined implantation of a small-aperture IOL and a segmental-refractive bifocal lens showed good results in far and intermediate distances and functional results at near distance, while causing minimal photic phenomena. [J Refract Surg. 2019;35(9):551-558.]., (© 2019 Son, Khoramnia, Yildirim, et al.)

Individualized Medicine, Electronic Patient File and Further Use of Health Data Abstract. The present article is dedicated to the two subject areas 'individualized medicine' and 'electronic patient file'. Both individualised medicine and the electronic patient file generate numerous different health data. The article focuses on the value of health data for human research and discusses the further use of the data according to the Human Research Act. At present, the Human Research Act does not provide for the possibility of making data from the electronic patient file accessible for research. However, efforts are being made to change this legal situation so that data from the patient file can be incorporated into human research for the benefit of (future) patients.

Milk production in Malawi is stagnating while steadily increasing in neighboring countries. We performed a literature review to identify factors negatively affecting milk production in Malawi and propose potential solutions to overcome the particular constraints. We structured the analysis along the value chain with three major components: upstream providers, on-farm production, and downstream providers. It is shown that the literature strongly focuses on on-farm constraints and most prominently on animal health, breed choice, and milk yield. The literature research did not reveal any reasons specific to Malawi that would hinder an increase in milk production as experienced in neighboring countries. We suggest a focus on crossbreeds accompanied by local breeding programs, which would reduce dependence on imported semen and strengthen the currently unreliable artificial insemination services. In addition, crossbreeds may be better suited than pure breeds for current feeding practices and climate conditions and thus come close to reaching their genetic potential. We conclude that cost advantages of crossbreeds clearly compensate for the lower revenues compared with exotic breeds such as Holstein-Friesian or Jersey.

We have built an ion-microbeam for studies of the nuclear topography and kinetics of double-strand break repair at the single cell level. Here, we show that a first and a second, delayed single ion exposure at different nuclear sites led to comparable accumulations of phospho-ATM, gamma-H2AX and Mdc1 at both earlier (e) and later (l) microirradiated sites. In contrast, accumulations of 53BP1 and the recombination protein Rad51 were strongly reduced at l-sites. This apparent competition effect is accompanied by a reduced amount of 53BP1 in undamaged areas of the irradiated nuclei. We suggest that a critically limited pool size combined with strong binding at irradiated sites leads to the exhaustion of unbound factors freely roaming the nuclear space. The undersupply of these factors at l-sites requires in addition a long-lasting binding at e-sites or a weaker binding at l-sites. The observed effects suggest that DNA damage response at individual nuclear sites depends on the time course of damage load. This may have implications for therapeutic radiation treatments.

Several proteins are known to form foci at DNA sites damaged by ionizing radiation. We study DNA damage response by immunofluorescence microscopy after microirradiation of cells with energetic ions. By using microirradiation, it is possible to irradiate different regions on a single dish at different time-points and to differentiate between cells irradiated earlier and later. This allows to directly compare immunofluorescence intensities in both subsets of cells with little systematic error because both subsets are cultivated and stained under identical conditions. In addition, by using irradiation patterns such as crossing lines, it is possible to irradiate individual cells twice and to differentiate between immunofluorescence signals resulting from the cellular response to the earlier and to the later irradiation event. Here, we describe the quantitative evaluation of immunofluorescence intensities after sequential irradiation.

A comprehensive analysis of the proteins found in human spermatozoa is essential for understanding the events leading up to, and including, fertilization and development. Proteomics offers a platform for investigating this process, provided that the dynamic range is relatively low. In this report, spermatozoa from a number of human sperm ejaculates were isolated in a pure state using discontinuous Percoll gradient centrifugation. Triton X-100 soluble and insoluble proteins were recovered and separated by SDS-PAGE. The separation lanes were dissected into 96 fractions and analyzed individually by LC-MS(n) . A comprehensive protocol, involving LC-MS/MS analysis eventually down to the ninth most intense peak found in the MS-survey scan, was performed. Analysis of purified human sperm populations resulted in the identification of 1056 gene products, of which approximately 8% have not previously been characterized. The data were supported by the large number of proteins represented by expressed sequence tags in the testis. Bioinformatic analysis demonstrated that 437 of the gene products were involved in various metabolic pathways including glycolysis and oxidative phosphorylation. The inventory of proteins present in the human sperm proteome includes a number of notable discoveries including the first description of a nicotinamide adenine dinucleotide phosphate oxidase, dual-oxidase 2, finally laying to rest any doubts about the presence of such enzymes in spermatozoa. Furthermore, a number of different classes of receptor have also been detected in these cells and are potential regulators of sperm function. This list includes at least six seven-pass transmembrane receptors, six tyrosine kinase receptors, a tyrosine phosphatase receptor, glutamate-gated ion channel receptors, transient receptor potential cation channels, and a non-genomic progesterone receptor. This is the first published list of identified proteins in human spermatozoa using LC-MS/MS analysis., (Copyright © 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The sorption of selenite and selenate to ettringite (3CaO x Al2O3 x 3CaSO4 x 32H2O), "monosulfate" (3CaO x Al2O3 x CaSO4 x 12H2O), and calcium silicate hydrate (C-S-H) was investigated in order to understand Se immobilization by cement in hazardous wastes. Sorption kinetics were fast with equilibrium between the minerals and Se species reached within 1 d. Selenite is suggested to sorb by surface reactions, and for ettringite, a sorption maximum of 0.03 mol kg(-1) was determined. Distribution ratios (Rd) for selenite were 0.18, 0.38, and 0.21 m3 kg(-1) for ettringite, monosulfate, and C-S-H, respectively. At high selenite additions, CaSeO3 was precipitated with a solubility product of Kso = 10(-7.27) (I = 0, 25 degrees C). Selenate sorbed only weakly to ettringite (Rd = 0.03), and no significant sorption to C-S-H was found. In contrast, sorption to monosulfate was strong (Rd = 2.06). With increasing selenate addition, XRD analyses revealed changes in the interlayer distance of monosulfate, in parallel with an increase of the ettringite fraction. Substitution of sulfate is suggested to be the relevant process. This indicates that selenate is sorbed more efficiently by monosulfate-rich cement, while the cement composition is of minor importance for selenite sorption.