A new histopathology scoring protocol reveals myopathy features in PSE-like pork.

Pale, Soft, and Exudative (PSE)-like pork defects are associated with fiber destruction and pale discoloration and have become a severe economic burden for the European meat sector. However, robust detection of PSE-like pork and its diverse features is challenging and makes studies into defect causation difficult. Implementation of histological examination may improve our knowledge about less-known features linked to PSE-like defects. Here we evaluate if a new histological protocol can reveal how myopathy in ham may be associated with visual and traditional physicochemical anomalies of PSE-like pork. We first created a list of pathological features, quantified them, and integrated them into a myodegeneration scoring scheme (MYO) for semimembranosus muscle sections. We then explored potential associations between overall MYO scoring and individual histology features with visual PSE-like defect scoring (DES) and with individual meat quality variables [pHu, color: L*, a*, b* (CIELAB), bioimpedance, and near-infrared spectroscopy (NIR)]. As the primary finding of this study, we show a significant association between overall myopathy (MYO) scoring and PSE-like defect (DES) scores. We also found associations of specific myopathy features with DES scores, and of overall MYO scoring with specific quality variables. In all, our data suggest links between signs of acute myodegeneration and PSE-like defects. Our data, hence, supports the implementation of semi-quantitative histopathological approaches for diagnosing PSE-like pork features and may help identify the underlying mechanisms behind these defects.

Structural and proteomic analyses reveal regional brain differences during honeybee aging.

Among insects, learning is particularly well studied in the fruit fly Drosophila melanogaster and the honeybee Apis mellifera. A senescence-dependent decline in classic pavlovian conditioning is demonstrated for both species. To understand how aging affects learning, genetic approaches used with Drosophila can benefit from complementary studies in Apis. Specifically, honeybees have a larger brain size allowing for compartment-specific approaches, and a unique life-history plasticity. They usually perform within-nest tasks early in life (nest bees) and later they collect food (foragers). Senescence of learning performance is a function of the bees' foraging duration but underlying causes are poorly understood. As cognitive aging is commonly associated with structural and biochemical changes in the brain, we hypothesized that brain areas implicated in learning change in synaptic and biochemical composition with increased foraging duration. First, we used synapse-specific immunohistochemistry and proteomics to screen for alterations in the calyx region of the mushroom body, a key structure for memory formation. Using proteomics, we next profiled the central brain, which comprises all higher-order integration centers. We show that, with increased foraging duration, levels of kinases, synaptic- and neuronal growth-related proteins decline in the central brain while the calyx region remains intact both in structure and biochemistry. We suggest that proteome-level changes within major anatomical sites of memory formation other than the calyx region could be central to learning dysfunction. These include large compartments of the central brain, such as the mushroom body's output regions and the antennal lobes. Our data provide novel information toward heterogeneity in the aging insect brain, and demonstrate advantages of the honeybee for invertebrate neurogerontological research.

Selective method for the analysis of perchlorate in drinking waters at nanogram per liter levels, using two-dimensional ion chromatography with suppressed conductivity detection.

The United States Environmental Protection Agency (EPA) collected drinking water occurrence data for perchlorate in the Unregulated Contaminant Monitoring Regulation (UCMR 1; 2001-2005) using EPA Method 314.0. To address the interest in increasing sensitivity and selectivity for the analysis of perchlorate, three new methods, EPA Methods 314.1, 331.0 and 332.0, were subsequently published by EPA for the analysis of perchlorate in drinking water. In 2006, an automated two-dimensional ion chromatography (2D-IC) method for measuring perchlorate with suppressed conductivity detection was developed. Two-dimensional IC is essentially an automated "heart-cutting", column concentration and matrix elimination technique. In the first dimension, a large sample volume is injected onto a first separation column and the separated matrix ions are diverted to waste while the analyte(s) of interest are selectively cut, trapped and concentrated in a concentrator column. In the second dimension, the contents from the concentrator column are eluted onto a second analytical column for separation and quantitation of the analyte(s) of interest. Incorporation of two columns with different affinities for the analyte(s) in a single analysis can provide comparable selectivity and superior sensitivity to a method using second column confirmation in a second separate analysis step. Use of this approach led to the development of a new, highly sensitive and selective 2D-IC, suppressed conductivity method with a Lowest Concentration Minimum Reporting Level (LCMRL) of 55 ng/L for perchlorate in drinking water samples. This new method has comparable sensitivity and selectivity and is simpler and more economical than IC-mass spectrometric (MS) or IC-MS-MS techniques. The method is now being prepared for publication as EPA Method 314.2.

Minute Impurities Contribute Significantly to Olfactory Receptor Ligand Studies: Tales from Testing the Vibration Theory.

Several studies have attempted to test the vibrational hypothesis of odorant receptor activation in behavioral and physiological studies using deuterated compounds as odorants. The results have been mixed. Here, we attempted to test how deuterated compounds activate odorant receptors using calcium imaging of the fruit fly antennal lobe. We found specific activation of one area of the antennal lobe corresponding to inputs from a specific receptor. However, upon more detailed analysis, we discovered that an impurity of 0.0006% ethyl acetate in a chemical sample of benzaldehyde-d5 was entirely responsible for a sizable odorant-evoked response in Drosophila melanogaster olfactory receptor cells expressing dOr42b. Without gas chromatographic purification within the experimental setup, this impurity would have created a difference in the responses of deuterated and nondeuterated benzaldehyde, suggesting that dOr42b be a vibration sensitive receptor, which we show here not to be the case. Our results point to a broad problem in the literature on use of non-GC-pure compounds to test receptor selectivity, and we suggest how the limitations can be overcome in future studies.

Perchlorate in water via US Environmental Protection Agency Method 331 Determination of method uncertainties, lowest concentration minimum reporting levels, and Hubaux-Vos detection limits in reagent water and simulated drinking water.

US Environmental Protection Agency (EPA) Method 331 determines perchlorate in drinking water using non-suppressed ion chromatography with tandem mass spectrometry. This study reports the results of calibration and recovery studies in reagent water, as well as of a recovery study in simulated drinking water (i.e., total dissolved solids are 500 mg/mL each of chloride, sulfate, and bicarbonate). The perchlorate concentrations in the study ranged from 0.05 to 64 ng/mL. At 95% confidence, the Hubaux-Vos detection limit (H-V DL) was 0.04 ng/mL for the calibration study and the simulated-drinking-water recovery study, and 0.03 ng/mL for the reagent-water recovery study. The lowest concentration minimum reporting level was 0.03 ng/mL for reagent water and 0.0 7 ng/mL for simulated drinking water, again at 95% confidence.

US Environmental Protection Agency Method 314.1, an automated sample preconcentration/matrix elimination suppressed conductivity method for the analysis of trace levels (0.50 microg/L) of perchlorate in drinking water.

Since 1997 there has been increasing interest in the development of analytical methods for the analysis of perchlorate. The US Environmental Protection Agency (EPA) Method 314.0, which was used during the first Unregulated Contaminant Monitoring Regulation (UCMR) cycle, supports a method reporting limit (MRL) of 4.0 microg/L. The non-selective nature of conductivity detection, combined with very high ionic strength matrices, can create conditions that make the determination of perchlorate difficult. The objective of this work was to develop an automated, suppressed conductivity method with improved sensitivity for use in the second UCMR cycle. The new method, EPA Method 314.1, uses a 35 mm x 4 mm cryptand concentrator column in the sample loop position to concentrate perchlorate from a 2 mL sample volume, which is subsequently rinsed with 10 mM NaOH to remove interfering anions. The cryptand concentrator column is combined with a primary AS16 analytical column and a confirmation AS20 analytical column. Unique characteristics of the cryptand column allow perchlorate to be desorbed from the cryptand trap and refocused on the head of the guard column for subsequent separation and analysis. EPA Method 314.1 has a perchlorate lowest concentration minimum reporting level (LCMRL) of 0.13 microg/L in both drinking water and laboratory synthetic sample matrices (LSSM) containing up to 1,000 microg/L each of chloride, bicarbonate and sulfate.

Performance evaluation of a method for the determination of bromate in drinking water by ion chromatography (EPA method 317.0) and validation of EPA method 324.0.

The potential carcinogenic nature of bromate has prompted global regulatory agencies, and industrial and academic institutions to publish several methods for the analysis of bromate in both drinking and bottled waters. The United States Environmental Protection Agency (EPA) has reported two methods capable of detecting bromate at or below the promulgated maximum contaminant level of 10.0 microg/l. These methods are EPA Method 300.1 and 317.0. Method 300.1 has been promulgated by EPA for compliance monitoring of bromate under Stage 1 of the Disinfectants/Disinfection By-Products Rule. Due to its sensitivity, selectivity and simplicity, Method 317.0 has been drafted and evaluated for potential use as a future compliance monitoring method. This manuscript describes the performance evaluation work with Method 317.0 and efforts completed at EPA's Technical Support Center that improved the sensitivity of Method 317.0, leading to the development of EPA Method 324.0

Eliminating the chlorite interference in US Environmental Protection Agency Method 317.0 permits analysis of trace bromate levels in all drinking water matrices.

A post-column reagent (PCR) method for bromate analysis in drinking water with a method detection limit (MDL) and method reporting limit (MRL) of 0.1 and 0.5 microg/l, respectively, has been developed by the United States Environmental Protection Agency (EPA) for future publication as EPA Method 317.0. The PCR method provides comparable results to the EPA's Selective Anion Concentration (SAC) method used to support the laboratory analysis of Information Collection Rule (ICR) low-level bromate samples and offers a simple, rugged, direct injection method with potential to be utilized as a compliance monitoring technique for all inorganic Disinfectants/Disinfection By-Products (D/DBPs). It has superior sensitivity for bromate compared to EPA Method 300.1, which was promulgated as the compliance monitoring method for bromate under Stage 1 of the D/DBP rule. This paper addresses elimination of the chlorite interference that was previously reported in finished waters from public water systems (PWSs) that employ chlorine dioxide as the disinfectant. An evaluation of Method 317.0 for the analysis of bromate in commercial bottled waters is also reported.

Analysis of 500-ng/l levels of bromate in drinking water by direct-injection suppressed ion chromatography coupled with a single, pneumatically delivered post-column reagent.

In July 1997, the US Environmental Protection Agency (EPA) began sampling and analyzing drinking water matrices from US municipalities serving populations greater than 100,000 for low-level bromate (> 0.20 microgram/l) in support of the Information Collection Rule (ICR) using the selective anion concentration (SAC) method. In September 1997, EPA published Method 300.1 which lowered the Method 300.0 bromate method detection limit (MDL) from 20.0 to 1.4 micrograms/l. This paper describes the research conducted at the EPA's Technical Support Center laboratory investigating a single post-column reagent, o-dianisidine (ODA), which has been successfully coupled to EPA Method 300.1 to extend the MDL for bromate. Initial studies indicate that this method offers a MDL which approaches the EPA's SAC method with the added benefit of increased specificity, shortened analysis time and reduced sample preparation. The method provides excellent ruggedness and acceptable precision and accuracy with a bromate MDL in reagent water of 0.1 microgram/l, and a method reporting limit of 0.50 microgram/l.

Review of the methods of the US Environmental Protection Agency for bromate determination and validation of method 317.0 for disinfection by-product anions and low-level bromate.

In recent years several methods have been published by the United States Environmental Protection Agency (EPA) which specify bromate as a target analyte. The first of these was EPA Method 300.0. As technological improvements in ion chromatographic hardware have evolved and new detection techniques have been designed, method detection limits for bromate have been reduced and additional procedures have been written, including EPA Method 300.1, 321.8 and, most recently, EPA Method 317.0. An overview of the evolution of these bromate methods since 1989 is presented. The focus is specific to each of these respective procedures, highlighting method strengths, weaknesses, and addressing how these methods fit into EPA's regulatory agenda. In addition, performance data are presented detailing the joint EPA/American Society for Testing and Materials multilaboratory validation of EPA Method 317.0 for disinfection by-product anions and low-level bromate.

Collaborative study of EPA Method 317.0 for the determination of inorganic oxyhalide disinfection by-products in drinking water using ion chromatography with the addition of a postcolumn reagent for trace bromate analysis.

The development of the U.S. Environmental Protection Agency (EPA) Method 317.0 is initiated to provide a sufficiently sensitive and fundamental technique for the compliance monitoring of trace levels of bromate in drinking water. After a comparative evaluation of Method 317.0 and elimination of a chlorite interference, this method is tested by a collaborative study in order to determine the precision and bias of the method and evaluate its potential role as a future compliance-monitoring method for inorganic disinfection by-products (DBPs) and trace bromate. This technique provides a practical method for future compliance monitoring for all of the inorganic oxyhalide DBPs including trace concentrations of bromate.

[Initial manifestation of thromboangiitis obliterans as necrotizing colitis]. / Erstmanifestation einer Thrombangiitis obliterans als nekrotisierende Kolitis.

Necrotizing colitis as primary manifestation of Buerger's disease. We report the disease process of a 41 year old woman, who was referred to our clinic with intermittent claudication of the leg. She has been an excessive smoker since early youth. Three years ago a hemicolectomy was carried out because of a necrotizing colitis. The clinical, angiographic and histologic findings are presented. Finally the frequency of intestinal Buerger's disease and the types of clinical course are discussed.

[Breast augmentation with autologous fat - experience of 96 procedures with the BEAULI-technique]. / Brustaugmentation mit autologem Fett - Erfahrungen aus 96 Operationen mit der BEAULITM-Methode.

Over the past 30 years, interest in the use of autologous fat for aesthetic body contouring, especially for breast augmentation has been continuously on the rise. The benefits of an autologous fat transplant include the absence of any inflammatory reaction to a foreign body, its harmonious appearance and a natural feeling. In earlier years, complications such as necrosis, infections or the formation of cysts, poor resorption rates as well as the difficulty of harvesting large amounts of fat within a reasonable amount of time provided grounds for criticism of the methodology of autologous fat transplantation. With the advent of the so-called BEAULI method, since 2007 a procedure is available for the efficient harvesting and processing of larger quantities of transplantable fat. The aim of the study is to describe the technique in detail and reproducibly and to present a detailed overview of autologous fat transfer due to the basis of our own clinical experience. Between 1 September 2010 and 30 June 2012 the author performed 96 fat transfer procedures on 84 patients. Patients aged 18-60 with a desire for a moderate augmentation of volume and shape of the breasts were selected for the procedure. The fat was harvested using water jet-assisted liposuction (Bodyjet) to flush out the fat cells and subsequent separation of the fat components with the Lipo-Collector. There were no occurrences of post-operative complications in any of the cases. The results were evaluated in the context of a check-up, a photographic comparison and with the completion of a questionnaire. With regard to the increase in size and/or shape enhancement of the breasts, 31% of the patients were very happy with the results, 45% indicated they were satisfied, 23% would have liked more volume, while 1% were dissatisfied. This study indicates that the autologous fat transplant into the female breast using the BEAULI method represents an excellent, safe method for the achievement of a moderate and harmonious breast size augmentation as well as sustainable and natural-looking contour improvements in selected patients. Additional studies with a larger number of cases and longer observation periods over several years as well as guidelines from the professional associations could contribute to the further perfection of the autologous fat transplant method in terms of resorption rate, efficiency and safety.

Role of autophagy in disease resistance and hypersensitive response-associated cell death.

Ancient autophagy pathways are emerging as key defense modules in host eukaryotic cells against microbial pathogens. Apart from actively eliminating intracellular intruders, autophagy is also responsible for cell survival, for example by reducing the deleterious effects of endoplasmic reticulum stress. At the same time, autophagy can contribute to cellular suicide. The concurrent engagement of autophagy in these processes during infection may sometimes mask its contribution to differing pro-survival and pro-death decisions. The importance of autophagy in innate immunity in mammals is well documented, but how autophagy contributes to plant innate immunity and cell death is not that clear. A few research reports have appeared recently to shed light on the roles of autophagy in plant-pathogen interactions and in disease-associated host cell death. We present a first attempt to reconcile the results of this research.

Ageing in a eusocial insect: molecular and physiological characteristics of life span plasticity in the honey bee.

Commonly held views assume that ageing, or senescence, represents an inevitable, passive, and random decline in function that is strongly linked to chronological age. In recent years, genetic intervention of life span regulating pathways, for example, in Drosophila as well as case studies in non-classical animal models, have provided compelling evidence to challenge these views.Rather than comprehensively revisiting studies on the established genetic model systems of ageing, we here focus on an alternative model organism with a wild type (unselected genotype) characterized by a unique diversity in longevity - the honey bee.Honey bee (Apis mellifera) life span varies from a few weeks to more than 2 years. This plasticity is largely controlled by environmental factors. Thereby, although individuals are closely related genetically, distinct life histories can emerge as a function of social environmental change.Another remarkable feature of the honey bee is the occurrence of reverted behavioural ontogeny in the worker (female helper) caste. This behavioural peculiarity is associated with alterations in somatic maintenance functions that are indicative of reverted senescence. Thus, although intraspecific variation in organismal life span is not uncommon, the honey bee holds great promise for gaining insights into regulatory pathways that can shape the time-course of ageing by delaying, halting or even reversing processes of senescence. These aspects provide the setting of our review.We will highlight comparative findings from Drosophila melanogaster and Caenorhabditis elegans in particular, and focus on knowledge spanning from molecular- to behavioural-senescence to elucidate how the honey bee can contribute to novel insights into regulatory mechanisms that underlie plasticity and robustness or irreversibility in ageing.

Prediction of regional myocardial blood flow in dogs.

Three indices of regional coronary flow were tested to determine if a good correlation between calculated driving pressure and flow could be found. Four different myocardial depths were examined. All three indexes were based on subtraction of a backpressure to flow. The first assumed backpressure to be equal to ventricular pressure; the second assumed backpressure to be twice ventricular pressure; and the third index assumed backpressure to be zero and thus was simply coronary pressure. Only the index that used left ventricular pressure as the backpressure to flow exhibited proportionality for the subendocardial samples. Similarly, only coronary pressure yielded a good fit for the subepicardial samples. Although subendocardial flow was always in proportion to the index, the index was poor at predicting the absolute value of that flow. The usefulness of such a directional predictor of subendocardial perfusion was illustrated by examining the phase shift resulting from partial occlusion of a coronary artery. The index predicted that subendocardial flow would be lower when coronary pressure was reduced by partially occluding the coronary vessel than when it was presented with an equivalent steady pressure. Microsphere measurements confirmed this prediction. We conclude that an index that assumes ventricular pressure to be the backpressure to flow is an accurate predictor of relative changes in subendocardial perfusion.

In vitro establishment of Marek's disease herpesvirus-transformed productive and nonproductive lymphoblastoid cell lines.

The establishment of two Marek's disease herpesvirus-transformed lymphoblastoid cell lines (JMV-1 and JM-1) is reported. Cocultivation with chicken embryo fibroblasts, attempts to rescue Marek's disease virus from kidney cell cultures, and immunofluorescent staining show that JMV-1 is a nonproductive line and JM-1 is a productive line.

[Hormonal therapy (medical orchiectomy) in prostatic carcinoma]. / Die hormonelle Therapie (medikamentöse Orchiektomie) des Prostatakarzinoms.

Hormonal therapy of prostate cancer is still the most effective systemic treatment. Possible alternatives to subcapsular orchiectomy are discussed, comprising the estrogens with their known cardiovascular side effects, the expensive LH-RH analogues, and/or the antiandrogens, which depend on the patient's compliance. Due to more intensive health care and more sensitive diagnostic methods, there is now an increase not only in the overall number of diagnosed prostate carcinomas, but also in the percentage of those which do not need treatment. The question when, and in which patient hormonal treatment should be started becomes increasingly important.