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The reversibility of bacterial resistance to antibiotics is poorly understood. Therefore, the aim of this study was to determine, over a period of five years, the effect of fluoroquinolone (FQ) use in primary care on the development and gradual decay of Escherichia coli resistance to FQ. In this matched case−control study, we linked three sources of secondary data of the Health Service of the Autonomous Province of Bolzano, Italy. Cases were all those with an FQ-resistant E. coli (QREC)-positive culture from any site during a 2016 hospital stay. Data were analyzed using conditional logistic regression. A total of 409 cases were matched to 993 controls (FQ-sensitive E. coli) by the date of the first isolate. Patients taking one or more courses of FQ were at higher risk of QREC colonization/infection. The risk was highest during the first year after FQ was taken (OR 2.67, 95%CI 1.92−3.70, p < 0.0001), decreased during the second year (OR 1.54, 95%CI 1.09−2.17, p = 0.015) and became undetectable afterwards (OR 1.09, 95%CI 0.80−1.48, p = 0.997). In the first year, the risk of resistance was highest after greater cumulative exposure to FQs. Moreover, older age, male sex, longer hospital stays, chronic obstructive pulmonary disease (COPD) and diabetes mellitus were independent risk factors for QREC colonization/infection. A single FQ course significantly increases the risk of QREC colonization/infection for no less than two years. This risk is higher in cases of multiple courses, longer hospital stays, COPD and diabetes; in males; and in older patients. These findings may inform public campaigns and courses directed to prescribers to promote rational antibiotic use.

The developmental effects of chemicals that co-occur in vulnerable populations with elevated psychological stress are of increasing concern to the public. To investigate these concerns, we developed a rodent model of co-occurring perinatal manipulations and conducted a series of cognitive assessments in male and female offspring. Manganese (Mn), a neurodevelopmental toxicant when exceeding physiological requirements, was delivered in the drinking water (0, 2, or 4 mg Mn/mL) of rats from gestational day (GD) 7 to postnatal day (PND) 22. A variable perinatal stress paradigm was applied to half of the animals from GD13 to PND9. Novel object recognition (NOR), Morris water maze (MWM), differential reinforcement of low-rates procedure (DRL) and cued and uncued choice reaction time (CRT) tests were used to assess cognitive functions in offspring. Mn (4 mg/mL) and stress impaired NOR in adolescent males but facilitated NOR performance in females. However, when stress and Mn were combined these effects were attenuated in both sexes. During training for the DRL, Mn (2 mg/mL) facilitated, while stress impaired, lever press learning in both sexes. Few effects related to the treatments were found on DRL or MWM. During cued CRT, Mn (2 and 4 mg/mL) and stress reduced accuracy in males, while stress and Mn (2 mg/mL) increased anticipatory responding and slowed decision time in both sexes. Stress combined with Mn (2 mg/mL) improved cued accuracy and decision time, and Mn attenuated the effect of stress on anticipatory responding in both sexes. Stress slowed female movement time but when combined with Mn (4 mg/mL) the effect of stress was attenuated. During uncued CRT, except for decision time (which replicated effects observed with the cued task), no other effects of Mn or its combination with stress occurred. Females remained negatively affected by stress in most uncued CRT performance measures, while stressed improved male uncued accuracy. Taken together these data do not support increased cognitive impairment produced by Mn when combined with stress. However, the effects of perinatal stress alone, on these cognitive functions may hinder the detection of effects due to chemical exposures and underscores the need to consider the psychological health and wellbeing of the mother and her environment in risk assessment for developmental neurotoxicity of chemicals., (Published by Elsevier Inc.)

Today, integrative taxonomy is often considered the gold standard when it comes to species recognition and delimitation. Using the Tetrix bipunctata complex, we here present a case where even integrative taxonomy may reach its limits. The Tetrix bipunctata complex consists of two morphs, bipunctata and kraussi , which are easily distinguished by a single character, the length of the hind wing. Both morphs are widely distributed in Europe and reported to occur over a large area in sympatry, where they occasionally may live also in syntopy. The pattern has led to disparate classifications, as on the one extreme, the morphs were treated merely as forms or subspecies of a single species, on the other, as separate species. For this paper, we re-visited the morphology by using multivariate ratio analysis (MRA) of 17 distance measurements, checked the distributional data based on verified specimens and examined micro-habitat use. We were able to confirm that hind wing length is, indeed, the only morphological difference between bipunctata and kraussi . We were also able to exclude a mere allometric scaling. The morphs are, furthermore, largely sympatrically distributed, with syntopy occurring regularly. However, a microhabitat niche difference can be observed. Ecological measurements in a shared habitat confirm that kraussi prefers a drier and hotter microhabitat, which possibly also explains the generally lower altitudinal distribution. Based on these results, we can exclude classification as subspecies, but the taxonomic classification as species remains unclear. Even with different approaches to classify the Tetrix bipunctata complex, this case is, therefore, not settled. We recommend continuing to record kraussi and bipunctata separately., (Valentin Moser, Hannes Baur, Arne W. Lehmann, Gerlind U. C. Lehmann.)

Research is lacking on the reversibility of antimicrobial resistance (AMR). Thus, we aimed to determine the influence of previous antibiotic use on the development and decay over time of third generation cephalosporin (3GC)-resistance of E. coli . Using the database of hospital laboratories of the Autonomous Province of Bolzano/Bozen (Italy), anonymously linked to the database of outpatient pharmaceutical prescriptions and the hospital discharge record database, this matched case-control study was conducted including as cases all those who have had a positive culture from any site for 3GC resistant E. coli (3GCREC) during a 2016 hospital stay. Data were analyzed by conditional logistic regression. 244 cases were matched to 1553 controls by the date of the first isolate. Male sex (OR 1.49, 95% CI 1.10-2.01), older age (OR 1.11, 95% CI 1.02-1.21), the number of different antibiotics taken in the previous five years (OR 1.20, 95% CI 1.08-1.33), at least one antibiotic prescription in the previous year (OR 1.92, 95% CI 1.36-2.71), and the diagnosis of diabetes (OR 1.57, 95% CI 1.08-2.30) were independent risk factors for 3GCREC colonization/infection. Patients who last received an antibiotic prescription two years or three to five years before hospitalization showed non-significant differences with controls (OR 0.97, 95% CI 0.68-1.38 and OR 0.85, 95% CI 0.59-1.24), compared to an OR of 1.92 (95% CI 1.36-2.71) in those receiving antibiotics in the year preceding hospitalization. The effect of previous antibiotic use on 3GC-resistance of E. coli is highest after greater cumulative exposure to any antibiotic as well as to 3GCs and in the first 12 months after antibiotics are taken and then decreases progressively.

Case Description: Osseointegration is a relatively new technique for prosthetic limb attachment that offers various improvements for patients with amputation and facilitates joint preservation. We present a case of implant loosening during rehabilitation in a patient with transtibial amputation that was successfully managed through a combination of measures, aiming to promote re-osseointegration of the implant., Objectives: Not much is known about structured management of adverse events after osseointegration. Septic or aseptic loosening is currently regarded as implant failure, prompting removal and possible re-implantation at a later stage. The objective of this case report was to evaluate the feasibility of salvaging a loosened implant., Study Design: Case report., Treatment: A novel treatment approach was employed to enable renewed osseointegration of the implant. First, the bone-implant interface was disrupted and renewed through axial rotation and distal repositioning of the implant. Afterwards, extracorporal shockwave therapy and antibiotic treatment were administered. Prosthetic rehabilitation was then started anew. Regular follow-up x-rays and clinical evaluations were conducted, including standardized outcome tests., Outcomes: These combined measures led to a successful re-osseointegration of the implant. In a 21-month follow-up, the patient regained a stable and secure gait pattern, using his prosthesis every day for 15 hours and scoring above average on standardized outcome measures., Conclusion: This represents the first report of implant salvage after failed primary osseointegration. As the associated risks of this novel treatment are very low, investigations are warranted to evaluate this approach on a larger scale.

The transverse cervical nerve (TCN) is a superficial cutaneous branch of the cervical plexus that innervates the skin of the anterolateral neck. Therefore, it is exposed to injury in anterolateral cervical surgery, which can cause neuropathic pain. To provide a method with which to relieve patients' pain, this study aimed to evaluate the possibility of visualization, diagnostic assessment and blockade of the TCN with high-resolution ultrasound (HRUS). HRUS with high-frequency probes (15-22 MHz), guided ink-marking and consecutive dissection on both sides in nine fresh cadaver necks (n = 18) was conducted. On both sides of 20 healthy volunteers (n = 40), the distances between the greater auricular nerve (GAN) and the TCN at the posterior border of the sternocleidomastoid muscle were measured. Finally, cases referred to HRUS examinations because suspected TCN lesions were assessed. The TCN was visible in all anatomic specimens and in healthy volunteers. Dissection confirmed HRUS findings in all anatomic specimens (100%). In healthy volunteers, the mean distance between the GAN and the TCN was 10.42 ± 3.20 mm. The median visibility, rated on a five-point Likert scale, was four, reflecting good diagnostic quality. There were six patients with visible abnormalities on HRUS. This study confirmed the reliable visualization of the TCN with HRUS in anatomic specimens, healthy volunteers and patients., Competing Interests: Conflict of interest disclosure The authors declare no competing interests., (Copyright © 2020 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Background and Aims: Carriage of rs738409:G in patatin-like phospholipase domain containing 3 (PNPLA3) is associated with an increased risk for developing alcohol-related cirrhosis and hepatocellular carcinoma (HCC). Recently, rs72613567:TA in hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13) was shown to be associated with a reduced risk for developing alcohol-related liver disease and to attenuate the risk associated with carriage of PNPLA3 rs738409:G. This study explores the risk associations between these two genetic variants and the development of alcohol-related cirrhosis and HCC., Approach and Results: Variants in HSD17B13 and PNPLA3 were genotyped in 6,171 participants, including 1,031 with alcohol-related cirrhosis and HCC, 1,653 with alcohol-related cirrhosis without HCC, 2,588 alcohol misusers with no liver disease, and 899 healthy controls. Genetic associations with the risks for developing alcohol-related cirrhosis and HCC were determined using logistic regression analysis. Carriage of HSD17B13 rs72613567:TA was associated with a lower risk for developing both cirrhosis (odds ratio [OR], 0.79; 95% confidence interval [CI], 0.72-0.88; P = 8.13 × 10 -6 ) and HCC (OR, 0.77; 95% CI, 0.68-0.89; P = 2.27 × 10 -4 ), whereas carriage of PNPLA3 rs738409:G was associated with an increased risk for developing cirrhosis (OR, 1.70; 95% CI, 1.54-1.88; P = 1.52 × 10 -26 ) and HCC (OR, 1.77; 95% CI, 1.58-1.98; P = 2.31 × 10 -23 ). These associations remained significant after adjusting for age, sex, body mass index, type 2 diabetes, and country. Carriage of HSD17B13 rs72613567:TA attenuated the risk for developing cirrhosis associated with PNPLA3 rs738409:G in both men and women, but the protective effect against the subsequent development of HCC was only observed in men (OR allelic , 0.75; 95% CI, 0.64-0.87; P = 1.72 × 10 -4 )., Conclusions: Carriage of variants in PNPLA3 and HSD17B13 differentially affect the risk for developing advanced alcohol-related liver disease. A genotypic/phenotypic risk score might facilitate earlier diagnosis of HCC in this population., (© 2019 by the American Association for the Study of Liver Diseases.)

In neonatal intensive care units (NICUs), 87.5% of alarms by the monitoring system are false alarms, often caused by the movements of the neonates. Such false alarms are not only stressful for the neonates as well as for their parents and caregivers, but may also lead to longer response times in real critical situations. The aim of this project was to reduce the rates of false alarms by employing machine learning algorithms (MLA), which intelligently analyze data stemming from standard physiological monitoring in combination with cerebral oximetry data (in-house built, OxyPrem)., Materials & Methods: Four popular MLAs were selected to categorize the alarms as false or real: (i) decision tree (DT), (ii) 5-nearest neighbors (5-NN), (iii) naïve Bayes (NB) and (iv) support vector machine (SVM). We acquired and processed monitoring data (median duration (SD): 54.6 (± 6.9) min) of 14 preterm infants (gestational age: 26 6/7 (± 2 5/7) weeks). A hybrid method of filter and wrapper feature selection generated the candidate subset for training these four MLAs., Results: A high specificity of >99% was achieved by all four approaches. DT showed the highest sensitivity (87%). The cerebral oximetry data improved the classification accuracy., Discussion & Conclusion: Despite a (as yet) low amount of data for training, the four MLAs achieved an excellent specificity and a promising sensitivity. Presently, the current sensitivity is insufficient since, in the NICU, it is crucial that no real alarms are missed. This will most likely be improved by including more subjects and data in the training of the MLAs, which makes pursuing this approach worthwhile.

This prospective study was conducted to investigate electrophysiological qualities and patient's satisfaction of a synovial gliding tissue flap in treating true recurring carpal tunnel syndrome. In 14 patients (11 women, three men), 15 median nerves were included in this retrospective study. For all 15 nerves, motor and sensory nerve conduction velocity, compound muscle action potential, a Visual Analogue Scale-score (VAS-score) questionnaire and an adapted Levine-Test were evaluated pre- and postoperatively. All participants underwent operative neurolysis of the median nerve, which was then enwrapped by a synovial gliding tissue flap. Eleven procedures were completed by integument enlargement. Follow-up period was 12 months. Postoperatively, distal latency decreased significantly by 15.6%. Compound muscle action potential and sensory nerve conduction velocity did not improve significantly. VAS score regarding pain reduced highly significantly with 74.1%. The adapted Levine-Test function score improved highly significantly with 39.2%. The synovial gliding tissue flap lead to an excellent patient's satisfaction for treating true recurring carpal tunnel syndrome. Primary wound closure should be completed with integument enlargement if needed., Competing Interests: The authors declare no conflict of interest.

Early disease diagnosis is key to the effective treatment of diseases. Histopathological analysis of human biopsies is the gold standard to diagnose tissue alterations. However, this approach has low resolution and overlooks 3D (three-dimensional) structural changes resulting from functional alterations. Here, we applied multiphoton imaging, 3D digital reconstructions and computational simulations to generate spatially resolved geometrical and functional models of human liver tissue at different stages of non-alcoholic fatty liver disease (NAFLD). We identified a set of morphometric cellular and tissue parameters correlated with disease progression, and discover profound topological defects in the 3D bile canalicular (BC) network. Personalized biliary fluid dynamic simulations predicted an increased pericentral biliary pressure and micro-cholestasis, consistent with elevated cholestatic biomarkers in patients' sera. Our spatially resolved models of human liver tissue can contribute to high-definition medicine by identifying quantitative multiparametric cellular and tissue signatures to define disease progression and provide new insights into NAFLD pathophysiology.

A deeper epigenomic understanding of spatial organization of cells in human tissues is an important challenge. Here we report the first combined positional analysis of transcriptomes and methylomes across three micro-dissected zones (pericentral, intermediate and periportal) of human liver. We identify pronounced anti-correlated transcriptional and methylation gradients including a core of 271 genes controlling zonated metabolic and morphogen networks and observe a prominent porto-central gradient of DNA methylation at binding sites of 46 transcription factors. The gradient includes an epigenetic and transcriptional Wnt signature supporting the concept of a pericentral hepatocyte regeneration pathway under steady-state conditions. While donors with non-alcoholic fatty liver disease show consistent gene expression differences corresponding to the severity of the disease across all zones, the relative zonated gene expression and DNA methylation patterns remain unchanged. Overall our data provide a wealth of new positional insights into zonal networks controlled by epigenetic and transcriptional gradients in human liver.

The measurement of peripheral oxygen saturation (SpO2) in neonatal intensive care units (NICUs) poses a significant challenge. Motion artifacts due to the patient's limb motion induce many false alarms, which in turn cause an additional workload for the medical staff and anxiety for the parents. We developed a reflectance pulse oximeter dedicated to be placed at the patient's forehead, which is less prone to such artifacts. We trained our algorithms for SpO2 estimation on 8 adult healthy volunteers participating in a controlled desaturation study. We then validated our SpO2 monitoring system on 25 newborn patients monitored in an NICU. We further evaluated the versatility and resilience to low signal-tonoise ratios (SNR) of our solution by testing it on signals acquired in a low-perfusion region (upper right part of the chest) of our adult volunteers. We obtained an SpO2 estimation accuracy ($A &#95;{\mathbf {rms}}$) of 1.9 % and 3.1 % at the forehead and the chest in our adult volunteers, respectively. These performances were obtained after automatic rejection of 0.1 % and 30.0 %, respectively, of low-SNR signals by our dedicated quality index. In the dataset recorded on newborn patients in the NICU, we obtained an accuracy of 3.9 % after automatic rejection of 11.7 % of low-SNR signals by our quality index. These analyses were carried out following the procedures suggested by the ISO 80601-2-61:2011 standard, which specifies a target $A &#95;{\mathbf {rms}} \le $ 4 % for SpO2 monitoring applications. These promising results suggest that reflectance pulse oximeters can achieve clinically acceptable accuracy, while being placed at locations less sensitive to limb motion artifacts - such as the forehead - thereby reducing the amount of SpO2-related false alarms in NICUs.

Purpose:  Neuropathy of the intermediate (IFCN) and medial femoral cutaneous nerve (MFCN) is a potential iatrogenic complication of thigh surgery and its diagnosis is limited. This study aimed to evaluate the possibility of the visualization and diagnostic assessment of the IFCN and MFCN with high-resolution ultrasound (HRUS)., Materials and Methods:  In this study, HRUS with high-frequency probes (15 - 22MHz) was used to locate the IFCN and the MFCN in 16 fresh cadaveric lower limbs. The correct identification of the nerves was verified by ink-marking and consecutive dissections at sites correlating to nerve positions (R1 - 3), namely, the origin, the mid portion, and the distal portion, respectively. 12 cases with suspected IFCN and MFCN lesions referred to our clinic for HRUS examinations were also assessed., Results:  Anatomical dissection confirmed the correct identification of the IFCN in 16/16 branches at all of the different locations (100 %). MFCN was correctly identified at R1 + 3, in all cases (16/16; 100 %), and in 14/16 cases (88 %) at (R2). 12 cases of patients with IFCN and MFCN pathologies (all of iatrogenic origin) were identified. 9 instances of structural damage were visible on HRUS, and all pathologies were confirmed by almost complete resolution of symptoms after selective HRUS-guided blocks with 0.5 - 1 ml lidocaine 2 %., Conclusion:  This study confirms that the IFCN and the MFCN can be reliably visualized with HRUS throughout the course of these nerves, both in anatomical specimens and in patients., Competing Interests: Dr. Riegler reports grants from Medical Scientific Fund of the Mayor of the City of Vienna, during the conduct of the study., (© Georg Thieme Verlag KG Stuttgart · New York.)

The potential for developmental neurotoxicity (DNT) of environmental chemicals may be evaluated using specific test guidelines from the US Environmental Protection Agency or the Organisation for Economic Cooperation and Development (OECD). These guidelines generate neurobehavioral, neuropathological, and morphometric data that are evaluated by regulatory agencies globally. Data from these DNT guideline studies, or the more recent OECD extended one-generation reproductive toxicity guideline, play a pivotal role in children's health risk assessment in different world areas. Data from the same study may be interpreted differently by regulatory authorities in different countries resulting in inconsistent evaluations that may lead to inconsistencies in risk assessment decisions internationally, resulting in regional differences in public health protection or in commercial trade barriers. These issues of data interpretation and reporting are also relevant to juvenile and pre-postnatal studies conducted more routinely for pharmaceuticals and veterinary medicines. There is a need for development of recommendations geared toward the operational needs of the regulatory scientific reviewers who apply these studies in risk assessments, as well as the scientists who generate DNT data sets. The workshops summarized here draw upon the experience of the authors representing government, industry, contract research organizations, and academia to discuss the scientific issues that have emerged from diverse regulatory evaluations. Although various regulatory bodies have different risk management decisions and labeling requirements that are difficult to harmonize, the workshops provided an opportunity to work toward more harmonized scientific approaches for evaluating DNT data within the context of different regulatory frameworks. Five speakers and their coauthors with neurotoxicology, neuropathology, and regulatory toxicology expertise discussed issues of variability, data reporting and analysis, and expectations in DNT data that are encountered by regulatory authorities. In addition, principles for harmonized evaluation of data were suggested using guideline DNT data as case studies., (Copyright © 2017. Published by Elsevier Inc.)

Background: Since the WHO study "Family Health Nursing" (FHN) the concepts have been established different within Europe. Due to this, the international project "Family Health Nursing in European Communities" was started in order to develop a standardised educational concept., Aims: The aim of this project was to gather the different requirements and to clarify problematic issues within the participating EU countries., Method: Thus, a literature review, a Delphi study in order to achieve consensus on definition of FHN, an analysis of competencies and requirements as well as an overview of available education/training with experts from the field of care was carried out., Results: The results of the review reflect the varying level of occupation within the countries included in this analysis. Over the time, various conceptual orientations and professional designations have been established. Within this project the preferred job title was "Family Nurse" (46,3 %). Health promotion and prevention (85,8 %) were seen as main tasks of the Family Health Nurse. Therefore, the respondent experts state the need for more specialized training at an undergraduate (81,5 %) level., Conclusions: The project outlines the countries' effort towards a high-level in familial care which could be supported in long term by a consistent FHN concept.

Clostridium difficile is the most common cause of antibiotic-associated diarrhoea in hospitals and other healthcare facilities. The elderly are particularly susceptible and at increased risk for adverse outcome as a result of C. difficile infection. The aim of this study was to determine the prevalence of C. difficile colonization among residents of nursing homes in Hesse and to compare it with the prevalence in the general population living outside long-term care facilities (LTCF). We assessed possible risk factors for C. difficile colonization and determined the genotype of circulating strains. C. difficile was isolated from 11/240 (4.6%) nursing home residents and 2/249 (0.8%) individuals living outside LTCF (p = 0.02). Ten of 11 (90.9%) isolates from nursing homes and one of two isolates from the population outside LTCF were toxigenic. The prevalence of C. difficile colonization varied from 0% to 10% between different nursing homes. Facilities with known actual or recent CDI cases were more likely to have colonized residents than facilities without known CDI cases. C. difficile PCR-ribotypes 014 and 001 were the most prevalent genotypes and accounted for 30% and 20% of toxigenic isolates in nursing homes, respectively. Interestingly, no individuals carried the epidemic strain PCR-ribotype 027. Our results suggest that residents of nursing homes in Germany are at high risk for colonization by virulent C. difficile strains. The high prevalence of C. difficile colonization in nursing homes underscores the importance of good adherence to standard infection control precautions even in the absence of a diagnosed infection. They also emphasize the need for specific programs to increase the awareness of healthcare professionals in LTCF for CDI.

Introduction: Management of the painful neuroma has been subject to controversy since the earliest descriptions of this disabling problem. Today, treatment is limited to resection of the neuroma and implantation of the nerve in a muscle at a location where it is safe from irritation and trauma. This however is not attainable in many cases and it is our clinical experience, that nerves without a target remain a source of constant discomfort and pain. Recently we reported of the feasibility of neuroma prevention through end-to-side neurorraphy into adjacent sensory and/or motor nerves to provide a target for axons deprived of their endorgan. Here we report of our first clinical experience with this method in sixteen patients with longstanding upper and lower extremity neuromas., Patients and Methods: 16 patients were included in this study. All had neuromas of different sensory nerves of both the upper and lower extremity. 11 were of iatrogenic origin, 5 were caused by different traumas. 8 had previous attempts to surgically treat the neuroma. Finally, all were treated by end-to-side neurorraphy into adjacent nerves. Postoperatively quantitative sensorymotor testing was performed to evaluate possible changes of nerve function of the recipient nerves. Pain was evaluated by visual analogue score and changes in pain medication., Results: In no patient a sensory or motor deficit or painful sensations were induced in the target area of the recipient nerve. Some had dysaesthesias for about 6 months, which finally subsided. All but 1 patient improved in their symptoms at a follow-up of more than 2 years., Conclusion: Previous experimental work and present clinical results suggest that axons of a severed peripheral nerve that are provided with a pathway and target through an end-to-side coaptation will either be pruned or establish some type of end-organ contact so that a neuroma can be prevented without inducing sensory or motor dysfunctions in the recipient nerve., (Georg Thieme Verlag KG Stuttgart New York.)

This study aims to assess the suitability of biodegradable membranes as transfer matrix materials for the culture of subconfluent fibroblasts and keratinocytes. The materials investigated were based on collagen, chitosan and enzyme-digestible cellulose. The proliferation and growth behaviour of human keratinocytes and dermal fibroblasts were analysed and morphology and distribution determined. Cultured fibroblasts exhibited no significant differences in proliferation for the different membrane types, whereas keratinocytes revealed significantly higher proliferation on collagen membranes compared with membranes based on cellulose and chitosan. Co-cultured fibroblasts and keratinocytes from the same donor on collagen membranes showed more homogenous cell distribution, but they segregated in heterologous co-cultures; this effect must be further investigated. Thus, collagen and collagen-coated chitosan membranes are suitable for the subconfluent transfer of human fibroblasts and keratinocytes.

The reliable detection, measurement, and interpretation of treatment-related developmental neurotoxicity (DNT) effects depend on appropriate study design and execution, using scientifically established methodologies, with appropriate controls to minimize confounding factors. Appropriate statistical approaches should be optimized for the specific endpoints in advance, analyzing effects across time and functional domains as far as possible. If available, biomarkers of exposure are useful to assess the bioavailability of toxicants to the dam and offspring in utero and after birth. Finally, "weight of evidence" principles are used to aid assessment of the biological significance of differences from concurrent controls. These effects should be interpreted in light of available information from historical controls, positive controls, maternal and offspring systemic toxicity, and other relevant toxicological data. This review provides a framework for the integration of all these types of information in the interpretation of DNT studies.

Background: Previous studies demonstrated, that cultured epithelial autografts (CEA) can be isolated and skin cell sprays can be produced for application on different types of wounds. The purpose of the present study was to determine which cell types can be isolated from the human scalp and whether these cells can be used for spray transplantation., Methods: Outer root sheath cells (ORS), keratinocytes, melanocytes, dermal papilla cells (DP), and dermal sheath cells (DSC) were isolated from human scalp tissue. Isolated cells were characterized, expanded and sprayed in an in vitro model. Growth behaviour, morphology and cell counts were compared with non-sprayed cells., Results: With acceptable time, equipment and laboratory personnel a sufficient amount of keratinocytes, ORS, melanocytes, DP cells and DSC cells could be achieved. The cells are sufficient for application as a cell spray. Cells, positive for Integrin alpha6, Cytokeratin 19, CD73 and CD105 were identified within the cultures., Conclusions: Human scalp is suitable to gain epidermal and dermal cells for the development of therapeutic cell spray transplantation. Further studies have to determine, whether these cells can be combined to produce wound specific skin substitutes.

Purpose/background: High-voltage burns represent a challenging surgical entity. Compared to conventional burns, these injuries are characterized by an increased morbidity and worse potential for rehabilitation. The aim of the present study was to analyse the management of high-voltage injuries during the early posttraumatic period with special emphasis on the surgical procedures., Patients/material and Method: We retrospectively evaluated the medical records of patients with electrical injuries treated from 1995 - 2007. A total of 61 patients (57 men, 4 females, mean age: 34 +/- 13 years) with high-voltage burns was included for analysis., Results: The majority of high-voltage burns was work-related (75 %). The mean total burn area was 35 % of the total body surface, with a mean of 29 % deep burns. An average of 4.8 +/- 4 operations were performed per patient (range: 1 - 23 operations). Surgical procedures included repeated debridement/necrectomy (100 % of all patients), early escharotomy/fasciotomy (47.5 %), and amputations (18 %). 14 patients (23 %) underwent reconstructive surgery using either local or free flaps. The mortality rate was 15 %., Conclusion: The surgical management of high-voltage burns is characterised by repetitive debridements and necrectomies. Despite an aggressive approach to remove necrotic tissue, the mortality in this type of injury is considerably high. Limb salvage may be achieved with the use of free microvascular flaps. However, an amputation of necrotic extremities must be considered in the copresence of septic complications.

Although midcarpal wrist arthrodesis is recognized as a standard procedure to treat scapholuate advanced collapse (SLAC) and scaphoid nonunion advanced collapse (SNAC) of the wrist, little has been reported about patients with bilateral involvement and the number, cause, and results of failed cases requiring conversion to total wrist arthrodesis. This study investigated the results of 20 patients with bilateral procedures and of 22 patients who underwent total wrist fusion after failed midcarpal arthrodesis out of an overall group of 907 patients treated by this method during a 12-year period. Of these, 16 bilateral and 20 converted cases were reexamined after an average of 48 months and 42 months, respectively. Patients after bilateral midcarpal arthrodesis experienced a pain reduction by an average of 54% of the preoperative pain values at rest and by 56% at stress on the visual analog scale (scale range: 0 to 100) and from intolerable (3.7) to pain only during stress (1.9) on the verbal scale (scale range: I to 4). A mean arc of wrist extension and flexion of 53 degrees on the right and 49 of the left wrist was preserved. The mean DASH score was 45 points and 70% of the patients felt impaired only during certain activities. Total arthrodesis reduced pain in 18 of 20 reexamined wrists by 67% of the previous values after the failed partial arthrodesis at rest and by 46% at stress on the visual analog scale andfrom intolerable pain (3.7) to pain only during stress (2.1) on the verbal scale. Seven of the 20 reexamined patients noted complete pain relief at rest and two also under stress conditions. The DASH score averaged 39 points. A mean Krimmer score of 46 points and a mean Buck-Gramcko and Lohman evaluation of 6 points represented a satisfactory result. Grip strength of the operated hand averaged 53% of the opposite side. Subjectively, 30% felt impaired only during certain activities, 55%felt considerably and 15% strongly limited in daily life. However, all but two patients were satisfied with the secondary total wrist fusion as pain was considerably reduced. Midcarpal arthrodesis reliably reduced pain and preserved valuable wrist mobility thus improving daily activity and quality of life also in bilateral carpal collapse. In the rare cases when midcarpal arthrodesis failed, total wrist arthrodesis markedly improved the complaints in most patients, but in contrast to other studies complete pain was seldom.

The protection of human health from the adverse effects of cumulative environmental exposure to chemical mixtures is an important issue. Of particular interest is the detection and characterization of interaction among chemicals in complex mixtures. Response surface methodology, often supported by factorial designs, is the classical statistical experimental approach. Fixed-ratio ray designs, which may include the use of single chemical data in addition to data along mixture ray(s), have been proposed as an alternative approach. Such designs permit a reduction in the amount of experimental effort when the region of interest can be restricted to exposure-relevant mixing ratios. A 'single chemicals required' (SCR) approach and a 'single chemicals not required' (SCNR) approach are both described. The methods are illustrated with a five-chemical mixture of organophosphorus pesticides-acephate (ACE), diazinon (DIA), chlorpyrifos (CPF), malathion (MAL) and dimethoate (DIM). Their relative proportions in the mixture were based on the relative dietary human exposure estimates of each chemical as projected by the U.S. EPA Dietary Exposure Evaluation Model (DEEM). Use of the SCR and SCNR methods for binary endpoints are demonstrated using a dichotomized gait score as an indication of toxicity. For both methods, the overall hypothesis of additivity was rejected, indicating significant departure from additivity when the five pesticides were combined at the specified mixing ratio. By comparison of the predicted response under additivity to the modeled response of the experimental mixture data this departure from additivity was characterized as synergy (greater than additive toxicity). To examine the influence of malathion in the mixture, it was removed from the five-pesticide mixture (full ray) and the remaining four chemicals (reduced ray) were combined at the same relative proportions used in the full fixed-ratio ray There was not a significant departure from additivity along the ray with the four remaining pesticides omitting malathion. Thus, although malathion was not dose-responsive alone, it significantly interacted with the other pesticides.

It was the aim of this study to improve our knowledge on thiolated polymers by the synthesis and in vitro characterization of a poly(acrylic acid)-homocysteine conjugate. Mediated by a carbodiimide, homocysteine was therefore covalently attached to poly(acrylic acid) via the formation of an amide bond. The isolated conjugate displayed 930 micromol +/- 83 micromol sulfur atoms per gram polymer. Of these thiol groups, 80.1% were oxidized to disulfide bonds during the coupling reaction. In aqueous solutions the conjugate was rapidly oxidized by the formation of disulfide bonds at pH 8, whereas it remained stable at pH 7 and below during the observation period of 4 hours. Due to the immobilization of thiol groups on the polymer, the mucoadhesive and cohesive properties of poly(acrylic acid) were strongly improved. Furthermore, the thiolated polymer exhibited a significantly (p < 0.05) improved permeation enhancing effect in comparison to the unmodified polymer. Because of these features the poly(acrylic acid)-homocysteine conjugate seems to represent a promising novel tool, which might be useful in particular for aqueous formulations based on thiomers.

Objective: Radiofrequency (RF) has been demonstrated as a relatively safe procedure in the treatment of hepatocellular carcinoma (HCC)., Methods: A 60-year-old man with liver cirrhosis presented with a large, infiltrating HCC close to the right colon. The patient was treated with internally cooled RF., Results: On the 8th day post-ablation, a complication of right colon perforation was diagnosed., Conclusion: RF can induce untoward thermal damage, and may not be indicated in large infiltrating lesions abutting the gastrointestinal tract.

Female C57BL6 mice were exposed to 0 or 800 ppm carbon disulfide (CS2), 6 h/d, 5 d/wk for 20 weeks. The neurologic function of all mice was assessed once at the end of exposures using a functional observational battery. General health effects included a decrease in body weight gain, piloerection, hunched body posture, and ptosis. Treatment-related effects included altered gait (uncoordinated placement of hind limbs and ataxia) and impaired function on an inverted screen test. In addition, rearing and locomotor movement were decreased in treated mice. Focal to multifocal axonal swelling was seen predominantly in the muscular branch of the posterior tibial nerve, and occasionally giant axonal swelling was detected in the lumbar segment of the spinal cord. Electron microscopic examination revealed swollen axons with massive accumulation of neurofilament proteins within the axoplasm. Covalent cross-linking of erythrocyte spectrin (surrogate protein to neurofilament protein) was demonstrated in mice exposed to CS2 but not in mice receiving filtered air. These data provide supportive evidence that covalent cross-linking of neurofilament proteins is a significant feature of the axonal swellings in mice produced by inhalation exposure to CS2.

When given to rats, O,O'-diethyl-O-[3,5,6-trichloro-2-pyridyl]- phosphorothionate (chlorpyrifos), a common insecticide, causes an unusually lengthy dose-dependent fall in the activity of brain acetylcholinesterase (AChE; EC 3.1.1.7). To determine whether the slow recovery involves impaired AChE synthesis, experiments were designed to measure AChE activity, immunoreactive AChE protein (AChE-IR) and AChE mRNA. Male, Long-Evans rats, maintained at 350 +/- 5 g, were dosed (s.c.) weekly for 4 weeks with 0, 15, 30, or 60 mg/kg chlorpyrifos in peanut oil. Brain tissue was harvested 1, 3, 5, 7 and 9 weeks after treatment began. AChE activity was measured by Ellman assay, and AChE-IR was estimated by two-site ELISA using monoclonal antibodies to rat brain AChE. While AChE activity fell significantly at all times and doses, AChE-IR increased at 3 and 5 weeks in the two higher dosage groups. Larger increases of AChE-IR were observed after chlorpyrifos was administered for 4 weeks by the oral route. Northern blots quantified with reference to cyclophilin were consistent with stable levels of AChE mRNA. Overall, it appears that chronically reduced brain AChE activity after chlorpyrifos reflects sustained enzyme inhibition, not loss of enzyme protein or suppression of AChE message.

Swedish civil servants, who reported considerable stress while working in larger government authorities, were randomized into either a work stress reduction intervention or a control group. Four work units constituted the intervention group (n = 94). and one unit formed the control group (n = 35). Participants in the two groups were of similar age and two thirds were women. Psychosocial, lifestyle, and physiological assessments were made before, during, and after the 8-month intervention period. A significant decrease in apolipoprotein B/apolipoprotein A I ratio (p < 0.05) occurred in the intervention group but not in the control group. No changes were seen in either group in smoking, eating. exercise, relative weight, or other lifestyle factors. which could explain the lipid profile changes. Stimulation from and autonomy over work increased in the intervention group (p < 0.01) hut remained the same in the control group. Results have implications for the efficacy of work stress interventions in decreasing coronary risk.

Systemically injected anti-acetylcholinesterase antibodies in rats cause selective lesions of preganglionic sympathetic neurons. Adult rats were examined up to four months after a single i.v. injection of murine monoclonal acetylcholinesterase antibodies or normal immunoglobulin G (1.5 mg). Within 4 h, antibody-treated rats developed ptosis, a sign of sympathetic dysfunction that was never reversed. Persistent pupillary constriction reflected preserved and unopposed parasympathetic function. Weight gain was depressed, but locomotor activity, excitability, and sensorimotor responses were normal, and gross neuromuscular performance was near normal. These findings were supported by biochemical evidence for selective sympathetic damage. Acetylcholinesterase activity was reduced for the whole period of observation in sympathetic ganglia and adrenal glands but fell only transiently in muscle and serum. At all times, choline acetyltransferase activity (a marker of presynaptic terminals) was unaffected in muscle but grossly depleted in ganglia. Light and electron microscopy showed that preganglionic sympathetic terminals of superior cervical ganglia were severely damaged while parasympathetic ganglia were less affected and motor endplates of skeletal muscle were apparently spared. Immunocytochemistry revealed punctate deposits of murine immunoglobulin G and complement component C3 in ganglionic neuropil 12 h after antibody injection. This finding was consistent with complement-mediated lysis of preganglionic terminals. Morphometric analysis of preganglionic neurons in the intermediolateral nucleus of the spinal cord showed progressive loss of cholinergic perikarya over several months. We conclude that antibody-induced destruction of ganglionic terminals leads to death of preganglionic sympathetic neurons and, hence, permanent dysautonomia.