In vitro characterization of recombinant factor VIII concentrates reveals significant differences in protein content, activity and thrombin activation profile.

Recombinant factor VIII (rFVIII) concentrates differ due to cell lines, culture conditions, presence of the B domain and authorized potency assays. This study characterizes three commercially available rFVIII concentrates: a second-generation full length (A), a third-generation full length (B) and a third-generation B domain-deleted (BDD) product (C). rFVIII concentrates were characterized for FVIII activity (FVIII:C) by one-stage clotting and chromogenic assays, FVIII antigen (FVIII:Ag), thrombin activation profile and FXa-generation assay. The rFVIII concentrates exhibited significant differences with regard to FVIII:C, FVIII:Ag and thrombin activation profile. Product A had significantly greater FVIII:C and FVIII:Ag relative to the measured values of products B and C. In addition, product A demonstrated faster and more complete activation by thrombin than the two others. BDD product C had the slowest measured thrombin activation rate. Product A exhibited a greater in vitro FXa generation than products B and C. We found no differences in FXa generation among all three products when FXa generation was normalized for FVIII:Ag. The greater FVIII:C and FVIII:Ag values for product A compared with that for products B and C are due to application of different authorized potency assays (one-stage assay for A vs. chromogenic assay for B and C). The variation in thrombin activation profiles may arise from differences in cell line-dependent posttranslational modifications of the various recombinant proteins.

Liver transplant from a donor with occult HEV infection induced chronic hepatitis and cirrhosis in the recipient.

Acute hepatitis E virus (HEV) infection is a self-limiting symptomatic or asymptomatic disease. However, as recently observed, it can manifest itself as chronic hepatitis in patients receiving solid organ transplants as well as in patients with HIV infection or severe hematologic disorders. Here, we describe the clinical course of a 73-year-old male patient in whom HEV transmission occurred after receiving a HEV-infected liver from a donor with occult HEV infection, whereby the patient had tested negative for HEV RNA and anti-HEV antibodies shortly before explantation. Anti-HEV IgG, IgM, and HEV RNA were detected in the first tested serum sample of the liver recipient obtained 150 days after liver transplantation and remained positive (earlier samples after OLT were not available). Liver cirrhosis developed within 15 months and the patient died of septic shock. Based on phylogenetic analyses of the donor and recipient's HEV strains, we were able to prove that the occult HEV infection was transmitted via the graft.

Building bridges between natural and social science disciplines: a standardized methodology to combine data on ecosystem quality trends.

Despite a growing interest in interdisciplinary research, systematic ways of how to integrate data from different disciplines are still scarce. We argue that successful resource management relies on two key data sources: natural science data, which represents ecosystem structure and processes, and social science data, which describes people's perceptions and understanding. Both are vital, mutually complementing information sources that can underpin the development of feasible and effective policies and management interventions. To harvest the added value of combined knowledge, a uniform scaling system is needed. In this paper, we propose a standardized methodology to connect and explore different types of quantitative data from the natural and social sciences reflecting temporal trends in ecosystem quality. We demonstrate this methodology with different types of data such as fisheries stocks and mangrove cover on the one hand and community's perceptions on the other. The example data are collected from three United Nations Educational Scientific and Cultural Organization (UNESCO) Biosphere reserves and one marine park in Southeast Asia. To easily identify patterns of convergence or divergence among the datasets, we propose heat maps using colour codes and icons for language- and education-independent understandability. Finally, we discuss the limitations as well as potential implications for resource management and the accompanying communication strategies. This article is part of the theme issue 'Nurturing resilient marine ecosystems'.

Randomized controlled clinical trial on the antiseptic efficacy of polihexanide 0.04% on acute traumatic wounds.

Prevention of wound infections is a challenge in clinical practice. The aim of this study was to assess the efficacy of polyhexamethylene biguanide (PHMB, polihexanide) 0.04% on acute traumatic wounds. It was a randomized, double-blind, placebo-controlled prospective trial which included 61 patients. The polihexanide group showed a significant decrease in log10 colony-forming units (cfu) (P < 0.001) after 60 min treatment in comparison to baseline cfu, whereas the Ringer solution group did not show a significant change in cfu during 60 min treatment. Treatment of polihexanide 0.04% resulted in a significant reduction of bacterial load in acute traumatic wounds.

Cytoplasmic overexpression of ALCAM is prognostic of disease progression in breast cancer.

BACKGROUND: Activated leucocyte cell adhesion molecule (ALCAM, CD166) is a cell surface member of the immunoglobulin superfamily. ALCAM expression has prognostic relevance in prostate and colon cancer. OBJECTIVE: To evaluate ALCAM protein expression in breast cancer by immunohistochemistry and to correlate expression levels with clinicopathological data. METHODS: 162 primary breast carcinomas with a mean clinical follow up time of 53 months were immunostained using a monoclonal ALCAM antibody. The staining was evaluated as an immunoreactive score (IRS) and grouped into low v high for both membranous and cytoplasmic staining. RESULTS: Intraductal and invasive carcinomas showed a higher ALCAM expression (median IRS 4 and 6 respectively) than normal breast tissue (IRS 2). In univariate survival analyses a significant association of high cytoplasmic ALCAM expression with shortened patient disease-free survival (mean (SD) five year non-progression rate, 69.4 (4.6)% v 49.4 (11.1)%, p = 0.0142) was found. In multivariate analyses of disease-free survival times, high cytoplasmic ALCAM expression (relative risk (RR) = 2.086, p = 0.026) and nodal status (RR = 2.246, p = 0.035) were significantly associated with earlier disease progression, whereas tumour grading (RR = 1.6, p = 0.052) was of borderline significance. CONCLUSIONS: The data suggest that strong cytoplasmic ALCAM expression in primary breast cancer, as detected by immunohistochemistry, might be a new marker for a more aggressive breast cancer biology.

Microplastics in personal care products: Exploring perceptions of environmentalists, beauticians and students.

Microplastics enter the environment as a result of larger plastic items breaking down ('secondary') and from particles originally manufactured at that size ('primary'). Personal care products are an important contributor of secondary microplastics (typically referred to as 'microbeads'), for example in toothpaste, facial scrubs and soaps. Consumers play an important role in influencing the demand for these products and therefore any associated environmental consequences. Hence we need to understand public perceptions in order to help reduce emissions of microplastics. This study explored awareness of plastic microbeads in personal care products in three groups: environmental activists, trainee beauticians and university students in South West England. Focus groups were run, where participants were shown the quantity of microbeads found in individual high-street personal care products. Qualitative analysis showed that while the environmentalists were originally aware of the issue, it lacked visibility and immediacy for the beauticians and students. Yet when shown the amount of plastic in a range of familiar everyday personal care products, all participants expressed considerable surprise and concern at the quantities and potential impact. Regardless of any perceived level of harm in the environment, the consensus was that their use was unnatural and unnecessary. This research could inform future communications with the public and industry as well as policy initiatives to phase out the use of microbeads.

Prevalence of adrenal and extra-adrenal Conn syndrome in hypertensive patients.

BACKGROUND: Primary aldosteronism (PA) is caused by an adrenal aldosterone-producing tumor (A-APT) or adrenal hyperplasia. An extra-adrenal APT (E-APT) as a cause of PA has been reported in 5 cases. Autopsy studies show a high incidence of ectopic adrenocortical tissue. We did a prospective study of the prevalence of A-APTs and E-APTs and the biochemical features of E-APTs in patients with PA. METHODS: Hypertensive patients (N = 3900) referred to our unit were screened for PA by measuring renin activity, urinary aldosterone-18-glucuronide, tetrahydroaldosterone, and 18-hydroxycorticosterone (18-OH-B). Primary aldosteronism was found in 257 cases. The differentiation between A-APTs and adrenal hyperplasia was based on the results of postural response of renin, plasma aldosterone, 18-OH-B, computed tomography, isotope scanning, or adrenal venous aldosterone. Ultrasound examination of the abdomen was used to screen for E-APT. RESULTS: The cause of PA was bilateral adrenal hyperplasia in 101 cases, unilateral adrenal hyperplasia in 2, an A-APT in 146, and an E-APT in 1. The site of aldosterone production was uncertain in 7 patients who had normal adrenal glands on computed tomography but refused to undergo isotopic scanning and adrenal venous catheterization. Ultrasound examination disclosed normal retroperitoneum in 4 of the 7 cases but could not rule out E-APT in 3 cases. The biochemical features of the patient with the E-APT were similar to classic A-APT, with low renin, high aldosterone, and high 18-OH-B values without appropriate response to posture or to short-term volume expansion. The excision of the E-APT in the right kidney resulted in normalization of blood pressure and renin, aldosterone, and 18-OH-B levels. CONCLUSION: Although E-APT is rare, it should be considered in the interests of specific therapy for PA because aldosterone-secreting malignant ovarian tumors also have been reported.

A new subset of mineralocorticoid hypertension with excess of 21-deoxyaldosterone and Kelly's-M1 steroid: clinical and morphological findings.

Ten cases of adrenal adenomas, one case with unilateral adrenal hyperplasia, and another case with apparent bilateral are reported, in whom an alternative pathway of aldosterone via 21-deoxyaldosterone is operative. They all manifested hypertension, low renin activity, low normal potassium values, as well as high urinary excretion rates of 21-deoxyaldosterone and its related metabolite Kelly's-M1 steroid. In all cases, urinary aldosterone metabolites (aldosterone-18-glucuronide and tetrahydroaldosterone) and aldosterone precursor 18-hydroxycorticosterone levels were normal. Hence, the adrenal lesions give rise to hyper-21-deoxyaldosteronism. 21-Deoxyaldosterone is a weak mineralocorticoid, and its elevated production in the presence of normal aldosterone can induce a pathological state of hypermineralocorticoidism. Adrenalectomy resulted in normalization of hypertension in six of eight and amelioration in two of eight cases. Six of seven adenoma cases examined as well as the case of unilateral adrenal hyperplasia were sensitive to ACTH. One of the seven adenomas and, as expected, the case with apparent bilateral hyperplasia were angiotensin responsive. Histologically and electron microscopically, the operated adenomas consisted predominantly of clear cells, characterized by mitochondria with tubulo-vesicular internal structure similar to those of the zona fasciculata (in contrast, our classical Conn's adenoma with normal 21-deoxyaldosterone excretion exhibited a more heterogenous histological appearance and were, in terms of ultrastructure, more similar to cells of the zona glomerulosa). Ultrastructurally and immunocytochemically, the clear cells of 21-deoxyaldosterone adenomas showed features of both the zona glomerulosa and the zona fasciculata and are, hence, considered to be hybrid cells. We conclude that the determination of 21-deoxyaldosterone and Kelly's-M1 should be considered in the diagnosis of mineralocorticoid-induced forms of hypertension, especially when an adrenal adenoma has been detected with an imaging procedure.

Expression analysis of genes at 3q26-q27 involved in frequent amplification in squamous cell lung carcinoma.

Gene amplifications are known to occur frequently in lung cancer. Recently, we identified gene amplifications at 3q26 in squamous cell lung carcinoma (SCC) using reverse chromosome painting. Here, our aim was to analyse the expression of genes which map within the amplified chromosomal region. The genes which were selected for their known function and their potential involvement in tumour development included the genes for ribosomal protein L22 (RPL22), butyrylcholinesterase (BCHE), glucose transporter 2 (SLC2A2), transferrin receptor (TFRC), thrombopoietin (THPO) and the phosphatidylinositol-3 kinase catalytic alpha polypeptide (PIK3CA). While five genes were expressed in the majority of the 17 samples of SCC, the gene for the glucose transporter 2 (SLC2A2) was expressed in only three cases, excluding SLC2A2 as the target gene of the amplification unit. For a subset of tumours, we determined the amplification status of the six genes. The TFRC, PIK3CA, BCHE, THPO and SLC2A2 genes were amplified in several cases, whereas the RPL22 gene was amplified in only one case. The combined amplification and expression data of this and our previous studies indicate that the amplified region at 3q26 contains several genes that are transcribed in SCC, providing the possibility that several amplified and functionally important genes at 3q26 may be involved in the pathogenesis of SCC.

DNA amplification on chromosome 7q in squamous cell carcinoma of the tongue.

Squamous cell carcinoma of the head and neck exhibit a highly variable picture of chromosomal aberrations. In the present study the clearly defined anatomical region of the tongue was analyzed for potentially specific patterns of chromosomal alterations. Fresh tumor samples from 18 patients afflicted by squamous cell carcinoma of the tongue constituted the clinical basis of the present investigation. The tumor samples were analyzed on the basis of comparative genomic hybridization (CGH), a molecular cytogenetic FISH-approach. Gains in DNA copy numbers were detected as the predominant imbalance on chromosomes 7q (9/18), 3q (48/18), 16p (7/18) and 20q (7/18). The regions of minimal overlap on these chromosomes were mapped to 7q11.2q11.3 and 3q26. A conspicuous finding was the frequent detection of amplifications in the 7q11 region. Gains in the 7q region have been rarely reported in CGH studies of tumors derived from different regions of the head and neck. Amplifications on 7q could thus be specifically linked with the tongue region and could correlate with specific clinical factors of this tumor entity.

Thin section arrays for I-FISH analysis of chromosome-specific imbalances in squamous cell carcinomas of the head and neck.

Thin section arrays of 20 head and neck squamous cell carcinomas were studied by I-FISH for gains (including amplification) and losses of specific genomic segments. These arrays allow the examination not only of a number of tumor sections but also of the surrounding margins and of inconspicuous control tissue in one experiment. All tumor sections examined significantly differed from the inconspicuous control tissues by containing more or less extensive cell populations with aberrant signal constitutions. In no case, however, did the aberrant population constitute the whole area of the section. Gains of signals were strikingly more frequent than were losses. All tumors showed significant gains of the segments examined, the highest differences between tumor and control sections were found for the segments 9q34 and 8q24, followed by 5p15.3 and 11q13. Amplifications were most frequently found of 11q13: 8 of the 20 tumors showed amplifications in more than 20% of the nuclei, while no nucleus with more than four signals was found in any of the control tissues (control: 0%). Amplifications of the target sequences on chromosomes 8 (14 tumors) and 9 (8 tumors) were observed in low but significant percentages of nuclei, no significant cell population was detected with an amplification of 5p15.3. Fourteen tumors exhibited a significant loss of 13q14, and only 8 tumors a significant loss at any other site. In the tumor margin sections, in most cases, the margins apparently were also affected by the one or the other of the genomic changes of the pertinent primary tumor. Nevertheless, there were, in some cases, also large differences depending on the way of analysis, but also on the specific signal constitution considered. Tumor stages T3 and T4 tended to have higher frequency of nuclei with gains of 5p15.3, 8q24, and 11q13 as compared to T2 tumors and less gains of 9q34 and loss of 13q14. With the exception of 8q24 and 13q14 alterations there was also a trend to higher percentages of aberrant nuclei in the margin of T3-4 tumors vs. T2 tumors.

Genetic imbalances in preinvasive tissue of hypopharynx provide evidence for cytogenetic heterogeneity.

Multiple chromosomal aberrations have been reported in head and neck squamous cell carcinoma (HNSCC). But less information is available on specific patterns of chromosomal amplifications which distinguish different areas of head and neck tumors. To elucidate genetic mechanisms causing the aggressive growth and high proliferation of hypopharyngeal squamous cell carcinoma (SCC), we performed reverse chromosome painting (RCP) on a total of eight hypopharyngeal SCC including invasive carcinoma and preinvasive tissue. Five hypopharyngeal invasive carcinomas showed amplifications on chromosome 3q. Furthermore, we detected gains on chromosomes 11q and 6p. Compared to the histologically classified preinvasive tissues, we found amplified alterations on chromosome 6p, 11q and 12q, but none of them showed gains on chromosome 3q. This observed heterogeneity in hypopharyngeal SCC might reflect a specific role of chromosome 3q as a late event in the highly invasive capacity of these SCC.

[MRI in testicular diagnosis: differentiation of seminoma, teratoma and inflammation using a statistical score]. / MRT in der Hodendiagnostik-Differenzierung von Seminomen, Teratomen und Entzündungen mit Hilfe eines statistischen Bewertungsmassstabs (Score).

PURPOSE: To differentiate scrotal pathology via MRI by means of a statistical score. METHODS: Between 1989 and 1995 MR images of 105 patients with scrotal pathology were rated retrospectively. In 69 cases linear discriminant analysis was used to differentiate seminoma, teratoma and inflammation. Six MRI attributes were found to be necessary and were weighted with a factor according to their importance. These factors were used to build a score. RESULTS: Criteria found to be important contained the distribution of the variate extensions of elements inside the pathological area and their maximal and minimal signal intensities. Furthermore, the contrast pattern of the pathological area and the visibility of healthy tissue in the pathological testicle was of importance. Seminoma was found to be homogeneous and well demarcated against healthy tissue. Teratoma was also well defined but characterised by inhomogeneous distribution of signal intensities. Inflammation showed diffuse signal increase of the pathological testicle, especially in T1-sequences. Using the score differentiation between tumors and inflammation succeeded in 94.2% between seminoma and teratoma in 89.7%. CONCLUSIONS: Compared to other studies using visual MR image analysis differentiation of scrotal diseases was improved by using a statistical score.

A second primary intestinal-type adenocarcinoma of the sinonasal tract induced by wood dust.

Intestinal type adenocarcinoma of the sinonasal tract is associated with exposure to wood dust. We report the case of an adenocarcinoma tumor of the left sinonasal area diagnosed in 1998. Nineteen years earlier (1979), an intestinal type adenocarcinoma of the right nasal cavity was diagnosed. The first tumor was treated in 1979 surgically followed by postoperative radiotherapy. The second tumor showed the same histological features as an intestinal type adenocarcinoma (papillary type). The patient had a history of a 10 year exposure to wood dust (furniture worker), and the latency time of this patient was 48 years in 1998. We suppose that the tumor of the left side is a second primary intestinal type adenocarcinoma. This case indicates the usefulness of a lifelong follow up of patients with adenocarcinoma of the nose because of the wide range of latency times of these tumors.

Cystadenoma of the parotid gland with unusual prominent lymphoid stroma.

A rare variant of the cystadenoma of salivary gland origin is presented, which occurred in the parotid of a 36-year-old, otherwise healthy female patient. The tumour showed a dense follicle-containing lymphoid stroma, resembling papillary cystadenoma lymphomatosum (Warthin's tumour). In contrast to that, the epithelial lining gave a more irregular impression and oncocytic metaplasia were completely absent. Light microscopic and immunohistochemical features of the tumour are described and compared with those of classical cystadenoma and papillary cystadenoma lymphomatosum (Warthin's tumour). Other relevant multicystic epithelial parotid lesions, which are commonly associated with a prominent lymphoid component are discussed.

Malignant odontogenic myxoma of the maxilla: case with cytogenetic confirmation.

An odontogenic myxoma of the maxilla with an aggressive clinical course is presented. The tumour arose in a 53-year-old patient, recurred two times after extended maxillectomy and ultimately caused the patient's death by uncontrollable local disease with infiltration of the cranial cavity. Microscopically, the tumour showed histological features of a low grade malignant myxosarcoma with cellular areas, enhanced mitotic activity and nuclear pleomorphism. Cytogenetic analysis revealed an unexpectedly aberrant hypertetraploid chromosome complement, that was considered as incompatible with the usual karyotypic patterns of benign tumours.

[Clinical anatomy of the interosseous arteries of the forearm]. / Klinische Anatomie der interossären Arterien des Unterarmes.

In recent years, several local flaps have been developed, based on the anterior and posterior interosseous arteries and their anastomoses at the wrist (e.g. posterior interosseous flap, pronator quadratus flap etc.). The anatomy of the arterial network supplying the flap is well established in both clinical and anatomical literature. Most authors agree in the constancy of the anastomoses between the interosseous arteries at the wrist and the absence of major anatomical variations excluding the use of the flap. In the present study, which is based on cadaver dissections of 60 preserved specimens, several gross vascular variations of the interosseous arteries have been found. The most frequent types were an additional anastomosis between the posterior interosseous artery and a perforating branch of the anterior interosseous artery in the middle third of the forearm, which was found in 20%. Ring-formations of the palmar and the dorsal branch of the anterior interosseous artery occurred in 5%. The point of perforation of the interosseous membrane by the dorsal branch of the anterior interosseous artery was found to vary in a larger extent, as described previously.

The views and knowledge of parents of children with autism spectrum disorder on a range of treatments

Background. Autism spectrum disorder (ASD) is a neurodevelopmental disorder that appears before the age of 3 years. Symptoms reflect delayed or abnormal social interaction and communication skills, with restricted or repetitive behaviour warranting the need for early intensive treatment.Methods. The aim of the study was to investigate the knowledge and views of parents regarding treatments for their children, aged between 5 and 9 years old with ASD, in eThekwini Metropolitan Municipality, South Africa. An embedded mixed methods research design was utilised. Nonrandom purposive sampling was used to select 46 parents of children with ASD. A 42-item questionnaire was used and the data were interpreted using descriptive statistics and thematic analysis.Results. More than half of the parents (53%) were unfamiliar with or had only heard of treatments in question, while 13.4% had a practical understanding of the treatments. Of all the treatments, parents rated their knowledge of speech-language therapy (SLT) most highly. The majority (68%) stated that they had difficulties accessing ASD treatment facilities and healthcare professionals, and perceived treatments as being costly. Even so, 74% of parents reported that they had a good relationship with their healthcare professional.Conclusion. The above findings should be viewed as motivation for health professionals to share information regarding the range of ASD treatments. They can assist parents in accessing appropriate facilities, recommend treatments that are supported by research, and update their knowledge on advances in ASD treatment

[Malignant neoplasms of the lacrimal sac: rarity among tear duct diseases]. / Malignome des Saccus lacrimalis : Seltenheit unter den Tränenwegserkrankungen.

BACKGROUND: Malignant neoplasms of the lacrimal sac are rare in the ophthalmic literature, for which there are only very limited diagnostic and therapeutic recommendations. We present five consecutive cases of our hospital from 2006 to 2009. METHOD: Retrospective analysis of all surgically treated patients with lacrimal duct diseases in the years 2006 to 2009, recording of malignant neoplasms and presentation of diagnostic and therapeutic approach. RESULTS: From January 2006 until October 2009 we performed 213 dacryocystorhinostomies at the Eye Clinic, Charité Campus Virchow Klinikum. In five patients intrasaccal malignancies were histologically proven by biopsy. None of the patients showed typical symptoms such as bloody epiphora. In two patients, a squamous cell carcinoma was seen, and one patient showed an adenocarcinoma. The other patients had a lymphoma and a malignant fibrous histiocytoma. The therapeutic approach consisted of surgical resection and radiotherapy. Systemic manifestations did not occur in any of the five patients. In the limited follow-up period no recurrences and no metastases were seen.