bluuebunny/crossref_metadata_2025 huggingface dataset:
Induction of apoptosis by synthetic ceramide analogues in the human keratinocyte cell line HaCaT
Authors: Bektas M. , Dullin Y. , Wieder T. , Kolter T. , Sandhoff K. , Brossmer R. , Ihrig P. , Orfanos C. E. , Geilen C. C.
Published: May 2007

Abstract: In contrast to extracellular, long chain ceramides which comprise a structural component of the epidermal water barrier, intracellular ceramides originating from sphingomyelin hydrolysis have been shown to inhibit proliferation and to induce apoptosis in different cell populations. To further elucidate the possible role of intracellular ceramides in human epidermis, two new cell‐permeable ceramide analogues, N‐thioacetylsphingosine (C2‐Cer=S) and 4‐dodecanoylamino‐decan‐5‐ol (FS‐5), were synthesized and tested for their ability to suppress cell growth and to induce apoptosis in immortalized human keratinocytes. It was shown that the well‐investigated ceramide analogue N‐acetylsphingosine (C2‐Cer=O), as well as the new compound C2‐Cer=S inhibited proliferation of HaCaT cells with half‐inhibitory concentrations (IC50) of 20 μg/ml and 10 μg/ml, respectively, whereas FS‐5 has been potent with an IC50>40 μg/ml. Overall, all three ceramide analogues induced apoptosis in HaCaT cells as assessed by DNA‐fragmentation using ELISA technique and in situ nick end labelling, thereby confirming the importance of ceramide signalling in keratinocytes.
Similarities of cutaneous and breats pathology in Cowden's Syndrome
Authors: Schreager Carolina A. , Schneider Douglas , Gruener Alexandra C. , Tsou Hui C. , Peacocke Monica
Published: May 2007

Abstract: Cowden's Syndrome (CS), or multiple hamartoma syndrome, is an autosomal dominant disorder characterized by mucocutaneous lesions, multiple benign tumors of internal organs and an increased risk of breast cancer. Here, we describe and illustrate in detail the benign breast pathology of 59 breast cases from 19 women with CS. Fibrosis is a significant characteristic of the breasts of patients with CS. Fibroaden‐ omas appear to hyalinize at an early age and are frequently complex. The specimens demonstrate a spectrum of dense hyalinization of both the lobule and the stroma, and hyaline nodules appear to be the most charac‐ teristic lesion. This hyalinization process shares striking similarities with keloids, as well as the sclerotic nodules seen in the skin of CS individuals. Ductal carcinoma in CS was common, and it appeared to be associated with stromal hyalinization. Other frequently found benign features are adenosis and cysts. Of interest, the features of the benign breast disease in CS show certain similarities with senescent lobules, fibrous mastopathy of diabetes mellitus, and mammary hamartomas. These observations provide a framework for pathologists to identify this underrecognized syndrome.
Retinoid regulation of heparin‐binding EGF‐like growth factor gene expression in human keratinocytes and skin*
Authors: Stoll S. W. , Elder J. T.
Published: May 2007

Abstract: Retinoic acid (RA) has profound effects on epidermal homeostasis; however, the molecular mechanisms by which retinoids regulate keratinocyte cell proliferation and differentiation are not well understood. Here we report that mRNA expression of heparin‐binding EGF‐like growth factor (HB‐EGF), a member of the EGF family of growth factors, is induced by RA in human keratinocytes and skin, and is overexpressed in the context of epidermal hyperplasia in vivo. Treatment of normal adult human keratinocytes with micromolar concentrations of RA significantly induced the expression of HB‐EGF. The response was efficiently blocked by specific inhibitors of ErbB tyrosine kinase activity, MAP kinase kinase (MEK), or p38 stress‐activated protein kinase. RA also enhanced the induction of HB‐EGF mRNA in human skin organ culture, an ex vivo model system displaying many similarities to wound healing in vivo. HB‐ EGF transcripts were markedly increased in human skin by topical treat‐ ment with RA under conditions known to provoke epidermal hyperplasia. HB‐EGF transcripts were also markedly overexpressed in the hyperplas‐ tic epidermis of psoriatic lesions, relative to normal skin. These results support the hypothesis that the effects of RA on epidermal hyperplasia are mediated at least in part by HB‐EGF, and suggest that signal transduction mechanisms other than or in addition to nuclear RA receptors contribute to this effect.
Mapping complex traits in diseases of the hair and skin
Authors: Aita V. M. , Christiano A. M. , Gilliam T. C.
Published: May 2007

The past decade has witnessed the ascendance of human genetics in modern medicine, and at the forefront of this movement is the identification of genetic factors underlying inherited diseases. The methods of genetic mapping and positional cloning have made the discovery of genes with alleles that cause simple Mendelian diseases commonplace. The elucidation of the genetic basis of such disorders has vitalized both human genetics and the entire medical community as the field has gained prominence. The fact remains, however, that diseases resulting from the action of alleles of a single gene comprise only a minor percentage of traits that are medically relevant to humanity. The majority of these are multifactorial “complex traits”, which result from the aggregate contribution of an unknown number of genes interacting with each other and with the environment. The current challenge has become one of parlaying successes in the mapping of Mendelian diseases into the discovery of genes whose alleles predispose the development of a complex disease. In light of this challenge, this review summarizes the methods and addresses some of the central issues of complex trait mapping, while using examples from dermatologically‐relevant complex traits such as psoriasis and alopecia. Additionally, current techinical and theoretical advances as well as the potential impact of the Human Genome Project will be discussed.
Detection of melanocortin‐1 receptor antigenicity on human skin cells in culture and in situ
Authors: Böhm M. , Metze D. , Schulte U. , Becher E. , Luger T. A. , Brzoska T.
Published: May 2007

The proopiomelanocortin (POMC) products α‐melanocyte stimulating hormone (α‐MSH) and adrenocorticotropin (ACTH) bind to specific receptors known as the melanocortin (MC) receptors. There is increasing evidence that the MC receptor subtype 1 (MC‐1R) is expressed in vitro by several other cutaneous cell types besides melanocytes and keratinocytes. Our knowledge on the MC‐1R expression in skin, however, remains fragmentary. In order to examine the expression of MC‐1R in human skin cells in vitro and in situ, we made use of a recently described antibody directed against the amino acids 2‐18 of the human MC‐1R. Flow cytometry analysis revealed the highest MC‐1R antigenicity in normal melanocytes and keratinocytes, followed by dermal fibroblasts, microvacualar endothelial cells and WM 35 melanoma cells. Little or no expression was detected in KB carcinoma cells and Fs4 fibroblasts. In normal human skin, immunoreactivity for the anti‐MC‐1R antibody was detected in hair follicle epithelia, sebocytes, secretory and ductal epithelia of weat glands, and periadnexal mesenchymal cells. Interfollicular epidermis was largely unreactive in adult skin as opposed to undifferentiated kertinocytes of fetal skin. Our findings form a framework within which MC‐1 receptor expression can be studied in various skin diseases.
Skin roughness and wrinkle formation induced by repeated application of squalenemonohydroperoxide to the hairless mouse
Authors: Chiba K. , Sone T. , Kawakami K. , Onoue M.
Published: May 2007

The present study examines the cumulative effects of sub‐erythema application of squalene‐monohydroperoxide (Sq‐OOH), the initial products of UV‐peroxidated squalene, to the skin of hairless mice. Sq‐OOH was isolated by the methanol extraction and preparative HPLC method. Repeated topical application of 10mM Sq‐OOH to hairless mice for 3 weeks induced definite skin roughness and crinkle formation. 3‐D surface parameter analysis revealed changes in all roughness parameters (number of furrows and crests, distance between a furrows and next crest, and irregularity) of the group treated with more than 3 mM Sq‐OOH compared to the control group. Theses skin surface changes were not induced by squalene, squalene‐monohydroxide (Sq‐OH) or organic hydroperxide such as tert‐butyl hydroperoxide and cumene ‐hydroperoxide at 10mM. Similarly, such changes were not induced by primary irritants, such as sodium lauryl sulfate and n‐tetradecane under the same experimental conditions. Skin conductance decreased, following application of 10 mM Sq‐OOH. Histological observation revealed that application of 10 mM Sq‐OOH induced slight hyperkeratosis, moderate epidermal thickening and slight hyperplasia of sebaceous glands.
Tumor necrosis factor‐alpha impairs contact hypersensitivity induction after ultraviolet B radiation via TNF‐receptor 2 (p75)
Authors: Kurimoto I. , Streilein J. W.
Published: May 2007

Acute, low dose ultraviolet B radiation (UVR) impairs induction of contact hypersensitivity (CH) in genetically susceptible mice. Polymorphic alleles at the TNF‐α locus dictate the susceptibility phenotype, and neutralizing anti‐TNF‐α antibodies restore CH induction in mice exposed to UVR. This circumstantial evidence strongly implicates TNF‐α in the pathogenesis of failed CH induction after UVR. Using mice genetically deficient in TNF‐receptor 1 (p55) or TNF‐receptor 2 (p75), we now report that the capacity of TNF‐α to impair CH induction after UVR required signaling via TNF‐receptor 2, rather than TNF‐receptor 1. Moreover, acting via the same receptor, TNF‐α altered the density and morphology of class II MHC‐bearing epidermal Langerhans cells. However, UVR retained its capacity to induce tolerance in both TNF‐receptor 1 and TNF‐receptor 2 deficient mice, indicating that TNF‐α plays no role in the systemic immune deficit created by UVR.
A recurrent frameshift mutation in exon 19 of the type VII collagen gene (COL7A1) in Mexican patients with recessive dystrophic epidermolysis bullosa
Authors: Mellerio J. E. , Salas‐Alanis J. C. , Amaya‐Guera M. , Tamez E. , Ashton G. H. S. , Mohammedi R. , Eady R. A. J. , McGrath J. A.
Published: May 2007

Abstract: Dystrophic epidermolysis bullosa (DEB) is an inherited blistering skin disorder caused by mutations in the type VII collagen gene (COL7A1). In this study, we determined the molecular basis of autosomal recessive DEB in a 19‐year‐old Hispanic‐Mexican woman by PCR amplification of genomic DNA, heteroduplex analysis, and automated sequencing of heteroduplex bandshifts. This approach revealed a homozygous frameshift mutation, 2470insG, in exon 19 of COL7A1 and resulted in attenuated basement membrane zone expression of type VII collagen, a reduced number of anchoring fibrils at the dermal‐epidermal junction, and a sub‐lamina densa level of blister formation. Clinically, the patient had widespread trauma‐induced skin fragility and complete loss of the nails, but had less pseudosyndactyly of the fingers and toes and milder mucosal involvement compared to most patients with the generalized form of this genodermatosis. We also screened 7 other Hispanic‐Mexican patients with recessive DEB, none of whom were known to be related to this individual, for the mutation 247OinsG using heteroduplex analysis and direct sequencing and detected this mutation on 7/14 alleles. Haplotype analysis using intragenic COL7A1 and flanking polymorphisms and microsatellite markers revealed that all the mutant alleles had arisen on similar allelic backgrounds, consistent with propagation of a common Hispanic‐Mexican ancestral haplotype. In view of the high allelic frequency of the mutation 247insG in the patients studies, we recommend initial screening for this mutation when attempting to identify the molecular pathology of recessive DEB in Hispanic‐Mexican patients.
Administration of acetylcholine and vasoactive intestinal polypeptide to atopic eczema patients
Authors: Rukwied R. , Heyer G.
Published: May 2007

Abstract: Responses to acetylcholine (ACh) and vasoactive intestinal polypeptide (VIP) were investigated in atopic eczema (AE) patients. To elucidate the involvement of histamine to ACh‐provoked vasoreactions and sensations, we applied a selective H1‐antagonist (cetirizine) 3 h prior to the ACh‐administration. Solutions of acetylcholine (ACh, 0.55 M) and vasoactive intestinal polypeptide (VIP, 1.5x10−5 M) were injected (10 μl) intracutaneously into the volar forearm of 14 healthy subjects and 14 atopic eczema (AE) patients. The substances were applied as single stimulus as well as in combination. Sensations evoked by the stimulation were recorded using 2 visual analog scales (VAS). Vasoreactions were analyzed with the new technique of computer assisted video image analysis. With this method we measured the dynamics of the flare development and the extension of the final flare size independent of the observer's assessment. In control subjects the development and extension of the observer's assessment. In control subjects the development and extension of the final flare size was almost similar, regardless whether ACh and VIP were applied in combination or separately. Compared to healthy controls, after injection of ACh, VIP and the combination of VIP and ACh smaller flare sizes were recorded in AE patients. After VIP was given, the control subjects reported pruritus, which was significantly augmented compared to AE patients. In contrast, controls reported a burning pain after the injection of ACh, whereas AE patients felt predominantly pruritus. Itch sensation after the combined application of VIP and ACh was significantly elevated in AE patients. Consequently, we assume that mediators of sudomotor neurons, i.e., VIP and ACh meet in AE patients apparently sensitized nociceptive primary afferents and induce exaggerated itch, pain and flare responses. When pretreated with the selective H1‐antagonist cetirizine before ACh was injected, pain and erythema due to ACh was diminished in healthy controls. In contrast, cetirizine did not influence the size of erythema and the magnitude of sensation in AE patients. We conclude, that the release of histamine is not involved in ACh‐induced erythema and pruritus in AE. These data provide evidence that pruritus can be elicited in atopic eczema by a cholinergic, histamine independent mechanism.
Human herpesvirus 8 (HHV‐8) in familial Kaposi's sarcoma
Authors: Kempf W. , Perniciaro C. , Adams V. , Mueller B. , Burg G.
Published: May 2007

Abstract: Human herpesvirus 8 (HHV‐8) has been detected in various epidemiological forms of Kaposi's sarcoma (KS). Since familial KS cases are exceedingly rare and the occurrence of familial KS in siblings has thought to depend rather on genetic factors than on a viral factor, familial KS has not been investigated for the presence of HHV‐8. To investigate whether HHV‐8 is present also in this rare form of KS, we examined tumor biopsies of 2 siblings with familial KS for the presence of HHV‐8 specific DNA sequences by a nested PCR protocol. HHV‐8 DNA sequences could be detected in KS specimens of both patients. Sequence analysis revealed an identical DNA sequence of HHV‐8 in KS tissue of both siblings, but the sequence in our cases differs in one base pair at position 67 from the previously published HHV‐8 KS330Bam fragment. The findings indicate that besides the yet poorly defined genetical factors involved in the pathogenesis of KS, HHV‐8 may act as a cofactor also in familial KS. In addition, our data demonstrate that HHV‐8 is found in all epidemiological forms of KS, including the rarely occuring familial KS. Familial KS may act as a further model to study the interaction of an oncogenic virus with genetic host factors in the context of a neoplastic disorder.
Epithelial stem cells in the skin: definition, markers, localization and functions
Authors: Cotsarelis G. , Kaur P. , Dhouailly D. , Hengge U. , Bickenbach J.
Published: May 2007

In recent years, cutaneous epithelial stem cells have attained a genuine celebrity status. They are considered the key resource for epidermal and skin appendage regeneration, and are proposed as a preferential target of cutaneous gene therapy. Follicular epithelial stem cells may also give rise to a large variety of epithelial tumors, and cutaneous epithelial stem cells likely are crucial targets for physical or chemical agents (including carcinogens) that damage the skin and its appendages. However, as this Controversies feature illustrates, few experts can agree on how exactly to define and identify these elusive cells, or on where precisely in the skin they are localized. Given their potential importance in skin biology, pathology and future dermatological therapy, it is, therefore, timely to carefully reconsider the basic questions: What exactly is a stem cell, and how can we reliably identify epithelial stem cells? How many different kinds are there, and how do they differ functionally? Where exactly in the skin epithelium is each of the putative stem cell subpopulations located, and can we selectively manipulate any of them?
Identification of a de novo glycine substitution in the type VII collagen gene in a proband with mild dystrophic epidermolysis bullosa
Authors: Cserhalmi‐Friedman P. B. , Grossman J. , Karpati S. , Ahmad W. , Horvath A. , Christiano A. M.
Published: May 2007

Abstract: The dystrophic forms of epidermolysis bullosa result from different types and combinations of mutations in the type VII collagen gene (COL7A1). We describe a novel glycine substitution arising as a de novo mutation in a proband with a clinically mild form of dystrophic epidermolysis bullosa and no family history of any blistering disease. This report underscores the predominance of glycine substitutions in the dominantly inherited forms of dystrophic form epidermolysis bullosa, and heightens our awareness of unusual modes of inheritance. This information is critical for accurate genetic counseling and determination of recurrence risk in families with dystrophic EB.
Squamous cell carcinoma in a family with dominant dystrophic epidermolysis bullosa: a molecular genetic study
Authors: Christiano A. M. , Crollick J. , Pincus S. , Uitto J.
Published: May 2007

Abstract: Squamous cell carcinoma (SCC) is a frequent complication in the severe, recessively inherited forms of dystrophic epidermolysis bullosa (RDEB), however, only rarely reported in dominant DEB. Although the SCCs in RDEB are frequently well‐differentiated by histopathology, they often have a poor prognosis due to multicentricity, rapid invasiveness, and development of distant metastases. In this study, we sought to determine the molecular basis of DDEB in a family with the unusual occurrence of SCCs. Specifically, a large DDEB family with 2 individuals being affected with SCC was analyzed for potential mutations in the type VII collagen gene (COL7A1) by heteroduplex scanning and direct nucleotide sequencing of PCR amplified segments of the gene. This mutation detection strategy disclosed a G A transition at nucleotide position 6,235 which resulted in substitution of a glycine by arginine within the collagenous region of COL7A1. This study establishes, for the first time, the molecular basis in a family with DDEB/SCC. Clinically, this study reemphasizes the importance of vigilance in surveying DEB patients, not only those with recessive but also with dominant inheritance, for SCC.
Expression of the 72‐kD heat shock protein is induced by ultraviolet A radiation in a human fibrosarcoma cell line
Authors: Trautinger F. , Kokesch C. , Klosner G. , Knobler R. M. , Kindas‐Mügge I.
Published: May 2007

Abstract: The 72‐kD heat shock protein (hsp72) belongs to a family of stress inducible proteins (heat shock proteins, hsp) and its expression is associated with increased survival of cells in culture following exposure to ultraviolet radiation (UV). Hsp72 can be induced by a number of stresses, including heat, cold, and toxic chemicals. The purpose of this study was to evaluate whether UV is able to activate transcription of hsp72. The human fibrosarcoma cell line HT1080 was used for these experiments because hsp72 is not detectable in these cells under normal culture conditions. Cells were exposed to UVA and UVB using a solar simulating source and hsp72 was determined in whole cell extracts by immunoblotting. For inhibition of mRNA and protein synthesis cordycepin (20 μg/ml) and cycloheximide (10 μg/ml) were added to the cultures, respectively. UVA‐induced lipid peroxidation was inhibited by α‐tocopherol and butylated hydroxytoluene (BHT). UVA but not UVB induced hsp72 with maximal expression at 40 J/cm2, 8–12 h after exposure. Induction was blocked by cordycepin as well as by cycloheximide indicating that both, mRNA and protein synthesis, are required for UVA‐induction of hsp72. Inhibition of cell lipid peroxidation with α‐tocopherol and BHT had no effect on hsp72 expression. These results suggest that induction of hsp72 is part of an adaptive response mechanism in human cells to UV‐related stress.
Alterations in ganglioside expression during the differentiation of human mast cells
Authors: Zuberbier T. , Guhl S. , Hantke T. , Hantke C. , Welker P. , Grabbe J. , Henz B. M.
Published: May 2007

Abstract: Gangliosides are physiological components of the outer cell membrane. In the present study, the role of ganglioside expression during differentiation of human mast cells was evaluated. After 11 days of culture in medium known to induce mast cell differentiation, 70% of peripheral blood mononuclear cells (PBMC) showed positive staining for the high affinity IgE receptor and tryptase on immunocytochemistry and an associated 20‐fold increase of ganglioside GM3 expression. Furthermore, exogenous addition of GM3 during cultivation of PBMC in medium containing low levels of growth factors induced an increase of mast cell specific tryptase. The association of ganglioside expression with mast cell differentiation was confirmed by experiments with the human mast cell line HMC‐1. FcɛRI‐positive cultured cells enriched with immunobeads exhibited a 3‐fold higher expression of GM3, compared to FcɛRI negative HMC‐1 cells. Furthermore, measurable amounts of the gangliosides GM2, GM1 and GD1a were found only in the FcɛRI positive cells. A corresponding transient increase of mRNA for GalNAcT, the key enzyme in the production of these latter gangliosides, could be detected preceding the expression of these gangliosides and the FcɛRI by RT‐PCR. Taken together, these data point to a functional role of gangliosides in the differentiation of human mast cells.
How realistic is cutaneous gene therapy?
Authors: Hengge U. R. , Taichman L. B. , Kaur P. , Rogers G. , Jensen T. G. , Goldsmith L. A. , Rees J. L. , Christiano A. M.
Published: May 2007

Recent progress with innovative, experimental gene therapy approaches in animals, and recent improvements in our understanding and manipulation of stem cells, gene expression and gene delivery systems, have raised plenty of hopes in essentially all branches of clinical medicine that hitherto untreatable or poorly manageable diseases will soon become amenable to treatment. Few other organ systems have received such enthusiastic reviews in recent years as to the chances and prospects of gene therapy as the skin, with its excellent accessibility and its pools of – seemingly – readily manipulated epithelial stem cells (cf. Cotsarelis et al., Exp Dermatol 1999: 8: 80–88).

However, as in other sectors of clinical medicine, the actual implementation of general gene therapy strategies in clinical practice has been faced with a range of serious difficulties (cf. Smith, Lancet 1999: 354 (suppl 1): 1–4; Lattime & Gerson (eds.), Gene Therapy of Cancer, Academic Press, San Diego, 1999). Thus, it is critically important to carefully distinguish unfounded hype from justified hope in this embryonal area of dermatologic therapy, to discuss in detail what can be realistically expected from cutaneous gene therapy approaches in the next few years, and importantly, what kind of promises should not be made to our patients at this time.
Anesthesia and intraoperative support of homeostasis in fast‐track rehabilitation for elective colonic surgery
Authors: HENSEL MARIO , STRACKE S. , KRIMPHOVE M. , BLOCH A. , VON HEYMANN CHRISTIAN , SPIES CLAUDIA
Published: May 2007

This review discusses the fast‐track rehabilitation protocol in use at the Charité University Hospital for patients undergoing elective colonic resection, as well as underlying mechanisms and clinical pathways. In particular, it examines the anesthesiological aspects of this multimodal approach, such as revised guidelines for preoperative fasting, modified pre‐ and postoperative fluid management and effective analgetic therapy, including epidural analgesia, short‐acting anesthetic agents and the avoidance of high doses of systemic opioids. Maintenance of normothermia and normovolemia is also discussed.
Postoperative care in fast‐track rehabilitation for elective colonic surgery
Authors: JUNGER MARC , SCHOENBERG MICHAEL HERMANN
Published: May 2007

The success of a ‘fast‐track’ rehabilitation program in colonic surgery depends on multiple factors. The major factor post surgery is the prevention of postoperative ileus (POI), resulting in a faster recovery and a shorter hospital stay. All fast‐track programs led to an accelerated intake of oral fluids and solid foods and normalization of intestinal function, thus reducing both the primary and overall hospital stay. Fortunately, the postoperative treatment was not accompanied by an increase in morbidity or mortality. Nevertheless, fast‐track surgery may mean a higher rate of hospital readmissions in the first 2 weeks after discharge. Besides the prevention of POI and postoperative nausea and vomiting through restricted intraoperative fluid management, early oral food intake, rigorous pain control and early mobilization, education – on the part of patient, nurses and other medical staff – is equally as important. Ensuring that the patient takes an active rather than a passive role in the recovery process empowers the patient and avoids needless suffering. The patient is thus surrounded by encouraging staff members who motivate him or her not as a patient but as a ‘partner’ in a precarious situation. Early oral food intake and early mobilization suggest to the patient (and, possibly, to the nursing staff) that the surgical procedure was less invasive than believed and that the postoperative phase need not be eventful or traumatic. In contrast to the conventional treatment with infusions, the typical postoperative diet consisting of tea and soup for days, the fast‐track protocol gives the patient the feeling that he or she is actually in good health and about to leave the hospital. All optimistic signals received during the postoperative phase undoubtedly have a very suggestive and positive influence psychologically on the patient. It is therefore reasonable to assume that the psychological effect of fast‐track rehabilitation, although not assessed in clinical trials, is very important and influences the recovery substantially. Good clinical practice should be based on an ongoing audit of the outcome. In order to ensure the success of a fast‐track rehabilitation program in surgery, it is essential to assess the outcome in terms of morbidity and mortality. Moreover, such an ambitious protocol has to take into account new clinical trial findings in order to introduce improvements to the postoperative phase. Besides an understanding of the pathophysiology and new therapeutic approaches to prevent POI and improve pain control, such measures include the development of an intradepartmental infrastructure and staff education initiatives.
Results of fast‐track rehabilitation in elective colonic surgery
Authors: JUNGHANS TIDO , JUNGHANS ELENA , SCHWENK WOLFGANG
Published: May 2007

Single centers have reported that the implementation of multimodal ‘fast‐track’ rehabilitation concepts improves postoperative convalescence after elective colonic surgery. A systematic review of the literature, including a hand search, for the keywords fast‐track and colo was performed. Fifteen (15) of the 27 submitted manuscripts offered low levels of evidence. Only two manuscripts compared a fast‐track group with a randomized control group. There was no evidence that surgical complications or lethality was increased with fast‐track rehabilitation. Patients received early enteral feeding, were mobilized and discharged 2–5 days after surgery. The maximum readmission rate was 20%. The advantages of early discharge need to be confirmed in high‐volume centers and additional controlled studies performed.
‘Fast‐track’ protocols in colorectal surgery
Authors: SENAGORE ANTHONY J.
Published: May 2007

A greater appreciation of newer concepts in healthcare process management, improved understanding of postoperative physiology, and economic pressures have combined to produce safe reductions of hospital stays after major gastrointestinal surgery. Preoperative and postoperative education is a key component of successful fast‐track care pathways. Effective treatment of pain, nausea and vomiting, ileus, and fatigue combined with minimization of mechanical factors all decrease stress‐induced organ dysfunction. An important factor leading to enhanced recovery is early ambulation, which maintains muscle strength, limits disruption of orthostatic reflexes, and plays a role in reducing ileus and pulmonary complications. A growing body of evidence supports the contention that laparoscopic colectomy may allow optimization of fast‐track care of the colectomy patients. Epidural anesthesia/analgesia appears to be an important component of the perioperative care plan in many fast‐track programs. Evaluation of an enhanced care plan called the ‘Controlled Rehabilitation and Early Ambulation and Diet’ showed that patients who were suitable for the program at the time of surgery stayed in hospital for 4.3 ± 1.6 days postoperatively. The presence of a defunctioning loop ileostomy, or a history of previous abdominal surgery did not affect time to discharge. Time to discharge from hospital was significantly delayed in patients who showed poor compliance with the protocol. The process of care was effective in patients undergoing colonic, rectal and reoperative surgery, not just the ‘optimal’ colectomy patient.
Application of Different Methods for the Diagnosis of Experimental Paratuberculosis in Goats
Authors: Munjal S. K. , Tripathi B. N. , Paliwal O. P. , Boehmer J. , Homuth M.
Published: May 2007

The diagnosis of subclinical paratuberculosis is still considered a major problem worldwide. As part of investigating diagnostic strategies for the paratuberculosis infection, sequential results of various diagnostic methods in a progressive experimental infection in goats were evaluated. Twenty‐three goat kids were divided into three groups: the infected, contact and control, comprising 10, five and eight goats respectively. Animals of the infected group were orally inoculated on seven occasions with 5 ml of inoculum containing 2 × 109Mycobacterium aviumssp.paratuberculosisper ml. Lymphoycte proliferation test using johnin PPD detected paratuberculosis infection from 60 days post‐infection (DPI) onwards. The johnin PPD was found to be a better antigen for the proliferative assays as compared with the sonicated antigen. The faecal smear examination with acid‐fast staining detected more goats as positive than bacterial culture and polymerase chain reaction (PCR). Lipoarabinomannan enzyme‐linked immunosorbent assay (ELISA) started detecting infected goats from 150 DPI onwards followed by indirect ELISA and agar gel immunodiffusion from 180 DPI onwards. Histological examination was confirmatory and detected five infected goats as positive.
Antibacterial Activity of Bovine Lactoferrin Hydrolysate against Mastitis Pathogens and Its Effect on Superoxide Production of Bovine Neutrophils
Authors: Kawai K. , Shimazaki K. , Higuchi H. , Nagahata H.
Published: May 2007

Antibacterial activity of bovine lactoferrin hydrolysates (LFH) on microorganisms isolated from bovine mastitis, and superoxide () production of bovine neutrophils were evaluated. Antibacterial effects of LFH were measuredin vitroagainstStaphylococcus aureus, coagulase‐negative staphylococci,Streptococci,Enterococci,Escherichia coli,Klebsiella pneumoniae, yeast‐like fungi andPrototheca zopfiiisolated from clinical cases of bovine mastitis. To compare susceptibilities against LFH, minimal inhibitory concentration (MIC) values were determined by a micro‐plate assay method. Most organisms were sensitive to LFH.Prototheca zopfiiwas highly sensitive to LFH; the growth of the microorganism was inhibited completely even at 1 μg/ml.Staphylococcus aureusandEscherichia coliwere resistant to LFH. The production ofby bovine neutrophils was used to evaluate the effect of LFH administration on functional activity. Increase inproduction by bovine neutrophils occurred upon addition of LFH to neutrophils. These results demonstrate that LFH possesses antibacterial activity against pathogens that cause mastitis and activates neutrophil superoxide production.
Comparison of a Commercial ELISA with the Modified Agglutination Test for the Detection ofToxoplasma gondiiInfection in Naturally Exposed Sheep
Authors: Klun I. , Djurković‐Djaković O. , Thulliez P.
Published: May 2007

Surprisingly few commercial ELISAs are available for the detection ofToxoplasma gondiiinfection in animals, and none for use in sheep have been evaluated. We thus compared the Bommeli Diagnostics ELISA Toxotest for the detection ofT. gondiiantibodies in ruminants with the reference modified agglutination test (MAT) in a series of 180 sheep sera. ELISA results were analysed at two cut‐off levels (30%, comprising both weakly positive and positive results, and 100%, comprising only positive results), and compared with MAT at three cut‐off levels (titre of 1 : 25, 1 : 50 and 1 : 100). The results showed a moderate agreement of ELISA at both cut‐offs (κ = 0.46 and 0.51) with MAT at a cut‐off titre of 1 : 100. However, the specificity and positive predictive value were above 95% only at an ELISA cut‐off of 100%, indicating its potential as a diagnostic test, particularly in areas with a high prevalence of infection. On the other hand, lower sensitivity and negative predictive value limit its value as a screening test. Thus, the ELISA Toxotest may be used for quick diagnosis ofT. gondiiinfection in sheep in the field, i.e. for the differential diagnosis of ovine abortion storms.
Seroprevalence of Antibodies toNeospora caninumin Urban and Rural Dogs in north‐west Italy
Authors: Ferroglio E. , Pasino M. , Ronco F. , Benà A. , Trisciuoglio A.
Published: May 2007

Sera were collected from 490 dogs from north‐west Italy. One hundred and eighty‐eight dogs were urban, while 302 dogs were rural. Among the latter, 190 were shepherd dogs and 112 were cattle farm dogs. Sera were tested for the presence of antibodies againstNeospora caninumusing theNeosporaagglutination test. Seroprevalence at 1/40, 1/80, 1/160 dilution titres was significantly higher in rural (36.4%, 19.5%, 9.9% respectively) than in urban dogs (20.2%, 10.6%, 4.8% respectively). Seroprevalence did not differ significantly in males and females. In shepherd dogs, prevalence increased according to dogs’ age, thus suggesting a post‐natal exposure by horizontal transmission. The observed higher seroprevalence in rural dogs suggests the importance of lifestyle and alimentary habits (i.e. aborted foetuses, placentas and small mammals) in the acquisition ofN. caninuminfection. Our results confirm that dogs are exposed toN. caninumand play an important role in the epidemiology ofN. caninum.
First Diagnosis of Encephalitozoon intestinalis and E. Hellem in a European Brown Hare (Lepus europaeus) with Kidney Lesions
Authors: De Bosschere H. , Wang Z. , Orlandi P. A.
Published: May 2007

Encephalitozoon intestinalis and Encephalitozoon hellem were diagnosed in the kidneys of a free – ranging European brown hare (Lepus europaeus) with multifocal wedge‐shaped chronic interstitial nephritis using real‐time PCR and microarray. This is the first description of these microsporidia species in a European brown hare, which are both potential zoonotic agents.
Comparison of Duplex PCR and Microscopic Techniques for the Identification of Babesia bigemina and Babesia bovis in Engorged Female Ticks of Boophilus microplus
Authors: Quintão‐Silva M. G. , Melo M. N. , Ribeiro M. F. B.
Published: May 2007

A single‐step duplex polymerase chain reaction (PCR) technique and traditional microscopic examination of haemolymph smears were used to detect Babesia bigemina and/or Babesia bovis infection in engorged female ticks of Boophilus microplus recovered from calves raised in an endemic area of the State of Minas Gerais, Brazil. In the PCR amplification of tick‐derived DNA, pairs of oligonucleotide primers specific for a 278‐bp sequence from B. bigemina and for a 350‐bp sequence from B. bovis were used conjointly. The microscopic examination of haemolymph revealed that 16.7% of the engorged ticks were infected with Babesia spp., although no significant differences (ρ > 0.05) were found in the infection rate of ticks collected from calves of different age groups. PCR analysis showed that 77.8% of the engorged ticks whose haemolymph contained sporokinetes were infected with B. bigemina, 7.8% with B. bovis and 14.4% with both protozoan species. However, the PCR assay further revealed that, amongst the engorged female ticks whose haemolymph was apparently negative for the presence of sporokinetes, 15.6% were infected with B. bigemina, 2.2% with B. bovis and 10.0% with both species. The duplex PCR method is thus more efficient and sensitive than the microscopic assay and also permits facile identification of the protozoa species present in engorged female ticks.
Immune Response to and Faecal Shedding of Mycobacterium avium ssp. paratuberculosis in Young Dairy Calves, and the Association Between Test Results in the Calves and the Infection Status of their Dams
Authors: Antognoli M. C. , Hirst H. L. , Garry F. B. , Salman M. D.
Published: May 2007

The objectives of this study were to: (i) evaluate the results of an intradermal skin test, a modified IFN‐γ test, and a commercial ELISA in commercially raised dairy calves at 2, 4, 6 and 8 months of age relative to faecal shedding of Mycobacterium avium ssp. paratuberculosis (MAP); (ii) determine the proportion of 8‐month‐old calves shedding MAP in faeces as detected by culture and One Tube Nested Polymerase Chain Reaction (OTSN‐PCR) and (iii) explore the association between results of tests described above in the calves and the Paratuberculosis (PTB) status of their dams as determined by faecal culture and/or serology. The study calves belonged to two dairy herds with different risk of exposure to MAP (high and low) and were enrolled based on their dam’s ELISA results prior to calving. Approximately 3% of the calves were shedding MAP in faeces at 8 months of age. No agreement was observed among the evaluated immunity‐based tests or between the immunity‐based tests and the detection of MAP in faeces. Although no association was observed between the infection status of the dam and the results from the IFN‐γ and skin tests on the calves, there is an indication that calves born from dams that were faecal shedders might be at a higher risk of testing positive to the IFN‐γ test at 8 months of age. The disagreement among all tests evaluated in this calf cohort suggests that the detection of MAP infection in young stock requires the use of combined multiple tests. The early detection of PTB in calves is a challenge that requires further exploration of new methods to confirm infection status. These new testing methods should be both affordable and compatible with regular husbandry practices.
Modification of fetal plasma amino acid composition by placental amino acid exchangers in vitro
Authors: Cleal Jane K. , Brownbill Paul , Godfrey Keith M. , Jackson John M. , Jackson Alan A. , Sibley Colin P. , Hanson Mark A. , Lewis Rohan M.
Published: May 2007

Fetal growth is dependent on both the quantity and relative composition of amino acids delivered to the fetal circulation, and impaired placental amino acid supply is associated with restricted fetal growth. Amino acid exchangers can alter the composition, but not the quantity, of amino acids in the intra‐ and extracellular amino acid pools. In the placenta, exchangers may be important determinants of the amino acid composition in the fetal circulation. This study investigates the substrate specificity of exchange between the placenta and the feto‐placental circulation. Maternal–fetal transfer of radiolabelled amino acids and creatinine were measured in the isolated perfused human placental cotyledon. Transfer of l‐[14C]serine or l‐[14C]leucine, and [3H]glycine, were measured in the absence of amino acids in the fetal circulation (transfer by non‐exchange mechanisms) and following 10–20 μmol boluses of unlabelled amino acids into the fetal circulation to provide substrates for exchange (transfer by exchange and non‐exchange mechanisms). The ability of fetal arterial boluses of l‐alanine and l‐leucine to stimulate release of amino acids from the placenta was also determined using HPLC in order to demonstrate the overall pattern of amino acid release. Experiments with radiolabelled amino acids demonstrated increased maternal–fetal transfer of l‐serine and l‐leucine, but not glycine, following boluses of specific amino acids into the fetal circulation. l‐[14C]Leucine, but not l‐[14C]serine or [3H]glycine, was transferred from the maternal to the fetal circulation by non‐exchange mechanisms also (P < 0.01). HPLC analysis demonstrated that fetal amino acid boluses stimulated increased transport of a range of different amino acids by 4–7 μmol l−1 (P < 0.05). Amino acid exchange provides a mechanism to supply the fetus with amino acids that it requires for fetal growth. This study demonstrates that these transporters have the capacity to exchange micromolar amounts of specific amino acids, and suggests that they play an important role in regulating fetal plasma amino acid composition.
Mechanisms of skeletal muscle injury and repair revealed by gene expression studies in mouse models
Authors: Warren Gordon L. , Summan Mukesh , Gao Xin , Chapman Rebecca , Hulderman Tracy , Simeonova Petia P.
Published: May 2007

Common acute injuries to skeletal muscle can lead to significant pain and disability. The current therapeutic approaches for treating muscle injuries are dependent on the clinical severity but not on the type of injury. In the present studies, the pathophysiology and molecular pathways associated with two different types of skeletal muscle injury, one induced by direct destruction of muscle tissue (i.e. FI) and the other induced by a contractile overload (more specifically high‐force eccentric contractions, i.e. CI) were compared side by side. Histopathological evaluation and measurements of muscle strength were accompanied by analyses of expression for 12 488 known genes at four time points ranging from 6 h to 7 days after injury. Real‐time RT‐PCR was used to confirm some of the injury type differences in the temporal profiles of gene expression. Our data revealed several pools of genes, including early induction of transcription, myogenic and stress‐responsive factors, common for both types of injury as well as pools of genes expressed specifically with one of the injury types. Only CI activated a set of genes associated with the repair of impaired proteins and structures including genes related to apoptosis, whereas FI uniquely activated gene sets involved in extensive inflammatory responses, tissue remodelling, angiogenesis and myofibre/extracellular matrix synthesis. In conclusion, knowledge of the sets of genes associated specifically with the nature of the injury may have application for development of new strategies for acceleration of the recovery process in injured skeletal muscle.
Adenosine modulates transmission at the hippocampal mossy fibre synapse via direct inhibition of presynaptic calcium channels
Authors: Gundlfinger A. , Bischofberger J. , Johenning F. W. , Torvinen M. , Schmitz D. , Breustedt J.
Published: May 2007

The modulation of synaptic transmission by presynaptic ionotropic and metabotropic receptors is an important means to control and dynamically adjust synaptic strength. Even though synaptic transmission and plasticity at the hippocampal mossy fibre synapse are tightly controlled by presynaptic receptors, little is known about the downstream signalling mechanisms and targets of the different receptor systems. In the present study, we identified the cellular signalling cascade by which adenosine modulates mossy fibre synaptic transmission. By means of electrophysiological and optical recording techniques, we found that adenosine activates presynaptic A1 receptors and reduces Ca2+ influx into mossy fibre terminals. Ca2+ currents are directly modulated via a membrane‐delimited pathway and the reduction of presynaptic Ca2+ influx can explain the inhibition of synaptic transmission. Specifically, we found that adenosine modulates both P/Q‐ and N‐type presynaptic voltage‐dependent Ca2+ channels and thereby controls transmitter release at the mossy fibre synapse.
The role of the actin cytoskeleton in oxytocin and vasopressin release from rat supraoptic nucleus neurons
Authors: Tobin Vicky A. , Ludwig Mike
Published: May 2007

Magnocellular neurons of the supraoptic nucleus (SON) can differentially control peptide release from the somato/dendritic and axon terminal compartment. Dendritic release can be selectively regulated through activation of intracellular calcium stores by calcium mobilizers such as thapsigargin (TG), resulting in preparation (priming) of somato/dendritic peptide pools for subsequent activity‐dependent release. As dynamic modulation of the actin cytoskeleton is implicated in secretion from synaptic terminals and from several types of neuroendocrine cells, we studied its involvement in oxytocin and vasopressin release from SON neurons. Confocal image analysis of the somata revealed that the normally continuous cortical band of F‐actin is disrupted after high potassium (K+, 50 mm) or TG (200 nm) stimulation. The functional importance of actin remodelling was studied using cell‐permeable actin polymerizing (jasplakinolide, 2 μm) or depolymerizing agents (latrunculin B, 5 μm) to treat SON and neural lobe (NL) explants in vitro and measure high K+‐induced oxytocin and vasopressin release. Latrunculin significantly enhanced, and jasplakinolide inhibited, high‐K+‐evoked somato/dendritic peptide release, while release from axon terminals was not altered, suggesting that high‐K+‐evoked release in the SON, but not the NL, requires depolymerization of the actin cytoskeleton. TG‐induced priming of somato/dendritic release was also blocked by jasplakinolide and latrunculin, suggesting that priming involves changes in actin remodelling.