Splenectomy modulates early immuno-inflammatory responses to trauma-hemorrhage and protects mice against secondary sepsis.

In polytrauma patients, the impact of splenectomy is equivocal, ranging from negative to protective. We investigated the impact of splenectomy on immune responses in the 1st-hit polytrauma alone and on survival in the post-traumatic sepsis (2nd hit). Female BALB/c mice underwent polytrauma (1st hit) consisting of either a) TH: femur fracture, hemorrhagic shock or b) TSH: splenectomy, femur fracture, hemorrhagic shock. Additionally, the polytrauma hit was followed by cecal ligation and puncture (CLP) 48 h later and compared to CLP alone. Splenectomy improved the 28-day survival in secondary sepsis to 92% (from 62%), while TH lowered it to 46% (p < 0.05). The improved survival was concurrent with lower release of inflammatory cytokines (IL-6, CXCL-1, MCP-1) and increase of C5a post-CLP. In the polytrauma hit alone, TSH induced stronger neutrophilia (1.9 fold) and lymphocytosis (1.7 fold) when compared to TH mice. Moreover, TSH resulted in a 41% rise of regulatory T-cells and reduced the median fluorescence intensity of MHC-2 on monocytes by 55% within 48 h (p < 0.05). Conversely, leukocyte phagocytic capacity was significantly increased by 4-fold after TSH despite a similar M1/M2 macrophage profile in both groups. Summarizing, splenectomy provoked both immuno-suppressive and immuno-stimulatory responses but was life-saving in secondary sepsis. Additionally, the polytrauma components in 2-hit models should be tested for their effects on outcome; the presumed end-effect of the 1st hit solely based on the common immuno-inflammatory parameters could be misleading.

Evaluation of small-group education on the shade determination ability of preclinical dental students-A controlled clinical trial.

OBJECTIVES: To evaluate the effect of small-group education including practical training on dental students' shade determination performance compared with a control group. METHODS: Sixty-three non-colour-blind preclinical students were asked to participate and belonged to an intervention (n = 31) and a control (n = 32) group, respectively. All students received a lecture on dental shade determination. The members of the intervention group were educated and practically trained in small groups in addition. Ten volunteers, each with unrestored upper right middle and lateral incisors, were recruited. The students of both the intervention and control groups were then asked to determine the colour of the volunteers' test teeth using the 3D-Master shade guide (Vita Zahnfabrik, Bad Säckingen, Germany) under artificial daylight illumination. For analytical purposes, the CIE L*a*b* coordinates were obtained using a spectrophotometer. Colour differences, ΔE, between all shade tabs selected by the students and the natural teeth were calculated. ΔE observed in the intervention and control groups were compared using t tests. In addition, a multi-level regression model was adjusted for age and gender. RESULTS: Mean (SD) ΔE between the shade tabs selected by the students and the natural teeth in the intervention and control groups were 3.8 (2.1) and 4.3 (2.3), respectively (P < .001). The effect of group membership was reproduced in multivariate analysis whereas age and gender did not reach statistical significance. CONCLUSIONS: With more intensive education and practical training the ability to perform clinical shade determinations improved for students without previous clinical experience. The results obtained support the idea of implementing thematic small-group education in the dental curriculum.

Clinical Manifestation, Histopathology, and Imaging of Traumatic Injuries Caused by Brazilian Porcupine (Sphiggurus villosus) Quills.

Injuries to humans caused by porcupines are rare. However, they may occur due to the proximity of urban areas and the animal's habitat in areas such as the Floresta da Tijuca in Rio de Janeiro. Outdoor sports and leisure activities in areas close to forests or in the rain forest are also relevant for incidents of this kind and a better knowledge of the local forest fauna would prevent such undesirable accidents. Porcupine quills have microscopic barbs at their tips which facilitate skin penetration, but hampering their removal. Once the spines are lodged in tissue, the microscopic backward-facing deployable barbs at the tips cause trauma if anyone tries to remove them. Local haemorrhage and an inflammatory response to the contaminated foreign body occur. Depending on the time lapse in removing the spines either septic or sterile foreign body reactions may occur. There is also the risk of migration of the spines, where fatal cases have been reported in human and veterinary medicine. Herein we report two unusual cases of accidents involving humans and the South American porcupine. The Sphiggurus villosus spines removed from scalp skin were also documented through Scanning Electron Microscopy.

Systemic inhibition and liver-specific over-expression of PAI-1 failed to improve survival in all-inclusive populations or homogenous cohorts of CLP mice.

BACKGROUND: The role of plasminogen activator inhibitor type-1 (PAI-1) in abdominal sepsis remains elusive. OBJECTIVES: To study the influence of inhibition and over-expression of PAI-1 upon survival in cecal ligation and puncture (CLP) sepsis. METHODS: (i) Mice underwent moderate CLP and received 10 mg kg(-1) of either monoclonal anti-PAI-1 (MA-MP6H6) or control (MA-Control) antibody intravenously at 0, 18 or 30 h post-CLP. The 30-h treatment group was additionally stratified into mice predicted to survive (P-SUR) or die (P-DIE) based on IL 6 measured at 24 h post-CLP. (ii) PAI-1 expression was induced with pLIVE.PAI-1 plasmid administered 72 h pre-CLP. Blood was sampled for 5 days and survival was monitored for 28 days. RESULTS: MA-MP6H6 effectively neutralized active PAI-1 and fully restored fibrinolysis while PAI-1 over-expression was liver-specific and correlated with PAI-1 increase in the blood. Without stratification, MA-MP6H6 co-/post-treatment conferred no survival benefit. Prospective stratification (IL-6 cut-off: 14 ng mL(-1) ) suggested increased mortality by MA-MP6H6 treatment in P-SUR that reached 30% difference (vs. MA-Control; P < 0.05) after a retrospective cut-off readjustment to 3.3 ng mL(-1) for better P-SUR homogeneity. Subsequent prospective anti-PAI-1 treatment in P-SUR mice with 3.3 ng mL(-1) cut-off demonstrated a negative but statistically insignificant effect: mortality was higher by 17% after MA-MP6H6 vs. MA-Control. Over-expression of PAI 1 did not alter post-CLP survival. Neither PAI-1 inhibition nor over-expression meaningfully modified inflammatory response and/or organ function. CONCLUSIONS: Restoration of fibrinolysis in early abdominal sepsis was not beneficial and it may prove detrimental in subjects with the lowest risk of death, while preemptive PAI-1 up-regulation at the current magnitude was not protective.

Factor structure of the Emotional Eating Scale in overweight and obese adults seeking treatment.

The purpose of this study was to examine the factor structure and anthropometric correlates of the Emotional Eating Scale in overweight and obese adults presenting for weight loss. Participants were 217 men and women with a mean body-mass index of 33.1 (±3.4) kg/m². Results indicated a four factor structure: depression, anger, anxiety, and somatic arousal. These factors demonstrated strong internal consistency, and together accounted for approximately 60% of the total variance. Women had significantly higher depression and total scores than did men. There were no significant correlations between the Emotional Eating Scale scores and anthropometric measures. This work begins to add to the literature base regarding the applicability of the original design of the Emotional Eating Scale for samples consisting of men and African Americans.

[Special pain conditions in urology. Summary, literature review and suggestions]. / Spezielle Schmerzzustände in der Urologie. Zusammenfassung, Literaturübersicht und Anregung.

The diagnosis of acute or chronic pain syndromes of the urogenital system can be verified and successfully treated by exact exploration of the patients medical history, the physical examination and a few image-guided or laboratory analyses Pain conditions which remain of unknown etiology even after extensive investigations are difficult to treat by the urologist alone. In cases where no causally determined morphological changes can be found or the alteration is disproportionate to the discomfort sensed by the patient, a multidisciplinary therapy strategy offers an approach to find a solution.

An international collaborative study to establish the 2nd World Health Organization International Standard for hepatitis B virus DNA nucleic acid amplification technology-based assays.

BACKGROUND AND OBJECTIVES: The aim of this study was to replace the 1st World Health Organization International Standard for hepatitis B virus DNA for nucleic acid amplification technique (NAT)-based assays (code 97/746) with a new International Standard. Two lyophilized preparations freeze dried from the same bulk were evaluated in the original collaborative study (coded 97/746 and 97/750, and termed AA and BB, respectively, in the original study). This present study re-evaluates these two preparations in terms of potency and real-time stability. MATERIALS AND METHODS: The 1(st) International Standard (97/746) and the second lyophilized preparation (97/750) were coded Samples 1 and 2, respectively, in the present study. The samples were distributed to six laboratories and assayed on four separate occasions. Accelerated thermal degradation samples of the two preparations were examined after long-term storage at 4 degrees C and 20 degrees C for more than 51 months. RESULTS: Data were returned from a total of nine different NAT-based assays, five in qualitative format and four in quantitative format. The results of this study confirm the results of the original collaborative study, with no significant differences being found in estimated international units (IU)/ml or polymerase chain reaction-detectable units/ml for the 1(st) International Standard (Sample 1 in this study) and the proposed replacement preparation, Sample 2 (97/750). Real-time and accelerated degradation studies indicate that both samples are very stable. Storage of both preparations at 20 degrees C for more than 51 months resulted in no detectable degradation. CONCLUSIONS: On the basis of the data presented in this collaborative study, Sample 2 (code 97/750) was established as the 2nd International Standard for hepatitis B virus DNA for NAT-based assays with a potency of 10(6) IU/ml (500,000 IU/vial).

Using skin models to assess the effects of a pre-work cream. Methodological aspects and perspective of the industry.

BACKGROUND: There is a basic necessity to understand the mechanisms of the protective effects of pre-work creams. Additionally a lot of workplace-related irritants cannot be tested with the existing in vivo methods due to their toxicological profile. As a consequence, there is a need for additional in vitro models for testing pre-work creams. OBJECTIVE: An in vitro skin model test was developed to evaluate the protective mechanism of a pre-work cream. METHODS: The efficacy of 3 products was assessed by an in vivo test (repetitive occlusive irritation test) and then 3-dimensional skin model tests were carried out. RESULTS: In vivo test results demonstrate that the best protection against sodium dodecyl sulfate is offered by a multiple emulsion. In the case of a skin model test, sodium dodecyl sulfate led to cell damage, an increase in proinflammatory markers and some barrier lipids. The pre-work cream increased the content of skin lipids, without inducing irritation or cell death. CONCLUSION: Skin models support the understanding of the interaction of irritants and pre-work creams. Because they are in vitro models, there are no limitations regarding the selection of irritants, which offers numerous opportunities to test a broad range of workplace irritants.

Using skin models to assess the effects of a protection cream on skin barrier function.

BACKGROUND: There is a basic necessity to understand the mechanisms of the protective effects of emulsions. This would promote the development of protective cosmetics and therefore improve the prevention and treatment of occupational skin diseases. However, for such studies, no reliable skin model is available. OBJECTIVE: An in vitro skin model test was developed to evaluate the protective mechanism of cosmetic ingredients. METHODS: The efficacy of three products was assessed by an in vivo test (Repetitive Occlusive Irritation Test) and then 3-dimensional skin model tests were carried out. RESULTS: In vivo test results demonstrate that the best protection against sodium dodecyl sulphate is offered by a multiple emulsion. In the case of a skin model test, sodium dodecyl sulphate led to cell damage, an increase in pro-inflammatory markers and some barrier lipids. The multiple emulsion increased the content of skin lipids, without inducing irritation or cell death. CONCLUSION: Skin models react similarly to sodium dodecyl sulphate compared to human skin and therefore they are suitable to study barrier repair after sodium dodecyl sulphate damage. It is likely that the superior protective effect of the multiple emulsion in vivo is based on the increased amount of skin barrier lipids.

Feasibility of surgical technique and evaluation of postoperative quality of life after laparoscopic treatment of intrathoracic stomach.

BACKGROUND: Because of the risk of life-threatening complications, the discovery of a complete intrathoracic stomach demands urgent surgery with the aim of repositioning the stomach and gastropexy, and secondarily, to improve life quality. In this study the feasibility of surgical technique and postoperative quality of life after laparoscopic treatment of complete intrathoracic stomach has been evaluated. METHODS: From June 1999 to December 2001 16 patients with an intrathoracic stomach (hiatus hernia Types IIB and III) were treated by laparoscopic techniques, including the repositioning of the stomach, hemi-fundoplication and anterior gastropexy. During the postoperative follow-up the recurrence rate and quality of life (Eypasch index) were evaluated. RESULTS: All operations were performed laparoscopically without conversion, with a mean operating time of 155 min. Pleural injuries occurred in 31% of patients and pleural effusions in 38%, which required puncture in three cases. Complete follow-up showed no recurrences at a median of 14 months. The median quality of life index was 84.6 preoperatively and had significantly improved to 117.8 after the operation. CONCLUSION: Laparoscopic access for the treatment of intrathoracic stomach represents a minimally invasive and safe treatment option for complete intrathoracic stomach, with a low level of perioperative morbidity and significant improvement in quality of life.

Occupational skin-protection products--a review.

Skin-protection products are used in the occupational field to protect the skin against hazards from the workplace. They are only to be used for non-toxic, non-cancerogenous and non-sensitizing low-grade irritants such as water, detergents, and cutting fluids and cannot replace other protective measures such as gloves. The recommendation of products mainly focuses on the physico-chemical properties of the irritant and the protective cream. More in vivo studies and intervention studies are needed to prove the efficacy of skin-protection products under real-life conditions.

Skin cleansers for occupational use: testing the skin compatibility of different formulations.

OBJECTIVES: Skin-compatible hand cleansing is crucial for the prevention of occupational dermatitis. In this article we give an overview of skin-cleansing products and their ingredients and investigate whether solvents or abrasives could be avoided in cleansers for severely soiled skin. METHODS: Comparison of skin compatibility, cleansing power and effectiveness of different formulations of skin cleansers was performed via Duhring chamber tests [visual score, trans-epidermal water loss (TEWL)] and standard hand-wash tests. RESULTS: The results show that, dependent on the grade of contamination, a range of skin cleansers that allow adequate skin cleansing, is necessary. Solvents could be avoided, by the use of powerful cleansing systems based on tenside/oil mixtures. Only for special contaminations may solvents be useful. Powerful cleansing activity could be performed without abrasives. CONCLUSIONS: The worker must be able to choose the skin cleanser that is adequate for the contamination that is present. Therefore, information and education of the workers are crucial for skin-compatible skin cleansing.

Master-slave synchronization in chaotic discrete-time oscillators.

In this paper, we study a simple discrete-time neural oscillator model that, in certain parameter regimes, exhibits periodic or chaotic dynamics. The present model with intrinsically chaotic dynamics is capable of spatiotemporal information processing: in response to constant external stimulation, the oscillator can switch into different chaotic states restricted to distinct parts of the phase space. Of particular interest is the processing of time-dependent input in a master-slave configuration of two coupled oscillators. Here, the response of an oscillator is studied by driving it with the signal of the other. Following the input, the response system adapts to the state of the drive. For a chaotic drive, we can observe generalized synchronization. The onset of adaptation to the drive state by the response is accompanied by on-off intermittency resulting in irregular bursting behavior.

A non-specific aminopeptidase from Aspergillus.

A fermentation broth supernatant of the Aspergillus oryzae strain ATCC20386 contains aminopeptidase activity that releases a wide variety of amino acids from natural peptides. The supernatant was fractionated by anion exchange chromatography. Based on the primary amino acid sequence data obtained from proteins in certain fractions, polymerase chain reaction (PCR) primers were made and a PCR product was generated. This PCR product was used to screen an A. oryzae cDNA library from which the full length gene was then obtained. Fusarium venenatum and A. oryzae were used as hosts for gene expression. Transformed strains of both F. venenatum and A. oryzae over-expressed an active aminopeptidase (E.C. 3.4.11), named aminopeptidase II. The recombinant enzyme from both fungal hosts appeared as smears on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After deglycosylation of the N-linked sugars, both samples were a sharp band at approximately 56 kDa and had identical N-terminal amino acid sequences. Aminopeptidase II is a metalloenzyme with, presumably, Zn in the active site. Using various natural peptides and para-nitroanilides (pNAs) of amino acids as substrates, the aminopeptidase was found to be non-specific. Only X-Pro bonds demonstrated resistance to hydrolysis catalyzed by this aminopeptidase. The optimal enzyme activity was observed at pH 9.5 and 55 degrees C. Among amino acid pNAs, Leu-pNA appears to have the highest value of bimolecular constant of 40 min(-1) mM(-1) (k(cat) = 230 min(-1); K(m) = 5.8 mM) at pH 7.5 and 21 degrees C. Among Xaa-Ala-Pro-Tyr-Lys-amide pentapeptides, the velocity of catalytic hydrolysis at pH 7.5 and 21 degrees C was in a decreasing order: Pro, Ala, Leu, Gly and Glu.

Multicentre study for the development of an in vivo model to evaluate the influence of topical formulations on irritation.

Although skin protective products to prevent irritant skin reactions are in wide use, neither standardized test models to prove differences in efficacy exist, nor has the quality or the reproducibility of results been evaluated in a multicentre approach. This should be mandatory when developing or testing skin care products. Therefore, we have designed a multicentre study in an approach to find a standardized test procedure for the evaluation of skin protective products. In this irritation study, a repeated short-time occlusive irritation test (ROIT) with a standardized protocol has been evaluated in 2 phases (12 days and 5 days protocol) in 4 (n=20) respectively 6 (n=33) skilled centres. The skin reaction was induced by 2 irritants (0.5% aq. SLS and toluene, 2x a day for 30 min). Its modification by 3 different cream bases with different hydrophilicity was analyzed. The irritation was monitored by bioengineering methods (TEWL measurement, colorimetry) and by clinical scoring. The evaluation showed that significant results could already be achieved with the 5-day protocol. Furthermore, in spite of the expected inter-centre variations due to heterogeneity of the individual threshold of irritation, interpretation of clinical score, and inter-instrumental variability, the ranking of the vehicles regarding reduction of the irritant reaction was consistent in all centres.

Polyglutamylation of atlantic cod tubulin: immunochemical localization and possible role in pigment granule transport.

In higher organisms, there is a large variety of tubulin isoforms, due to multiple tubulin genes and extensive post-translational modification. The properties of microtubules may be modulated by their tubulin isoform composition. Polyglutamylation is a post-translational modification that is thought to influence binding of both structural microtubule associated proteins (MAPs) and mechano-chemical motors to tubulin. The present study investigates the role of tubulin polyglutamylation in a vesicle transporting system, cod (Gadus morhua) melanophores. We did this by microinjecting an antibody against polyglutamylated tubulin into these cells. To put our results into perspective, and to be able to judge their universal application, we characterized cod tubulin polyglutamylation by Western blotting technique, and compared it to what is known from mammals. We found high levels of polyglutamylation in tissues and cell types whose functions are highly dependent on interactions between microtubules and motor proteins. Microinjection of the anti-polyglutamylation antibody GT335 into cultured melanophores interfered with pigment granule dispersion, while dynein-dependent aggregation was unaffected. Additional experiments showed that GT335-injected cells were able to aggregate pigment even when actin filaments were depolymerized, indicating that the maintained ability of pigment aggregation in these cells was indeed microtubule-based and did not depend upon actin filaments. The results indicate that dynein and the kinesin-like dispersing motor protein in cod melanophores bind to tubulin on slightly different sites, and perhaps depend differentially on polyglutamylation for their interaction with microtubules. The binding site of the dispersing motor may bind directly to the polyglutamate chain, or more closely than dynein.

Deletion of the trichodiene synthase gene of Fusarium venenatum: two systems for repeated gene deletions.

The trichodiene synthase (tri5) gene of Fusarium venenatum was cloned from a genomic library. Vectors were created in which the tri5 coding sequence was replaced with the Neurospora crassa nitrate reductase (nit3) gene and with the Aspergillus nidulans acetamidase (amdS) gene flanked by direct repeats. The first vector was utilized to transform a nitrate reductase (niaD) mutant of F. venenatum to prototrophy, and the second vector was utilized to confer acetamide utilization to the wild-type strain. Several of the transformants lost the capacity to produce the trichothecene diacetoxyscirpenol and were shown by hybridization analysis to have gene replacements at the tri5 locus. The nit3 gene was removed by retransformation with a tri5 deletion fragment and selection on chlorate. The amdS gene was shown to excise spontaneously via the flanking direct repeats when spores were plated onto fluoroacetamide.

Transepithelial transport processes at the intestinal mucosa in inflammatory bowel disease.

Crohn's disease (CD) and ulcerative colitis (UC) are inflammatory bowel diseases (IBD) of unknown etiology. Oral absorption studies have shown an increased intestinal permeability for various sugar molecules in patients with IBD and their healthy relatives as a possible pathogenetic factor. However, the various transport pathways through the mucosal barrier have not yet been examined. This study therefore investigated whether antigens pass the epithelial barrier by a transcellular or a paracellular pathway. Mucosa of freshly resected specimens from CD (n = 10) or UC (n = 10) patients was investigated by immunoelectron microscopy and compared with healthy mucosa. Epithelial transport was studied with the antigens ovalbumin and horseradish peroxidase after defined incubation. Labeling density of subunit c of ATP synthetase was determined in mitochondria of enterocytes of all specimens. In all specimens epithelial transport of OVA and HRP was principally transcellular through enterocytes with normal ultrastructure, although some tight junctions in CD and UC were dilated. Antigens were transported within vesicles to the basolateral membrane 2.5 min after incubation. The level of enterocytes with electron-lucent cytoplasm containing a high amount of antigens was higher in CD and UC than in healthy mucosa, depending on the grade of inflammation. ATP synthetase was significantly decreased in electron-lucent cytoplasm of CD and UC to normal ultrastructure of healthy mucosa. Our study shows that ovalbumin and horseradish peroxidase taken up by the apical membrane reach the paracellular space by vesicular transport in healthy and IBD enterocytes within a few minutes. Transcellular pathway is affected in both CD and UC, which is indicated by a high level of antigens within the cytosol. We speculate that increased intestinal permeability in IBD results substantially from enhanced transcellular transport.

A small-size neural network for computing with strange attractors.

A small-size model for a chaotic neural network is proposed using strange attractors for computation. This network has a chaotic ground state and is capable of responding to external stimuli by constraining the network dynamics to specific parts of the ground state attractor. For parameter optimization, bifurcation diagrams are evaluated.