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Summary Nucleosome-to-protamine exchange during mammalian spermiogenesis is essential for compaction and protection of paternal DNA. It is interesting that, depending on the species, 1% to 15% of nucleosomes are retained, but the generalizability and biological function of this retention are unknown. Here, we show concordantly in human and bovine that nucleosomes remained in sperm chromatin predominantly within distal intergenic regions and introns and associated with centromere repeats and retrotransposons (LINE1 and SINEs). In contrast, nucleosome depletion concerned particularly exons, 5′-UTR, 3′-UTR, TSS, and TTS and was associated with simple and low-complexity repeats. Overlap of human and bovine genes exhibiting nucleosome preservation in the promoter and gene body revealed a significant enrichment of signal transduction and RNA- and protein-processing factors. Our study demonstrates the genome-wide uniformity of the nucleosome preservation pattern in mammalian sperm and its connection to repetitive DNA elements and suggests a function in preimplantation processes for paternally derived nucleosomes.

The insulin-like growth factor 2 receptor (IGF2R) is essential for prenatal growth regulation and shows gene dosage effects on fetal weight that can be affected by in-vitro embryo culture. Imprinted maternal expression of murine Igf2r is well documented for all fetal tissues excluding brain, but polymorphic imprinting and biallelic expression were reported for IGF2R in human. These differences have been attributed to evolutionary changes correlated with specific reproductive strategies. However, data from species suitable for testing this hypothesis are lacking. The domestic cow (Bos taurus) carries a single conceptus with a similar gestation length as human. We identified 12 heterozygous concepti informative for imprinting studies among 68 Bos taurus fetuses at Day 80 of gestation (28% term) and found predominantly maternal IGF2R expression in all fetal tissues but brain, which escapes imprinting. Inter-individual variation in allelic expression bias, i.e. expression of the repressed paternal allele relative to the maternal allele, ranged from 4.6−8.9% in heart, 4.3−10.2% in kidney, 6.1−11.2% in liver, 4.6−15.8% in lung and 3.2−12.2% in skeletal muscle. Allelic bias for mesodermal tissues (heart, skeletal muscle) differed significantly (P, PLoS ONE, 8 (4), ISSN:1932-6203

For advanced out-of-hospital airway management, skilled personnel and adequate equipment are key prerequisites. There are little data on the current availability of airway management equipment and standards of medical staff on Helicopter Emergency Medical System (HEMS) helicopters in central Europe.An internet search identified all HEMS helicopters in Austria, Switzerland and Luxembourg. We identified 15 HEMS helicopter bases in Switzerland, 28 in Austria and three in Luxembourg. A questionnaire was sent to all bases, asking both for the details of the clinical background and experience of participating staff, and details of airway management equipment carried routinely on board.Replies were received from 14 helicopter bases in Switzerland (93%), 25 bases in Austria (89%) and all three bases in Luxembourg. Anaesthesiologists were by far the most frequent attending physicians (68-85%). All except one bases reported to have at least one alternative supraglottic airway device. All bases had capnometry and succinylcholine. All bases in the study except two in Austria had commercial pre-packed sets for a surgical airway. All helicopters were equipped with automatic ventilators, although not all were suitable for non-invasive ventilation (NIV; Switzerland: 43%, Austria: 12%, Luxembourg: 100%). Masks for NIV were rarely available in Switzerland (two bases; 14%) and in Austria (three bases; 12%), whereas all three bases in Luxembourg carried those masks.Most HEMS helicopters carry appropriate equipment to meet the demands of modern advanced airway management in the pre-hospital setting. Further work is needed to ensure that appropriate airway equipment is carried on all HEMS helicopters.

Bovine trophoblast cells release interferon-τ (IFNT), a type I IFN, as the pregnancy recognition signal. Since type I IFNs exert growth inhibitory and proapoptotic actions, the effect of the conceptus on components of the apoptosis pathways was determined in the bovine endometrium during the periimplantation period. Uteri of Simmental heifers were flushed post mortem at days 12, 15, and 18 of cycle or pregnancy for the recovery of conceptuses and the sampling of ipsilateral endometrial tissue at slaughter for quantitative RT-PCR, immunohistochemistry, caspase activity and TUNEL assays. Endometrium samples of pregnant animals revealed increased transcript levels for the proapoptotic genes XAF1 (day 15: 2.9-fold; day 18: 15.1-fold; p = 0.005) and CASP8 (day 18: 2.4-fold; p = 0.007). The mRNA expression increased significantly with the day of the cycle for the proapoptotic genes FASLG, TNFSF10, TNF and TNFSF1A (p = 0.004, p = 0.006, p = 0.001 and p = 0.007) and the antiapoptotic gene BIRC4 (p = 0.03). We detected high amounts of FASLG transcripts in day 18 conceptuses (16-fold higher than day 18 endometria). This finding was validated at the protein level by immunohistochemistry. To further analyse the endometrial activation of the caspase cascade, the activities of initiator caspase 8 and effector caspases 3/7 were determined luminometrically. No difference between pregnant and cyclic animals was found for either caspase activity. Additionally, a TUNEL assay showed no increase of apoptotic cells in the pregnant endometrium. In conclusion, although the bovine conceptus induces the expression of proapoptotic genes, neither an activation of a caspase cascade nor an increase of apoptotic cells was noticed. These results suggest inhibitory mechanisms preventing endometrial cells from programmed cell death.

A mediastinitis or intraoperative bleeding are the most feared complications of the transoral laser assisted diverticulotomy of Zenker's diverticulum. To reduce these complications we developed a new endoscopic surgical technique and compared the results of this new method with the results of patients who we treated earlier.At the department of Otorhinolaryngology, Head and Neck Surgery, University of Luebeck 68 patients suffering from a hypopharyngeal diverticulum have been treated since 1987. In 64 patients we performed an endoscopic diverticulotomy using a CO2-Laser and the Weerda distending diverticuloscope. Four of the Zenker's diverticulua had to be removed by an external approach. From 1987 to 1994 a five millimeter bar between pouch and hypopharynx after diverticulotomy was left and sealed with fibrin glue in the first 38 patients. From 1995 to 1999 the spur of the hypopharyngeal pouch of the other 26 patients was completely separated. Afterwards we sutured the mucosa between the pouch and the hypopharynx to close the opened mediastinal space and sealed the former spur with fibrin glue.Comparing both endoscopic methods we had to observe concerning our first method one mediastinitis, a rise of body temperature in 29 patients (76%) and we had to perform a revision diverticulotomy in three patients (8%). Our modified technique didn't cause any mediastinitis, a rise of body temperature did only occur in five patients (18%) and no patient had to undergo revision surgery.The transoral complete separation of the spur of the hypopharyngeal pouch with suturing the mucosa of the pouch and the hypopharynx causes good functional results and means a further reduction of complications and a further improvement of the endoscopic diverticulotomy. Our new instruments (Fa. K. Storz, Tuttlingen) will be demonstrated.

One of the most important criteria of malignancy of head and neck cancer are the cervical lymph metastases. Being significant for the therapeutical plan is how tumor depending parameters like T-stage, degree of differentiation and tumor localisation will influence the N-stage and therefore the extension of neck dissection.To evaluate the pattern of formations of metastases and the success of therapy a retrospective study was performed on 405 patients with carcinoma of the oral cavity (n = 47), the oropharynx (n = 117), the hypopharynx (n = 47) and the larynx (n = 193).By the time of surgery carcinoma of the hypopharynx were most frequently accompanied by cervical metastases (80%), followed by carcinoma of the oropharynx (70%), the oral cavity (52%) and the larynx (26%). Occurrence and extension of regional lymph node metastases correlated well with T-stages and degree of differentiation. After surgical therapy locoregional recurrence could be observed in 5.2% of the patients. Five-year-survival rate was reduced to 50% on patients with positive lymph nodes. The different tumour sites showed preferred patterns of metastatic spread, without complete avoidance of certain levels.For the decision on indication and extent of neck dissection the preoperative diagnostic (ultrasound, CT-scan, MRI), localisation of tumour, T-stage, degree of differentiation and the knowledge of typical metastatic spread must be considered.

This study explored the personal characteristics and the health and health-related concerns reported by members of the local homeless population in order to design population-specific health programming. The study also examined whether there were significant differences between homeless who are shelter residents and those who are not. An exploratory descriptive design was used to analyze retrospective data collected by a local County Health Department in interviews of 132 homeless adults. The demographic characteristics found reflect many common patterns: marked over-representation of males, mean age in the mid-thirties, education levels comparable to similar socio-economic groups, high unemployment rates, and low health insurance rates. One third of the sample reported self-assessed health statuses of fair or poor. The most frequently identified physical health issue was joint problems, followed by cardiovascular disease. Depression was mentioned most frequently as a self-identified mental health problem. Loneliness was the number one fear identified. Chi Square analysis showed that homeless who did not stay in shelters were significantly longer term residents (p < 0.0001) of the community and reported fear of loneliness significantly more frequently (p < 0.01). This study identifies health concerns that local homeless people themselves find important and provides direction for development of sound population-specific health programming.

The intranasal allergen Provocation test (INT) is the standard procedure to verify the effect of an allergen when a nasal allergy is suspected. It may be especially useful if results from skin tests or in vitro tests conflict with those from the history. In the clinical and research setting, INT may be used to study the efficacy of different treatment modalities such as pharmacotherapy or immunotherapy. The technique of INT has not been well standardized in the past. Variations exist in planning the optimum time for testing, especially in seasonal allergies.We examined influences of different test dates with regard to the pollen season on the result of INT. A total of 19 patients (eight females, 11 males, aged 26.1 +/- 4.8 years) with a history of at least two years of allergic rhinitis to birch pollen, positive skin prick test, and RAST (or = CAP class II) to birch pollen. INT was performed during the winter-period after approximately six months without natural pollen exposition (V1), during the birch pollen season (V2) and six weeks after the birch pollen season (V3). Allergens were applied using a pump spray (0.1 ml) in concentrations of 10, 50, 100, 500, 1000, 5000, 10,000, 50,000, and 100,000 SQ units per ml (SQ/ml).The threshold concentrations were significantly lower at V2 (100 SQ/ml) and V3 (500 SQ/ml) if compared to V1 (1000 SQ/ ml).Seasonal influences should be taken into account when performing INT in birch pollen allergy.

In private accident insurance, a degree of disability established for the insured and the insuring party for the first time in a binding manner, cannot be established as a new fact if more than 3 years have elapsed since the accident occurred. In private accident insurance for children, the right to confirm the disability as a new fact, is limited to a period of 5 years, but not beyond the age of 18. If post-traumatic disturbances in young children (following injuries of the epiphyseal cartilage, bone and articular infections etc..) do not supply sufficient pointers towards the possible or probable findings at the termination of growth, the physician writing the expertise should draw attention to this fact and should state that as far as the medical situation is concerned, determination of the first occurrence cannot be made now and is possible at the stage of termination of growth only.