Acute administration of antibiotics modulates intestinal capillary perfusion and leukocyte adherence during experimental sepsis.

Antibiotic treatment represents a mainstay of therapy for clinical sepsis. Distinct from their antimicrobial effects, antibiotics may impact the inflammatory process in sepsis, e.g. within the intestinal microcirculation. The impact of seven antibiotics relevant to clinical sepsis on intestinal leukocyte recruitment and capillary perfusion was studied in rats with colon ascendens stent peritonitis (CASP)-induced sepsis or after endotoxin [lipopolysaccharide (LPS)] challenge. The following antibiotics were included: daptomycin; erythromycin; imipenem; linezolid; tigecycline; tobramycin; and vancomycin. The number of rolling and adherent leukocytes in intestinal submucosal venules and the functional capillary density (FCD) in three layers of the intestinal wall were assessed using intravital microscopy. CASP-induced sepsis reduces the intestinal FCD by 30-50%. Single administration of daptomycin, tigecycline or linezolid increased the intestinal FCD. CASP sepsis increased the number of rolling leukocytes by 4.5-fold, which was reduced by erythromycin but increased by vancomycin. The number of adherent leukocytes increased 3-fold in rats with CASP sepsis. It was reduced following administration of daptomycin, tigecycline (in V1 and V3 venules), erythromycin and linezolid (in V1 venules). However, following tobramycin and vancomycin, leukocyte adhesion was further enhanced. Administration of tigecycline and linezolid reduced the LPS-induced increase in the number of adherent leukocytes by 50%. However, imipenem did not affect leukocyte adherence. In conclusion, this work highlights the beneficial impact of the antibiotics daptomycin, tigecycline, erythromycin and linezolid in that they improve intestinal capillary perfusion and/or reduce leukocyte recruitment, whilst the antibiotics imipenem, tobramycin and vancomycin do not exert these properties.

Histological changes and changes in the myosin mRNA content of the porcine masticatory muscles after masseter treatment with botulinum toxin A.

OBJECTIVES: Botulinum toxin A (Botox) is increasingly used for treatment of muscle hyperfunction. For a better understanding of the possible morphologic and chewing changes in patients induced by a therapy with Botox, muscle fiber and myosin heavy chain (MyHC) mRNA alterations were examined in this animal study. MATERIALS AND METHODS: The investigation was carried out on 14-week-old pigs (seven treated animals, eight controls; calculated animal size with a power of 0.5). To initialise the total immobilisation of the right masseter, the Botox injection was distributed into ten areas. After a 56-day period, muscle tissue was taken from the left and right side of the masseter (three regions), temporal (two regions), medial pterygoid and geniohyoid muscles using a standardized method. The muscle fiber cross sections were examined immunohistochemically. Fiber staining was accomplished with antibodies to specific MyHC isoforms. The MyHC mRNA changes were analysed using real-time RT-PCR. RESULTS: Muscles adapt to such stress by changing fiber types and MyHC mRNA content. Paralysed masseters display atrophic changes while other masticatory muscles show hypertrophic changes. The results indicated that the typical distributions of type IIa und IIb fiber types in masticatory muscles were increased in the masseter muscles due to Botox application. On the other hand, the masseters without Botox in the treated group showed a significant increase of type I MyHC. CONCLUSIONS: Application of Botox may lead to uncontrolled structural changes in affected and unaffected muscles. CLINICAL RELEVANCE: Treatment of muscle hypertrophy with Botox may cause muscle imbalance.

Comparison of reference points in different methods of temporomandibular joint imaging.

PURPOSE: Conventional radiography is a well-established method for imaging of the temporomandibular joint (TMJ) structures. However, the dental computer tomography becomes more important for the visualization of teeth in the jaw-bone. The applicability of dental computer tomography for the visualization of the TMJ it not yet been proven. The aim of the study was to identify TMJ structures using reference points with the magnetic resonance imaging (MRI) and the computed tomography (CT). METHODS: In order to compare the visualization and measurement of the TMJ a total of eight human cadaver heads was examined with CT and MRI and analysed using reference points. RESULTS: In both imaging techniques the selected reference points and distances are well definable and allow objective evaluation of anatomical structures. The CT images display a clearly better contrast to noise ratio than the MR images. The distance measurement of different width and length showed significant correlation of both images techniques. CONCLUSIONS: In TMJ diagnostics, maximum information could be obtained using both imaging techniques together due to synergistic effects.

Changes in insulin like growth factors, myostatin and vascular endothelial growth factor in rat musculus latissimus dorsi by poly-3-hydroxybutyrate implants.

The present study aimed at researching the synergistic effect between an ectopic bone substitute and surrounding muscle tissue. To describe this effect, changes of insulin like growth factors (IGF1, IGF2), myostatin (GDF8) and vascular endothelial growth factor (VEGF) mRNA content of 12 Wistar-King rats musculus latissimus dorsi with implanted poly-3-hydroxybutyrate (PHB) scaffold were examined after 6 and 12 weeks. At each time interval six rats were killed and implants and surrounding tissues prepared for genetic evaluation. Eight rats without any implants served as controls. RNA was extracted from homogenized muscle tissue and reverse transcribed. Changes in mRNA content were measured by Real-Time PCR using specific primers for IGF1, IGF2, GDF8 and VEGF. Comparing the level of VEGF mRNA in muscle after 6 and 12 weeks to the controls, we could assess a significant increase of VEGF gene expression (p<0.05) whereas the level of mRNA expression was higher after 6 than after 12 weeks of treatment. Expression of IGF1 gene was also significantly increased as compared to the controls over the observed period of time (p<0.05). In the case of the IGF2 gene, the expression was significantly elevated after 6 weeks (p<0.05), but not significantly increased after 12 weeks (p>0.05). We observed a significantly decreased GDF8 gene expression (p<0.05) both after retrieval of implants after 6 as well as after 12 weeks. Moreover, mRNA level of GDF8 after 6 and 12 weeks were comparable the same. Our results show that PHB implants in rat musculus latissimus dorsi interact with the surrounding muscle tissue. This interaction works itself on growth potential of the muscle.

Skeletal muscle relaxants inhibit rat tracheal smooth muscle tone in vitro.

Neuromuscular blocking drugs (NMBD) can inhibit not only nicotinic but also muscarinic (M) receptors and thereby affect not only skeletal but also smooth muscle (SM) tone. A selective postjunctional muscarinic inhibition would relax, while prejunctional inhibition of muscarinic M2 receptor might hasten SM contraction thereby increasing the risk of bronchospasm. In rat tracheal rings in vitro we evaluated the effects of cumulative concentrations of some NMBD and M receptor blocking agents for their effects on tracheal smooth muscle (TSM) tone pre-contracted with carbachol (CARB; 5 x 10(-7)M or 10(-6)M), pilocarpine (PILO; 5 x 10(-6)M), or by electrical field stimulation. The NMBDs produced relaxation in the preparations precontracted with CARB or PILO. The order of potency after CARB (10(-6)M) was (EC(50)): 4-DAMP (9.8) >atropine (9.2) >methoctramine (6.4) >pancuronium (6.0) >mivacurium (5.8) >cisatracurium (5.6) >gallamine (5.2) >rocuronium (4.8) >succinylcholine (2.9); NMBDs also partially prevented contraction elicited by the electrical field stimulation. We demonstrated that the clinically used NMBD that were examined produced rat TSM relaxation, probably by predominantly blocking postjunctional muscarinic receptors.

Glutamine and alanyl-glutamine dipeptide reduce mesenteric plasma extravasation, leukocyte adhesion and tumor necrosis factor-α (TNF-α) release during experimental endotoxemia.

Glutamine (GLN) appears to be an essential nutrient during organism development and critical illness. The aim of our study was to evaluate the effects of GLN and its generic preparation alanyl-glutamine-dipeptide (DIP) on the microcirculation in endotoxemia in rats and its effects on tonus or aortal rings in vitro. Male Lewis rats (n=40) were separated in 4 groups. Group 1 (CON) served as healthy control group while the other groups received an endotoxin bolus i.v. (5 mg/kg lipopolysaccharide, LPS i.v.). In group 3 (LPS+GLN) 0.75 g/kg-1 GLN i.v. before LPS challenge was administered. In group 4 (LPS+DIP) DIP containing 0.75 g/kg GLN was given. Leukocyte-endothelial interactions and mesenteric plasma extravasation were determined at 0, 1 and 2 hours during the experiment by intravital fluorescence microscopy (IVM). Cytokine release (TNF-alpha, IL-1 beta, IL-6, IL-10) was measured by ELISA. GLN treatment reduced leukocyte adherence (-49.7% vs. LPS group, p<0.05) and plasma extravasation (-12.3% vs. LPS group, p<0.05) significantly during endotoxemia compared to untreated LPS animals. In group 4 (DIP+LPS), a decrease of leukocyte adherence (-56.0%) and mesenteric plasma extravasation (-18.8% vs. LPS group, p<0.05) was also found. TNF-alpha levels were reduced in both GLN and DIP (p<0.05). In vitro experiments demonstrated that glutamine agents could attenuate the response to contracting agents in presence of the vascular endothelium, implying nitric oxide pathway. In vivo, GLN as well as DIP pre-treatment diminish the detrimental impact of endotoxemia on the mesenteric microcirculation and the TNF-alpha release, the effects whose clinical importance should be further examined.

Perfusion culture promotes differentiation of oral keratinocytes in vitro.

In order to reconstruct the mucosal lining of the oral cavity tissue engineered autologous mucosa grafts could be of great benefit. In conventional stagnant cultures cells often tend to dedifferentiate. Perfusion culture has been demonstrated to reestablish differentiation in various epithelial cell types. Thus, in secondary cultures of human oral keratinocytes from ten patients conventional stagnant culture versus perfusion culture technique was compared. Proliferation and state of differentiation as expressed morphologically and immunohistochemically were assessed. After 14 days oral keratinocytes in a perfusion culture system tend to be further differentiated. They build up a thicker epithelium (3.4+/-1.0 vs. 2.4+/-0.4 layers), form microridges, express cytokeratins 1, 2, 10, 11, 19 but not 13 in all cells and cytokeratins 5, 6 particularly in cells attached to the carrier membrane. In contrast, oral keratinocytes in stagnant culture do not form microridges and rather express cytokeratins 13, 14, and 19 in mattering amounts.

Morphological evaluation of bone defect regeneration after treatment with two different forms of bone substitution materials on the basis of BONITmatrix.

In the design of biomaterials for therapeutic application the evaluation of cellular/tissue responses play a key role. In this study, the in vivo bone-regenerative capacity and resorption of granular BONITmatrix and a paste-like bone substitution material on the basis of BONITmatrix were investigated in a rat cranial defect model. The results obtained with both biomaterials were compared to each other. For these, the paste-like composite and the granular BONITmatrix were implanted in adult male WOK-W rats, the skulls were harvested after eight weeks, and histopathological examinated. The comparison of the both tested biomaterials showed that the paste-like composite is much better to handle, the resorption of the material and the ossification process is much faster than those of granular BONITmatrix. The amount of newly formed bone was also measured and more bone formation was found in bone defects filled with the paste-like composite compared to those with granular BONITmatrix. The present study showed that both biomaterials could stimulate bone regeneration, but the paste-like composite leads in comparison to granular BONITmatrix to an accelerated more comprehensive bone regeneration.

The factors associated with body mass index in adults from the study of health in Pomerania (SHIP-0), Germany.

OBJECTIVE: To investigate the relation between dental status, BMI and systemic diseases and to evaluate the risk factors for having a higher BMI. MATERIALS AND METHODS: Apopulation based cross sectional study of 6248 subjects aged 18-80 years (response of 68.8%, n=4310) was conducted in the Study of Health in Pomerania (SHIP-0). Socio-demographic, medical and oral health information was recorded by 5 dentists at two similarly equipped medical/dental services in the cities of Greifswald and Stralsund. Bivariate statistics, multivariate statistics, linear and logistic regression models were performed to assess the relationship between following covariates: gender, educational level, family status, social activities, income, quality of life (SF-12), smoking, alcohol abuse, diabetes, renal disease, high blood pressure, dental status and high physical activity. RESULTS: Significant risk factors for subjects having a higher BMI were: high blood pressure (OR=2.28), diabetes (OR=2.10), educational level (low: OR=1.49; medium OR=1.27), male (OR=1.32) and former smoker (OR=1.20). whereas young age, being single and being dentate (natural teeth, replaced teeth or fixed teeth) was shown to be protective for having a "high" BMI. CONCLUSION: The most important predictors of BMI were shown to be social and medical factors. Dental factors are most significantly influenced by social factors and also exhibit an important impact on BMI.

Free DIEP-flap reconstruction of tumour related defects in head and neck.

The free deep inferior epigastric perforator flap (DIEP) is a well-established therapy for plastic reconstruction of the breast or defects of the lower extremity without distinct donor site morbidity. Because of its particular qualities we started to apply the DIEP-flap also in reconstruction of defects in the cranio-maxillofacial area. A series of 10 consecutive patients, who received a DIEP-flap for reconstruction of large soft tissue defects after ablative tumour surgery, was reviewed. Nine of the 10 flaps survived and uneventfully healing was observed in 8 of the 10 flaps. Primary layered closure of the abdominal wall was achieved in all cases and no complications at the donor site were observed. In our experience the DIEP may serve as a well considerable alternative to the rectus abdominis flap and the latissimus dorsi flap for bridging extensive reconstructions in the cranio-maxillofacial region. It offers the possibility for flap elevation simultaneously to the surgical procedures in the head and neck area. A special advantage of the DIEP-flap is the very low donor site morbidity.

Mandible, maxilla and cervical spine--a functional unit?

The motion patterns of mandibular points were recorded in vivo in closed, free movements of the mandible parallel to the sagittal-vertical plane. The points ran along loops which were valued by their area and length. All points whose loops included the same area under regarding the sense of circulation formed a straight line. Lines belonging to different areas were parallel. When the absolute areas of the oops were plotted for particular points a hollow depression with two minima resulted. The point that showed the lowest minimum in the depression corresponded to the position of the neuromuscular mandibular axis of rotation. The points running along equal loop lengths formed elliptical lines with a minimum below the condyle. The lines of constant loop area and loop length were overlaid with lateral radiographs, to match the patterns of motion with anatomical structures. The mandibular axis of rotation lay mostly cranial anterior of the condyle whereas the point with the shortest path lay mainly below this axis point, inside the bony structures. The row of teeth in the maxilla was found to be located below the line of minimal loop lengths. The cervical spine was arranged along the depression of the minimal absolute areas.

Evaluation of shape and size changes of bone and remodelled bone substitute after different fixation methods.

Suitable tissue fixation is indispensable to histological analysis. This investigation, therefore, sought to evaluate changes of shape and size of bone specimens and remodelled bone substitute material following different fixation methods. Mandibular bones of 9 pigs (Sus scrofa domesticus) served as specimens. Two mandibular premolars were extracted respectively and the extraction alveoli were filled with synthetic bone substitute material. The samples were collected after 70 days. Fixation of 6 specimens respectively was done for 7 days in 4% formalin (formaldehyde), 70% ethanol and glycerol at 18 degrees C room temperature. The samples were radiographically examined before and after fixation using a reference specimen and subsequently underwent histological analysis. After fixation in formalin, the samples showed no size changes. After fixation in glycerol, morphological analysis revealed minor shape changes. Fixation in ethanol causes shrinking of the tissue specimens. Histological inspection of the tissues shows no morphological changes except slight shrinking. In conclusion there is no universal fixative that could met all requirements and permited proper examination without affecting tissues or bone specimens.

A novel post-and-core restoration system--results of three years of clinical application of the "Wuerzburg Post".

The Wuerzburg Post is a new post-and-core restoration system designed to eliminate the weak parts of post-and-core restorations and the associated problems, respectively. In contrast to conventional posts, the Wuerzburg Post is a short and thick post, which no longer relies on cementation or luting for retention in the root, but on stress-free positive locking, which it achieves by means of a post which can be spread into a predefined and form-congruent undercut cavity. The second key feature is an annular groove which runs in the dentin, girded by a corresponding structure, ensuring regular force transmission and stress dissipation, as opposed to the classic ferrule design. There are two versions: one with a machined core which can be prepared like a classic build-up to support crowns and bridges, and another one with a 2.25 mm ball end to connect to common dies which can be integrated into removable prostheses. As the system utilizes prefabricated parts made from Titanium, a precise fit is ensured, enabling the user to restore teeth quickly and easily. Over the course of the past three years, 129 posts were inserted, most commonly on upper and lower incisors and canines. The main application was restoration of fractured telescopes. During the observation period, five failures were observed. Two of the failiures did not cause significant damage to the tooth, and were subsequently immediately repairable. The survival rate amounts to over 95% after three years under risk.

Geochemistry of Bulgarian soils in villages affected and not affected by Balkan endemic nephropathy: a pilot study.

Balkan endemic nephropathy (BEN) is a chronic tubulointerstitial disease whose mosaic-like distribution throughout the Balkans has not changed significantly since its initial description. In this study, we explored the hypothesis that the occurrence of BEN is linked with the environmental geochemistry of villages. Soil samples were collected from BEN and non-BEN villages in the Vratza region of Bulgaria. Samples were digested in nitric acid and analyzed for 22 elements by hexapole, inductively coupled plasma, and mass spectrometry. Selected results are that: 1) absolute concentrations from both types of villages were not enriched above "background" concentrations; and 2) copper, molybdenum, lead, and cadmium concentrations were higher in BEN than in non-BEN soils, while selenium concentrations were lower. Although geochemical differences between BEN and non-BEN villages were found, not all differences were statistically significant, in part due to a limited number of samples.

[Synthesis and antibacterial and antitumoral activity of some methylhydrazonium salts of pyrimidine bases and their analogues (author's transl)]. / Synthese, antibakterielle und geschwulsthemmende Wirkung einiger Methylhydrazoniumsalze von Pyrimidinbasen und deren Analoga.

On reacting pyrimidine metabolites containing a carboxyl or sulfhydryl group with methylhydrazine or N-benzyl-N'-methylhydrazine, various methylhydrazonium salts of these metabolites were synthetized, e.g., the salts of 5-fluororotic acid, 5-azaorotic acid, 2-thiouracil-5-carboxylic acid and 2-thio-6-azathymine. Some of these salts exhibited a more marked antibacterial activity against St. aureus and several of its mutants as well as a greater antitumoral activity against different transplantable tumours than their single components.

A new substance effective against transplantable tumors in vivo: L-cystine-bis-(N,N-beta-chloroethyl)-hydrazide.

It is shown that L-cystine-bis-(N,N-beta-chloroethyl)-hydrazide-hydro-bromide possesses strong (50-100%) inhibitory effect in vivo against myeloma P-8, carcinosarcoma Walker, lymphosarcoma Pliss, sarcoma Yoshida, sarcoma Jensen and sarcoma 180 in doses 5-12 mg/kg/day. No suppression of the growth of Ehrlich ascites tumor was observed. The acute toxicity (LD50) of this substance on mice and rats is 71 mg/kg and 47 mg/kg respectively.

Antitumor effect of 2-thio-4-hydrazinouracil on transplantable tumors.

The cytostatic activity of 2-thio-4-hydrazinouracil (THU) on some transplantable tumors has been studied. A strong effect of this compound (between 60% and 100% suppression) has been found in the case of Myeloma P-8 (MOPC-21) and Sarcoma 180 (Crocker). A less pronounced effect has been observed on Yoshida sarcoma, while the development of Jensen sarcoma is not influenced.