Autoantibody against N(epsilon)-(carboxymethyl)lysine: an advanced glycation end product of the Maillard reaction.

Prolonged incubation of proteins with reducing sugar produces advanced glycation end products (AGEs), which are implicated as factors for aging and diabetic complications. We previously demonstrated the presence of N(epsilon)-(carboxymethyl)lysine (CML), one of the main AGE structures, in human and animal tissues using a monoclonal anti-CML antibody (6D12). These findings suggest that CML structures present in vivo could serve as immunogens to generate autoantibodies. This suggestion was tested in the present study. First, plasma samples from diabetic rats reacted positively with AGE bovine serum albumin (BSA). These reactivities increased with the duration of diabetic states and were inhibited specifically by CML-BSA. Second, a fraction purified from plasma of diabetic patients, which bound to AGE-BSA, showed a positive reaction to CML-BSA and furthermore also to human lens proteins, which are known to undergo CML modification in vivo. Finally, patients with renal failure caused by diabetes or nondiabetic pathologies had a higher autoantibody activity against CML structure than that in normal subjects or diabetic patients without renal failure. These results indicate that CML accumulated in vivo serves as an immunological epitope to generate an autoantibody specific for CML that might be used as a potential marker for diabetic nephropathy or chronic renal failure.     

Expression of desmin in normal and laser (HeNe) irradiated regenerating skeletal muscles in the toad (Bufo viridis) and the white rat

The expression of desmin was investigated using immunohistochemical methods in normal and low-energy laser (HeNe) irradiated regenerating rat and toad gastrocnemius muscles following partial excision in the former and cold injury in the latter. During the initial stages of regeneration, presumptive myoblasts immunoreacted to desmin antibodies while other mononucleated cells remained unstained. At a later stage the cytoplasm of the myotubes was intensely stained with anti-desmin. At all time intervals there were more mature myogenic structures in the injured zones of the laser irradiated rat or toad muscles than non-irradiated muscles as indicated by the positive immunoreactivity to desmin. It is concluded that desmin immunostaining provides additional information on the role of histological structures during the regeneration process. The process of skeletal muscle regeneration follwing injury is markedly promoted by low-energy direct laser irradiation during the regeneration process. The results also indicate that the rate of differentiation of myoblasts from undifferentiated stem cells is enhanced by the laser irradiation.     

Acceptance of computer-assisted surgery planning in visceral (abdominal) surgery

The acceptance of computer assisted surgical planning was studied by polling 500 German surgeons. 500 questionnaires were sent, from which 105 have been returned, from which 102 could be analysed. The questionnaire consisted of 14 questions relating to the current practice of surgery planning as well as to desired improvements. In general, the computer assisted surgical planning was assessed as a promising tool in the future. 82 % of the surgeons expected a benefit by computer assisted surgical planning. Better assessment of resectability, better training of residents and more safety of surgical procedures were mentioned in the majority. Computer assisted surgical planning should focus on 3D visualization of spatial relations between lesions and vascular structures, on facilities for the preoperative trial of resection strategies and on the documentation of the planned procedure. The current version of a research prototype (Surgery-Planner) is presented which is dealing with these goals.     

Mechanotransduction and tensegrity (I)

The current review deals with mechanotransduction by means of an architectural model of cell function called tensegrity. This concept was introduced by D. E. Ingber in order to frame more focused mechanisms of mechano-transduction, i.e. different signaling pathways, which are less able to predict global cellular behaviour in response to stress, into a coherent mechano-chemical theory of cell function. Tensegrity structures are made and held up by interconnecting a continuous series of tension elements with a discontinuous series of compression resistant struts, in a simple "stick and string" model. These structures develops an intrinsic stabilizing tension called prestress and reacts by global rearrangements of their configuration to a local action of a mechanical stress. The only requirement of a tensegrity network is that tension is continuous and compression is local. At the cellular level the theory assumes that membrane, nucleus and all the organelles are hard-wired by the insoluble cytoskeletal (CSK) scaffold. More than that, the interconnection between CSK and the extracellular matrix (ECM) provides an efficient mechanical couple responsible for changes in cell shape and movement. Cell shape, in turn, regulates cellular function. Focal adhesion complexes, which mediate the CSK-ECM interaction are viewed as integrative devices for both mechanical signaling and soluble factor-dependent signaling. Furthermore, the tensegral molecular network is considered to be a solid-state regulatory system of all cell functions. The way tensegrity influences molecular mechanics, cellular response tuning and coordinated behaviour of large scale structures are also discussed. Besides its conceptual importance, the tensegrity model has multiple applications, being used in combined approach of cell biology, bioengineering, architecture, and biomechanics in order both to improve cell culture and its subsequent applications and the open the way to a more complex technology of tissular engineering.     

Bilateral Anterior Thoracotomy (Clamshell Incision) Is the Ideal Emergency Thoracotomy Incision: An Anatomic Study

Background

Emergency thoracotomy (ET) is a procedure that provides rapid access to intrathoracic structures for thoracic trauma patients arriving at the hospital in extremis. This study assesses the accessibility of intrathoracic structures provided by six different ET incisions. We hypothesize that the bilateral anterior thoracotomy (“clamshell” incision) provides the most rapid and definitive accessibility to intrathoracic structures.

Methods

Six ET incision types (left anterolateral thoracotomy, right anterolateral thoracotomy, left 2nd intercostal space incision, left 3rd intercostal space incision, median sternotomy, and bilateral anterior thoracotomy) were performed multiple times on eight cadavers. The critical intrathoracic structures were assessed for rapid accessibility and control, and they were characterized as “readily accessible,” “accessible,” and “inaccessible” on anatomic accessibility maps.

Results

Median sternotomy provided better access to intrathoracic structures than left and right anterior thoracotomies. Definitive control of the origin of the left subclavian artery was difficult with left 2nd or 3rd intercostal space incisions. Bilateral anterior thoracotomy, the clamshell incision, was easy to perform and gave superior access to all intrathoracic structures.

Conclusions

In severe thoracic trauma, specific injuries are unknown, even if they can be anticipated. The best incision is therefore one that provides the most rapid and definitive access to all thoracic structures for assessment and control. While the right and left anterolateral incisions may be successfully employed by surgeons with extensive experience in ET, the clamshell incision remains the superior incision choice.

Level of Evidence II

Observational study.     

Isolated bowel segment (Iowa Model 1): technique and histological studies

An isolated bowel segment (IBS) is a viable loop of bowel that is completely free of its mesenteric attachments. We created an IBS by staged procedures: (1) myoenteropexy between the undersurface of the abdominal wall muscle and a jejunal loop that is exteriorized at both ends as mucous fistulae; and (2) division of the IBS mesentery several weeks later. Viability of the IBS is preserved by vascular collaterals that develop at the myoenteropexy during the interval between these two procedures. In this study, histological observation of the IBS was performed to retrospectively determine the optimal interval required for adequate collateral circulation to develop. Twenty-eight rats were subdivided into seven groups of four rats each; each group underwent mesenteric division of the IBS at successive 1-week intervals after myoenteropexy (1 to 7 weeks). The bowel wall structures were histologically examined under light microscopy for each group after mesenteric division. Ischemic changes were observed in the groups in which the intervals were shorter than 6 weeks. With the time interval longer than 6 weeks, no ischemic changes were observed in the intramural ganglia and muscle layers and minimal changes were noted in the mucosa. This study concluded that an IBS can be safely created in the rat that preserve normal bowel structures when its mesentery is divided 7 weeks after myoenteropexy.     

Role of membrane-bound heparin-binding epidermal growth factor-like growth factor (HB-EGF) in renal epithelial cell branching

Role of membrane-bound heparin-binding epidermal growth factor-like growth factor (HB-EGF) in renal epithelial cell branching.BACKGROUND: The developing metanephros is characterized by growth and differentiation of the ureteric bud and the surrounding mesenchymal tissue. These processes can be influenced by several growth factors, including epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha). We examined whether another member of the EGF family of growth factors, heparin-binding epidermal growth factor (HB-EGF), might act as a morphogen in renal epithelial tubulogenesis.METHODS: Expression of HB-EGF mRNA and immunoreactive protein were examined in fetal, neonatal and adult rat kidneys. For in vitro studies of tubulogenesis, a rat renal epithelial cell line (NRK52E) stably transfected with proHB-EGF (NRKproHB-EGF) was treated with TPA for 30 minutes, washed with 2 mol/L NaCl to remove soluble HB-EGF trapped by cell surface heparan sulfate proteoglycan and replated onto plastic dishes in the absence of fetal calf serum. In further experiments, NRKproHB-EGF were suspended in a type I collagen gel in serum-free media.RESULTS: Northern blot analysis indicated that HB-EGF was strongly expressed in embryonic rat kidney (embryonic days 18-20) and was still increased in the neonatal kidney (day 10), compared to the low basal levels in adult kidney. Immunohistochemical analysis confirmed that immunoreactive HB-EGF expression in the fetal rat kidney was localized predominantly to the ureteric bud. When NRKproHB-EGF were plated onto plastic substrata, they became progressively flattened and enlarged and exhibited filopoidia. By 10 hours after plating, NRKproHB-EGF began to migrate and subsequently developed cell-cell contact and fully established tubular-like structures. Immunoelectron microscopy revealed that the initial recovery of cellular proHB-EGF was localized predominantly to areas of cell-cell attachment. No tubule-like structures were observed in similarly treated NRK52E cells transfected with the vector alone. In collagen gels, NRKproHB-EGF developed short tubule-like structures in the absence of TPA treatment, but with simultaneous TPA treatment, longer and more arborized structures developed. MMP-1 mRNA and immunoreactive protein increased in the TPA-treated cells, suggesting that protein kinase C-mediated collagenase activity was important for the observed tubulogenesis. However, inhibition of EGF receptor tyrosine kinase with AG 1478 significantly blunted the TPA-induced tubulogenesis by NRKproHB-EGF grown in collagen gels.CONCLUSIONS: These results indicate that membrane-bound HB-EGF can mediate both epithelial cell branching and cell motility. Localization of proHB-EGF to the site of cell-cell contact and development of tubule-like structures in collagen gels suggests that proHB-EGF may be an important morphogen for renal epithelial cells.     

Role of alpha(3)beta(1) integrin in tubulogenesis of Madin-Darby canine kidney cells

BACKGROUND: We isolated several Madin-Darby canine kidney (MDCK) subclones that exhibit different degrees of branching tubulogenesis in lower concentrations of collagen gel. The M634 clone formed cell aggregates in 0.3% collagen gel, but developed branching tubules vigorously in 0.1% collagen gel. In contrast, the Y224 clone formed cysts in 0.3% collagen gel and displayed fewer branching structures in 0.1% collagen gel. Morphologically, M634 cells exhibited higher levels of cell scattering as well as collagen-induced cell migration than Y224. We conducted this study to delineate the underlying mechanism of branching tubulogenesis in M634 cells. METHODS: Components of the focal contact machinery were analyzed in both cell lines, including the extracellular matrix glycoproteins fibronectin, laminin, and vitronectin; cytoskeleton-associated elements alpha-actinin, talin, and vinculin; and receptors for extracellular matrix and alpha(2), alpha(3), alpha(5), alpha(v), beta(1), and beta(3) integrins. Furthermore, we established several stable transfectants of alpha(3) integrin antisense RNA in M634 cells to examine the role of alpha(3)beta(1) integrin in branching morphogenesis directly. RESULTS: There were no obvious differences in levels of the focal adhesion complex proteins between M634 and Y224 cells, except that the content of the alpha(3) and beta1 integrins were 1.2- and 0.6-fold higher in M634 cells, respectively. The expression of alpha(3) integrin antisense RNA significantly lowered the levels of alpha(3) integrin mRNA and protein. The potential of cell scattering, migration, and branching tubulogenesis in M634 cells was inhibited according to the decrease in alpha(3) integrin expression. CONCLUSION: Our data indicate that expression of alpha(3)beta(1) integrin regulates cell scattering, migration, and branching tubulogenesis of MDCK cells, possibly via adhesion to or serving as a signaling molecule for type I collagen.     

Repair of spinal dural defects with vicryl (polyglactin 910) mesh

This study examined and compared the effectiveness of woven vicryl (polyglactin 910) mesh and lyophilized cadaver dura (Lyodura) for the repair of spinal dural defects. A woven vicryl mesh was used to repair spinal dural defects in 16 mongrel dogs. As an internal control, all animals had a separate dural incision that was closed with 9-0 vicryl suture. Animals were killed, and results were evaluated at 4, 8, 12, and 24 weeks. The repair of dural defects was achieved in all animals, and there were no cases of pseudo-meningocele formation, cerebrospinal fluid leakage, or postoperative infection. The vicryl mesh served as a latticework for formation of a neodural membrane. Inflammatory or reactive response to vicryl mesh was minimal, and no adhesion to underlying neural structures was noted. The neodural membrane formed in the animals in which lyophilized dura was employed was usually thicker than those in which vicryl mesh was used and was associated with thick arachnoid-neural adhesions in two of eight animals. We believe vicryl mesh is a suitable dural substitute and offers promise for use in clinical situations.     

Myoid (Muscular) Hamartoma of the Breast: Case Report and Review of the Literature

BACKGROUND: Myoid (muscular) hamartoma (MH) of the breast is a rare benign tumour-forming lesion composed of differentiated mammary glandular and stromal structures, fatty tissue, and areas of smooth muscle. It is considered to be a variant of mammary hamartoma. CASE REPORT: We report the case of a 46-year-old woman with MH, and provide a literature review explaining the origin of smooth muscle cells. Histologically, the tumour consisted of fibrolipomatous stroma containing ductal and lobular structures of the mammary gland located mainly at the tumour borders. The glandular structures showed signs of micro- and macrocystic changes, apocrine metaplasia, and adenosis. The dominant feature was the presence of a fascicular formation of spindle cells, predominantly in central parts, with incursion between glandular structures. Immunohistochemically, foci of smooth muscle tissue were positive for desmin, smooth muscle actin, and h-caldesmon. Oestrogen and progesterone receptors (PR) showed positive expression which was markedly higher for PR. There was negative expression of CD34, S-100 protein, and CD10. CONCLUSIONS: The origin of smooth muscle cells in MH is unknown. However, it is presumed to be derived from hormonally responsive breast stromal cells by smooth muscle metaplasia, based on evidence of hormone receptor expression in the lesion.     

Rifampicin Carrying Poly (D, L-Lactide)/Poly(Ethylene Glycol) Microspheres: Loading and Release

Abstract: The aim of this study is to prepare rifampicinloaded poly ( d , l -lactide)/poly(ethylene glycol) (PDLLA/PEG) copolymer microspheres as an injectable drug delivery system. PDLLA homopolymers with three different molecular weights (9,760, 14,540, and 23,050 daltons) were synthesized and then transesterified with PEG (with a molecular weight of approximately 3,300-4,000 daltons). By changing the ratio of PEG to PDLLA, block copolymers with different chain structures were synthesized. PDLLA and PDLLA/PEG microspheres in the size range of 2–10 μm were prepared by a modified solvent evaporation technique with the use of methylene chloride as the solvent and methyl cellulose as the emulsifier within the aqueous dispersion medium. Rifampicin was loaded within the microspheres during particle formation. Effects of the solvent/polymer and drug/polymer ratios, PDLLA molecular weight, and PEG content on drug loading and release were investigated. High drug loadings up to 100 mg rifampicin/g polymer were achieved. Both size and drug loadings were decreased by an increase in the solvent/polymer ratio and PEG content and by a decrease in the drug/polymer ratio and PDLLA molecular weight. High release rates were observed in the first 5 days after which constant and slow release rates were noted. Drug release was decreased by a decrease in the solvent/polymer ratio and PEG content and by an increase in the drug/polymer ratio and PDLLA molecular weight.     

Brain nitric oxides synthase in major pelvic ganglia of aged (LETO) and diabetic (OLETF) rats

This study was conducted to evaluate the effects of aging and diabetes mellitus (DM) on brain nitric oxide synthase (bNOS) expression in major pelvic ganglia (MPG) of rats. Otsuka Long Evans Tokushima Fatty rats (12, 30, and 70 weeks old), which are genetic models with non-insulin-dependent DM (NIDDM), and age-matched nondiabetic Long Evans Tokushima Otsuka controls were used. The MPG of all rats in this study were subjected to cryo-sectioning and staining with bNOS polyclonal AB and rhodamine-conjugated rabbit IgG. Fluorescence intensities of the stained neurons were assessed in randomly selected fields per each specimen. Animals of both groups revealed significant decline in the staining intensity of their neurons with aging and the progress of DM, but diabetic rats showed more decline than controls. In conclusion, both aging and NIDDM could decrease bNOS expression in rat MPG. However, NIDDM has a more evident effect than aging on that expression. The decrease in bNOS may cause a disturbance in functions of the target pelvic structures of these ganglia under both conditions.     

Image-guided transsphenoidal surgery for pituitary lesions using Mehrkoordinaten Manipulator (MKM) navigation system.

Twenty-five patients with pituitary lesions were operated on by image-guided transsphenoidal surgery (TSS) using the Mehrkoordinaten Manipulator (MKM) navigation system. The cases included 21 cases of pituitary adenomas, 2 cases of craniopharyngioma and 2 cases of Rathke's cleft cyst. All operations were performed through the sublabial approach under an operative microscope. In some cases, an endoscope was used for the observation of the residual tumor and surrounding structures. The tumors and surrounding important structures such as the internal carotid arteries, the basilar artery, and the optic nerves were precisely localized, and mechanical error was less than 2 mm in almost all cases. In 3 early cases of pituitary adenoma, the patient's head was moved slightly during the insertion of the nasal speculum; in these cases, the resulting error was more than 2 mm. In evaluating the procedures, we determined that the most useful benefit of the MKM system compared with other systems is that the navigation information is not only displayed on the monitor, but also presented in the operative field under the microscope. Therefore, the surgeon can obtain the navigation information without removing his eyes from the operative field under the microscope. The most important drawback to the system is its bulky size.     

Die Akademie der Unfallchirurgie (AUC)

At the beginning of this century the German Trauma Society (DGU) became extensively active with an initiative on quality promotion, development of quality assurance and transparency regarding treatment of the severely injured. A white book on “Medical care of the severely injured” was published, focusing on the requirements on structural quality and especially procedural quality. The impact of the white book was immense and a trauma network with approved trauma centers, structured and graded for their individual trauma care performance, was developed. In order to monitor and document the required quality of care, a registry was needed. Furthermore, for cooperation within the trauma networks innovative methods for digital transfer of radiological images and patient documents became necessary. Finally, the auditing criteria for trauma centers had and still have to be completed with advanced medical education and training programs. In order to realize the implementation of such a broad spectrum of economically relevant and increasingly complex activities the Academy of Trauma Surgery (AUC) was established as a subsidiary of the DGU in 2004. The AUC currently has four divisions: 1) networks and health care structures, 2) registries and research management, 3) telemedicine, 4) medical education and training, all of which serve the goal of the initiative. The AUC is a full service provider and management organization in compliance with the statutes of the DGU. According to these statutes the business operations of the AUC also cover projects for numerous groups of patients, projects for the joint society the German Society for Orthopedics and Trauma (DGOU) as well as other medical institutions. This article describes the success stories of the trauma network (TraumaNetzwerk DGU®), the TraumaRegister DGU®, the telecooperation platform TKmed®, the new and fast-growing orthogeriatric center initiative (AltersTraumaZentrum DGU®) and the division of medical education and training, e.g. advanced trauma life support (ATLS®) and other training programs including the innovative interpersonal competence (IC) course.     

椎动脉V2段与周围结构关系的解剖学观察

目的 了解椎动脉V2段与周围结构的解剖关系,探讨颅外椎动脉供血不足的解剖学机制。方法 在20例头部尸体标本,解剖观察椎动脉与周围结构的关系;其中17例制成干骨,测量C3－C7有关横突孔参数。另在2例新鲜尸体标本取材行组织学切片观察椎动脉周围组织。结果 延续自颈椎、横突孔、关节突等骨膜组织形成明显的纤维鞘膜样结构,将椎动脉、椎静脉及神经根包裹在内。椎动脉外膜与钩突之间有纤维这样组织相连,限制椎动脉。横突孔内未见明显骨性狭窄。结论 椎动脉周围的骨性吸软组织性因素对椎动脉起到限制固定作用,在颈椎运动或不稳时,椎动脉易受波及而出现供血不足。椎动脉牵系机制在椎动脉供血不足发生中可能起着重要的作用。     

En bloc (non-chest wall) resection for bronchogenic carcinoma with parietal fixation. Factors affecting survival

Seventy-three patients (57 men and 16 women) underwent en bloc resection of lung and attached parietes between 1970 and 1982. All patients had documented malignant pleural invasion. Chest wall parietal pleura was invaded in 33 patients, pericardium in 14, phrenic or vagus nerve in nine, left atrium in five, superior vena cava in four, esophagus in two, diaphragm in one, and multiple structures in five. No patient underwent chest wall resection. Parietal pleurectomy was performed in all patients with involvement of the chest wall parietal pleura; 37 lobectomies and 36 pneumonectomies were performed. Operative mortality was 12.3%. The actuarial overall 5 year survival rate (Kaplan-Meier method) was 39.7%. We conclude that en bloc resection for primary bronchogenic carcinoma with invasion of adjacent intrathoracic structures, although associated with a significant mortality, can be performed with a reasonable likelihood of long-term survival.     

Percutaneous endoscopic external ring (PEER) hernioplasty

This pilot study was conducted to determine if percutaneous endoscopic external ring (PEER) hernioplasty would be a viable alternative to the conventional and laparoscopic methods of tension-free repair. The procedure consists of (1) a 2.0–2.5-cm incision over the external inguinal ring to reach the emerging spermatic cord structures, and ligation and excision of the hernia sac and (2) insertion of an endoscope-attached retractor through the external ring, into the inguinal canal for visualization, dissection of posterior inguinal wall, and placement of mesh to complete tension-free repair. PEER hernioplasty was used to treat 48 patients with 60 primary hernias (bilateral in 12 patients) between January 1993 and December 1994. Median follow-up was 12 months and ranged from 5 to 22 months. All patients were discharged within 24 h after surgery except for one. All patients resumed their normal activity within 2–3 weeks. Only three complications were encountered (two scrotal hematomas and one inguinal seroma). To date, there has been recurrence of two hernias in one patient. We conclude that PEER hernioplasty is an effective method of repair of primary hernias that is less invasive than the conventional approach and both less invasive and more cost-effective than laparoscopic approaches.     

Isolated dorsal midtarsal (Chopart) dislocation: a case report

A case of isolated dorsal midtarsal (Chopart) dislocation resulting from blunt trauma is reported. The mechanism of injury, management, and long-term outcome are discussed. A 45-year-old inebriated man fell from a 4-m height and landed on his feet. He had immediate bilateral foot and back pain. Radiographs showed a burst fracture of the second lumbar vertebra, a left Pott's fracture, and an isolated dorsal dislocation of the right midtarsal (Chopart) joint. No neurovascular deficits or signs of compartment syndrome were noted. The image intensifier showed the unstable midtarsal joint with dorsal translation of the navicular and cuboid from the talus and calcaneum, respectively. Closed reduction of the midtarsal joint was performed with 2 transfixing 3.5-mm AO cortical screws. The right foot was immobilised in a short leg cast for 6 weeks; screws were removed 14 weeks later. At 76-month follow-up, the patient had returned to work as a construction worker, but still had right midfoot pain on prolonged walking or standing, because of persistent plantar opening up and dorsal ossification of the midtarsal joints and degenerative change at the talonavicular joint. With hindsight, open reduction may have led to a better outcome through repair of the plantar ligamentous structures, especially the plantar calcaneonavicular (spring) ligament.     

Antitumor effect of combined use of purified human natural tumor necrosis factor (N-TNF) and cis-diamminedichoroplatinum (II) (cisplatin) on the implanted bladder carcinoma (MBT-2) in mice]

The antitumor effect of tumor necrosis factor (TNF) in combination with Cis-diammine-dichloroplatinum (II) (Cisplatin) upon implanted bladder carcinoma (MBT-2) in mice was analysed together with the toxicity. Mice were implanted into the bladder wall with about 10(4) cells MBT-2 viable cells. TNF was the purified human natural tumor necrosis factor. The antitumor effects were evaluated by the volume of the tumor thus implanted and the drug-toxicity by the body weight of mice and microscopic as well as macroscopic findings of the main organs. As a result, 1) The toxicity of TNF to mice was not enhanced in a dose-dependent fashion when combined with Cisplatin. 2) The antitumor effect by combination of TNF and Cisplatin was significantly increased compared to control group. 3) The effects of TNF was enhanced in a dose-dependent fashion when combined with Cisplatin. 4) The antitumor effects was confirmed by pathological findings. These results suggest that the combined use of n-TNF and Cisplatin is promising as a clinically effective treatment against bladder cancer in human.