Topical application of vitamin A reverses metaplasia of rat vaginal epithelium: a rapid and efficient approach to improve mucosal barrier function.

Retinyl palmitate and its metabolites retinol and retinoic acid control growth and epithelial differentiation. Systemic or local vitamin A deficiency induced by malnutrition, continuous chemical irritation or locally induced by inflammation causes squamous metaplastic changes in the epithelium of mucous membranes. We demonstrate that in an animal model topically applied retinyl palmitate can be taken up by the mucosal cells independently from the systemic supply. Under in vivo conditions metabolic changes in vaginal epithelium of rats were shown to be reversed by treatment with topical retinyl palmitate. - After only two days treatment squamous metaplastic vaginal epithelium in rats shows a reversal of the epithelium into a normal phenotype which continues after cessation of the treatment for 7 to 11 days. Higher concentrations and longer retention times lead to a statistically significant (p = 0.025) increase in the protection time. These data demonstrate that squamous changes induced by vitamin A deficiency can be totally reversed with topically applied retinyl palmitate.

Intermittent activity-induced hemobilia caused by liver hemangioma.

BACKGROUND: Intestinal bleeding of unknown origin can lead to a difficult workup. Abdominal colic, melena/hematemesis, and jaundice represent the pathognomonic triad for hemobilia, but clinical presentation and etiology of this entity are varying. Seldom all of these symptoms are present, and rarely does hemobilia cause melena or hematemesis. Often the correct diagnosis is missed. Patients frequently have a long history of complaints and inadequate therapy. CASE REPORT: We report on a patient who complained of repeated, severe epigastric pain and massive melena induced by exercise activity. After 2 years of complaints and an unnecessary operation, ultrasound detected a liver hemangioma. It was supposed that the hemangioma was causing hemobilia during strenuous physical activity. The patient underwent a partial liver resection to eliminate the hemangioma. All complaints resolved, and the patient remained asymptomatic postoperatively. CONCLUSIONS: Physicians should be aware of hemobilia as a rare cause of upper gastrointestinal bleeding, especially if esophagogastroduodenoscopy cannot demonstrate any bleeding source. Ultrasound is able to visualize many diseases leading to hemobilia and should be integrated into the early workup of unclear intestinal bleedings.

Vascular injuries within the hepatoduodenal ligament: recognition by laparoscopic color Doppler ultrasound.

BACKGROUND: Lesions of vascular structures are rare but serious complications of laparoscopic cholecystectomy. The purpose of this blind randomized animal study was to investigate the possibility of detecting different vascular lesions within the hepatoduodenal ligament using laparoscopic color Doppler ultrasound (LCDU). METHODS: Twenty-four lesions of the hepatic artery and portal vein were created laparoscopically in six farmer pigs using titanium clips. The following injuries were studied: (a) partial occlusion of the hepatic artery (eight cases), (b) complete occlusion of the hepatic artery (eight cases), (c) partial occlusion of the portal vein (eight cases). There were also eight cases without lesions of the vascular vessels. The order in which the injuries were created was randomly assigned. The study was performed in a blind fashion. Recognition of the injuries was attempted with LCDU. RESULTS: All injuries were recognized correctly by LCDU. There were no false positive results. The clips were reliably located. Using color Doppler imaging, partial occlusions of the hepatic artery and portal vein were visualized by changes of the blood flow from laminar to turbulent behind the clip. Complete occlusion of the hepatic artery was recognized as a complete cessation of the colored blood flow. CONCLUSION: LCDU is a very efficient tool for visualizing vascular structures and evaluating the bloodstream. Partial or complete vascular occlusion by clips that may occur as a result of difficult dissection during laparoscopic cholecystectomy can be visualized reliably using this technique.

Laparoscopic ultrasonography reliably visualizes bile-duct injuries - a blind randomized porcine study.

INTRODUCTION: Despite use of intraoperative cholangiography (IOC), only one-third or less of bile duct injuries that occur during laparoscopic cholecystectomy are detected at the primary operation. Therefore, there is a need to investigate other intraoperative diagnostic methods, especially laparoscopic ultrasound, for detecting bile-duct injuries. MATERIALS AND METHODS: Detection of different bile-duct injuries by laparoscopic intraoperative ultrasonography was evaluated in a blind, randomized animal study. Fifty bile-duct injuries were created using laparoscopic techniques in 23 pigs. The number of single lesions and the order in which the injuries were created was randomly assigned. The lesions created were: (1) partial occlusion by a clip, (2) complete occlusion by a clip, (3) partial laceration, (4) transection without clips, (5) transection between clips and (6) excision between clips. In addition, there were cases without lesions. Different types of clips were used. A screening of the injuries by laparoscopic ultrasound using a flexible probe was attempted. The study was performed in a blind fashion. RESULTS: Complete ultrasonographical imaging of the hepatic and common bile duct between its bifurcation and its insertion into the duodenum was accomplished in all 23 animals before surgery was performed. Eight cases without lesions were correctly recognized. In 48 of 49 cases, the injuries were detected (sensitivity 98%). In one instance, the injury was obvious without the use of ultrasound. Forty-four cases were correctly diagnosed and, in 12 cases, the injuries were equivocally differentiated (specificity of 100%, overall accuracy of 98.3%). There was only one false diagnosis. The different types of clips were also reliably differentiated. CONCLUSIONS: We conclude that laparoscopic ultrasonography can reliably visualize most relevant bile-duct injuries. A standard for doing the ultrasound examination is proposed. Direct and indirect ultrasound signs of bile-duct injuries are explained.

Expression patterns of complex glycoconjugates and endogenous lectins during fetal development of the viscerocranium.

Experimental evidence suggests that carbohydrates and their corresponding receptors (endogenous lectins) decode biological information. Therefore, the expression of complex oligosaccharides--the potential ligand part of this recognition system--during chondrogenesis and osteogenesis was determined in the viscerocranium of fetal rats by mapping the staining patterns of exogenous lectins. Results were compared with the expression of bone- and/or cartilage-specific core proteins and the binding profiles of neoglycoconjugates. These synthetic tools make possible the localization of sugar-ligand-binding sites. The spatial and temporal distribution patterns of glycoconjugates were highly dynamic and demonstrated a clear correlation with characteristic morphological modifications. The glycobiological characterization of precartilage mesenchymal cells revealed distinct differences compared to prospective bone anlagen. Especially the binding of the exogenous lectin from Griffonia simplicifolia II, that selectively visualized prechondral aggregations, reveals that regulation of early chondral growth is at least phenomenologically correlated with a relatively atypical oligosaccharide composition terminating with N-acetylglucosamine.

Prospective randomized comparison of laparoscopic ultrasonography using a flexible-tip ultrasound probe and intraoperative dynamic cholangiography during laparoscopic cholecystectomy.

BACKGROUND: We performed a prospective randomized comparison of laparoscopic intraoperative ultrasonography (LIOU) and dynamic intraoperative cholangiography (IOC) during laparoscopic cholecystectomy (LC). METHODS: LIOU and IOC were attempted in 518 consecutive patients scheduled for laparoscopic cholecystectomy. The order in which the diagnostic procedures were performed was randomly assigned. RESULTS: LIOU failed in two patients (0.4%), and there were 41 (7.9%) failed IOC. The common bile duct (CBD) was visualized reliably with both methods. Our patients showed sensitivities of 83.3% and 100% and specificities of 100% and 98.9%, with an overall accuracy of 99.2% and 98.9% for LIOU as compared to IOC for identifying unsuspected common bile duct stones. The time necessary for the examination was significantly shorter in LIOU than in IOC (7 versus 16 min). CONCLUSION: LIOU performed by experienced surgeons is a good and effective method to assess the CBD, including the neighboring structures of hepatoduodenal ligament. Using powerful, flexible-tip ultrasound probes, CBD exploration can be done in a longitudinal fashion, which is necessary for good anatomical clarity. A lack of adverse effects, shorter examination times, and lower costs are some of the advantages of this method. The most important advantage is the possibility of unlimited repetition, especially if there is difficulty identifying anatomic structures. In addition, there are some indications that LIOU has the potential to recognize major iatrogenic bile duct injuries.

Impaired responses to sweet taste in vitamin A-deficient rats.

In brief-exposure, two-choice preference tests (sucrose solution v water), vitamin A-deficient (VAD) rats exhibited a decreased preference for sucrose relative to control rats. There was no difference in total fluid intake from both choices between the two groups, nor was any significant difference found in circumvallate taste papilla keratin size. It is concluded that the impaired preference for sucrose in VAD rats is due to a specific impairment in taste sensation rather than general malaise.

[Online infection recording within the scope of total quality management]. / Online-Infektionserfassung im Rahmen des Total Quality Management.

The continuous and exact recording of infections is a condition sine qua non for total quality management. Therefore, at the Diakoniekrankenhaus Rotenburg (Wümme) a program which offers the possibility of online recording of data was integrated into the hospital intranet. The recording is done with network clients. The recording of data is supported by a series of plausibility controls. The advantage consists in immediate evaluation of up to date and extensive statistics of infections. By integration into the existing system, relevant influences and consequences such as the prolongation of hospitalisation, use of material, and frequency of reoperation can by recognized and immediately considered with high validity.

Necrotizing cutaneous lesions caused by interferon beta injections in a patient with multiple sclerosis.

We report a patient with multiple sclerosis who was receiving subcutaneous injections of recombinant interferon beta. During the third month of treatment, painful necrotizing cutaneous lesions appeared at the injection sites. The possible pathogenesis of interferon-induced skin necrosis is discussed.

Cutaneous malakoplakia on the forehead.

A case of cutaneous malakoplakia in an 81-year-old woman in reported. A reddish slowly growing plaque had appeared on her forehead 13 years previously. Histopathology revealed a dense dermal granulomatous infiltrate consisting of lymphocytes and numerous histiocytes containing Michaelis-Gutmann bodies. Ciprofloxacin treatment resulted in partial involution of the lesion.

Interaction of vitamins E and K: effect of high dietary vitamin E on phylloquinone activity in chicks.

To determine the influence of vitamin E on phylloquinone activity, one day-old chicks were raised on a masch diet supplemented with different amounts of vitamin E for 31 days. In chicks fed a diet high in vitamin E (4000 mg allrac-alpha-tocopheryl acetate/kg) but adequate in vitamin K (0.14 mg phylloquinone/kg) a threefold increase in prothrombin time and an increase in mortality rate (five out of twelve animals died from increased bleeding tendency) was observed. The inhibiting effect of high dietary vitamin E on procoagulant factors could be prevented by increasing dietary phylloquinone supplementation. Weight development, and feed utilization were insignificantly different in chicks fed different amounts and ratios of vitamins E and K1. Plasma and liver alpha-tocopherol levels correlated with dietary amounts of vitamin E. Increased phylloquinone levels in the diet did not significantly influence alpha-tocopherol concentrations in plasma and liver, but coagulopathy caused by high vitamin E intake could be reversed.

Stabilization of rat heart mitochondria by alpha-tocopherol in rats.

alpha-Tocopherol (alpha-T) acts by different mechanisms: scavenging free oxygen radicals (FOR) and stabilizing membranes; both may contribute to the prevention of the pathogenesis of diseases like cardiovascular events. The present work shows that alpha-T breaks the vicious cycle generated by damaged mitochondria. The latter stimulate the immune system increasing FOR formation that leads to more damaged mitochondria. alpha-T inhibits lipid peroxidation induced by recombinant human tumor necrosis factor (rh-TNF). Polymorphonuclear neutrophils (PMN) are activated by TNF to produce FOR causing lipid peroxidation of cardiomyocyte membranes. Since TNF is used as a therapeutic agent against certain tumors, treating patients with alpha-T could protect them against systemic side effects of TNF.

Biodiscrimination of the eight alpha-tocopherol stereoisomers results in preferential accumulation of the four 2R forms in tissues and plasma of rats.

According to the USP, 2R,4'R,8'R-alpha-tocopheryl acetate (RRR-alpha-TAc) is 1.36 times more active than all-rac-alpha-tocopheryl acetate (all-rac-alpha-TAc). The all-rac form contains 12.5% each of the stereoisomers RRR, RRS, RSR, RSS, SSS, SSR, SRS and SRR, which display different biopotencies. In the present study, female rats fed a vitamin E-deficient diet were administered 0.82 mg of all-rac-alpha-TAc or 0.60 mg of RRR-alpha-TAc daily for up to 90 d. alpha-Tocopherol concentrations in liver, brain, adipose tissue and plasma were not significantly different among groups on treatment d 64 and 90. Thus, equipotent dosages of all-rac-alpha-TAc or RRR-alpha-TAc resulted in equimolar alpha-tocopherol plasma and tissue concentrations. A comparison with rats administered tocopherol-free placebo showed that plasma and tissue alpha-tocopherol of alpha-TAc-treated rats represented alpha-tocopherol uptake during the repletion period. The eight individual alpha-tocopherol stereoisomers in tissues and plasma were determined by chiral HPLC and capillary gas chromatography. Rats treated with all-rac-alpha-TAc preferentially accumulated the four 2R alpha-tocopherol stereoisomers (15-22% each, sum of all 2R = 70-86%) in tissues and plasma. The remaining 14-30% were 2S stereoisomers with a predominance of the SRS form. In conclusion, all-rac-alpha-TAc administration led to the presence of all eight alpha-tocopherol stereoisomers in rat liver, brain, adipose tissue and plasma. The four 2R stereoisomers including RRR-alpha-tocopherol were equally and significantly enriched. This confirmed that the configuration at C-2 of the alpha-tocopherol molecule has a major impact on stereoisomer biodiscrimination. Furthermore, the results are in agreement with the hypothesis that for alpha-tocopherol stereoisomers, biopotency differences are related to corresponding differences of alpha-tocopherol concentrations.

Laparoscopic transabdominal preperitoneal hernioplasty: results of 1000 consecutive cases.

From October 1992 until September 1995, 859 patients with 1000 inguinal hernias underwent a transabdominal preperitoneal mesh repair (TAPP) in the First Department of Surgery of the Diakonie-Hospital Rotenburg/Wümme, Germany. One hundred and seventeen of these repairs were for recurrences after previous open hernia surgery. There were 76 (7.6%) peri- and postoperative complications, but most of them were minor without necessity for surgical intervention. There were no implant infections or any adverse reaction of the mesh. During the postoperative follow-up period, which ranged from 2-36 months, 11 early recurrences were observed after TAPP, but none were in the recurrent hernia group. Our results support other studies and randomized trials reported in the literature and demonstrate that TAPP is an effective and safe operative procedure especially for the repair of recurrent groin hernias.

Recognition of laparoscopic bile duct injuries by intraoperative ultrasonography.

BACKGROUND: The purpose of this study was to investigate the possibility of detecting bile duct injuries using laparoscopic intraoperative ultrasound (LIOU). METHODS: Fifty bile duct injuries were created using laparoscopic techniques in ten farmer pigs. The lesions created were: (1) partial occlusion, (2) complete occlusion (1 clip), (3) complete occlusion (2 clips), (4) transection between clips, (5) excision between clips. RESULTS: All injuries were easily visualized using LIOU. The relation of clips impinging upon or occluding the bile duct was readily visualized on LIOU in all cases. In some cases it was difficult to distinguish between partial and complete occlusion. It was also difficult to distinguish between transection and excision due to retraction of the severed bile duct. CONCLUSIONS: In cases of iatrogenic occlusion of the bile duct involving hemoclips (including transection or excision between clips), LIOU is a potentially useful modality that may allow earlier recognition and repair. Further studies are needed to evaluate the efficacy of LIOU in detection of bile duct injuries that do not involve hemoclips.

Modulation of the glycoconjugate expression in the tracheo-bronchial epithelium during sustained hypovitaminosis A.

The aim of this study was to determine the influence of sustained marginal vitamin A deficiency on the morphology of glycoconjugate expression in the tracheobronchial epithelium of guinea pigs. The distribution of oligosaccharide chains was investigated by applying a panel of 24 lectins. Glycosaminoglycans were detected by histochemical techniques. Number as well as morphology of ciliated cells showed no significant alterations in hypovitaminosis A. In contrast, the quantity of goblet cells was constantly decreased. A considerable reduction of secretory granules was also observed in these cells. Cytomembranes of ciliated cells (especially in the area of ciliar extensions) showed constant alterations in the patterns of lectin binding in vitamin A-depleted guinea pigs. Our results demonstrate a significant augmentation of accessibility of fucosyl molecules in proximal domains of glycoconjugates of ciliary membranes, whereas the presence of mannose structures seemed unchanged. In distal bronchioli, terminal N-acetylgalactosamine molecules were expressed. During marginal vitamin A deficiency, ciliary cells were specially labelled by GSA I(B), indicating presentation of terminal galactose molecules in alpha-position. Additionally, the cytoplasm of epithelial cells demonstrated enhanced concentrations of polyantennary oligosaccharide core structures. Staining of epithelial cells by VVA was restricted to control specimens. Abundance of N-acetylglucosamine residues on the non-reducing terminus of oligosaccharides was significantly enhanced in the connective tissue of depleted animals as demonstrated by the binding patterns of GSA II. We suggest that altered oligosaccharide patterns may contribute to enhanced predisposition to tracheobronchial infection in marginal vitamin A deficiency.

Long-term administration of high dose vitamin A to rats does not cause fetal malformations: macroscopic, skeletal and physicochemical findings.

A rat model was used to investigate whether high oral doses of vitamin A lead to fetal malformations and to what extent retinyl esters (RES) are transferred from the mother to the fetuses. Retinol and RES concentrations in plasma behave similarly in rats and humans. When high concentrations of vitamin A are administered, plasma retinol concentrations remain relatively constant, whereas plasma RES increased in parallel with the dose. To achieve an elevation from approximately 150 to > 1525 nmol x L(-1) in the experimental group before mating, female Ibm: RORO (spf) rats were fed a maintenance diet enriched with 15.2 x 10(3) retinol equivalents (RE) x kg(-1) at the start and increased stepwise to 52.5 x 10(3) for a total of 8 mo. A parallel subgroup was maintained to measure progress in experimental rats without interference by blood taking. Rats of the control group received the basal diet analyzed to contain 4.5 x 10(3) RE x kg(-1). Before mating the mean body weights of experimental and control rats were not significantly different. All-trans, 13-cis, 4-oxo-all-trans and 5,6-epoxy-all-trans retinoic acid (RA) concentrations were determined in maternal and fetal plasma. With high vitamin A intake, 4-oxo- and 5,6-epoxy RA concentrations were significantly higher in the fetuses than in their mothers. Although these high intakes of vitamin A by the rat dams resulted in high maternal and fetal plasma concentrations of vitamin A and its metabolites, fetal malformations were not observed. This may be due to the fact that circulating RES are not teratogenic and that after crossing the placental barrier, they are stored mainly in fetal liver.

Glycoconjugate expression of chondrocytes and perichondrium during hyaline cartilage development in the rat.

Alterations in the expression of glycoconjugate structures during cartilage development in the chondrocranium, nasal skeleton, Meckel's cartilage, limb buds, vertebral bodies and ribs were investigated comparatively in 13 to 21-d-old rat embryos. The binding patterns of 24 biotinylated lectins were analysed in serial sections and compared with results obtained using histochemical methods. Proteoglycan distribution, assessed by conventional staining procedures, was not associated with lectin binding sites. During early fetal development, hyaluronate concentrations were enhanced in areas of prospective chondrogenesis. With few exceptions, the lectins showed a general increase in intensity of binding to mesenchymal structures. Con A (Canavalia ensiformis), DSL (Datura stramonium), and WGA (Triticum vulgare) displayed a ubiquitous distribution of binding sites. After incubation with LCA (Lens culinaris), PSA (Pisum sativum), STL (Solanum tuberosum), and VAA (Viscum album), characteristic differences in binding intensity between focal areas of developing mesenchyme were seen. DBA (Dolichus biflorus), ECL (Erythrina cristagalli), GSL I (Griffonia simplicifolia), LTA (Lotus tetragonobolus), SJA (Saphora japonica), UEA I (Ulex europaeus) and VVL (Vicia villosa) consistently failed to bind. During chondrogenesis a general reduction of lectin staining was detected. In early stages of development GSL II (Griffonia simplicifolia) was a specific marker of the prechondral blastema in the viscerocranium. PNA (Arachis hypogaea) selectively labelled the prevertebral blastema. In contrast, condensing mesenchyme of limb buds and viscerocranium was not stained. Using RCA (Ricinus communis), it was possible to distinguish chondroblasts from mature cells. All chondrocytes were stained by PSA, PHA-E, PHA-L (Phaseolus vulgaris E and L), and WGA, whereas Con A, LCA, and GSL II detected distinct differences between cartilage with different localisations. Cartilage matrix was constantly negative. Applying GSL II it was possible to distinguish specific segments of the perichondrium. From our results we conclude that especially high mannose oligosaccharides are amplified during development. Terminal sialic acid molecules, branched intralaminar glucose and/or mannose, respectively, internal galactose-(beta 1,4)-N-acetylglucosamine sequences as well as galactose-(beta 1,3)-N-acetylgalactosamine sequences in a preterminal position are diffusely distributed in mesenchymal tissue. In contrast, no evidence for the presence of terminal GlcNAc(beta 1,4)GlcNAc sequences and terminal alpha-fucosyl residues in (1,2) or (1,3)-linkage was obtained. Chondrogenesis appears to be correlated with a general reduction in the extent of expression of oligosaccharide structures. No proof of terminal N-acetylgalactosamine and alpha-galactose moieties was found, whereas our staining results document the expression of terminal beta-galactose structures in restricted areas of the developing mesenchyme.(ABSTRACT TRUNCATED AT 400 WORDS)