Osteoanabolic effect of alendronate and zoledronate on bone marrow stromal cells (BMSCs) isolated from aged female osteoporotic patients and its implications for their mode of action in the treatment of age-related bone loss.

SUMMARY: In the present study, we evaluated the potential for aminobisphosphonates to enhance the development of bone-forming osteoblasts from progenitor cells isolated from aged female osteoporotic patients. The aminobisphosphonates tested significantly enhanced osteoblast formation and thus lend further insights into their possible mode of action in the treatment of osteoporosis. INTRODUCTION: The primary aim of this study was to evaluate the influence of aminobisphosphonates on the osteogenesis of human bone marrow stromal cells (hBMSCs) and mineralization of differentiating bone-forming cells isolated from osteoporotic patients. METHODS: The influence of aminobisphosphonate treatment on hBMSC osteogenesis was assessed by the quantitative measurement of alkaline phosphatase (ALP) activity, in addition to quantitative reverse transcription polymerase chain reaction and Western blot analysis of known osteogenic markers. Mineralized matrix formation by hBMSC-derived osteoblasts was visualized and quantified using Alizarin red staining. RESULTS: hBMSC cultures treated with osteogenic medium supplemented with zoledronate demonstrated a significant increase in Alizarin red staining after 3 weeks as compared to cells cultured in osteogenic medium alone. Similarly, cultures of differentiating hBMSCs isolated from patients receiving alendronate treatment also demonstrated an increased propensity for mineralization, even in the absence of further in vitro stimulation by zoledronate. The stimulatory effects of aminobisphosphonate treatment on hBMSC-derived osteoblast-mediated mineralization were independent of any alterations in ALP activity, although significant decreases in the expression levels of osteopontin (SPP1) were evident in hBMSCs following exposure to aminobisphosphonates. Further analysis including Western blotting and loss-of-function studies revealed osteopontin as having a negative influence on the mineralization of differentiating osteoporotic bone-forming cells. CONCLUSIONS: The results presented here demonstrate for the first time that aminobisphosphonate treatment of osteoporotic hBMSCs enhances their capacity for osteoblast formation and subsequent mineral deposition, thus supporting the concept of aminobisphosphonates as having an osteoanabolic effect in osteoporosis.

Factors affecting long-term outcomes after thromboembolectomy for acute lower limb ischemia.

AIM: The aim of this study was to analyze factors influencing outcomes of surgical management for lower limb acute ischemia. METHODS: A retrospective analysis of 490 thromboembolectomies performed in 468 patients was conducted. Perioperative and follow-up results were analyzed. Univariate and multivariate analysis of clinical variables and patients' characteristics for the risk of reocclusion, amputation and mortality at 2 years were performed. Statistical significance was defined as a P value <0.05. RESULTS: Cumulative reocclusion, amputation and mortality rates at 24 months were 22.6%, 14.3% and 42.8%, respectively. At univariate analysis, the factors associated with increased 2-year reocclusion rate were severity of clinical presentation, current smoking habit, arterial thrombosis rather than embolism, atrial fibrillation and the avoidance of completion angiography. All these factors except clinical presentation maintained significance at multivariate analysis. Factors associated with increased 2-year amputation rate at univariate analysis included severity of clinical presentation, smoke, arterial thrombosis, atrial fibrillation and valvulopathy. All these factors except clinical presentation and valvular defects maintained significance at multivariate analysis. Factors associated with increased 2-year mortality rate at univariate analysis included age >80 years, arterial thrombosis, history of peripheral arterial disease and antiplatelet drugs. The same factors, except antiplatelet treatment, were found to be significant at multivariate analysis. CONCLUSION: Surgical intervention for lower limb ischemia is associated with high 2-year mortality but offers good 2-year limb salvage. The pattern of risk factors for reocclusion and amputation rates is quite different from those affecting mortality. Only thrombotic aetiology is a significant risk factor for all the three outcomes.

Contrast media-induced nephropathy in patients undergoing angiography prior to or during vascular surgery: a systematic review.

Nephropathy induced after administration of contrast medium is an acute and severe complication that is of particular concern in vascular surgery. While patients undergoing coronary procedures have been extensively studied, there is a paucity of data on pre- and intraoperative prevention of contrast-induced nephropathy in vascular surgery patients. There is lack of a robust analysis exploring the additive effect of pre- or intraoperative administration of drugs such as N-acetylcysteine, dopamine and sodium bicarbonate in addition to adequate hydration in patients undergoing angiography prior to or after completion of vascular surgery. A systematic review of contrast-induced nephropathy after angiography in patients undergoing vascular surgery was carried out. Eligible trials were sought by multiple methods, and the pooled odds ratios for contrast-induced nephropathy were computed under a random effects model. Twenty-one publications were identified for screening and 6 studies were included for systematic review. All 6 studies investigated preoperative angiography-related contrast-induced nephropathy; one study also investigated completion angiography. The overall frequency of contrast-induced nephropathy in patients undergoing vascular surgery was 9.2% (79/862). Risk factors for contrast-induced nephropathy identified were age >70 years, high contrast volume, pre-existing renal disease and antihypertensive medication. Two studies found that administration of N-acetylcysteine prior to angiography does not provide added benefit in preventing contrast-induced nephropathy. Advanced age and pre-existing renal and vascular risk factors such as arterial hypertension expose vascular surgery patients to increased risk of contrast-induced nephropathy. Those undergoing completion angiography appear to be at even higher risk, particularly if severely azotemic. Further randomized clinical trials analyzing strategies for preventing contrast-induced nephropathy are needed.

Lower extremity traumatic vascular injury at a level II trauma center: an analysis of limb loss risk factors and outcomes.

AIM: The objectives were to review in our series the risk factors, management and outcomes of patients who sustained vascular injuries in the lower limbs and to determine the effect of risk factors and treatment on the outcome of the injured extremity. METHODS: Fifty-six patients submitted to surgical treatment were retrospectively reviewed. Results were analysed in terms of type of operation and reconstruction, intraoperative and 30 day complications, reconstruction occlusion, major amputation and mortality. RESULTS: The mechanism of trauma was blunt in 30.4% and penetrating in 69.6%. The overall primary amputation rate was 5.4%, the overall secondary amputation rate was 1.8%. The overall intraoperative and postoperative mortality were 1.8% and 5.4% respectively. At univariate analysis, the presence of compartment syndrome and ischemia time >6 hours were associated with a significantly higher risk of early reconstruction thrombosis (both P=0.03). It showed also that the number of patent vessels (P=0.0000) and the presence of a MESS score >7 (P=0.0000) significantly affected primary amputation, and that the occurrence of postoperative deep wound infection or sepsis (P=0.0000), of tibio-peroneal trunk injury (P=0.003) and of a MESS score >7 (P=0.004) significantly affected secondary amputation. CONCLUSION: The number of patent arteries (0-1), the presence of a MESS score >7, the incidence of tibio-peroneal trunk injury and the occurrence of postoperative deep wound infection are significant independent factors for limb loss. The presence of compartment syndrome and of ischemia time >6 hours are associated with a significantly higher risk of early reconstruction thrombosis.

Late endosomal traffic of the epidermal growth factor receptor ensures spatial and temporal fidelity of mitogen-activated protein kinase signaling.

Mitogen-activated protein kinase (MAPK) signaling is regulated by assembling distinct scaffold complexes at the plasma membrane and on endosomes. Thus, spatial resolution might be critical to determine signaling specificity. Therefore, we investigated whether epidermal growth factor receptor (EGFR) traffic through the endosomal system provides spatial information for MAPK signaling. To mislocalize late endosomes to the cell periphery we used the dynein subunit p50 dynamitin. The peripheral translocation of late endosomes resulted in a prolonged EGFR activation on late endosomes and a slow down in EGFR degradation. Continuous EGFR signaling from late endosomes caused sustained extracellular signal-regulated kinase and p38 signaling and resulted in hyperactivation of nuclear targets, such as Elk-1. In contrast, clustering late endosomes in the perinuclear region by expression of dominant active Rab7 delayed the entry of the EGFR into late endosomes, which caused a delay in EGFR degradation and a sustained MAPK signaling. Surprisingly, the activation of nuclear targets was reduced. Thus, we conclude that appropriate trafficking of the activated EGFR through endosomes controls the spatial and temporal regulation of MAPK signaling.

Carotid endarterectomy: comparing anesthesia in awakened and intubated patients with general anesthesia.

AIM: The purpose of this study was to determine utility and efficacy of anesthesia with remifentanil in awakened and intubated patients in comparison with general anesthesia and routine shunting. METHODS: Hundred patients were randomized for general anesthesia (group A) or anesthesia with remifentanil (group B). The two groups were compared in terms of postoperative outcome, neurological complications, arterial pressure during intervention and in during the following 36 hours. Group B patients answered a proposed questionnaire about intraoperative and postoperative degree of comfort, fear, pain and discomfort for the orotracheal tube. RESULTS: The two groups differed significantly only for age (group B>A); 10% of patients was not considered fit for the anesthesia with remifentanil (initial experience only). The two groups did not differ in terms of central neurologic complications (0%), general complications or technical problems in terms of intra-operative arterial pressure problems (the anesthesia with remifentanil caused less hypertensive problems (P>0,19) and (not technique-dependent) peripheral neurologic deficits. All patients accepted the method and did not report fear or discomfort. Only 3 patients (6.6%) described light pain during the intervention. In 2 cases (4.4%) the surgeon complained about the restlessness of the patient; this did not cause a change in procedure. CONCLUSIONS: General anesthesia with remifentanil produces an association between the advantage of locoregional anesthesia and those of general anesthesia, probably lowering the risk for the patients.

Extracellular matrix mimicking scaffold promotes osteogenic stem cell differentiation: A new approach in osteoporosis research.

BACKGROUND: Osteoporosis is a common metabolic disease, with mesenchymal stem cells discussed to play an important role in its pathomechanism. For in vitro osteoporosis studies, selection of adequate culture conditions is mandatory so as to preserve cell properties as far as possible. A suitable cell culture surface would ideally provide reproducible experimental conditions by resembling those in-vivo. OBJECTIVE: Generating an improved growth surface for osteogenic differentiation of human bone marrow derived mesenchymal stem cells (hBMSCs). METHODS: We modified electrospun gelatine meshes with hydroxyapatite nanopowder. The potential beneficial impact of the ensuing culture conditions were evaluated by cultivating and comparing the growth of cells from osteoporotic and non-osteoporotic donors on either hydroxyapatite-gelatine (HA) meshes, pure gelatine meshes, or 2D standard tissue culture surfaces. RESULTS: After 21 days of differentiation, cells grown on pure or HA-gelatine meshes showed significantly higher mineralization levels compared to cells cultured in standard conditions. The amount of mineralization varied considerably in hBMSC cultures of individual patients but showed no significant difference between stem cells obtained from osteoporotic or non-osteoporotic donors. CONCLUSIONS: Overall, these results indicate that the use of HA-gelatine meshes as growth surfaces may serve as a valuable tool for cultivation and differentiation of mesenchymal stem cells along the osteogenic lineage, facilitating future research on osteoporosis and related issues.

Vitamin A acid in lichen planus.

Since 1971, Vitamin A acid (VAA) has been used for topical treatment of lichen planus (L.p.) at the 2nd Department of Dermatology in Vienna. During this period, 43 patients have been treated in the following way: for a period up to 3 weeks, a 0.05% or 0.1% VAA cream or solution was applied twice daily to the affected skin or mucosa. In the first days of this therapy, a characteristic local reaction of the L.p. lesions could be observed; first an edematous swelling appeared, followed by scaling and regression of efflorescences. Finally, an inflammatory skin reaction also developed. At this juncture, VAA was abandoned and the treatment was continued with steroid-containing ointments. Occlusive dressings were used only in cases of hypertrophic L.p. The same therapy was performed in L.p. of mucous membranes. By this method it was possible to achieve good therapeutical results in all treated cases. However, the frequency of recurrences of L.p. could not be diminished.

[Results of systemic treatment of lichen planus with a new aromatic retinoid (RO 10-9359) (author's transl)]. / Erfahrungen mit der systemischen Retinoidbehandlung (RO 10-9359) bei Lichen ruber planus.

24 patients suffering from generalized lichen planus were treated systemically with a aromatic retinoid (RO 10-9359). Initially a dose of 100 mg retinoid per day was administered for 2 to 3 weeks, then the daily intake was reduced slowly. The average duration of treatment was 72 days. This therapy produced a significant improvement in all patients. Lesions recurred during reduction of administered retinoid in 3 patients and after stopping treatment in 5 cases. Initially side effects consisted of cheilitis and exfoliation of the skin over the palmar and plantar regions; later 50% of the patients developed an effluvium. The results of the present study indicate that the aromatic retinoid can be considered as an alternative drug to systemic steroids in the treatment of lichen planus.

[Wild silk-induced asthma. A contribution to the knowledge of inhalation allergies caused by wild and tussah silk-filled bed quilts]. / Wildseidenasthma. Ein Beitrag zur Kenntnis der Inhalationsallergien verursacht durch Wild- bzw. Tussahseide-gefüllte Bettdecken.

In the silk industry occupational IgE-mediated hypersensitivities to different allergens are well known. Since the use of silk waste for the filling of bed quilts a great number of patients suffering especially from silk-asthma could be observed. Immunological investigations showed clearly that in this context sericin and also antigens from an insect of the genus anthrenus in the silk material are of allergenic importance. On the basis of typical case histories from the occupational and private scene the diagnostic difficulties of silk allergy are discussed.

[A comparative study of the value of skin tests and radio-allergo-sorbent tests in inhalation allergies (author's transl)]. / Untersuchungen über die Korrelation von Hauttest- und RAST-Ergebnissen bei Inhalationsallergien

Skin tests were performed and the specific IgE levels determined by means of the radio-allero-sorbent test (RAST) in fifty patients with allergic rhinitis/conjunctivitis and extrinsic bronchial asthma. An overall correlation between in vitro and in vivo diagnosis was found in 87% of cases. The results of this study indicate that in such allergies the RAST might well be of value as a screening test. In addition it is a valuable supplement to the skin tests, particularly in the elucidation of the diagnosis in problematic cases.

[Leg ulcer and allergic eczematous contact dermatitis incidence of contact allergies induced by topical therapy (author's transl)]. / Ulcus crurus und allergisches Kontaktekzem. Untersuchungen über die Häufigkeit lokalterapeutisch induzierter Kontaktallergien

In the IInd Department of Dermatology, Vienna University, 200 patients (143 female, 57 male) suffering from leg ulcers due to chronic venous insufficiency underwent patch testing with routine allergens, vehicles and their ingredients, as well as topical antibiotics. 114 patients (57%) had contact allergy. Balsam of Peru heads the list of allergens with postive reactions in 22%, closely followed by oil/water emulsions such as Drointin (21%), Ultrasicc (20%) and Ung. emulsificans aqu. (15,5%). The causative agents in these preparations were emulsifiers on the one hand (Lanette O, E, N) and preservatives (parabens) on the other hand. In comparison, allergic reactions to water/oil emulsions were less common. The low index of sensitivity to local antibiotics was also surprising. The results of the present study, in accordance with other reports in the literature, underline the need (1) for full label diclosure of cosmetics and topically-applied drugs and (2) for patch-testing every patient suffering from leg ulcers of long duration.

[Some observations on farmer's lung (author's transl)]. / Beitrag zur Kenntnis der "Farmerlunge".

Sera from 72 patients--29 females, 43 males--with symptoms suggestive of farmer's lung were tested by the Ouchterlony double diffusion technique using 10 different antigen solutions (extracts from mycetes and thermophilic actinomycetes, pigeon serum, extracts from pigeon droppings and Sitophilus granarius). In 23 cases precipitating antibodies were observed and Micropolyspora faeni was the most common antigen responsible for positive reactions. In 17 seropositive patients the diagnosis of exogen allergic alveolitis was established by clinical and laboratory findings. Measurements of immunoglobulins in these sera showed pronounced elevation of IgG and slightly elevated levels of IgA; IgM, IgE and C 3 c levels were in the normal range. The results are discussed with regard to the classification of farmer's lung as an occupational disease in Austria since December 30th, 1980.

[Comparative studies of the effectiveness of specific immunotherapy in house dust mite allergy]. / Vergleichende Untersuchungen zur Wirksamkeit einer spezifischen Immuntherapie bei Hausstaubmilben-Allergie.

In a prospective study, 60 patients with allergic rhinoconjunctivitis and/or asthma due to house dust mites were chosen for hyposensitization treatment with Migen (M) or Pharmalgen (P). Immunotherapy stretched over a whole year and every 3 months clinical results were evaluated by the patient's symptom score, by results of skin prick and conjunctival provocation tests, as well as by RIA and ELISA regarding the total and specific IgE and also specific IgG and IgG4 levels. Out of 30 patients of the M group, 15 were followed up over the whole therapeutic regimen, 4 of whom showed a very good, 7 a good to moderate clinical outcome and 4 showed no improvement at all. In the P group, 17 out of 30 patients were followed up whereby 9 showed a very good and 8 a good to moderate response. In both groups of patients a statistically significant decrease in skin and conjunctival sensitivity to mite allergens was observed after 12 months of therapy. However, there was no correlation between this observation and the failure or success of immunotherapy. Furthermore, in both groups there was significant increase in total and specific IgE (with a slight decrease after 6 to 12 months) and also in specific IgG and IgG4 (especially in the P group), but again these changes in antibody levels gave no indication of a good or bad clinical outcome. Hence, we believe other reasons than the usually presented thesis of inducing "blocking antibodies" by immunotherapy to be responsible for the well-known effects of hyposensitization.

[The implication of trace metals in household products in the causation of contact dermatitis (author's transl)]. / Zur allergologischen Bedeutung von Metallspuren in Haushaltsprodukten.

The 6 most common fluid and powder detergent brands in use in Austria were analyzed as to their nickel and chromium content. The analysis was carried out by means of atomic absorption spectroscopy. 1. The above-mentioned metals were found to be present in each of the products analysed. On average, the fluid detergents contained ten times less nickel and chromium than the powders. 2. Repeated investigations revealed a considerable variation in metal ion content in different batches of the same detergent. Thus, presumably the raw products already contain nickel and chromium and it is unlikely that these ions get into the products during the manufacturing process. 3. In the appropriately diluted detergent powder solutions, as recommended by the producer, the content of nickel was found to lie between 0.4 ppm and 0.717 ppm and the chromium content ranged from 0.733 to 0.917 ppm. Since powder detergents also contain complex-producing agents, it can be assumed that these concentrations of metal ions do not suffice for sensitization. In the fluid detergents the nickel and chromium contents were found to be below the safety limit for the elicitation of an excematous reaction.

[Extrinsic allergic alveolitis: comparison of 2 methods (Ouchterlony gel precipitation and ELISA) for antibody detection in routine diagnosis]. / Exogen-allergische Alveolitis: Vergleich zweier Methoden (Gelpräzipitation nach Ouchterlony und ELISA) für den Antikörpernachweis in der Routinediagnostik.

Sera from 58 patients with suspected extrinsic allergic alveolitis (EAA) were tested in parallel by the double diffusion test and the "enzyme-linked immunosorbent assay (ELISA)" using 10 different antigen solutions (extracts from thermophilic actinomycetes, various mycetes, pigeon serum, pigeon droppings and the wheat weevil sitophilus granarius). The ELISA technique was more sensitive than the double diffusion test in detecting antibodies to the panel of antigens used. However, by using several antigen dilutions the number of sera positive sera with the double diffusion test increased and an overall correlation of 93.1% with the ELISA was achieved. The results of the study are discussed with regard to the advantages and disadvantages of both test systems for the in vitro diagnosis of EAA.

Incidence and costs of osteoporosis-associated hip fractures in Austria.

This study is an investigation into the epidemiologic and socioeconomic impact of osteoporosis-associated hip fractures in Austria. We determined age- and gender-specific incidence rates of osteoporosis-associated hip fractures for all patients treated in hospitals in 1995 and calculated mortality rates, hospitalization days and direct costs of hospitalization. The data were obtained from the hospital discharge statistics for all general hospitals and for all hospitals of the General Austrian Accident Insurance. To calculate the portion of hip fractures attributable to osteoporosis in a given age-group, a basic, non-osteoporotic incidence of hip fractures was determined for ages 20-39, using gender-specific regression models. 11,379 patients with osteoporotic hip fractures underwent treatment in Austrian hospitals in 1995, accounting for 79 percent of all hip fracture patients treated. 82 percent of those were female, with the highest incidence among women aged 95 years and older with a rate of 3,000/100,000. For male patients the highest incidence was observed for the age-group of 90-94 years with 1,743/100,000. International comparisons indicate these incidence rates to be similar to those reported for the Swiss population. In 1995, 778 patients or 6.8 percent of all patients with osteoporotic hip fractures died during hospitalization. Hospital care of patients with osteoporotic hip fractures required an overall 250,268 bed-days with an age-group-specific length of stay between 8.5-27 days for female and 16-23 days for male patients. The total cost of hospital treatment of osteoporotic hip fractures in Austria was ATS 1,043,379,000 (US$ 103,509,800), with average costs per patient of ATS 91,700 (US$ 9,097). Due to the aging of the population in the years to come, an increase of osteoporotic hip fractures among individuals aged 50 years and older must be expected. The economic importance of this development and its impact on the health care system must be considered as significant.

[Quality control in the allergy laboratory--4 years "Allergy Ring Trial" in Austria]. / Qualitätskontrolle im Allergielabor--4 Jahre "Ringversuch Allergie" in Osterreich.

In 1988 the Austrian Society for Allergology and Immunology initiated an external quality control program for the in vitro allergy diagnosis. In 12 mailings, 62 sera from allergic patients were sent to selected laboratories in order to determine total and antigen specific IgE according to the laboratory-specific methods. The values for total IgE varied considerably (73.9% were within +/- 1 SD, 94.3% within +/- 2 SD and 99.1% within +/- 2 SD, but only 0.9% beyond that). Sources of error were mainly attributable to inappropriate equipment and low quality reagents, but also bad test performance, with respect to personnel and the routines. In contrast, antigen specific IgE against pollen, mites, moulds, insect venoms, animal danders, drugs, parasites, environmental and food antigens revealed quite homogeneous results. Out of 1492 data, only 46 (3.1%) had to be declared as "wrong", and the variability of the RAST-classes was low. Whereas the quality of the reagents from all the different suppliers was not absolutely reliable at the beginning of the study, it improved considerably with time, as consequence of our complaints. The comparability of the methods for detecting total-IgE were non-satisfactory, whereas those for antigen specific IgE were generally good. The variety of methods employed (radio- and enzyme immunologic, fluoro- and nephelometric methods, etc.) should entail appropriate consequences, especially critical comparisons within one and the same laboratory; in addition, international standardization of the "normal" values should replace company-standards, and quality control programs for each test system before it is marketed should be mandatory. Our results confirm, that external control should be obligatory for any laboratory.(ABSTRACT TRUNCATED AT 250 WORDS)

[Oral hyposensitization (author's transl)]. / Orale Hyposensibilisierung.

A review of literature concerning the theoretical and practical aspects of oral hyposensitization is presented. This method is in the moment the most recommendable in the treatment of type-I allergy of children.

Clinical value of cervical mediastinoscopy in the staging of bronchial carcinoma.

Whether mediastinoscopy is indicated in the preoperative staging of bronchogenic carcinoma is still a controversial issue. It may be performed routinely (to exclude locally inoperable patients from surgery), selectively, or it may be regarded as superfluous (in centers which prefer an extended lympho adenectomy at the time of thoracotomy). We regard mediastinoscopy as indicated for the following purposes: 1) staging of NSCLC and SCLC; 2) diagnostic (mediastinal masses or lung tumors without previous histology); 3) restaging after primary chemotherapy; 4) assessment of prognosis in patients with borderline operability. The indication for 224 mediastinoscopies performed at our institution in the period from September 1991 through March 1999 was mainly for staging (59.2%) or diagnostic (30.6%). Eight (5.4%) patients underwent mediastinoscopy for the assessment of operability, and 7 (4.8%) after primary chemotherapy for the restaging of loco-regionally advanced lung cancer. Sensitivity and specificity rates were 87% and 100%, respectively, with an accuracy of 93% for the mediastinoscopy performed for the staging of lung cancer at all stages. If we consider the N2 tumors (42 cases) alone, the sensitivity was 76.7% and the specificity 100%, with an accuracy of 83.3%. Overall positive and negative predictive value resulted 100% and 87%, respectively, according to the data reported in literature. Our data confirm the role of mediastinoscopy as the gold standard for regional staging of lung cancer.