Rapid Onset and Sustained Efficacy (ROSE) study: results of a randomised, multicentre trial comparing the effect of zoledronic acid or alendronate on bone metabolism in postmenopausal women with low bone mass.

SUMMARY: The ROSE study compared a once-yearly intravenous dose of zoledronic acid with a once-weekly oral dose of alendronate in postmenopausal women. Once-yearly zoledronic acid showed a greater and faster reduction in the levels of two markers of bone turnover and may be an effective option for the treatment of osteoporosis. INTRODUCTION: The open-label Rapid Onset and Sustained Efficacy (ROSE) study was designed to compare a once-yearly intravenous (iv) dose of zoledronic acid with a once-weekly oral dose of alendronate with respect to markers of bone turnover in approximately 600 postmenopausal women in Germany. METHODS: Levels of N-telopeptide of collagen type I (NTx) and procollagen 1 C terminal extension peptide (P1NP) were assessed during the study. The primary objective was to assess if zoledronic acid was superior to alendronate in reducing serum NTx levels after 12 months' treatment. RESULTS: A significantly greater reduction in NTx levels from baseline to month 12 (as determined by the area under the curve) was observed in patients treated with zoledronic acid (n = 408) versus those receiving alendronate (n = 196; 0.282 ng/mL vs. 0.270 ng/mL; P = 0.012). The reduction in levels of P1NP after 1 year was also significantly greater in patients treated with zoledronic acid compared with those receiving alendronate (28.21 vs. 25.53 ng/mL; P = 0.0024). The overall incidence of adverse events was similar between groups; both treatments were generally well tolerated. Although post-dose symptoms, including the incidence of influenza-like symptoms, were higher with zoledronic acid than alendronate initially, the incidence was similar between groups from days 4-360. Gastrointestinal symptoms were more frequent with alendronate than zoledronic acid throughout the study. CONCLUSION: In this study, once-yearly iv zoledronic acid provided a greater and faster reduction in the levels of NTx and P1NP versus once-weekly oral alendronate.

A syringe adapter to facilitate aspiration at amniocentesis.

BACKGROUND: Syringes were developed for injection, not aspiration. We describe an adapter that attaches to a standard syringe and aspirates by advancement rather than withdrawal. TECHNIQUE: The adapter consists of a syringe barrel extension and an attachable finger grip. The adapter attached to a standard syringe allows aspiration by advancement rather than withdrawal, with the same hand motion used as with injection. EXPERIENCE: In a comparison of timed in vitro fluid aspiration by 10 practitioners, mean times for aspiration of 15 mL were 32.4 seconds (range 24-45) with standard technique and 25.3 seconds (range 24-30) with the adapted syringe (P < .003, paired t test). Thirteen of 14 physicians who used the adapted syringe in vivo reported that much less effort was required, and one physician reported that a little less effort was required. Overall, eight practitioners reported that the adapted syringe made amniocentesis much easier and six found no difference between the two devices. CONCLUSION: This adapter converts a standard syringe to one permitting aspiration by advancement rather than withdrawal. Practitioner feedback was positive with regard to ease of use, degree of effort, and overall satisfaction.

Activation and enzyme characteristics of a DNA-restrained phosphatase in chromatin-associated complexes.

DNA-bound polypeptide complexes composed of several non-histone polypeptides that resisted harsh DNA deproteinization procedures were characterized. The three major polypeptides of these complexes have molecular masses of 62, 52, and 40 kDa. They constitute supramolecular structures that reside on isolated DNA in dense clusters. The supramolecular complexes were released from DNA as globular 12.8 +/- 0.8-nm particles; these particles were gradually disassembled to form smaller supramolecular structures. The DNA-bound complexes comprise of an encrypted adenosinetriphosphatase/phosphatase activity, which is a minor but intrinsic component of the complexes. The enzyme remained inactive as long as the complexes were bound to DNA. However, the enzyme was activated concomitantly with the progression of DNA digestion, which indicated that DNA was involved in the downregulation of the enzyme. The inactive DNA-restrained complex could not be restored in vitro, which indicated its non-trivial nature. Once released from DNA, the enzyme was inactivated over a period of several hours. However, in the DNA-associated complexes its potential to become activated during DNA digestion was conserved for several months. In the activated state, the enzyme showed an optimum activity at pH 9.5, was stimulated by Mg2+, inhibited by vanadate and EDTA, but was not significantly inhibited by okadaic acid. The active enzyme, which consists of two subunits of 56 kDa and 59 kDa, can be released from the supramolecular structures by agarose gel electrophoresis. A regulatory mechanism therefore exists for the downregulation of this phosphatase by DNA.

Detection of oestrus in dairy cows: a new approach to an old problem.

Fertility indices, such as non-return rate and number of services per conception, have worsened significantly over the past few years. Because oestrus detection is one of the major factors influencing these indices, this study was performed to determine whether, how long, and at what time dairy cows show signs of oestrus in their behaviour. Two high producing herds with about 40 open cows were observed for 30 minutes every 2 hours for 6 weeks. The various signs of oestrus were recorded. The milk progesterone concentration was monitored daily to detect ovulation. A scale (Table 2) was composed according to the frequency of these symptoms during and between oestrous periods. This scale was used in order to determine which cows were in oestrus. Each oestrous symptom was given a score and the scores were summed for a 24-hour period. If a score of > 100 points was reached within a 24-h period, the cow was considered to be in oestrus. As used, the oestrus detection scale proved to be very convenient. Because standing oestrus was observed in only 37% of the oestri, this criterion had a very low sensitivity for oestrus detection. With the use of this scale and 12 observations per day, we reached a detection rate of 100%. If a more practical observation scheme would have been used (e.g. observing the herd twice daily, for 30 min) lower threshold values would have been obtained. This observation scheme resulted in a detection rate of 74% with no incorrect assessments.

Graft-versus-leukaemia activity can be predicted by natural cytotoxicity against leukaemia cells.

We have investigated graft-versus-leukaemia (GVL) effects after allogeneic bone marrow transplantation (BMT), using three murine leukaemia models, A20 (B lymphocytic), WEHI-3 (myelomonocytic) and PU5-1R (myeloid). Injection of leukaemia cells in a high number (10(6) cells) into syngeneic Balb/c mice (H-2d) invariably led to death with a median survival time of 22 d (A20), 18 d (WEHI-3) and 45 d (PU5-1R). A lower tumour load of A20 (5 x 10(5) cells) was used in some experiments resulting in a leukaemic death rate of 94%. Lethal total-body irradiation followed by syngeneic BMT prolonged survival (P<0.05) for animals bearing the leukaemia A20 and WEHI-3 but was unsuccessful for animals injected with cells from the monocytic leukaemia PU5-1R. Graft-versus-host (GVH)-nonreactive marrow of (C57 x Balb/c)F1 mice (H2bxd) exerted a significant GVL-effect with reduced relapse rate and improved survival in mice receiving the leukaemia cell line A20. In animals with low tumour load a significant reduction of the relapse rate from 82% following syngeneic BMT to 47% following allogeneic, GVH-nonreactive BMT could be achieved. Depletion of natural killer (NK) cells from the GVL-reactive semi-allogenic bone marrow graft enhances the relapse rate of the leukaemia A20 to 65%. In mice bearing the leukaemias WEHI-3 or PU5-1R allogeneic GVH-nonreactive BMT did not improve survival compared to syngeneic BMT. Transplantation of GVH-reactive bone marrow from DBA mice (MHC identical to Balb/c, minor difference) caused only a limited and insignificant reduction of relapse rate for animals with the leukaemia A20. These in vivo data are in close correlation with in vitro natural killer cell (NK) activity of the donor strains against the respective leukaemia targets. Depletion of NK cells from the GVL-reactive (C57 x Balb/c)F1 bone marrow resulted in a significant loss of GVL activity. We conclude that NK cells are involved in graft-versus-leukaemia effects independent of graft-versus-host disease (GVHD).

Multiple organ manifestations in thromboangiitis obliterans (Buerger's disease). A case report.

Thromboangiitis obliterans (TAO) occurs almost exclusively in young male smokers. Its involvement of the small and medium-sized arteries and veins leads to ischemic complaints and/or changes in the extremities. The possibility of organ involvement is a matter of controversy. The authors report a case of TAO with multiple organ involvement, including myocardial, splenic, and cerebral infarctions; pulmonary embolisms; and probable intestinal ischemia during a twenty-three-year course.

Baseline transepidermal water loss in patients with acute and healed irritant contact dermatitis.

To examine the skin barrier function of patients with acute and healed irritant contact dermatitis (n = 80) baseline transepidermal water loss (TEWL) was quantitatively measured using an evaporimeter. Healthy subjects served as controls (n = 40). Test areas were the forearm and the thigh. A significant increase in TEWL was observed in the patients with acute and with healed irritant contact dermatitis (ICD) as compared to healthy volunteers (P < or = 0.01). TEWL values in both test areas were comparable and markedly correlated (P < or = 0.01) with each other in every group. Thus, it is possible that basal TEWL depends more on the intrinsic skin barrier function of the subjects rather than the 2 anatomical regions examined. TEWL at the forearm with acute ICD was significantly higher (P < or = 0.01) than that of the group with healed ICD, but not for TEWL at the thigh suggesting that ICD may aggravate the barrier function of the adjacent uninvolved skin. It is assumed, that increased basal TEWL in patients with ICD may reflect a constitutional deviation of epidermal barrier function. This event seems to be comparable with the well-known symptom of atopic individuals. Using a detailed atopic scoring system in such a study may clarify the question of whether a proportion of patients with hand ICD may indeed be atopic individuals.

Influence of donor lymphocytes on the incidence of primary graft failure after allogeneic bone marrow transplantation in a murine model.

We used a murine model to determine the impact of donor lymphocyte subsets on the incidence of primary marrow graft failure after transplantation of lymphocyte-depleted bone marrow. After lethal irradiation with 7.5 Gy, Balb/c mice received 1 x 10(5) to 4 x 10(7) GvH-nonreactive (C57 x Balb)F1 or GvH-reactive C57Bl/6 marrow cells. Pretreatment with anti-Thy-1.2, anti-CD4/CD8, anti-asialo-GM1 or L-leucyl-L-leucine methyl ester (Leu-Leu-OMe) was employed to eliminate T lymphocytes and/or natural killer cells. Primary graft failure was defined as death with neutrophils < 0.5 x 10(9)/l. To assess long-term chimaerism, the percentage of H-2b-positive spleen cells was determined. Pretreatment with anti-Thy-1.2, anti-CD4/CD8 or Leu-Leu-OMe successfully eliminated GvHR-induced mortality. Graft failure rates gradually declined from 88% after transplantation of 1 x 10(5) cells to 0% after transplantation of 4 x 10(7) C57Bl/6 cells. The incidence of graft failure, however, was not altered by T-cell depletion, provided that the unspecific loss of marrow cells was compensated for. After transplantation of GvH-nonreactive (C57 x Balb)F1 bone marrow, neither ex-vivo treatment with anti-Thy-1.2 and anti-asialo GM1 nor addition of 1 x 10(7) donor thymocytes to the allograft significantly influenced engraftment. The data obtained in our animal model suggest that the total number of marrow cells is of critical importance for successful marrow engraftment and not the presence or absence of T cells, NK cells or GvHR.

Graft-versus-leukemia activity after bone marrow transplantation does not require graft-versus-host disease.

Clinical data have suggested that graft-versus-host disease (GVHD) plays a crucial role in the antileukemic effects of bone marrow grafts. We investigated (a) whether bone marrow cells unable to induce GVHD can effect graft-versus-leukemia (GVL) activity and (b) whether such antileukemic capacity depends on the presence of T lymphocytes in the graft. Balb/c mice were inoculated with A20 cells, a B-cell lymphoma/leukemia of Balb/c origin. Four weeks after tumor inoculation the animals were lethally irradiated and received a bone marrow graft. Cells from (Balb/c x C57) F1 or (C3H x Balb/c) F1 hybrids were transplanted into parental-strain Balb/c mice. Since lymphocytes from F1 hybrids are unable to cause graft-versus-host reactivity against a parental-strain animal, we used this experimental setting to explore GVL effects in a GVHD-free system. In vitro incubation with monoclonal anti-Thy-1.2 antibody plus complement was used to eliminate Thy-1+ cells. After syngeneic transplantation, the death rate due to leukemia remained unchanged (91%) compared with that among untreated animals (86%). Following transplantation of F1 marrow cells of either (C57 x Balb/c) F1 or (C3H x Balb/c) F1 origin, death rates of 40% and 50% were observed; these were significantly lower. Depletion of Thy 1+ cells from bone marrow graft caused only a slight increase in the leukemic death rate after transplantation of bone marrow of (C57 x Balb/c) F1 hybrid origin (50%), but a high leukemic death rate was seen after transplantation of (C3H x Balb/c) F1 bone marrow (100%). Additional experiments with fully allogeneic, T-cell-depleted C57 bone marrow transplantation suggest an antileukemic effect that is comparable to that seen after transplantation of unmanipulated F1 bone marrow. Taken together, our results indicate that GVL activity can be dissociated from graft-versus-host reaction.

Influence of cell dose and graft-versus-host reactivity on rejection rates after allogeneic bone marrow transplantation.

The number of cells transplanted and their capacity to induce graft-versus-host reactivity (GvHR) are two factors that are suspected to influence the engraftment of allogeneic bone marrow. We have investigated their impact on graft rejection rates in busulfan-treated LEW rats. In a series of experiments, we varied (1) the number of marrow cells transferred (1, 5, 10, 20, 30, and 40 x 10(7)), (2) the degree of pretransplant immunosuppression (1.5, 3.0, and 4.5 Gy of total body irradiation [TBI]; 0, 30, 60, 90, 120, and 180 mg/kg cyclophosphamide), and (3) the ability of the marrow graft to induce classical GvHR against major histocompatibility complex (MHC) antigens [semiallogeneic (CAP x LEW)F1 or CAP rats as marrow donors]. Reducing either the immunosuppressive pretreatment or the number of cells transplanted resulted in a stepwise increase in rejection rates. However, every reduction in the size of the marrow inoculum was compensated by increased immunosuppression and vice versa. While 60 mg/kg cyclophosphamide was sufficient to prevent rejections after grafting of 40 x 10(7) cells, 90 mg/kg was necessary to ensure 100% engraftment after transplantation of 20 x 10(7) cells, 120 mg/kg after 10 x 10(7) cells, and 180 mg/kg after 1 x 10(7) cells. Since CAP marrow leads to GvHR-mediated immunosuppression in LEW recipients, in contrast to (CAP x LEW)F1 marrow, we had supposed that lower cell numbers or cyclophosphamide doses are sufficient to achieve engraftment of CAP marrow. Although severe GvHR was present in all animals receiving escalating doses of CAP cells, the rejection rates were the same as for (CAP x LEW)F1 marrow. In conclusion, we have demonstrated that there is a sensitive balance between the immunosuppression of the host and the number of marrow cells transferred. We were not able to number of marrow cells transferred. We were not able to detect a beneficial effect of classical GvHR against MHC antigens on the engraftment of allogeneic marrow. Thus, our results do not support the hypothesis that the loss of GvHR-mediated immunosuppression is responsible for higher rejection rates following transplantation of T-cell-depleted bone marrow.

Effect of post-transplant methotrexate, cyclosporin A and prednisolone on graft rejection after allogeneic bone marrow transplantation.

Methotrexate, cyclosporin A and prednisolone have been shown to improve graft survival rates in solid organ transplantation. However, little is known concerning their capacity to promote lasting engraftment of allogeneic bone marrow. Therefore, we tested these agents in LEW rats receiving MHC-mismatched marrow after pretreatment with a myeloablative dose of busulfan plus different doses of total body irradiation (TBI). To avoid mortality due to graft-versus-host reaction (GHVR), F1(CAP x LEW) marrow was transferred. Hematological parameters were determined twice weekly to monitor engraftment and rejection. The pretreatment was lethal but not sufficiently immunosuppressive to ensure lasting engraftment in all animals. Thus, post-transplant immunosuppressive protocols could be evaluated for their capacity to improve engraftment rates. Standard clinical doses of methotrexate (0.25 mg/kg i.p. day 1, 3, 6, 11, 18, 25), cyclosporin A (10 mg/kg orally day 0-28) and prednisolone (1 mg/kg i.p. day 0-28) were administered and proved to be of nearly equivalent toxicity in our system. All three agents failed to allow engraftment after busulfan alone. After additional conditioning with 1.5 Gy of TBI, methotrexate and cyclosporin A reduced the rejection rate from 100% to 59% and 70%, respectively. When 3 Gy of TBI were added to busulfan, cyclosporin A and prednisolone were able to reduce the rejection rate from 67% to 33% and 39%, respectively, whereas 0.12 and 0.25 mg/kg methotrexate completely prevented graft rejections. After cessation of cyclosporin A therapy, late secondary rejections were frequently observed. These results demonstrate that postgrafting immunosuppression with protocols conventionally used for the prophylaxis of GVHR is able to facilitate lasting engraftment of MHC-mismatched bone marrow.(ABSTRACT TRUNCATED AT 250 WORDS)

Crystalline and amorphous gold in chrysiasis.

Skin biopsy specimens from five patients (three females and two males) treated parenterally with gold were investigated using transmission electron microscopy. X-ray microanalysis and electron diffraction were used to determine the dermal heavy metal content. Additional sections were stained for light microscopic examination. The amount of elemental gold administered to the patients over a period of years to alleviate rheumatoid arthritis lay between a minimum of 4.0 g and a maximum of 10.0 g. In one and the same patient dermal histiocytic gold aggregations in sun-exposed areas of skin displayed a different pattern and divergent physiochemical states from the gold deposits in non-UV-exposed skin, where aurosome-like amorphous formations are found in the cells of the upper dermis. Additional spherical particles are associated predominantly with phagolysosomes in melanophages beneath solar-irradiated epidermis. Convergent beam electron diffraction proves the crystalline nature of the spherical auriferous deposits. The occurrence of skin rash was not related to different physicochemical states of the precious metal.

Exercise performance of hemodialysis patients during short-term and prolonged exposure to altitude.

The work capacity of patients on maintenance hemodialysis is impaired even at normal inspiratory oxygen pressure. A further restriction can be expected when these patients are exposed to hypoxia at altitude, since most of the usual compensatory mechanisms required to adjust to this environment are impaired or even missing. We tested the tolerance of hemodialysis patients to hypoxia and measured work capacity, hematological, and cardiovascular parameters at rest and during incremental bicycle ergometry during 3-hour exposure to altitudes of 2,000 m and 3,000 m, and during 2 weeks of exposure to an altitude of 2,000 m and compared these data with prealtitude values or with data evaluated in a control group, respectively. In control tests the patients reached work loads at exercise termination of about 66% of age and sex-matched healthy controls, the reduction correlated well with the degree of anemia. During short-term altitude exposure to 2,000 m peak work performance remained unchanged in comparison to prealtitude tests, whereas at 3,000 m it was reduced by about 12%. During the 2-week stay at 2,000 m peak work loads increased significantly by 17% accompanied by an increase in peak oxygen uptake (+15%), blood lactate, heart rates (+10 min-1), and systolic blood pressure (+20 mmHg), whereas the diastolic pressure was comparable to prealtitude values. In another group of hemodialysis patients studied at low altitude under similar experimental conditions none of these parameters was changed. Our data show that during acute exposure to altitudes up to 2,000 m maximal work of hemodialysis patients is not reduced, but is restricted at altitudes higher than that.(ABSTRACT TRUNCATED AT 250 WORDS)

Tobacco-free schools in Minnesota.

Since 1985, Minnesota schools have participated in an intensive, statewide effort to reduce tobacco use among adolescents through a variety of prevention efforts. This article describes the "Tobacco-Free Schools" project, one component of the effort that encourages school districts to adopt policies which prohibit tobacco use for students, staff, and visitors. Tobacco-free policies contribute to comfortable, healthy working and learning environments for students and staff. In addition, tobacco use prevention education is more likely to be effective when health education programs, adult models, and school policies offer the consistent message that tobacco use is unhealthy and unacceptable. From January 1987 to January 1988, the percentage of tobacco-free school districts in Minnesota increased from 3% to 43%, demonstrating that tobacco-free policies can have mass appeal to local schools and communities.

Differential inhibition of host cell RNA synthesis in several picornavirus-infected cell lines.

Patterns of cellular RNA synthesis during poliovirus (types 1,2 and 3) and mengovirus infections in HeLa, HEp-2, Vero and L-cells were investigated by means of high-resolution autoradiography, kinetics of 3H-uridine incorporation, and sucrose gradient centrifugation. Only mengovirus-infected L-cells showed an early and exponential inhibition of hnRNA and rRNA synthesis. The other cell lines tested, whether infected with polio- or mengovirus, exhibited a slowly and linearly decreasing cellular RNA synthesis, with hnRNA production suppressed completely, although somewhat later than in L-cells, and rRNA synthesis continuing at a gradually reduced rate during the entire viral growth cycle. Locations of cleavage steps of rRNA precursor molecules are discussed.