Antimicrobial Susceptibility Patterns in Urinary Tract Infections in Dogs (2010-2013).

BACKGROUND: Urinary tract infections (UTIs) are common in dogs. The responsible bacterial populations have evolved with increasing resistance to many antimicrobials. OBJECTIVE: To characterize the antimicrobial susceptibility patterns of canine urinary tract isolates over a 51-month period. ANIMALS: One thousand six hundred and thirty-six bacterial isolates from 1,028 dogs. METHODS: Aerobic bacterial isolate growth and susceptibility data from urine cultures of dogs were identified, retrospectively. Medical records were reviewed to obtain signalment, comorbidities, and antimicrobial use in the previous 30 days. The UTIs were further categorized as uncomplicated, complicated, or pyelonephritis. RESULTS: Common bacterial isolates identified were Escherichia coli (52.5%), Staphylococcus spp. (13.6%), and Enterococcus spp. (13.3%). In vitro susceptibility among all isolates varied for commonly prescribed antimicrobials (amoxicillin [59%], amoxicillin/clavulanic acid [76%], cephalexin [66%], enrofloxacin [74%] and trimethoprim-sulfamethoxazole [86%]). For all antimicrobials tested (except aminoglycosides), in vitro susceptibility was higher in uncomplicated versus complicated infections (P < .05). Uncomplicated infection isolate susceptibility rates remained ≤90% for PO administered antimicrobials. Administration of amoxicillin, doxycycline, and enrofloxacin, but not amoxicillin/clavulanic acid in the previous 30 days was associated with resistance to that antimicrobial. Multidrug resistant isolates of E. coli and Staphylococcus spp. were more common in dogs with complicated than uncomplicated UTIs (36% versus 21%, P < .0001). CONCLUSIONS AND CLINICAL IMPORTANCE: In vitro susceptibility was highly variable and no PO administered antimicrobial had >90% efficacy among isolates tested. Multidrug resistance was frequent among isolates tested suggesting that routine culture and susceptibility testing is indicated. Previously prescribed antimicrobials may affect empirical choices made pending susceptibility testing.

Accuracy of potassium supplementation of fluids administered intravenously.

BACKGROUND: Potassium (K+) supplementation of isotonic crystalloid fluids in daily fluid therapy is commonly performed, yet its accuracy in veterinary medicine is undetermined. OBJECTIVE: To investigate the accuracy of K+ supplementation in isotonic crystalloid fluids. ANIMALS: None. METHODS: Observational study. 210 bags of fluid supplemented with KCl being administered to hospitalized dogs and cats intravenously (IV) were sampled over a 3-month period. Measured K+ concentration ([K+]) was compared to the intended [K+] of the bag. In a second experiment, 60 stock fluid bags were supplemented to achieve a concentration of 20 mmol/L K+, mixed well and [K+] was measured. In another 12 bags of 0.9% NaCl, K+ was added without mixing the bag, and [K+ ] of the delivered fluid was measured at regular time points during constant rate infusion. RESULTS: The measured [K+] was significantly higher than intended [K+] (mean difference 9.0 mmol/L, range 6.5 to >280 mmol/L, P < .0001). In 28% of clinical samples measured [K+] was ≥5 mmol/L different than intended [K+]. With adequate mixing, K+ supplementation of fluids can be accurate with the mean difference between measured and intended [K+] of 0.7 (95% CI -0.32 to 1.7) mmol/L. When not mixed, K(+) supplementation of 20 mmol/L can lead to very high [K+] of delivered fluid (up to 1410 mmol/L). CONCLUSIONS AND CLINICAL IMPORTANCE: Inadequate mixing following K+ supplementation of fluid bags can lead to potentially life threatening IV infused [K+]. Standard protocols for K+ supplementation should be established to ensure adequate mixing.

Trials of miscellaneous interventions to wean from mechanical ventilation.

We found eight randomized controlled trials (RCTs) of miscellaneous interventions that were designed to facilitate the process of weaning from mechanical ventilation. The two RCTs of high-fat/low-carbohydrate enteral nutrition found favorable physiologic effects on CO2 production and respiratory quotient, rendering this type of nutrition potentially useful in patients with impaired ventilatory reserve; however, no conclusions can be made about the outcomes of the duration of ventilation and weaning success. The two RCTs of postextubation use of noninvasive ventilation are conflicting, showing potential short-term physiologic benefit in one study, but no benefit in terms of reintubation rates or other morbidity. These RCTs are less promising than other applications of noninvasive ventilation such as those in patients with COPD exacerbations. One RCT showed no improvement in success of weaning with exogenous growth hormone administration. In the setting of very frequent baseline blood gas analyses, one RCT of oximetry and capnography was associated with significantly fewer blood gas analyses. Biofeedback to enhance safe and rapid weaning showed a dramatically lower duration of ventilation in one RCT that did not report the weaning methods used. One RCT of preextubation acupuncture showed lower rates of laryngospasm in the acupuncture group. Overall, these studies were underpowered for clinically important outcomes. Multidisciplinary, patient-centered, holistic, and non-pulmonary approaches to weaning may provide additional safe, effective adjunctive methods of hastening liberation from mechanical ventilation.

Could plasmid-mediated gene transfer into the myocardium be augmented by left ventricular guided laser myocardial injury?

Early studies have indicated no correlation between the amount of mechanical injury and the level of myocardial gene expression following direct plasmid vector injection. Recently, however, evidence suggests that combined laser myocardial injury and plasmid-based gene delivery exert synergistic effects on gene expression and activity. The purpose of the study was to determine whether laser-induced myocardial injury followed by transendocardial gene transfer increases gene expression compared to gene transfer alone. We assessed the ability of a plasmid vector to express its transgene after injection into porcine ischemic myocardium with and without preceding laser myocardial injury. Thirteen animals had transendocardial injections of the luciferase reporter gene in a plamid vector using a catheter-based injection system. Injections (0.5 mg per animal, 50 microg per injection site) were divided into 10 sites in the ischemic territory. Eight animals underwent transendocardial laser injury of the ischemic region (2 Joule per pulse x 10 sites) prior to gene delivery. In five animals, gene injection sites were dispersed between laser channels, and in three animals laser and gene delivery were applied in close proximity (< 5 mm) or at the same location. Luciferase activity was measured at 3 and 7 days. Luciferase expression in ischemic zones was markedly elevated at day 3 and 7, and similar whether animals were pretreated using laser injury followed by gene transfer compared to gene transfer alone. Neither same-spot injection nor dispersed gene delivery were associated with augmented gene expression compared to gene transfer alone. Using the above-described catheter-based approach to combine localized laser injury and injection of naked DNA into ischemic myocardium, laser injury did not augment gene expression above levels present with gene transfer alone.

Effect of the interaction of parathyroid hormone and cyclosporine a on bone mineral metabolism in the rat.

Cyclosporine A (CsA) induces high turnover osteopenia in the rat and there is evidence for this in humans. Recent studies suggest that increases in parathyroid hormone (PTH) may be involved in posttransplantation bone loss. However, human studies are difficult to interpret since transplant patients usually receive a cocktail of immunosuppressants and have underlying disease. Our aim was to try to resolve the influence of the absence or presence of PTH on CsA-induced bone disease. Male Sprague Dawley rats aged 7-9 months, either sham operated or parathyroidectomized (PTX), were randomly divided into vehicle and CsA groups. All PTX rats were given oral calcium supplementation ad libitum. The rats were divided into groups: basal, sham/vehicle, sham/CsA, PTX/vehicle, and PTX/CsA. Serial biochemistry was performed 0, 14, and 28 days after the start of the experimental period; bone histomorphometry was performed 28 days after the start of the experimental period. Statistical analysis consisted of group comparisons and factorial analyses. The results showed that CsA alone produced a high turnover osteopenia consistent with previous studies. In the PTX animals there was an increase in bone mass. PTX also decreased osteoblast activity and recruitment, and serum 1,25OH2D levels. Serum levels of osteocalcin (BGP) were unaffected by PTX. The combination group (PTX/CsA) did not differ statistically from the controls in most of the histomorphometric parameters measured, with the exception of reduced mineral apposition and bone formation rates, reflecting the effects of PTX. Serum BGP and 1,25OH2D levels did not differ, but PTH was reduced from the control. Explanations for these results are (1) CsA and PTX exert their effects via separate mechanisms, negating each other; (2) in the absence of PTH, CsA managed to cause bone loss, and thus PTH may not be essential for CsA-induced bone loss; or (3) the profound accelerated bone loss produced by CsA in normal rats requires PTH. These findings may help explain the discrepancies found in clinical studies where bone loss occurs with either elevated or normal PTH levels.

Immunosuppressant use without bone loss--implications for bone loss after transplantation.

Cyclosporine A (CsA) is associated with posttransplantation bone disease. Immunosuppressant drugs such as sirolimus (SRL), which are more potent and less deleterious than CsA, are being developed. Previous experiments have shown that SRL although immunosuppressive, is relatively bone sparing. The use of low doses of CsA and SRL in combination has displayed in vivo synergism. This study was initiated to examine the effect of low-dose CsA and SRL on bone metabolism, thereby hopefully providing a bone sparing immunosuppressive regimen for transplant recipients. One hundred and nineteen rats were divided into groups: basal, vehicle, CsA high dose, CsA low dose, SRL low dose, and combination low-dose CsA and SRL. The basal group was killed on day 0 for histomorphometry. The experimental groups were weighed and bled on days 0, 28, 56, and 84 and were killed on day 84 for histomorphometry. Serial assays for blood urea nitrogen (BUN), creatinine, and osteocalcin were performed. Osteocalcin was raised on days 28 and 56 in the high dose CsA group. Histomorphometry showed osteopenia with high-dose CsA. Low-dose CsA was relatively bone sparing, while low-dose SRL and combined low-dose CsA did not cause bone loss. In conclusion, the synergistic combination of low-dose CsA and SRL has the potential of providing both bone sparing and immunosuppressive benefits.

Interferon-alpha, unlike interferon-gamma, does not cause bone loss in the rat.

Interferons (IFN) are a group of related glycoproteins. IFN-gamma, in vitro, has been shown to inhibit resorption; however, an in vivo experiment showed that it had the opposite effect, resulting in bone loss that was comparable to that caused by cyclosporine A. IFN-alpha has numerous clinical applications but is used most extensively in the treatment of chronic hepatitis B and chronic hepatitis C. Research into the effects of IFN-alpha on bone mineral metabolism has been very sparse, and the majority of studies reflect in vitro models. Like IFN-gamma, there exists discordance between in vitro and in vivo studies on IFN-alpha. Both in vivo and in vitro studies demonstrate that IFN-alpha decreases bone resorption, whereas osteoblasts may or may not be affected in vivo. This study was designed to provide information on the in vivo effects of IFN-alpha in the rat model, because we feel that, given its widespread clinical use, this is an extremely important issue. Rats were given low dose IFN-alpha (1.6 x 10(6) IU/m2), intermediate dose IFN-alpha (5.35 x 10(6) IU/m2), and high dose IFN-alpha (30 x 10(6) IU/m2) three times per week for 28 days. Serum osteocalcin (bone gla protein, or BGP) and parathyroid hormone (PTH) were measured serially and, after double labeling, the bones were examined histomorphometrically. IFN-alpha did not alter any of the histomorphometric parameters measured and did not affect PTH. However, it produced a disparate BGP response. Low dose IFN-alpha resulted in a statistically significant increase in serum BGP on days 14 and 28, whereas intermediate and high doses of IFN-alpha did not. Overall, these results provide no evidence of a deleterious effect of IFN-alpha on bone metabolism and confirm the limited clinical study.

Arm training reduces the VO2 and VE cost of unsupported arm exercise and elevation in chronic obstructive pulmonary disease.

BACKGROUND: Patients with severe chronic obstructive pulmonary disease (COPD) may develop dyspnea with minimal arm activity, thoracoabdominal dyssynchrony with unsupported arm exercise (UAEX) and increased oxygen uptake (VO2), and minute ventilation (VE) with simple unsupported arm elevation (UAE) and UAEX. We investigated whether unsupported arm training, as the only form of exercise, could decrease the VO2 and VE cost (percentage increase from resting baseline) associated with unsupported arm elevation and exercise, respectively. METHODS: Twenty-six patients with severe COPD were randomized to 21-24 sessions of unsupported arm (ARMT) or low-intensity resistive breathing (RBT) training as the only form of exercise. Patients were studied before and after training using a metabolic cart and esophageal and gastric pressures to evaluate metabolic and respiratory muscle function. RESULTS: After ARMT, the VO2 (58% vs 38% increase, P < 0.05) and VE (41% v. 21% increase, P < 0.05) cost for UAEX at exercise isotime decreased and endurance time increased. Similarly the VO2 (25% vs 18% increase, P < 0.05) cost decreased and VE no longer increased in response to 2 minutes of UAE after ARMT. The RBT group showed no such change. No improvement in ventilatory load or respiratory muscle function could be identified to explain the physiologic changes observed. After ARMT, mean inspiratory flow (VT/TL), a measure of central respiratory drive, was reduced during UAEX and the expected increase during UAE did not occur. CONCLUSION: We conclude that arm training reduces the VO2 and VE cost of UAE and UAEX, possibly through improved synchronization and coordination of accessory muscle action during unsupported arm activity.

Raloxifene analog (LY117018 HCL) ameliorates cyclosporin A-induced osteopenia in oophorectomized rats.

Cyclosporin A (CsA) administered to the oophorectomized (Ox) rat exacerbates the high turnover osteopenia associated with estrogen deficiency. 17 beta-estradiol replacement therapy prevent this bone loss. The aim of this study was to see whether an estrogen-like compound, Raloxifene analog (LY117018 HCL, Ral) could likewise ameliorate CsA-induced osteopenia in the Ox rat. Sixty 6-month-old Sprague-Dawley rats, divided into five groups, underwent oophorectomy. One group acted as a basal group and the others received either vehicle (group B), CsA 15 mg/kg/day (group C), Ral 3 mg/kg/day (group D), or CsA 15 mg/kg/day and Ral 3 mg/kg/day (group E) for 28 days by gavage. A sixth sham operated group of 12 rats received vehicle only (group A). Rats were weighed and bled on days 0, 14, and 28 for measurement of ionized calcium, glucose, osteocalcin (BGP), 17 beta-estradiol, and 1,25-dihydroxyvitamin D3 (1,25[OH]2D3). Tibiae were removed on day 28 for bone histomorphometry after double tetracycline and calcein labeling. Oophorectomy caused a significant gain in weight in groups B and C which was prevented by Ral in groups D and E. Randomized blood glucose levels and 1,25(OH)2D3 levels were elevated in both CsA-treated groups. Blood ionized calcium levels were lower in vehicle (group B) compared with sham (group A) on day 28. Ox (group B) had significantly higher serum BGP levels compared with sham-operated rats. Serum BGP levels were further elevated in group C compared with vehicle and were lowered in both Ral-treated groups to vehicle levels by day 28. Bone histomorphometry revealed a high turnover osteopenia with increased parameters of bone formation and resorption and loss of cancellous bone volume postoophorectomy (group B). CsA (group C) exacerbated the effects of oophorectomy. Ral (group D) completely prevented the high turnover osteopenia caused by oophorectomy and was able to attenuate substantially the effects of CsA in the Ox rat (group E). Ral therapy ameliorated CsA-induced osteopenia in the Ox rat and might prove a useful agent in preventing bone loss in postmenopausal women receiving CsA.

Effect of tamoxifen on lumbar spine bone mineral density in postmenopausal women after 5 years.

BACKGROUND: Because adjuvant tamoxifen citrate is given to women with early-stage breast cancer for long periods, it is important to know how it affects risk factors for osteoporotic bone fractures, particularly since rates of bone fracture increase rapidly with age in postmenopausal women. In a 2-year randomized placebo-controlled toxicity study in 140 subjects, we demonstrated that tamoxifen was associated with preservation of bone mineral density (BMD), a major risk factor for fractures, in the lumbar spine. METHODS: Five years after entry on this study we reexamined 62 of the original subjects with lumbar spine BMD and serum osteocalcin measurements. These were women available for study because they had not suffered major illnesses and had continued to receive (1) tamoxifen or (2) the no-tamoxifen regimen that they had originally been randomized to receive for the entire 5 years. RESULTS: For lumbar spine BMD at baseline, the 30 subjects in the long-term tamoxifen group and the 32 subjects in the long-term no-tamoxifen group were not significantly different (P = .26). During the first 2 years of follow-up, the 30 subjects in the long-term tamoxifen group showed the same BMD pattern as the entire 70-patient tamoxifen cohort, and similarly the 32 subjects in the long-term no-tamoxifen group showed the same pattern as the entire 70-patient cohort who received placebo. Five-year mean BMD measurements for each long-term follow-up group showed no significant changes from their respective 2-year levels. However, 5-year BMD measurements between the two groups differed (tamoxifen group, +0.8%; placebo group, -0.7%) (P = .06), and the mean regression lines for the changes in BMD over 5 years differed significantly between the two groups (P = .0005). Adjustment for differences in body mass index, reported exercise, and calcium supplementation between these two groups did not change these results. Osteocalcin levels, also comparable at baseline in the two groups, were significantly lower in tamoxifen-treated subjects at 5 years (P = .0005). CONCLUSIONS: While remodeling of bone may be lower, resorption of lumbar spine bone mineral is also lower, and tamoxifen preserves BMD in the lumbar spine over 5 years of treatment in postmenopausal women. Over longer periods, this preservation of BMD might be expected to be associated with lower fracture rates.

Lack of change of cancellous bone volume with short-term use of the new immunosuppressant rapamycin in rats.

Immunosuppressants have adverse effects on bone mineral metabolism in animal and human studies, with corticosteroids producing low-turnover osteopenia, and cyclosporin-A (CsA) producing high-turnover osteopenia. Rapamycin (RAPA) is a new immunosuppressant reported to be at least 10 times more potent than CsA, and acts via a different pathway to CsA and the other new immunosuppressant FK506. This study investigated the effects of RAPA on bone mineral metabolism in the rat. Forty-two, 10-week-old, male Sprague Dawley rats were divided into three groups, and treated according to the following protocol: group A (control) received RAPA vehicle by daily gavage for 14 days (n = 12); group B (high dose RAPA) received RAPA 2.5 mg/kg/day by daily gavage for 14 days (n = 15); group C (low dose RAPA) received RAPA 1.25 mg/kg/day by daily gavage for 14 days (n = 15). Rats were weighed and bled on days 0, 7, and 14 for measurement of blood ionized calcium, bone Gla protein (BGP), parathyroid hormone (PTH), and 1,25(OH)2D. Tibial bone histomorphometry was determined on day 14 after double-calcein labeling. Weight gain was similar in the two groups treated with RAPA compared with control animals. High-dose RAPA (group B) transiently depressed serum BGP levels on day 7, with elevated blood ionized calcium levels on day 7, and lowered 1,25(OH)2D levels on day 14. Serum PTH levels were unchanged. Low-dose RAPA (group C) did not affect calciotropic hormones.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of the national cancer program and proposed reforms.

A statement by some 68 prominent national experts in industrial medicine, carcinogenesis, epidemiology, and public health, released at a February 4, 1992 press conference in Washington, D.C., charged that the National Cancer Institute (NCI) has confused the public by repeated claims of winning the war against cancer. In fact, age standardized incidence rates have increased sharply over recent decades, while ability to treat and cure most cancers has not materially improved. Furthermore, the NCI has minimized evidence for increasing cancer rates which are largely attributed to smoking and to diet. In so doing, NCI trivializes the importance of occupational carcinogens as non-smoking-attributable causes of lung and other cancers, and ignores the tenuous and inconsistent evidence for the causal role of diet per se and also the important role of carcinogenic dietary contaminants. Reflecting this near exclusionary blame-the-victim theory of cancer causation, with support from the American Cancer Society and industry, the NCI discounts the role of avoidable involuntary exposures to industrial carcinogens in air, water, food, the home, and the workplace. The NCI has also failed to provide scientific guidance to Congress and regulatory agencies on fundamental principles of carcinogenesis and epidemiology, and on the critical need to reduce avoidable exposures to environmental and occupational carcinogens. Contrary to NCI, analysis of their $2 billion budget reveals very limited allocations for research on primary cancer prevention, and for occupational cancer which receives only $19 million annually, 1% of NCI's total budget. Problems of professional mindsets in NCI leadership--fixation on diagnosis, treatment, and basic research (much of questionable relevance) and the neglect of cancer prevention--are exemplified by the composition of the Executive President's Cancer Panel and the National Cancer Advisory Board. Contrary to the explicit mandate of the National Cancer Act, the Board is virtually devoid of recognized authorities in occupational and environmental carcinogenesis. These problems are further compounded by institutionalized conflicts of interest reflected in the composition of past Cancer Panels, and of the current Board of Overseers of the Memorial Sloan Kettering Cancer Center, NCI's prototype comprehensive cancer center, with their closely interlocking financial interests with the cancer drug and other industries. Comprehensive reforms of NCI policies and priorities are overdue. Implementation of such reforms is, however, unlikely in the absence of further support from industrial medicine professionals, which is here solicited, besides action by Congress and concerned citizen groups.

Effect of antihypertensive treatment on endothelium-dependent vascular relaxation in patients with essential hypertension.

OBJECTIVES: This study was designed to determine whether antihypertensive treatment can restore the impaired endothelium-dependent vasodilation of patients with essential hypertension. BACKGROUND: The endothelium regulates vascular tone through the release of vasoactive agents that act on the underlying vascular smooth muscle. This endothelial function is impaired in certain cardiovascular conditions, including essential hypertension. METHODS: The vascular responses to acetylcholine (endothelium-dependent vasodilator, 7.5, 15 and 30 micrograms/min) and sodium nitroprusside (direct dilator of smooth muscle, 0.8, 1.6, 3.2 micrograms/min) were studied in 15 patients (11 men and 4 women with a mean age of 54.1 +/- 12 years) on two occasions: after withdrawal of medications, when the patients were hypertensive, and during the medical treatment that reduced blood pressure to within normal limits in each patient. The results were compared with those obtained in 15 normal control subjects (10 men and 5 women with a mean age of 52.3 +/- 7 years). Drugs were infused into the brachial artery, and forearm blood flow response was measured by strain gauge plethysmography. RESULTS: The blood flow and vascular resistance responses to acetylcholine were significantly reduced in the hypertensive patients (p < 0.0001); maximal forearm flow (ml/min per 100 ml) was 7.0 +/- 4 ml in the patients and 16.7 +/- 5 in the control subjects (p < 0.005). However, no significant differences between groups were observed in the responses to sodium nitroprusside. In patients with essential hypertension, the vascular responses to acetylcholine and sodium nitroprusside were not modified by medical therapy. Maximal forearm flow with acetylcholine (ml/min per 100 ml) was 7.2 +/- 2 during antihypertensive therapy and 7.0 +/- 4 after medication withdrawal. CONCLUSIONS: Clinically effective antihypertensive therapy does not restore the impaired endothelium-dependent vascular relaxation of patients with essential hypertension. This indicates that such endothelial dysfunction is either primary or becomes irreversible once the hypertensive process has become established.

Evaluation of the national cancer program and proposed reforms.

A statement by 68 prominent national experts in cancer prevention, carcinogenesis, epidemiology, and public health, released at a February 4, 1992, press conference in Washington, D.C., charged that the National Cancer Institute (NCI) has misled and confused the public by repeated claims of winning the war against cancer. In fact, age-standardized incidence rates have escalated to epidemic proportions over recent decades, while the ability to treat and cure most cancers has not materially improved. Furthermore, the NCI has minimized evidence for increasing cancer rates, which are largely attributed to smoking, trivializing the importance of occupational carcinogens as non-smoking attributable causes of lung and other cancers, and to diet per se, in spite of tenuous and inconsistent evidence and ignoring the important role of carcinogenic dietary contaminants. Reflecting this near exclusionary blame-the-victim theory of cancer causation, with lockstep support from the American Cancer Society and industry, the NCI discounts the role of avoidable involuntary exposures to industrial carcinogens in air, water, food, the home, and the workplace. The NCI has also failed to provide any scientific guidance to Congress and regulatory agencies on fundament principles of carcinogenesis and epidemiology, and on the critical needs to reduce avoidable exposures to environmental and occupational carcinogens. Analysis of the +2 billion NCI budget, in spite of fiscal and semantic manipulation, reveals minimal allocations for research on primary cancer prevention, and for occupational cancer, which receives only +19 million annually, 1 percent of NCI's total budget. Problems of professional mindsets in the NCI leadership, fixation on diagnosis, treatment, and basic research, much of questionable relevance, and the neglect of cancer prevention, are exemplified by the composition of the National Cancer Advisory Board. Contrary to the explicit mandate of the National Cancer Act, the Board is devoid of members authoritative in occupational and environmental carcinogenesis. These problems are further compounded by institutionalized conflicts of interest reflected in the composition of past executive President's Cancer Panels, and of the current Board of Overseers of the Sloan-Kettering Memorial Cancer Center, the NCI's prototype comprehensive cancer center, with their closely interlocking financial interests with the cancer drug and other industries. Drastic reforms of NCI policies and priorities are long overdue. Implementation of such reforms is, however, unlikely in the absence of further support from industrial medicine professionals, besides action by Congress and concerned citizen groups.

In vitro effects of a recombinant toxin targeted to the fibroblast growth factor receptor on rat vascular smooth muscle and endothelial cells.

The dominant mechanism responsible for restenosis after angioplasty is believed to be the activation of medial smooth muscle cells (SMCs), leading to their proliferation, migration to the subintima, and further proliferation. To develop novel strategies that might inhibit or prevent restenosis, we previously used a chimeric toxin composed of transforming growth factor-alpha (which targets the epidermal growth factor receptor) and mutated Pseudomonas exotoxin to preferentially recognize and kill rapidly proliferating, versus quiescent, vascular SMCs. We have recently cloned and expressed a recombinant gene encoding Pseudomonas exotoxin with a mutated (nonfunctional) cell recognition domain fused with the ligand acidic fibroblast growth factor, termed aFGF-PE66(4Glu)KDEL; thus, this recombinant toxin targets the fibroblast growth factor receptor. In the present study, we evaluated the relative effects of this chimeric toxin on quiescent versus rapidly proliferating vascular SMCs and also determined whether aFGF-PE66(4Glu)KDEL exerted different effects on SMCs versus endothelial cells. Rapidly proliferating SMCs (grown in 10% fetal bovine serum) were very sensitive to the cytotoxic effects of aFGF-PE66(4Glu)KDEL, whereas cytotoxicity was significantly less when the SMCs were in a quiescent state (grown in medium supplemented with 0.5% fetal bovine serum). The chimeric toxin was also significantly less cytotoxic against endothelial cells. Competition studies using excess acidic fibroblast growth factor indicated that the cytotoxic effects are specifically mediated by the fibroblast growth factor receptor. Thus, the present studies suggest a potentially expanded role of recombinant toxin therapy in restenosis: multiple receptors can be targeted, and cytotoxic effects, at least in vitro, can be preferentially directed to rapidly proliferating vascular SMCs, with relative sparing of vascular endothelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of a new vitamin D analog, 22-oxa-1 alpha,25(OH)2D3, on bone mineral metabolism in normal male rats.

The in vivo effects of 22-oxa-1 alpha,25 dihydroxyvitamin D3 (OCT), on bone mineral metabolism were investigated in normal male Sprague-Dawley rats. The rats were administered either vehicle (control), low-dose OCT (25 ng/100 g body weight), or high-dose OCT (250 ng/100 g body wt). High-dose OCT increased serum ionized calcium (P less than 0.05) and decreased serum parathyroid hormone (PTH) (P less than 0.05) at all time points and increased serum bone Gla protein on days 7 and 28 (P less than 0.05) compared with controls. Low-dose OCT decreased serum PTH at all the time points (P less than 0.05) compared with controls. Tibial bone histomorphometry showed no significant differences between the two doses of OCT and controls. We found that OCT has minimal direct effects on bone metabolism in normal male rats in contrast to 1,25 dihydroxyvitamin D3. This property may be advantageous in the treatment with OCT of cell-proliferative diseases.

Effects of tamoxifen on bone mineral density in postmenopausal women with breast cancer.

BACKGROUND AND METHODS: Tamoxifen, a synthetic antiestrogen, increases disease-free and overall survival when used as adjuvant therapy for primary breast cancer. Because it is given for long periods, it is important to know whether tamoxifen affects the skeleton, particularly since it is used extensively in postmenopausal women who are at risk for osteoporosis. Using photon absorptiometry, we studied the effects of tamoxifen on the bone mineral density of the lumbar spine and radius and on biochemical measures of bone metabolism in 140 postmenopausal women with axillary-node-negative breast cancer, in a two-year randomized, double-blind, placebo-controlled trial. RESULTS: In the women given tamoxifen, the mean bone mineral density of the lumbar spine increased by 0.61 percent per year, whereas in those given placebo it decreased by 1.00 percent per year (P less than 0.001). Radial bone mineral density decreased to the same extent in both groups. In a subgroup randomly selected from each group, serum osteocalcin and alkaline phosphatase concentrations decreased significantly in women given tamoxifen (P less than 0.001 for each variable), whereas serum parathyroid hormone and 1,25-dihydroxyvitamin D concentrations did not change significantly in either group. CONCLUSIONS: In postmenopausal women, treatment with tamoxifen is associated with preservation of the bone mineral density of the lumbar spine. Whether this favorable effect on bone mineral density is accompanied by a decrease in the risk of fractures remains to be determined.

Cytotoxic effects of a recombinant chimeric toxin on rapidly proliferating vascular smooth muscle cells.

BACKGROUND: Restenosis after percutaneous transluminal coronary angioplasty is associated with activation of medial smooth muscle cells (SMCs); they proliferate, migrate to the subintima, and narrow the vessel lumen. Cancer cells often express more cell surface receptors than do normal cells. This has allowed tumor cells to be specifically targeted using cytotoxic agents. We have examined whether a similar concept can be applied to rapidly proliferating but nontransformed SMCs. Pseudomonas exotoxin (PE; MW, 66 kDa) is a potent toxin that kills cells by inhibiting protein synthesis; its toxicity is diminished when its cell recognition domain is deleted to produce a 40-kDa protein (PE40). METHODS AND RESULTS: A complementary DNA encoding transforming growth factor alpha (TGF alpha) was ligated to that encoding PE40 and the chimeric toxin TGF alpha-PE40, which is cytotoxic to cancer cells displaying epidermal growth factor (EGF) receptors, was expressed in Escherichia coli. The ability of this toxin to kill proliferating SMCs was tested. When cells were seeded at low density (2,500 cells/cm2) and grown in medium supplemented with 10% fetal bovine serum, they were found to be rapidly proliferating; these cells were very sensitive to the cytotoxic effects of TGF alpha-PE40 (ID50, 4.0 +/- 0.17 ng/ml). In contrast, cytotoxicity was 30-fold less (ID50, 125 +/- 23 ng/ml; p less than 0.0004) when cells were in a quiescent state (grown in medium supplemented with 0.5% fetal bovine serum). CONCLUSIONS: Competition studies using excess EGF indicated that the cytotoxic effects of TGF alpha-PE40 are specifically mediated by the EGF receptor. EGF receptor binding analysis demonstrated that rapidly proliferating SMCs display 10-fold more EGF receptors than do quiescent SMCs in vitro. Thus, a chimeric toxin targeted toward the EGF receptor can selectively kill rapidly proliferating SMCs. Whether this toxin or other chimeric toxins directed against other cell surface receptors will effectively inhibit SMCs proliferating in vivo or be useful in preventing restenosis remains to be determined.

Osteoporosis associated with rheumatoid arthritis: pathogenesis and management.

Rheumatoid arthritis is associated with both localized and generalized osteoporosis. Localized osteoporosis can be considered to be caused by local disease mechanisms, including the generation of factors from activation of the cytokine pathway. The etiology of generalized osteoporosis has been difficult to elucidate, particularly because of the lack of sensitive techniques to measure bone mineral density. The introduction of single- and dual-photon absorptiometry and quantitative computed tomography has allowed more accurate assessment of bone mineral density. In general, bone mineral density loss at appendicular sites does not correlate well with axial bone density loss. Corticosteroid treatment exaggerates the development of osteoporosis in up to 40% of patients with rheumatoid arthritis. Sex hormone status, physical activity, disease duration, and functional class are all significant predictors for the development of osteoporosis. Current therapy for prevention and treatment is based largely on theoretical considerations. Physical activity should be encouraged once acute joint inflammation has settled. Postmenopausal women and amenorrheic premenopausal women will benefit from cyclical estrogen replacement. Patients with low serum 1,25-dihydroxy vitamin D3 levels, and males with low serum testosterone levels, are candidates for replacement therapy with the appropriate hormones. In patients who are receiving corticosteroids the dose should be limited, and oral calcium supplements are of benefit. The use of the newer corticosteroid deflazacort, and disease-modifying immunosuppressive drugs, are discussed. Other therapeutic options which should be considered, although published trials are scarce, are calcitonin and the diphosphonates. Further studies are awaited concerning the optimum prevention and treatment of osteoporosis associated with rheumatoid arthritis.(ABSTRACT TRUNCATED AT 250 WORDS)