BACKGROUND AND PURPOSE:The purpose of this study was to examine the effects of D-amphetamine (D-amph) and physical therapy separately or combined on fine motor performance, gross motor performance and cognition after middle cerebral artery thromboembolization in rats.METHODS:Seventy-four rats were trained in appropriate cognitive and motor behaviours. Thirteen animals were sham-operated and fifty-nine animals were embolized in the right carotid territory. Animals were randomly assigned to five groups: 1) SHAM (non-embolized, saline), 2) CONTROL (embolized, saline), 3) D-AMPH (embolized, D-amph), 4) THERAPY (embolized, saline + physical therapy) and 5) D-AMPH + THERAPY (embolized, D-amph + physical therapy). Rats of the groups 4-5 underwent d-amph or saline treatment on days 1, 3, 5 and 7 after surgery and were re-trained for 1 h starting 60 min after each treatment. During this time, rats were allowed to voluntarily engage in suitable cognitive or motor behaviours in order to obtain food. Animals from all groups were re-tested during days 21-28 after surgery.RESULTS:No differences in infarct volumes were observed between the groups of embolized animals. When evaluating performances on days 21-28 after surgery, rats of the SHAM and THERAPY groups had better fine motor performance than those of the CONTROL (P < 0.05), whereas rats of SHAM and D-AMPH groups achieved better cognitive performance than CONTROL rats (P < 0.05). No significant differences were observed between any groups regarding gross motor performance.CONCLUSIONS:After embolization, physical therapy improved fine motor performance and D-amph accelerated rehabilitation of cognitive performance as observed in the rats of the THERAPY and D-AMPH groups. As a result of the administration of a high dose of D-amph, the rats of the D-AMPH + THERAPY combination group failed to engage in physical therapy during D-amph intoxication, thereby limiting any promotion of rehabilitation by combining physical therapy and D-amph.

OBJECTIVES:The effects of pentasaccharide (PENTA), given alone or combined with thrombolysis using recombinant tissue plasminogen activator (rt-PA), on infarct size and clinical outcome were evaluated in a rat embolic stroke model.MATERIALS AND METHODS:Ninety-two rats were embolized unilaterally and assigned to: (i). controls, (ii). rt-PA 6 mg/kg, (iii). PENTA 0.5 mg/kg, (iv). PENTA 0.5 mg/kg and rt-PA 6 mg/kg. After 2 days animals were killed, the brains removed and evaluated microscopically.RESULTS:The median infarct size measured in percentage of the affected hemisphere was 25% in the control group, 4% (P < 0.01, Mann Whitney) in group 2, 19% (n.s.) in group 3, and 10% (P < 0.05) in group 4. rt-PA, and rt-PA combined with PENTA also promoted functional recovery.CONCLUSION:The present study found no effect of 0.5 mg/kg PENTA treatment. Compared with rt-PA treatment alone, 0.5 mg/kg PENTA alone or combined with rt-PA did not significantly increase mortality or tendency for hemorrhage.

OBJECTIVES:To evaluate how soon after stroke the diagnosis of hypertension could be established.METHODS:In a prospective study including 1192 patients with acute stroke within 6 h, blood pressure was measured serially at 2-h intervals during the first 24 h. Results are presented as mean arterial blood pressure (MAP). The Scandinavian Stroke Scale (SSS) assessed the neurological deficit.RESULTS:In 779 patients with mild to moderate ischaemic stroke or transient ischaemic attack (TIA) and SSS > 25, MAP was 118 mmHg (CI 95%: 116-119 mmHg) on admission and 109 mmHg (CI 95%: 108-110 mmHg) 4 h later (paired t-test, P < 0.001). No such early decrease was observed in 228 patients with severe cerebral infarction (CI). In mild to moderate ischaemic stroke or TIA, MAP at 24 h was not different from MAP at 3 months in paired t-test.CONCLUSIONS:Blood pressure 24 h after admission in patients with mild to moderate CI or TIA was representative of the patient's blood pressure 3 months after stroke. A diagnosis of arterial hypertension can be established a few days after stroke.

In pharmacodynamic studies using focal ischaemia models, the size of the infarct measured by quantitative histology is the most important outcome measure. Precise, unbiased and reproducible assessment of infarct volume is of foremost importance. A frequent problem in interventional stroke models is the evaluation of infarcts in animals found dead, where instant post‐mortem fixation of the brain cannot be performed. The purpose of this study was to investigate possible bias from perfusion, immediate and 3‐h post‐mortem delayed immersion fixation on the measured volumes of cerebral infarction, oedema and hemispheres in a rat embolic stroke model. Thirty‐six male Sprague–Dawley rats were thromboembolized into the internal carotid artery. After survival for 24 h, the animals were divided into three groups: group 1 – immediate perfusion fixation; group 2 – immediate immersion fixation of the brain; and group 3 – animals left dead for 3 h at room temperature before removal of the brain for immersion fixation. Following histological preparation and evaluation, the volumes of the hemispheres and infarction were measured by quantitative histology and planimetry. Brains fixed by immersion were 7% larger than the perfusion‐fixed brains. Delaying the immersion fixation for 3 h may increase hemisphere volume by a further 12%. Independent of the fixation procedure, the size of infarction was ∼40% of the ipsilateral hemisphere, and the oedema was ∼11% of the size of the infarct. The used planimetric technique was accurate with measured values within ± 2% of the factual value. In conclusion, sizes of hemispheres, infarction and oedema in absolute volume measures are influenced by the effect of unwanted variation of brain size caused by biological factors and artificial shrinkage caused by fixation, dehydration and heat treatment of the specimens. Infarction and oedema expressed relatively in per cent of hemisphere and infarct, respectively, are robust measures independent of the investigated fixation procedures.

Background and Purpose-We sought to evaluate the effects of the combination of cytidine-5'-diphosphocholine (citicoline) and thrombolysis on infarct size, clinical outcome, and mortality in a rat embolic stroke model.Methods-Eighty-three Sprague-Dawley rats were embolized in the carotid territory with a single fibrin embolus and randomly assigned to the following treatment groups: (1) control (saline), (2) citicoline 250 mg/kg, (3) citicoline 500 mg/kg, (4) recombinant tissue plasminogen activator (rtPA) 5 mg/kg, (5) rtPA 5 mg/kg plus citicoline 250 mg/kg? and (6) rtPA 5 mg/kg plus citicoline 500 mg/kg. rtPA was administered as a continuous intravenous infusion over 45 minutes starting 45 minutes after embolization; citicoline was given intraperitoneally 30 minutes and 24, 48, and 72 hours after embolization. At 96 hours, the brains were fixed and stained by hematoxylin-eosin, and infarct volumes were measured. Neurological scores were determined daily.Results-The median infarct size, measured as percentage of the affected hemisphere, in the control group was 37% (interquartile range, 26% to 69%) compared with 22% (5% to 52%; P=NS) in group 2, 11% (5% to 34%; P=NS) in group 3, 24% (12% to 31%; P=NS) in group 4, 11% (3% to 22%; P=0.02) in the combined group 5, and 19% (9% to 51%; P=NS) in group 6. The infarct size was significantly reduced in the combined citicoline +rtPA-treated groups to a median of 13% (5% to 30%; P<0.01). Citicoline 500 mg/kg and citicoline combined with rtPA also promoted functional recovery.Conclusions-These results demonstrate that the combination of low-dose citicoline and rtPA significantly reduced infarct size in this focal ischemia model.

The purpose of this study was to investigate if cerebral blood flow (CBF) was influenced by the volume of the bolus using the intracarotid 133Xenon method in anesthetized rats, and to evaluate the effect of intracarotid embolization on hemispheric CBF. In 18 male Sprague-Dawley rats hemispheric CBF was calculated from recorded clearance of an intracarotid injected 50 microliters bolus of 133Xenon. A second clearance recording was done after the injection of a 200 microliter bolus of 133Xenon. Then the 18 animals were divided in 3 subsets each of 6 animals. In the first subset of 6 animals a bolus of 200 microliters 133Xenon was given, clearance was recorded before and after the injection of a suspension of microemboli. In all the above experiments within 90 sec after the initial bolus of 133Xenon, 20 microliters 133Xenon remaining in the carotid catheter was flushed into the brain by saline or the suspension of emboli. In the second subset of 6 rats the distribution of 133Xenon in various parts of the brain and head was examined after killing the animals 35 sec after injection of a 200 microliter bolus of 133Xenon. The third subset of 6 rats was treated as the second subset, except for the size of the bolus which was 50 microliters of 133Xenon. The calculated CBF values were independent of the volume of the bolus. At the post-mortem examination 76% of the radioactivity was located in the right hemisphere and majority of the remainder in the head outside the brain. The clearance curve was not monoexponential, but flattened out within the initial 30 sec when clearance from areas with CBF dominated the initial 15-second period, then clearance form areas with lower CBF values were more prominent in the following 15-second period. Embolization significantly reduced right hemispheric CBF to 0.26 (0.09-0.50) of the value immediately before embolization. The findings demonstrate applicability of CBF measurements using the intra-arterial 133Xenon injection method during embolization of the rat brain, provided a second bolus of 133Xenon is given before the embolization.

In animal stroke models, treatment with mild hypothermia (30-34 degrees C) for 3-4 hours may reduce the size of cerebral infarction if started within three hours of the initiation of cerebral ischaemia. The mechanism by which hypothermia exerts its neuroprotective effect is unknown, but experimental studies have shown the release of neurotoxic excitatory amino acids and free oxygen radicals to be reduced during hypothermic ischaemia. In patients with acute stroke, body temperature above 37.5 degrees C are associated with poor outcome, and temperatures below 36.5 degrees C with improved outcome, compared to normothermic patients. Due to the unpleasantness of cooling and side effects as shivering, hypothermia may not be tolerated by stroke patients without sedation of light anaesthesia which may increase the risk of hypotension and respiratory complications. However, lowering body temperature by 1-2 degrees C may suffice to improve functional outcome in acute stroke patients, and such mild hypothermia should be tested in randomized controlled clinical trials.

OBJECTIVESTo assess bone metabolism following bladder substitution with the ileal Kock reservoir.PATIENTS, SUBJECTS AND METHODSThe investigation comprised two separate studies, one with baseline measurements before and after surgery, and the second after surgery only, of bone mass, made using single-photon absorptiometry and dual-energy X-ray absorptiometry, biochemical variables of bone turnover, plasma analyses and measurements of renal calcium and phosphate excretion. After inclusion, both groups of patients were observed longitudinally for 2 years. The post-surgery study included 25 patients who had undergone bladder substitution (median age 67 years, range 44-75), with a median post-operative follow-up of 1.0 year (range 0.3-3.7), and 16 control subjects (either healthy or with other minor urological complaints; median age 62 years, range 34-80), and the pre-surgery study comprised seven patients who had undergone bladder substitution (median age 57 years, range 42-68).RESULTSTotal body, forearm and spinal bone mineral contents were similar in patients with an ileal bladder substitute measured 1 year after surgery and in control subjects. There were equivalent significant changes in both the patients and control subjects during the 2-year observation period, with a 2-3% decrease in total body and forearm bone mineral content. The values were similar in patients with and without a mild metabolic acidosis. Plasma calcium, phosphate, total alkaline phosphatase, intact parathyroid hormone, vitamin D and osteocalcin were normal in both patients and control subjects. Renal excretion of calcium and phosphate was also similar in patients and in control subjects.CONCLUSIONSIleal urethral Kock bladder substitution does not lead to accelerated bone mineral loss in elderly men, despite a mild metabolic acidosis in half of the patients.

We present baseline bone densitometry from the Early Postmenopausal Interventional Cohort study (EPIC, sponsored by Merck, Sharp & Dohme) for the first time, in which 1609 women from England, Oregon, Hawaii and Denmark are participating to investigate the efficacy of daily oral alendronate to prevent early postmenopausal bone loss. We compared radiographic absorptiometry (RA) of the phalanges for bone mineral density (BMD) measurement with single-energy X-ray absorptiometry (SXA) of the distal forearm, and dual-energy X-ray absorptiometry (DXA) of the lumbar spine, proximal femur and distal forearm. In a random subgroup of 308 women, aged 45–60 years, on average 6 years since menopause (YSM), bone densitometry was measured once at baseline by RA of the phalanges besides the mandatory measurements by DXA. Bone densitometry was furthermore measured by SXA at the Danish site (89 women). Sixty-eight of the women had duplicate measurements performed within 1–3 weeks to evaluate the short-term precision error (CV%). One hundred and one healthy premenopausal women, aged 25–48 years, were recruited at the Danish and Hawaiian sites to establish a reference group. The precision error was 1.5% for RA of the phalanges and in the range 1.0–2.2% for SXA and DXA. BMD by RA correlated with BMD measured by SXA and DXA in the range 0.45<r<0.72 (p<0.001). In conclusion, bone densitometry by RA of the phalanges is highly correlated with bone densitometry by SXA and DXA. RA of the phalanges has a short-term precision error comparable to that of SXA and DXA.

In 979 healthy women, aged 30-75 years, bone mass was measured by DXA in the lumbar spine and proximal femur, and by SXA in the distal forearm. Bone turnover was assessed by urinary CrossLaps (CrossLaps ELISA), a new assay which measures type I collagen degradation products in urine and by osteocalcin (two-site N-Mid hOsteocalcin ELISA), a new assay which measures the N-terminal-mid fragment (1-43) as well as the intact (1-49) osteocalcin (OCN-Mid) in serum. For comparison data on urinary hydroxyproline (fU Hpr/Cr) and serum, total alkaline phosphatase were included (AP). In premenopausal women below 50 years of age, the concentrations of the biochemical markers were stable with age. At menopause CrossLaps and OCN-Mid increased abruptly to a level 60% and 35% above the premenopausal mean values (p < 0.001). Premenopausal women in the highest quartiles, stratified according to the concentration of CrossLaps and OCN-Mid corrected for height and weight, had 6%-11% lower bone mass in all regions (p < 0.01) as compared to women in the lowest quartiles. CrossLaps and OCN-Mid corrected for height and weight correlated with bone mass in the spine and proximal femur, r = -0.13 to r = -0.28, p < 0.05. In postmenopausal women, the difference in bone mass between the highest and lowest quartiles was 8%-14% (p < 0.001). CrossLaps and OCN-Mid correlated with bone mass measured in all regions, r = -0.14 to r = -0.32, p < 0.05. The correlation between bone mass and AP and Fu Hpr/Cr was lower; r = -0.06 to r = -0.20 for premenopausal women, NS to p < 0.01, and r = -0.01 to r = -0.23, NS to p < 0.001 for postmenopausal women. In conclusion, the present data indicate that high bone turnover is associated with a significantly lower bone mass in not only postmenopausal, but interestingly also in premenopausal women. In consistence with previous results, we found that bone turnover increased perimenopausally and in the early menopause.

In a double-blind, placebo-controlled, randomized group comparison, new and specific biochemical markers for bone resorption as follow-up parameters on the therapeutic response to nasal salmon calcitonin (sCT) were evaluated. Evaluation took place at an outpatient clinic where osteoporosis was being researched. The subjects included 208 women aged 68–72 treated for 2 years with either 50 IU, 100 IU, or 200 IU of nasal sCT or placebo; all groups received a daily calcium supplementation of 500 mg. Only 164 women fulfilled the study as valid completers. Markers were applied to frozen urine samples of a previously published intervention study of a new fasting urinary (fU) biochemical marker for bone resorption (CrossLapsTM, ELISA) and the urinary excretion of cross-links (pyridinoline and deoxypyridinoline) was measured, all corrected for creatinine. Bone mineral density of the lumbar spine and rates of vertebral and peripheral fractures were measured after 2 years of treatment. The creatinine corrected urinary pyridinoline, deoxypyridinoline, and CrossLaps showed maximum decreases of 10–43% (95% confidence interval-29.5% to 9.6% and -75.1% to 9.3%;P < 0.01-0.001) after 6–9 months, after which the response leveled off. A significant difference among the four treatment groups was seen in fU CrossLaps(P < 0.01). The changes in spinal bone mass were significantly related to the decreases in fU CrossLaps: women with the highest response in spinal bone mass had decreases in fU CrossLaps of 44% (-83.5% to 7.4%) and women without response of 5% (-57.6% to 99.9%)P 0.001). In women who fractured during the 2-year period, fU CrossLaps remained unchanged, whereas decreases of 30% (-75.1% to 44.7%) were seen in women who did not fracture(P = 0.002). The results suggest that biochemical markers can be used to determine the optimum treatment regimen of nasal sCT. The response of the new marker, fU CrossLaps, significantly reflects the responses in bone mass of the spine and fracture rates.